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CLINICAL CHEMISTRY

REAGENT GUIDE

Beckman Coulter, Inc., 250 S. Kraemer Blvd.,Brea, CA 92821, USA.

EC REP Beckman Coulter Biomedical Limited, Lismeehan, O' Callaghan's Mills, Co. Clare, Ireland, Tel: ++353 65 6831100.

EN -i–
This guide is intended for use with:
BECKMAN COULTER AU400
BECKMAN COULTER AU600/600IVD
BECKMAN COULTER AU640
BECKMAN COULTER AU2700
BECKMAN COULTER AU5400
BECKMAN COULTER AU680
BECKMAN COULTER AU480

Version .11
Revision date: 2010-06

- ii –
CONTENTS

1. OVERVIEW

2. INSTRUCTIONS

2. 1 Guide Format.
2. 2 Guide Updates.

3. SYMBOLS

4. GENERAL PRECAUTIONS AND WARNINGS

5. NOTES

APPENDIX 1 PRODUCT GROUPINGS

APPENDIX 2 TABLE OF CALIBRATORS AND CONTROLS

APPENDIX 3 GLOSSARY **

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- iv –
1. OVERVIEW
This guide is designed to provide specific chemistry information for the Beckman Coulter clinical
chemistry systems.

Please note that the latest revision of the Instructions For Use (IFU) and Setting Sheets can be
found on the Beckman Coulter website at; www.beckman coulter.com.
Access to this website requires registration; therefore it is recommended that you register as soon
as possible.

The guide does not include all the requirements governing the safe and effective operation of the
Beckman Coulter clinical chemistry systems.

Refer to the analyser specific User Guide and your specific laboratory procedures governing
other areas, such as corrective actions, QC policies, or hazardous waste disposal that are not
addressed in this guide.

Beckman Coulter logo is a trademark of Beckman Coulter, Inc. and is registered in the USPTO.

2. INSTRUCTIONS

2.1 Guide Format:


This guide contains the Instructions For Use (IFU), which describe how the reagent is to be
used, and Setting Sheets for the Beckman Coulter range of reagent kits, which describe the
required Beckman Coulter Chemistry Analyser settings to use with the specified reagents.

The outer label on the Beckman Coulter reagent kit identifies the Part No. for the reagent and the
applicable revision code for the IFU and the Setting Sheet. Please refer to this each time a
kit is used – see below. Additionally, please ensure the use of the appropriate Setting
Sheet for application with the biological fluid under investigation.

Example: REF: OSR6287

BLOSR6x87.01 Î IFU reference and revision code.


BSOSR6x87.01 Î Setting sheet reference and revision code.

If the indicated revision or a higher revision is not present, then please contact your local
Beckman Coulter support organisation. Alternatively, you may access this document on the
Beckman Coulter website as described in Section 1.

Note: The ‘x’ in the reference code denotes a multiple kit format. For reference code
BLOSR6x87 incorporates OSR6187, OSR6287 and OSR6587.

2.2 Guide Updates:


Updates to IFUs and Setting Sheets will be delivered to you as and when these become
available. These updates should be inserted into the guide and the older version removed
(unless specifically required) so that the latest information is available at all times.

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3. SYMBOLS
The following symbols are used in the labelling of Beckman Coulter System Reagents.

Manufacturer Biological Risk

In Vitro Diagnostic Medical Device Contents

Consult Instructions for Use Protect from Light

Material safety data sheet available to


Temperature Limitation professional users on request.

Catalogue Number Serial Number

Keep Upright Caution, consult accompanying documents

Batch code Product conforms to the IVD directive

Use by:
For IVD performance evaluation only (YYYY-MM-DD or YYYY-MM)

Authorized Representative in the


European Community

4. GENERAL PRECAUTIONS AND WARNINGS


The following precautions and warnings are valid for all tests.
• There are literature reports of very rare cases where gammopathy, especially monoclonal IgM
(Waldenström’s macroglobulinemia), may cause protein precipitation when mixed with reagents giving rise
to turbidity. This may occasionally lead to unreliable results when processing such samples using
photometric methods. Further background information can be obtained from the following paper and its
references: “Berth M, Delanghe J. Protein precipitation as a possible important pitfall in the clinical
chemistry analysis of blood samples containing monoclonal immunoglobulins: 2 case reports and a review
of the literature. Acta Clin Belg. 2004;59:263-73.”
Due to the idiosyncratic nature of this interference it can theoretically occur with any liquid reagent. Where
Beckman Coulter has received reports of this type of interference, or for those reagents considered at
highest risk, a warning statement is included in the Interfering Substances section of the appropriate
reagent.
• Please note that recovery of non-Beckman Coulter controls may vary with reagent lots of immunoassay
products, due to the use of non-human materials in the controls.
• Reagents from different containers should not be intermixed.
• When using serum or plasma collection tubes, follow the tube manufacturer’s processing instructions.
Sample collection devices should be assessed for suitability and regularly monitored. Contact your
collection device supplier for further details.
• Samples containing precipitates must be centrifuged before performing the assay. Patient samples should
be homogeneous. Patient samples should not contain clots or air bubbles.
• Biological materials of human origin contained in these products were tested for Anti-HCV, HbsAg and
Anti-HIV 1/2 on a single donor basis and were found to be non-reactive. As there is no known test method
that can offer complete assurance that products derived from human blood will not transmit infectious
agents, these products should be handled as potentially infectious materials.
• Handle all patient samples as potentially infectious and follow universal precautions as dictated by local or
national regulations (for further details refer to CLSI GP17-A2, ISO15190 or 29CFR1910.1030).
• Handle samples in closed containers to avoid contamination and evaporation.

5. NOTES
* Control Material with values determined by this Beckman Coulter system may be used for this application.
** This is not required for the English Reagent Guide.

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APPENDIX 1 PRODUCT GROUPINGS
IFU Reference Setting Sheet Reference
Enzyme Code Code
OSR6x01 ACP BLOSR6x01.01 BSOSR6x01.01
OSR6x03 ALP BLOSR6x03.01 BSOSR6x03.01
OSR6x04 ALP BLOSR6x04.01 BSOSR6x04.01
OSR6x07 ALT BLOSR6x07.01 BSOSR6x07.02
OSR6x06 α-Amylase BLOSR6x06.01 BSOSR6x06.01
OSR6x82 α-Amylase BLOSR6x82.01 BSOSR6x82.01
OSR6x09 AST BLOSR6x09.01 BSOSR6x09.02
OSR6x14 Cholinesterase BLOSR6x14.01 BSOSR6x14.01
OSR6x79 CK (NAC) BLOSR6x79.01 BSOSR6x79.01
OSR6x155 CK-MB BLOSR6x155.01 BSOSR6x155.01
OSR6x20 GGT BLOSR6x20.01 BSOSR6x20.01
OSR6x29 HBDH BLOSR6x29.01 BSOSR6x29.01
OSR6x26 LDH BLOSR6x26.01 BSOSR6x26.01
OSR6x28 LDH BLOSR6x28.01 BSOSR6x28.01
OSR6x30 Lipase BLOSR6x30.01 BSOSR6x30.01

Metabolite
OSR6x02 Albumin BLOSR6x02.01 BSOSR6x02.01
OSR6x90 Bicarbonate BLOSR6x90.01 BSOSR6x90.01
OSR6x11 Direct Bilirubin BLOSR6x11.01 BSOSR6x11.01
OSR6x12 Total Bilirubin BLOSR6x12.01 BSOSR6x12.01
OSR6x13 Calcium oCPC BLOSR6x13.01 BSOSR6x13.01
OSR6x117 Calcium Arsenazo III BLOSR6x117.02 BSOSR6x117.01
OSR6x16 Cholesterol BLOSR6x16.02 BSOSR6x16.01
OSR6x78 Creatinine BLOSR6x78.02 BSOSR6x78.01
OSR6x204 Creatinine (Enzymatic) BLOSR6x204.01 BSOSR6x204.02
OSR6x21 Glucose BLOSR6x21.02 BSOSR6x21.02
OSR6x40 Glucose - STAT BLOSR6x40.01 BSOSR6x40.01
OSR6x87 HDL-Cholesterol BLOSR6x87.01 BSOSR6x87.01
OSR6x22 Inorganic Phosphorous BLOSR6x22.01 BSOSR6x22.01
OSR6x86 Iron BLOSR6x86.01 BSOSR6x86.01
OSR6x93 Lactate BLOSR6x93.01 BSOSR6x93.01
OSR6x83 LDL-Cholesterol BLOSR6x83.02 BSOSR6x83.01
OSR6x89 Magnesium BLOSR6x89.01 BSOSR6x89.01
OSR6x32 Total Protein BLOSR6x32.01 BSOSR6x32.01
OSR6x118 Triglyceride BLOSR6x118.01 BSOSR6x118.01
OSR6x24 UIBC BLOSR6x24.01 BSOSR6x24.01
OSR6x205 UIBC BLOSR6x205.01 BSOSR6x205.01
OSR6x34 Urea BLOSR6x34.01 BSOSR6x34.02
OSR6x41 Urea - STAT BLOSR6x41.02 BSOSR6x41.02
OSR6x98 Uric Acid BLOSR6x98.01 BSOSR6x98.01
OSR6x70 Urinary/CSF Protein BLOSR6x70.01 BSOSR6x70.01

Specific Protein
OSR6x62 α-1 Acidglycoprotein BLOSR6x62.01 BSOSR6x62.01
OSR6x63 α-1 Antitrypsin BLOSR6x63.01 BSOSR6x63.01
OSR6x42 Apo A1 BLOSR6x42.01 BSOSR6x42.01
OSR6x43 Apo B BLOSR6x43.01 BSOSR6x43.01
OSR6x94 ASO BLOSR6x94.01 BSOSR6x94.02
OSR6x51 ß-2 Microglobulin BLOSR6x51.01 BSOSR6x51.01
OSR6x59 C3 BLOSR6x59.01 BSOSR6x59.01
OSR6x60 C4 BLOSR6x60.01 BSOSR6x60.01
OSR6x64 Ceruloplasmin BLOSR6x64.01 BSOSR6x64.01
OSR6x47 CRP BLOSR6x47.01 BSOSR6x47.01
OSR6x99 CRP Latex BLOSR6x99.01 BSOSR6x99.01
OSR6x135 D-Dimer BLOSR6x135.01 BSOSR6x135.01
OSR6x50 Ferritin BLOSR6x50.01 BSOSR6x50.01
OSR6x138 Ferritin BLOSR6x138.01 BSOSR6x138.01
OSR6x203 Ferritin BLOSR6x203.02 BSOSR6x203.02

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IFU Reference Setting Sheet Reference
Specific Protein
Code Code
OSR6x65 Haptoglobin BLOSR6x65.01 BSOSR6x65.01
OSR6x92 HbA1c (Hemoglobin A1c) BLOSR6x92.02 BSOSR6x92.02
HbA1c APT (Hemoglobin A1c,
OSR6x177 BLOSR6x177.01 BSOSR6x177.01
Whole Blood Application)
OSR6x171 IgA BLOSR6x171.01 BSOSR6x171.01
OSR6x172 IgG BLOSR6x172.02 BSOSR6x172.02
OSR6x173 IgM BLOSR6x173.01 BSOSR6x173.01
OSR6x67 Microalbumin BLOSR6x67.01 BSOSR6x67.01
OSR6x68 Myoglobin BLOSR6x68.01 BSOSR6x68.01
OSR6x75 Prealbumin BLOSR6x75.01 BSOSR6x75.01
OSR6x105 RF Latex BLOSR6x105.01 BSOSR6x105.01
OSR6x52 Transferrin BLOSR6x52.01 BSOSR6x52.01

Therapeutic Drug Monitoring (TDM)


OSR6414 Carbamazepine BLOSR6414.01 BSOSR6414.01
OSR6403 Digitoxin BLOSR6403.01 BSOSR6403.01
OSR6404 Digoxin BLOSR6404.01 BSOSR6404.01
OSR6420 Gentamycin BLOSR6420.01 BSOSR6420.01
OSR61202 Paracetamol BLOSR6x202.01 BSOSR6x202.01
OSR6413 Phenobarbital BLOSR6413.01 BSOSR6413.01
OSR6411 Phenytoin BLOSR6411.01 BSOSR6411.01
OSR6412 Theophylline BLOSR6412.01 BSOSR6412.01
OSR6415 Valproic Acid BLOSR6415.01 BSOSR6415.01

Drugs of Abuse in Urine (DAU)


OSR6323 Amphetamines / Ecstasy BLOSR6323.01 BSOSR6323.01
OSR6315 Barbiturates BLOSR6315.01 BSOSR6315.01
OSR6316 Benzodiazepines BLOSR6316.01 BSOSR6316.01
OSR6317 Cocaine BLOSR6317.01 BSOSR6317.01
OSR6318 EDDP BLOSR6318.01 BSOSR6318.01
OSR6319 Methadone BLOSR6319.01 BSOSR6319.01
OSR6320 Opiates BLOSR6320.01 BSOSR6320.01
OSR6322 THC BLOSR6322.01 BSOSR6322.01

ISE
66320 ISE Buffer BL66320.02
66316 ISE High Serum Standard BL66320.02
66314 ISE Internal Reference BL66320.02
66317 ISE Low Serum Standard BL66320.02
66315 ISE Low/High Urine Standard BL66320.02
66319 ISE Mid Standard BL66320.02
+ +
66313 ISE Na /K Selectivity Check BL66320.02
66318 ISE Reference BL66320.02

Calibrator
ODC6413 Antibiotic TDM Multi-Calibrator BLODC6413.01
ODR3005 Apo A1 & B Calibrator BLODR3005.01
ODR3022 Apo A1 & B Calibrator BLODR3022.01
ODR3013 ASO Calibrator BLODR3013.01
ODC0019 Bicarbonate Calibrator BLODC0019.01
ODR30034 CK-MB Calibrator BLODR30034.01
ODC6411 Core TDM Multi-Calibrator BLODC6411.01
ODC0027 CRP Latex Calibrator Highly Sensitive Set BLODC0027.01
ODC0026 CRP Latex Calibrator Normal Set BLODC0026.01
ODR3033 D-Dimer Calibrator BLODR3033.01
ODC6326 DAU Low Intermediate Multi-Drug Calibrator BLODC6326.01 BSODC6326.01
ODC6319 DAU Methadone Cut-off Calibrator BLODC6319.01
ODC6320 DAU Methadone Intermediate Calibrator BLODC6319.01
ODC6321 DAU Methadone High Calibrator BLODC6319.01
ODC6315 DAU Primary Cut-off Multi-Drug Calibrator BLODC6315.01

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IFU Reference
Calibrator Setting Sheet Reference Code
Code
ODC6316 DAU Secondary Cut-off Multi-Drug Calibrator BLODC6315.01
ODC6317 DAU Intermediate Multi-Drug Calibrator BLODC6315.01
ODC6318 DAU High Multi-Drug Calibrator BLODC6315.01
ODC6314 DAU Negative Calibrator BLODC6314.01
ODC6322 DAU THC 25 Calibrator BLODC6322.01
ODC6323 DAU THC 50 Calibrator BLODC6322.01
ODC6324 DAU THC 75 Calibrator BLODC6322.01
ODC6325 DAU THC 100 Calibrator BLODC6322.01
ODC6403 Digitoxin Calibrator BLODC6403.01
ODC6404 Digoxin Calibrator BLODC6404.01
ODR3032 HbA1c Calibrator BLODR3032.01
ODC0011 HDL-Cholesterol Calibrator BLODC0011.01
ODC0012 LDL-Cholesterol Calibrator BLODC0012.01
ODR30037 MC Cal A BLODR30037.01
ODR3024 Microalbumin Calibrator BLODR3024.01
ODR3025 Myoglobin Calibrator BLODR3025.01
ODR3029 Prealbumin Calibrator BLODR3029.01
ODC0028 RF Latex Calibrator BLODR0028.01
ODR3021 Serum Protein Multi-Calibrator BLODR3021.01
ODR3023 Serum Protein Multi-Calibrator 2 BLODR3023.01
66300 System Calibrator BL66300.01
ODC0025 Urine Calibrator BLODC0025.01

Control
ODR2041 CK-MB Control Serum BLODR2041.01
ODR30035 CK-MB Control Level 1 BLODR30035.01
ODR30036 CK-MB Control Level 2 BLODR30036.01
ODC0003 Control Serum 1 BLODC0003.01
ODC0004 Control Serum 2 BLODC0004.01
ODC0013 CRP (Latex) Control Serum BLODC0013.01
ODC0029 D-Dimer Control BLODC0029.01
ODC0006 DAU Multi-Drug Control BLODC0006.01
ODC0007 DAU Speciality Control BLODC0007.01
ODC0008 DAU THC 25 Control BLODC0008.01
ODC0009 DAU THC 50 Control BLODC0008.01
ODC0022 HbA1c Control BLODC0022.01
ODC0005 HDL/LDL-Cholesterol Control Serum BLODC0005.01
ODC0014 ITA Control Serum 1 BLODC0014.01
ODC0015 ITA Control Serum 2 BLODC0015.01
ODC0016 ITA Control Serum 3 BLODC0016.01

Miscellaneous
ODR20067 Cleaning Solution BLODR20067.01
OE66039 Cleaning Solution BLOE66039.01
OSR0004 Hemoglobin Denaturant BLOSR0004.01
OSR62166 LIH BLOSR6x166.01 BSOSR6x166.01
OSR0001 Wash Solution BLOSR0001.01
ODR2000 Wash Solution BLOSR0001.01

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-x–
APPENDIX 2 CALIBRATORS AND CONTROLS
Parameter name as
Code appears on kit Calibrator Control
Enzyme
OSR6x01 ACP 66300 ODC0003, ODC0004
OSR6x03 ALP 66300 ODC0003, ODC0004
OSR6x04 ALP 66300 ODC0003, ODC0004
OSR6x07 ALT 66300 ODC0003, ODC0004
Serum/Plasma: 66300 Serum/Plasma: ODC0003, ODC0004
OSR6x06 α-Amylase
Urine: ODC0025 Urine: Biorad Liquichek Cat. No: 397 & 398
Serum/Plasma: 66300 Serum/Plasma: ODC0003, ODC0004
OSR6x82 α-Amylase
Urine: ODC0025 Urine: Biorad Liquichek Cat. No: 397 & 398
OSR6x09 AST 66300 ODC0003, ODC0004
OSR6x14 Cholinesterase 66300 ODC0003, ODC0004
OSR6x79 CK (NAC) 66300 ODC0003, ODC0004
OSR6x155 CK-MB ODR30034 ODR30035, ODR30036
OSR6x20 GGT 66300 ODC0003, ODC0004
OSR6x29 HBDH 66300 ODC0003, ODC0004
OSR6x26 LDH 66300 ODC0003, ODC0004
OSR6x28 LDH 66300 ODC0003, ODC0004
OSR6x30 Lipase OSR6x30 or 66300 ODC0003, ODC0004

Metabolite
OSR6x02 Albumin 66300 ODC0003, ODC0004
OSR6x90 Bicarbonate ODC0019 *
OSR6x11 Direct Bilirubin 66300 ODC0003, ODC0004
OSR6x12 Total Bilirubin 66300 ODC0003, ODC0004
Serum/Plasma: 66300 Serum/Plasma: ODC0003, ODC0004
OSR6x13 Calcium oCPC
Urine: ODC0025 Urine: Biorad Liquichek Cat. No: 397 & 398
Serum/Plasma: 66300 Serum/Plasma: ODC0003, ODC0004
OSR6x117 Calcium Arsenazo III
Urine: ODC0025 Urine: Biorad Liquichek Cat. No: 397 & 398
OSR6x16 Cholesterol 66300 ODC0003, ODC0004
Serum/Plasma: 66300 Serum/Plasma: ODC0003, ODC0004
OSR6x78 Creatinine
Urine: ODC0025 Urine: Biorad Liquichek Cat. No: 397 & 398
Serum/Plasma: 66300 Serum/Plasma: ODC0003, ODC0004
OSR6x204 Creatinine (Enzymatic)
Urine: ODC0025 Urine: Biorad Liquichek Cat. No: 397 & 398
Serum/Plasma: 66300 Serum/Plasma: ODC0003, ODC0004
OSR6x21 Glucose
Urine: ODC0025 Urine: Biorad Liquichek Cat. No: 397 & 398
OSR6x40 Glucose-STAT 66300 ODC0003, ODC0004
OSR6x87 HDL-Cholesterol ODC0011 ODC0005
Serum/Plasma: 66300 Serum/Plasma: ODC0003, ODC0004
OSR6x22 Inorganic Phosphorous
Urine: ODC0025 Urine: Biorad Liquichek Cat. No: 397 & 398
OSR6x86 Iron 66300 ODC0003, ODC0004
OSR6x93 Lactate 66300 ODC0003, ODC0004
OSR6x83 LDL Cholesterol ODC0012 ODC0005
Serum/Plasma: 66300 Serum/Plasma: ODC0003, ODC0004
OSR6x89 Magnesium
Urine: ODC0025 Urine: Biorad Liquichek Cat. No: 397 & 398
OSR6x32 Total Protein 66300 ODC0003, ODC0004
OSR6x118 Triglyceride 66300 ODC0003, ODC0004
OSR6x24 UIBC 66300 ODC0003, ODC0004
OSR6x205 UIBC 66300 ODC0003, ODC0004
Serum/Plasma: 66300 Serum/Plasma: ODC0003, ODC0004
OSR6x34 Urea
Urine: ODC0025 Urine: Biorad Liquichek Cat. No: 397 & 398
OSR6x41 Urea-STAT 66300 ODC0003, ODC0004
Serum/Plasma: 66300 Serum/Plasma: ODC0003, ODC0004
OSR6x98 Uric Acid
Urine: ODC0025 Urine: Biorad Liquichek Cat. No: 397 & 398
OSR6x70 Urinary/CSF Protein OSR6x70 *

- xi –
Specific Protein
OSR6x62 α-1 Acidglycoprotein ODR3023 ODC0014, ODC0015, ODC0016
OSR6x63 α-1 Antitrypsin ODR3023 ODC0014, ODC0015, ODC0016
OSR6x42 Apo A1 ODR3005 (3 pt) / ODR3022 (5 pt) ODC0003, ODC0004

OSR6x43 Apo B ODR3005 (3 pt) / ODR3022 (5 pt) ODC0003, ODC0004


OSR6x94 ASO ODR3021 or ODR3013 / ODR30037 ODC0014, ODC0015, ODC0016
OSR6x51 β-2 Microglobulin ODR3023 ODC0014, ODC0015, ODC0016
OSR6x59 C3 ODR3021 / ODR30037 ODC0014, ODC0015, ODC0016
OSR6x60 C4 ODR3021 / ODR30037 ODC0014, ODC0015, ODC0016
OSR6x64 Ceruloplasmin ODR3023 ODC0014, ODC0015, ODC0016
OSR6x47 CRP ODR3021 ODC0014, ODC0015, ODC0016
ODC0014, ODC0015, ODC0016
ODC0026 (Normal), (Normal, Highly sensitive),ODC0003
OSR6x99 CRP Latex
ODC0027 (Highly sensitive) (Normal)
ODC0013 (Highly sensitive)
OSR6x135 D-Dimer ODR3033 ODC0029
OSR6x50 Ferritin ODR3021 ODC0014, ODC0015, ODC0016
OSR6x138 Ferritin ODR3021 ODC0014, ODC0015, ODC0016
OSR6x203 Ferritin ODR3021 ODC0014, ODC0015, ODC0016
OSR6x65 Haptoglobin ODR3023 ODC0014, ODC0015, ODC0016
OSR6x92 HbA1c (Hemoglobin A1c) ODR3032 ODC0022
HbA1c APT (Hemoglobin A1c,
OSR6x177 ODR3032 ODC0022
Whole Blood Application)
OSR6x171 IgA ODR3021 ODC0014, ODC0015, ODC0016
OSR6x172 IgG ODR3021 ODC0014, ODC0015, ODC0016
OSR6x173 IgM ODR3021 ODC0014, ODC0015, ODC0016
OSR6x67 Microalbumin ODR3024 *
OSR6x68 Myoglobin ODR3025 *
OSR6x75 Prealbumin ODR3029 ODC0014, ODC0015, ODC0016
OSR6x105 RF Latex ODC0028 ODC0014, ODC0015, ODC0016
OSR6x52 Transferrin ODR3021 / ODR30037 ODC0014, ODC0015, ODC0016
TDM
OSR6414 Carbamazepine ODC6411 *
OSR6403 Digitoxin ODC6403 *
OSR6404 Digoxin ODC6404 *
OSR6420 Gentamycin ODC6413 *
OSR6x202 Paracetamol OSR6x202 *
OSR6413 Phenobarbital ODC6411 *
OSR6411 Phenytoin ODC6411 *
OSR6412 Theophylline ODC6411 *
OSR6415 Valproic Acid ODC6411 *
DAU
Amphetamines/ Qualitative: ODC6315 (1000 µg/L) or ODC6316 (500 µg/L). 1000 µg/L Cut off: ODC0006
OSR6323
Ecstasy Semi Quantitative: ODC6314-ODC6318 500 µg/L Cut-off: ODC0007
Qualitative: ODC6315 (300 µg/L) or ODC6316 (200 µg/L). 300 µg/L Cut-off: ODC0006
OSR6315 Barbiturates Semi Quantitative: ODC6314-ODC6318 200 µg/L Cut-off: ODC0007
Qualitative: ODC6315 (300 µg/L) or ODC6316 (200 µg/L). 300 µg/L Cut-off: ODC0006
OSR6316 Benzodiazepines Semi Quantitative: ODC6314 ODC6318 200 µg/L Cut-off: ODC0007
Qualitative: ODC6315 (300 µg/L) or ODC6316 (150 µg/L). 300 µg/L Cut off: ODC0006
OSR6317 Cocaine Semi Quantitative: ODC6314-ODC6318 150 µg/L Cut-off: ODC0007
Qualitative: ODC6315 or ODC6316 (both 100 µg/L).
OSR6318 EDDP Semi Quantitative: ODC6314, ODC6315 or ODC6316, ODC0006 or ODC0007
ODC6317, ODC6318
Qualitative: ODC6319 (300 µg/L).
OSR6319 Methadone Semi Quantitative: ODC6314, ODC6319-ODC6321 ODC0006

Qualitative: ODC6316 (300 µg/L).


OSR6320 Opiates Semi Quantitative: ODC6314, ODC6316-ODC6318 ODC0007

Qualitative: ODC6322 (25 µg/L) or ODC6323 (50 µg/L). 25 µg/L Cut-off: ODC0008
OSR6322 THC Semi Quantitative: ODC6314, ODC6322-ODC6325 50 µg/L Cut-off: ODC0009

- xii –
ACP
OSR6101 2 x 15 mL R1-1 ACP-T
2 x 15 mL R1-2 ACP-NP
4x R1-S ACP-S
1 x 10 mL ACP-St
Intended Use
Kinetic colour test for the quantitative determination of acid phosphatase, EC 3.1.3.2 (ACP), in human serum on Beckman Coulter analysers. For in vitro
diagnostic use only.

Summary1,2
The term acid phosphatase refers to a group of similar enzymes which cleave phosphate esters optimally at a pH below 7.0. These enzymes are present
in the lysosomes of all body cells with the highest activities in the liver, spleen, bone marrow, prostate, erythrocytes and platelets. Because the activities
from the prostate are inhibited by tartrate, the tartrate inhibited proportion of total ACP is also referred to as prostatic ACP. Elevations of the enzymatic
activity of prostatic ACP (and thus, generally, of total ACP activity) may be found in the sera of men with metastatic prostate cancer. The frequency of such
findings in a population of men with prostate cancer increases as the cancer progresses, thus prostatic ACP is useful in the staging of prostate carcinoma.
Serum prostatic ACP measurement is useful in monitoring remission or relapse of a prostatic malignancy and in assessing the effectiveness of various
treatment regimens.
Other diseases which can be associated with elevations of tartrate inhibited ACP include leukemia, polycythemia vera, primary thrombocythemia and
megaloblastic anemia.The majority of the normally low ACP activity of serum is of a tartrate-resistant type and probably originates mainly in osteoclasts.
Activities of this fraction are physiologically increased in growing children and pathologically in conditions of increased osteolysis and bone remodelling
such as Paget's disease, hyperparathyroidism, multiple myeloma, osteosarcoma, osteogenesis imperfecta and renal osteodystrophy. Other diseases
associated with elevated tartrate resistant ACP are Gaucher's disease and Niemann-Pick disease.

Test Principle3
Hillmann reaction
ACP
1-Naphthyl phosphate + H2O Phosphate + 1-Naphthol

1-Naphthol + FRTR salt Azo dye

Contents, Reagent Composition in the Test


Final concentration of reactive ingredients:
Total ACP method
Citrate buffer (pH 4.8) 140 mmol/L
1-Naphthyl Phosphate 11 mmol/L
Fast red TR salt 0.6 mmol/L
Pentanediol 210 mmol/L
Non-Prostatic ACP method
Tartrate 125 mmol/L
Citrate buffer (pH 4.8) 140 mmol/L
1-Naphthyl Phosphate 11 mmol/L
Fast Red TR salt 0.6 mmol/L
Pentanediol 210 mmol/L

Precautions and Warnings


Hazard Warnings and Risk Phrases:
R1-S (ACP substrate). Harmful, contains Fast Red TR salt. R68: Possible risk of irreversible effects.
Safety Phrases:
S36/37, S60. Wear suitable protective clothing and gloves. This material and its container must be disposed of as hazardous waste.
Exercise the normal precautions required for handling all laboratory reagents.
Dispose of all waste material in accordance with local guidelines.
Refer to Safety Data Sheets for further information.

Reagent Preparation
R1-1 (Total ACP)
To perform a total ACP (ACP-T) assay dissolve the contents of one R1-S vial (ACP substrate) completely with the contents of one bottle of R1-1 (Total
ACP buffer).
R1-2 (Non-Prostatic-ACP)
To perform a non-prostatic ACP (ACP-NP) assay dissolve the contents of one R1-S vial (ACP substrate) completely with the contents of one bottle of
R1-2 (Non-prostatic ACP buffer).
Attach the adaptor to the buffer bottle then connect the substrate powder vial to the other end. Gently mix the linked bottles until the powder is completely
dissolved. The solution can then be collected in the buffer bottle and placed on board the instrument.

Storage and Stability


The reagents are stable, unopened, up to the stated expiry date when stored at 2…8°C. Once prepared, reagents stored on board the instrument are
stable for 14 days.

Specimen
Serum. Do not use plasma.
Serum should be separated as soon as possible and 1 drop of stabiliser (ACP-St) added to 1mL of the sample.
Separated serum, without stabiliser, is stable for 15 min when stored at 15…25°C and 3-4 hours when stored at 2…8°C. Stabilised serum can be stored
4
for 8 days when stored at 2…25°C.
Lipemic, icteric and haemolysed samples should be avoided.

EN.01 BLOSR6x01.01 Enzyme


2009-08
Test Procedure
Refer to the appropriate User’s Guide and the accompanying Instrument Setting Sheet for analyser-specific assay instructions.

Calibration
The test is run in MB-mode, two analyser specific MB-factors (ACP-T & ACP-NP) are required. To provide a robust approach to generate the analyser
specific MB factor, it is recommended that 5 separate calibration events should be used. A fresh vial of calibrator, utilising System Calibrator Cat No. 66300
in the AB calibration mode, should be used for each of these runs. When calculating the mean factor from the separate runs the data should be examined
for obvious outliers which should be repeated and replaced. For the AU2700/AU5400 this procedure needs to be performed for each ring. Quality control
procedures should be undertaken immediately following calibration in accordance with good laboratory practice.
The calibrator value is traceable to a Beckman Coulter Master Calibrator.
Re-establishment of the analyser specific MB factor is recommended when a critical part of the analyser is replaced.
Reagent blank measurement is recommended when changing to a new lot of reagent.

Quality Control
controls Cat. No. ODC0003 and ODC0004 or other control materials with values determined by this Beckman Coulter system may be used.
Each laboratory should establish its own control frequency however good laboratory practice suggests that controls be tested each day patient
samples are tested and each time calibration/blanking is performed.
The results obtained by any individual laboratory may vary from the given mean value. It is therefore recommended that each laboratory
generates analyte specific control target values and intervals based on multiple runs according to their requirements. These target values should
fall within the corresponding acceptable ranges given in the relevant product literature.
If any trends or sudden shifts in values are detected, review all operating parameters.
Each laboratory should establish guidelines for corrective action to be taken if controls do not recover within the specified limits.

Calculation
The Beckman Coulter analysers automatically compute the ACP-T and ACP-NP activity of each sample. The activity of prostatic ACP can be generated by
the analyser when set as a calculated test:
Prostatic-ACP = Total-ACP – Non-Prostatic-ACP.

Reference Intervals1
Adults 37°C-Total ACP
Male ≤ 6.6 Pentanediol activation U/L (0.11 µkat/L)
Female ≤ 6.5 Pentanediol activation U/L (0.11 µkat/L)
Adults Tartrate-inhibited ACP (Prostatic ACP)
≤ 3.0 Pentanediol activation U/L (0.05 µkat/L)
Values obtained with different prostatic acid phosphatase assays cannot be used interchangeably. Before changing assays, laboratories must confirm
baseline values for patients being serially monitored.
Expected values may vary with age, sex, sample type, diet and geographical location. Each laboratory should verify the transferability of the expected
values to its own population, and if necessary determine its own reference interval according to good laboratory practice. For diagnostic purposes, results
should always be assessed in conjunction with the patient's medical history, clinical examinations and other findings.

Specific performance characteristics


Data contained within this section is representative of performance on Beckman Coulter systems. Data obtained in your laboratory may differ from these
values.
Linearity
The test is linear within an enzyme activity range of 0 - 100 U/L (0 - 1.67 µkat/L) for Total ACP and 0 - 30 U/L (0 - 0.5 μkat/L) for Prostatic ACP.
Precision
The following data was obtained on an AU600 using 3 serum pools analysed over 10 days.
Total ACP
n = 60 Within Run Total
Mean U/L SD CV% SD CV%
7.10 0.08 1.07 0.13 1.82
17.55 0.15 0.87 0.23 1.31
34.28 0.28 0.83 0.48 1.39
Prostatic ACP
n = 60 Within Run Total
Mean U/L SD CV% SD CV%
5.68 0.10 1.76 0.40 7.01
7.46 0.17 2.24 0.23 3.05
15.77 0.32 2.01 0.42 2.69
Sensitivity
The lowest detectable level on an AU600 analyser was estimated at 0.05 U/L for Total ACP and 0.10 U/L for Prostatic ACP.
The lowest detectable level represents the lowest measurable level of ACP that can be distinguished from zero. It is calculated as the absolute mean plus
three standard deviations of 20 replicates of an analyte free sample.
Method Comparison
Patient serum samples were used to compare this Total ACP OSR6101 assay on the AU600 against another commercially available Total ACP assay.
Results of linear regression analysis were as follows:
y = 0.944x – 0.271 r = 0.999 n = 116 Sample range = 0.58 – 58.51 U/L
Patient serum samples were used to compare this Prostatic OSR6101 assay on the AU600 against another commercially available Prostatic ACP assay.
Results of linear regression analysis were as follows:
y = 0.915x – 0.060 r = 0.966 n = 93 Sample range = 0.19 – 3.57 U/L

Interfering Substances
Results of studies conducted to evaluate the susceptibility of the Total ACP method to interference were as follows:
Icterus: Interference less than 10% up to 0.6 mg/dL or 10.3 µmol/L bilirubin
Ascorbate: Interference less than 10% up to 8 mg/dL ascorbate
Haemolysis: Interference less than 10% up to 4 g/L haemoglobin
®
Lipemia: Interference less than 10% up to 300 mg/dL Intralipid

Enzyme BLOSR6x01.01 EN.01


2009-08
Results of studies conducted to evaluate the susceptibility of the Prostatic ACP method to interference were as follows:
Icterus: Interference less than 10% up to 1 mg/dL or 17.1 µmol/L bilirubin
Ascorbate: Interference less than 10% up to 2 mg/dL ascorbate
Haemolysis: Interference less than 10% up to 3 g/L haemoglobin
®
Lipemia: Interference less than 10% up to 600 mg/dL Intralipid
5
Refer to Young for further information on interfering substances.

Limitations
Prostatic acid phosphatase results may be influenced by a number of factors other than malignancy and should always be interpreted in conjunction with
the patients clinical history and other diagnostic procedures. Do not use plasma.

Setting Sheet Footnotes


‡ For determination of Prostatic ACP the above parameters must be entered twice using test names ACPT (Total ACP) and ACPNP (Non-Prostatic
ACP). Set the test as a CALCULATED TEST in the INTER TESTS menu. See leaflet.
# User defined ¤ Analyser default value
§ For use in AB mode only, refer to leaflet for further instruction. Ensure that the correct calibrator value is entered for ACPT and ACPNP as
appropriate. Set factor ranges for Total ACP 800-1200* and Non-Prostatic ACP 1000-1550
* Values set for working in U/L. To work in SI units (µkat/L) divide by 60.
ж Set this test as COMMON TEST PARAMETER TEST NAME in CALCULATED TEST. Enter the formula (A-B), where A = ACPT and
B = ACPNP in Parameters Specific Test Parameters CALCULATED TEST.

BIBLIOGRAPHY
1. Thomas L. Saüre Phosphatase (SP). In:Thomas L, ed. Labor und Diagnose 6 Auflage:TH-Books Verlagsgesellschaft mbH Frankfürt/Main, 2005:118-120.
2. Moss DW, Henderson RA. Clinical enzymology. In: Burtis CA, Ashwood ER, eds. Tietz textbook of clinical chemistry. Philadelphia:WB Saunders Company,
1999;711-14.
3. Hillmann G. Fortlaufende photometrische messung der sauren prostataphosphatase-aktivität. Z Klin Chem u Klin Biochem 1971;9(3):273-274.
4. Young DS. Effects of preanalytical variables on clincal laboratory tests, 2nd ed.Washington: AACC press 1997:3-5pp.
5. Young DS. Effects of drugs on clinical laboratory tests, 5th ed. AACC Press, 2000.

EN.01 BLOSR6x01.01 Enzyme


2009-08
ACP, AU400/AU640 Serum Application ACP, AU600 Serum Application
System Reagent: OSR6101 System Reagent: OSR6101
Reagent ID: TOTAL (ACPT) 001, NON-PROSTATIC (ACPNP) 008 Reagent ID: TOTAL (ACPT) 001, NON-PROSTATIC (ACPNP) 008
Specific Test Parameters Specific test parameters
General LIH ISE Range
Test No # Test name ACP‡ ∇ Sample type Ser ∇ Page 1/2
Test Name: ACP‡ ∇ < > Type: Serum ∇ Operation: Yes ∇
Sample vol. 10 Dil. vol 0 μL Min. OD Max. OD
Sample: Volume 10 μL Dilution 0 μL Pre-Dilution Rate: 1 Reagent 1 vol 150 Dil. vol 0 μL L 0 H 2.5
Reagents: R1 Volume 150 μL Dilution 0 μL Min OD Max OD Reagent 2 vol 0 Dil. vol 0 μL Reagent OD limit
R2 Volume 0 μL Dilution 0 μL L 0 H 2.5 Fst. L -0.1 Fst. H 0.3
Reagent OD limit: Wave Main 410 Sub 800 ∇ Lst. L -0.1 Lst. H 0.3
Wavelength: Pri. 410 ∇ Sec. 800 ∇ First L -0.1 First H 0.3 Method RATE ∇ Dynamic range
Method: RATE ∇ Last L -0.1 Last H 0.3 Reaction + ∇ L 0* H 100*
Reaction slope: + ∇ Dynamic Range: Point 1 Fst 12 Lst 27 Correlation factor A 1
Measuring Point 1: First 12 Last 27 L 0* H 100* Point 2 Fst Lst B 0
Measuring Point 2: First Last Correlation Factor: Sample Pre-dil. Rate ¤
Linearity : 15 % A 1 B 0
No Lag Time: NO ∇ On-board stability period: 14 Linearity Fst 15 % Sec %
No lag time NO ∇ On-board stability period 14
Select using Space key, or select from list displayed by Guide key
Specific Test Parameters
General LIH ISE Range Test No # Test name ACP‡ ∇ Sample type Ser ∇ Page 2/2
Test Name: ACP‡ ∇ < > Type: Serum ∇ Level L Level H
Value/flag # ∇ # #
Value/Flag: # ∇ Level L: # Level H: # Normal range
Normal Ranges: Age L Age H Sex Age L Age H L H
Sex Year Month Year Month L H 1 # ∇ # Y # M # Y # M→ # #
ο 1. # ∇ # # # # # # 2 # ∇ # Y # M # Y # M→ # #
ο 2. # ∇ # # # # # # 3 # ∇ # Y # M # Y # M→ # #
ο 3. # ∇ # # # # # # 4 # ∇ # Y # M # Y # M→ # #
ο 4. # ∇ # # # # # # 5 # ∇ # Y # M # Y # M→ # #
ο 5. # ∇ # # # # # # 6 # ∇ # Y # M # Y # M→ # #
ο 6. # ∇ # # # # # # 7 Non select # #
7. None Selected # # 8 Out of range # #
8. Out of Range L H # # L H
Panic Value: # # Unit: U/L* Decimal places: # Panic value # #
Select the function using the Function key or the Mouse

SERUM APPLICATION
Calibration specific
Calibration Specific
General ISE Test No # Test name ACP‡ ∇
Test Name: ACP‡ ∇ < > Type Serum ∇ Cal type 1 MB ∇ Count #
Formula 1 Y=AX+B ∇ Process Conc ∇
Calibration Type: MB ∇ Formula: Y=AX+B ∇ Counts: # Process: CONC ∇
Selection calibrator
Cal No OD Conc Factor/OD-L Factor/OD-H
Cal. No. OD CONC Factor/OD-L Factor/OD-H
Point 1 § ∇ § § §
Point 1: § § § §
Point 2 ∇
Point 2:
Point 3: Point 3 ∇
Point 4: Point 4 ∇
Point 5: Point 5 ∇
Point 6: Point 6 ∇
Point 7: Point 7 ∇
1-Point Cal. Point: ο with CONC-0 Slope Check: None ∇ Advanced Calibration: # ∇ 1-point cal. point
MB type factor #
MB Type Factor: # Calibration Stability Period: Calibrator stability period
Select the function using the Function key or the Mouse
‡ For determination of Prostatic ACP the above parameters must be entered twice using test names ACPT (Total ‡ For determination of Prostatic ACP the above parameters must be entered twice using test names ACPT (Total
ACP) and ACPNP (Non-Prostatic ACP). Set the test as a CALCULATED TEST in the INTER TESTS menu, See leaflet. ACP) and ACPNP (Non-Prostatic ACP). Set the test as a CALCULATED TEST in the INTER TESTS menu. See leaflet.
# User defined # User defined ¤ Analyser default value
§ For use in AB mode only. Refer to IFU for further instruction. Ensure that the correct calibrator value is entered for § For use in AB mode only. Refer to IFU for further instruction. Ensure that the correct calibrator value is entered for
ACPT and ACPNP as appropriate. Set factor ranges for Total ACP 800-1200* and Non-Prostatic ACP 1000-1550* ACPT and ACPNP as appropriate. Set factor ranges for Total ACP 800-1200* and Non-Prostatic ACP 1000-1550*
* Values set for working in U/L. To work in SI units (µkat/L) divide by 60 * Values set for working in U/L. To work in SI units (µkat/L) divide by 60
Enzyme BSOSR6x01.01
2009-08
ACP, AU2700/AU5400 Serum Application
System Reagent: OSR6101
Reagent ID: TOTAL (ACPT) 001, NON-PROSTATIC (ACPNP) 008
Specific Test Parameters
General LIH ISE Range

Test Name: ACP‡ < > Type: Serum ∇ Operation: Yes ∇

Sample: Volume 8 μL Dilution 0 μL Pre-Dilution Rate: 1


Reagents: R1 Volume 120 μL Dilution 10 μL Min OD Max OD
R2 Volume 0 μL Dilution 0 μL L 0 H 2.5
Reagent OD limit: 0.3
Wavelength: Pri. 410 ∇ Sec. 800 ∇ First L -0.1 First H 0.3
Method: RATE ∇ Last L -0.1 Last H 0.3
Reaction slope: + ∇ Dynamic Range:
Measuring Point 1: First 12 Last 27 L 0* H 100*
Measuring Point 2: First Last Correlation Factor:
Linearity : 15 % A 1 B 0
No Lag Time: NO ∇ On-board stability period: 14

Specific Test Parameters


General LIH ISE Range

Test Name: ACP‡ ∇ < > Type: Serum ∇

Value/Flag: # ∇ Level L: # Level H: #


Normal Ranges: Age L Age H
Sex Year Month Year Month L H
ο 1. # ∇ # # # # # #
ο 2. # ∇ # # # # # #
ο 3. # ∇ # # # # # #
ο 4. # ∇ # # # # # #
ο 5. # ∇ # # # # # #
ο 6. # ∇ # # # # # #
7. None Selected # #
8. Out of Range L H # #
Panic Value: # # Unit: U/L* Decimal places: #

SERUM APPLICATION
Calibration Specific
General ISE

Test Name: ACP‡ ∇ < > Type Serum ∇

Calibration Type: MB ∇ Formula: Y=AX+B ∇ Counts: # Process: CONC ∇

Cal. No. OD CONC Factor/OD-L Factor/OD-H


Point 1: # § § §
Point 2:
Point 3:
Point 4:
Point 5:
Point 6:
Point 7:
1-Point Cal. Point: ο with CONC-0 Slope Check: None ∇ Advanced Calibration: # ∇
MB Type Factor: # Calibration Stability Period:

‡ For determination of Prostatic ACP the above parameters must be entered twice using test names ACPT (Total ACP)
and ACPNP (Non-Prostatic ACP). Set the test as a CALCULATED TEST in the INTER TESTS menu. See eaflet.
# User defined.
§ For use in AB mode only. Refer to IFU for further instruction Ensure that the correct calibrator value is entered for
ACPT and ACPNP as appropriate. Set factor ranges for Total ACP 800-1200* and Non-Prostatic ACP 1000-1550*
* Values set for working in U/L. To work in SI units (µkat/L) divide by 60

Enzyme BSOSR6x01.01
2009-08
ACP, AU680/AU480 Serum Application
System Reagent: OSR6101 Reagent ID: TOTAL (ACPT) 001, NON-PROSTATIC (ACPNP) 008
Parameters Specific Test Parameters
General LIH ISE HbA1c Calculated Test Range

Test Name: ACPж ∇ < > Type: Serum ∇ Operation Yes ∇

Sample Volume 8 μL Dilution 0 μL OD Limit


Pre-Dilution Rate 1 ∇ Min.OD 0 Max.OD 2.5
Rgt. Volume R1(R1-1) 120 μL Dilution 10 μL Reagent OD Limit
First Low -0.1 High 0.3
Last Low -0.1 High 0.3
R2(R2-1) 0 μL Dilution 0 μL
Dynamic Range Low 0* High 100*
Common Rgt. Type NoneΦ Name ф Correlation Factor A 1 B 0
Wavelength Pri 410 ∇nm Sec. 800 ∇nm Factor for Maker A 1 B 0
Method RATE ∇
Reaction Slope + ∇ Onboard Stability Period 14 Day 0 Hour
Measuring Point1 First 12 Last 27 LIH Influence Check # ∇
Measuring Point2 First Last Lipemia +++++ ∇
Linearity Limit 15 % Icterus + ∇
Lag Time Check NO ∇ Hemolysis ++++ ∇

Parameters Specific Test Parameters


General LIH ISE HbA1c Calculated Test Range

Test Name: ACPж ∇ < > Type: Serum ∇

Value/Flag: # ∇ Low High


Level # # Panic Value
Specificl Ranges: From To Low High
Sex Year Month Year Month Low High # #
ο 1. # ∇ # # # # # #
ο 2. # ∇ # # # # # #
ο 3. # ∇ # # # # # #
ο 4. # ∇ # # # # # #
ο 5. # ∇ # # # # # #
ο 6. # ∇ # # # # # #
7. No demographics # #
8. Not within expected values # #
Unit U/L* Decimal Places #
Parameters Calibration Parameters
Calibrators Calibration Specific STAT Table Calibration

SERUM APPLICATION
General ISE

Test Name: ACPж ∇ < > Type Serum ∇ ο Use Serum Cal.

Calibration Type: MB ∇ Formula: Y=AX+B ∇ Counts: # ∇


<Calibrator Parameters> Range
Calibrator OD Conc Low High Slope Check None ∇
Point 1: # ∇ § § §
Point 2: ∇ Allowance Range Check
Point 3: ∇
Point 4: ∇ ο Reagent Blank
Point 5: ∇ ο Calibration
Point 6: ∇
Point 7 ∇ Advanced Calibration
Point 8 ∇ Operation # ∇
Point 9 ∇
Point 10 ∇ Interval (RB/ACAL) # ∇
<Point Cal. For No. of Correction Points ∇ Use Master Curve ∇ ο Lot Calibration
Master Curve> OD Range ж Set this test as Common Test Parameter test name in CALCULATED TEST. Enter the formula (A-B) , where A=ACPT
Calibrator OD Conc Low High Stability and B=ACPNP in Parameters Specific Test Parameters CALCULATED TEST
Point-1 ∇ Reagent Blank Day Hour # User defined.
Point-2 ∇ Calibration Day Hour § For use in AB mode only. Refer to IFU for further instruction Ensure that the correct calibrator value is entered for
MB Type Factor: # 1-Point Calibration Point ∇ ο with Conc-0 ACPT and ACPNP as appropriate. Set factor ranges for Total ACP 800-1200* and Non-Prostatic ACP 1000-1550*
* Values set for working in U/L. To work in SI units (µkat/L) divide by 60
Ф AU680

Enzyme BSOSR6x01.01
2009-08
ALP
OSR6103 4 x 30 mL R1
4 x 30 mL R2
Intended Use
Kinetic colour test for the quantitative determination of alkaline phosphatase, EC 3.1.3.1 (ALP), in human serum and plasma on Beckman Coulter
analysers. For in vitro diagnostic use only.
Summary1,2
Alkaline phosphatase (ALP) is present in almost all body tissues, located at or in cell membranes. It occurs at particularly high levels in interstitial
epithelium, kidney tubules, bone (osteoblasts), liver and placenta. The precise metabolic function of ALP has not yet been fully elucidated, however the
enzyme is associated with intestinal lipid transport and bone calcification. ALP originates in approximately equal proportions from the liver and the skeletal
system. Approximately 25% of healthy individuals also have intestinal ALP which accounts for approximately 10% of the total ALP in a fasting sample.
Increases in total ALP are either due to physiological causes, or are caused by diseases of the liver or bone. Physiological increases in ALP are found in
pregnancy from the 2nd trimester onwards due to placental ALP, in growing children due to bone ALP and postprandially in individuals with blood groups
B and O, who are secretors of blood group substance H (intestinal ALP). The most common cause of elevated ALP is hepatobiliary disease, with
pathological ALP levels found in approximately 60% of patients with disease of the liver or biliary tract. ALP levels may also be elevated in primary bone
diseases, such as osteomalacia, osteogenesis imperfecta, vitamin D intoxication and primary bone tumours. ALP levels may also be increased in
secondary bone diseases, such as skeletal metastases, and in diseases such as multiple myeloma, acromegaly, renal insufficiency, hyperthyroidism,
ectopic ossification, sarcoidosis, bone tuberculosis and healing fractures. In bone diseases such as Paget’s disease, vitamin D deficiency rickets and
metastatic bone disease, ALP activity is a good indicator of bone activity, in the absence of co-existing chronic liver disease. Total ALP is only occasionally
elevated in some metabolic bone diseases such as hyperparathyroidism, osteopenia or osteoporosis. Reduced levels of ALP are found in familial
hypophosphatasia, hypoparathyroidism, achondroplasia, adynamic bone disease in dialysis patients, pituitary dwarfism, chronic radiation sickness and
malnutrition.
Test Principle3
Method based on the recommendations of the “German Society for Clinical Chemistry” (GSCC).
Alkaline phosphatase activity is determined by measuring the rate of conversion of p-Nitrophenyl phospate (pNPP) to p-Nitrophenol (pNP) in the presence
of magnesium ions and diethanolamine as phosphate acceptor at pH 9.8.
The rate of increase in absorbance due to the formation of pNP is measured at 410/480 nm and is directly proportional to the ALP activity in the sample.
Reaction Principle
ALP
p-Nitrophenyl phosphate + H2O Phosphate + p-Nitrophenol
2+
Mg
Contents, Reagent Composition in the Test
Final concentration of reactive ingredients:
Diethanolamine buffer, pH 9.8 1.0 mol/L
Magnesium chloride 0.5 mmol/L
p-Nitrophenyl phosphate 10 mmol/L
Preservative
Precautions and Warnings
Hazard Warnings and Risk Phrases:
R1 reagent: Harmful, contains 2,2’-iminodiethanol and a mixture of: 5-chloro-2-methyl-4-isothiazolin-3-one [EC No 247-500-7] and 2-methyl-4-isothiazolin-
3-one [EC No 220-239-6] (3:1). R22, R38, R41, R43, R48/22. Harmful if swallowed. Irritating to skin. Risk of serious damage to eyes. May cause
sensitisation by skin contact, Harmful: danger of serious damage to health by prolonged exposure if swallowed.
R2 reagent: Irritant, contains a mixture of: 5-chloro-2-methyl-4-isothiazolin-3-one [EC No 247-500-7] and 2-methyl-4-isothiazolin-3-one [EC No 220-239-6]
(3:1).R43. May cause sensitisation by skin contact.
Safety Phrases:
S24, S26, S37/39, S60.Avoid contact with skin, In case of contact with eyes, rinse immediately with plenty of water and seek medical advice. Wear suitable
gloves and eye/face protection. This material and its container must be disposed of as hazardous waste.
During the reaction p-nitrophenol is produced. This is harmful when inhaled, swallowed or absorbed through skin.
Dispose of all waste material in accordance with local guidelines.
Refer to Safety Data Sheets for further information.
Reagent Preparation
The reagents are ready for use and can be placed directly on board the instrument.
Storage and Stability
The reagents are stable, unopened, up to the stated expiry date when stored at 2…8°C.
Absorption of atmospheric CO2 by the reagent on board the analyser can impair its stability. This effect will vary depending upon the rate of use. Bottle
replacement is recommended when one of the following conditions are encountered:
14 days have elapsed on board the analyser,
Significant shift in control values (>7%) following local QC procedures,
Major preventative maintenance was performed on the analyser or a critical part was replaced.
Specimen
4
Serum and heparinised plasma. Complexing anticoagulants such as citrate, oxalate and EDTA should be avoided.
Haemolysed samples should be avoided.
5
Stable in serum and plasma for 7 days when stored at 2…25°C.
Test Procedure
Refer to the appropriate User Guide and Setting Sheet for analyser-specific assay instructions for the sample type as listed in the Intended Use statement.
The paediatric application is suitable for use with small volume serum/plasma samples.

EN.01 BLOSR6x03.01 Enzyme


2009-08
Calibration
The test is run in MB-mode. To provide a robust approach to generate the analyser specific MB factor, it is recommended that 5 separate calibration events
should be used. A fresh vial of calibrator, utilising System Calibrator Cat No. 66300 in the AB calibration mode, should be used for each of these runs.
When calculating the mean factor from the separate runs the data should be examined for obvious outliers which should be repeated and replaced. For the
AU2700/AU5400 this procedure needs to be performed for each ring. Quality control procedures should be undertaken immediately following calibration in
accordance with good laboratory practice.
The calibrator value is traceable to a Beckman Coulter Master Calibrator
Re-establishment of the analyser specific MB factor is recommended when a critical part of the analyser is replaced.
Reagent blank measurement is recommended when changing to a new lot of reagent.
Quality Control
Controls Cat. No. ODC0003 and ODC0004 or other control materials with values determined by this Beckman Coulter system may be used.
Each laboratory should establish its own control frequency however good laboratory practice suggests that controls be tested each day patient samples
are tested and each time calibration/blanking is performed.
The results obtained by any individual laboratory may vary from the given mean value. It is therefore recommended that each laboratory generates analyte
specific control target values and intervals based on multiple runs according to their requirements. These target values should fall within the corresponding
acceptable ranges given in the relevant product literature.
If any trends or sudden shifts in values are detected, review all operating parameters.
Each laboratory should establish guidelines for corrective action to be taken if controls do not recover within the specified limits.
Calculation
The Beckman Coulter analysers automatically compute the ALP activity of each sample.
Reference Intervals6,7
Women 64 - 300 U/L (1.0 – 5.0 µkat/L)
Men 80 – 300 U/L (1.3 – 5.0 µkat/L)
Children up to 15 years Up to 640 U/L (10.6 µkat/L)
Children 15-17 years Up to 480 U/L (8.0 µkat/L)
Expected values may vary with age, sex, sample type, diet and geographical location. Each laboratory should verify the transferability of the expected
values to its own population, and if necessary determine its own reference interval according to good laboratory practice. For diagnostic purposes, results
should always be assessed in conjunction with the patient's medical history, clinical examinations and other findings.
Specific Performance Characteristics
Data contained within this section is representative of performance on Beckman Coulter systems. Data obtained in your laboratory may differ from these
values.
Linearity
The test is linear within an enzyme activity range of 5 - 1500 U/L (0.1 – 25.0 µkat/L).
Precision
The following data was obtained on an AU600 using 3 serum pools analysed over 10 days.
n = 60 Within Run Total
Mean U/L SD CV% SD CV%
38 0.60 1.55 1.68 4.36
362 2.26 0.62 9.99 2.76
1088 5.81 0.53 23.40 2.15
Sensitivity
The lowest detectable level on an AU600 analyser was estimated at 1 U/L.
The lowest detectable level represents the lowest measurable level of ALP that can be distinguished from zero. It is calculated as the absolute mean plus
three standard deviations of 20 replicates of an analyte free sample.
Method Comparison
Patient serum samples were used to compare this ALP OSR6103 assay on the AU600 against another commercially available ALP assay. Results of
linear regression analysis were as follows:
y = 0.947x - 12 r = 0.999 n = 120 Sample range = 67 – 1404 U/L
Interfering Substances
Results of studies conducted to evaluate the susceptibility of the method to interference were as follows:
Icterus: Interference less than 5% up to 40 mg/dL or 684 µmol/L bilirubin
Haemolysis: Interference less than 10% up to 1 g/L haemoglobin
®
Lipemia: Interference less than 3% up to 1000 mg/dL Intralipid
8
Refer to Young for further information on interfering substances.
Setting Sheet Footnotes
# User defined ¤ Analyser default value
* Values set for working in U/L. To work in SI units (µkat/L) divide by 60.
§ For use in AB mode only, refer to leaflet for further instruction.
‡ Depends on usage pattern in the laboratory.
BIBLIOGRAPHY
1. Moss DW, Henderson RA. Clinical Enzymology. In: Burtis CA, Ashwood ER, eds. Tietz textbook of clinical chemistry. Philadelphia:WB Saunders Company,
1999; 676-684.
2. Thomas L. Alkaline phosphatase (ALP). In: Thomas L, ed. Clinical laboratory diagnostics. Use and assessment of clinical laboratory results. Frankfurt/Main:
TH-Books Verlagsgesellschaft, 1998:36-46.
3. Empfehlungen der Deutschen Gesellschaft für Klinische Chemie. Z Klin Chem u Klin Biochem 10 Jg 1972,182-192.
4. Moss DW, Henderson RA, Kachmar JF. Enzymes In: Tietz NW, ed. Fundamentals of clinical chemistry. Philadelphia:WB Saunders Company, 1987:387pp.
5. Ehret W, Heil W, Schmitt Y, Töpfer G, Wisser H, Zawta B, et al. Use of Anticoagulants in Diagnostic Laboratory Investigations and Stability of Blood, Plasma and
Serum Samples. WHO/DIL/LAB/99.1 Rev.2:21pp.
6. Schlebusch H, Rick W, Lang H, Knedel M. Standards in the activities of clinically important enzymes. Dtsch Med Wochenschr 1974;Apr 12;99(15):765-6.
7. Rick W, ed. Klinische chemie und mikroskopie, 6th ed. Berlin: Springer-Verlag,1990: 294pp.
8. Young DS. Effects of drugs on clinical laboratory tests, 5th ed. AACC Press, 2000.

Enzyme BLOSR6x03.01 EN.01


2009-08
ALP, AU400/AU640 Serum/Plasma Application ALP, AU600 Serum/Plasma Application
System Reagent: OSR6103 Reagent ID: 003 System Reagent: OSR6103 Reagent ID: 003

Specific Test Parameters Specific test parameters


General LIH ISE Range
Test No # Test name ALP03 ∇ Sample type Ser ∇ Page ½
Test Name: ALP03 ∇ < > Type: Serum ∇ Operation: Yes ∇
Sample vol. 3 Dil. Vol 0 μL Min. OD Max. OD
Sample: Volume 3 μL Dilution 0 μL Pre-Dilution Rate: 1 Reagent 1 vol 60 Dil. Vol 60 μL L -0.1 H 2.5
Reagents: R1 Volume 60 μL Dilution 60 μL Min OD Max OD Reagent 2 vol 60 Dil. Vol 60 μL Reagent OD limit
R2 Volume 60 μL Dilution 60 μL L -0.1 H 2.5 Fst. L -0.1 Fst. H 0.8
Reagent OD limit: Wave Main 410 Sub 480 ∇ Lst. L -0.1 Lst. H 0.8
Wavelength: Pri. 410 ∇ Sec. 480 ∇ First L -0.1 First H 0.8 Method RATE ∇ Dynamic range
Method: RATE ∇ Last L -0.1 Last H 0.8 Reaction + ∇ L 5* H 1500*
Reaction slope: + ∇ Dynamic Range: Point 1 Fst 13 Lst 19 Correlation factor A 1
Measuring Point 1: First 13 Last 19 L 5* H 1500* Point 2 Fst Lst B 0
Measuring Point 2: First Last Correlation Factor: Sample Pre-dil. Rate ¤
Linearity : 15 % A 1 B 0
No Lag Time: NO ∇ On-board stability period: 14 Linearity Fst 15 % Sec %
No lag time NO ∇ On-board stability period 14
Select using Space key, or select from list displayed by Guide key
Specific Test Parameters
General LIH ISE Range Test No # Test name ALP03 ∇ Sample type Ser ∇ Page 2/2
Test Name: ALP03 ∇ < > Type: Serum ∇ Level L Level H
Value/flag # ∇ # #
Value/Flag: # ∇ Level L: # Level H: # Normal range
Normal Ranges: Age L Age H Sex Age L Age H L H
Sex Year Month Year Month L H 1 # ∇ # Y # M # Y # M→ # #
ο 1. # ∇ # # # # # # 2 # ∇ # Y # M # Y # M→ # #
ο 2. # ∇ # # # # # # 3 # ∇ # Y # M # Y # M→ # #
ο 3. # ∇ # # # # # # 4 # ∇ # Y # M # Y # M→ # #
ο 4. # ∇ # # # # # # 5 # ∇ # Y # M # Y # M→ # #
ο 5. # ∇ # # # # # # 6 # ∇ # Y # M # Y # M→ # #
ο 6. # ∇ # # # # # # 7 Non select # #
7. None Selected # # 8 Out of range # #
8. Out of Range L H # # L H
Panic Value: # # Unit: U/L* Decimal places: # Panic value # #
Select the function using the Function key or the Mouse
Calibration specific
Calibration Specific
General ISE Test No # Test name ALP03 ∇
Test Name: ALP03 ∇ < > Type Serum ∇ Cal type 1 MB ∇ Count #

SERUM/PLASMA APPLICATION
Formula 1 Y=AX+B ∇ Process ∇
Calibration Type: MB ∇ Formula: Y=AX+B ∇ Counts: # Process: ∇
Selection calibrator
Cal No OD Conc Factor/OD-L Factor/OD-H
Cal. No. OD CONC Factor/OD-L Factor/OD-H
Point 1 § ∇ § §4300* §7000*
Point 1: § § §4300* §7000*
Point 2 ∇
Point 2:
Point 3: Point 3 ∇
Point 4: Point 4 ∇
Point 5: Point 5 ∇
Point 6: Point 6 ∇
Point 7: Point 7 ∇
1-Point Cal. Point: ο with CONC-0 Slope Check: None ∇ Advanced Calibration: # ∇ 1-point cal. Point
MB type factor #
MB Type Factor: # Calibration Stability Period: ‡ ‡
Calibrator stability period
Select the function using the Function key or the Mouse
# User defined
# User defined ¤ Analyser default value
* Values set for working in U/L. To work in SI units (µkat/L) divide by 60
* Values set for working in U/L. To work in SI units (µkat/L) divide by 60
§ For use in AB mode only, refer to leaflet for further instruction
§ For use in AB mode only, refer to leaflet for further instruction
‡ Depends on usage pattern in the Laboratory
‡ Depends on usage pattern in the Laboratory

Enzyme BSOSR6x03.01
2009-08
ALP, AU2700/AU5400 Serum/Plasma Application ALP, AU680/AU480 Serum/Plasma Application
System Reagent: OSR6103 Reagent ID: 003 System Reagent: OSR6103 Reagent ID: 003
Parameters Specific Test Parameters
Specific Test Parameters General LIH ISE HbA1c Calculated Test Range
General LIH ISE Range
Test Name: ALP03 ∇ < > Type: Serum ∇ Operation Yes ∇
Test Name: ALP03 ∇ < > Type: Serum ∇ Operation: Yes ∇

Sample: Volume 1.8 Dilution 0 Pre-Dilution Rate: 1 Sample Volume 1.8 μL Dilution 0 μL OD Limit
μL μL
Pre-Dilution Rate 1 ∇ Min.OD -0.1 Max.OD 2.5
Reagents: R1 Volume 36 μL Dilution 36 μL Min OD Max OD
Rgt. Volume R1(R1-1) 36 μL Dilution 36 μL Reagent OD Limit
R2 Volume 36 μL Dilution 36 μL L -0.1 H 2.5
First Low -0.1 High 0.8
Reagent OD limit:
Last Low -0.1 High 0.8
Wavelength: Pri. 410 ∇ Sec. 480 ∇ First L -0.1 First H 0.8
R2(R2-1) 36 μL Dilution 36 μL
Method: RATE ∇ Last L -0.1 Last H 0.8
Dynamic Range Low 5* High 1500*
Reaction slope: + ∇ Dynamic Range: Common Rgt. Type Noneф Name ф Correlation Factor A 1 B 0
Measuring Point 1: First 16 Last 27 L 5* H 1500* Wavelength Pri 410 ∇nm Sec. 480 ∇nm Factor for Maker A 1 B 0
Measuring Point 2: First Last Correlation Factor: Method RATE ∇
Linearity : 15 % A 1 B 0 Reaction Slope + ∇ Onboard Stability Period 14 Day 0 Hour
No Lag Time: NO ∇ On-board stability period: 14 Measuring Point1 First 16 Last 27 LIH Influence Check # ∇
Measuring Point2 First Last Lipemia +++++ ∇
Linearity Limit 15 % Icterus +++++ ∇
Specific Test Parameters Lag Time Check NO ∇ Hemolysis ++ ∇
General LIH ISE Range
Parameters Specific Test Parameters
Test Name: ALP03 ∇ < > Type: Serum ∇ General LIH ISE HbA1c Calculated Test Range

Value/Flag: # ∇ Level L: # Level H: # Test Name: ALP03 ∇ < > Type: Serum ∇
Normal Ranges: Age L Age H
Sex Year Month Year Month L H Value/Flag: # ∇ Low High
ο 1. # ∇ # # # # # # Level # # Panic Value
Specificl Ranges: From To Low High
ο 2. # ∇ # # # # # #
Sex Year Month Year Month Low High # #
ο 3. # ∇ # # # # # # # # # # # # #
ο 1. ∇
ο 4. # ∇ # # # # # # ο 2. # ∇ # # # # # #
ο 5. # ∇ # # # # # # ο 3. # ∇ # # # # # #
ο 6. # ∇ # # # # # # ο 4. # ∇ # # # # # #
7. None Selected # # ο 5. # ∇ # # # # # #
8. Out of Range L H # # ο 6. # ∇ # # # # # #
Panic Value: # # Unit: U/L* Decimal places: # 7. No demographics # #
8. Not within expected values # #
Unit U/L* Decimal Places #
Parameters Calibration Parameters
Calibration Specific
Calibrators Calibration Specific STAT Table Calibration
General ISE
General ISE

SERUM/PLASMA APPLICATION
Test Name: ALP03 ∇ < > Type Serum ∇ Test Name: ALP03 ∇ < > Type Serum ∇ ο Use Serum Cal.
Calibration Type: MB ∇ Formula: Y=AX+B ∇ Counts: # Process: ∇ Calibration Type: MB ∇ Formula: Y=AX+B ∇ Counts: # ∇
<Calibrator Parameters> Range
Cal. No. OD CONC Factor/OD-L Factor/OD-H Calibrator OD Conc Low High Slope Check None ∇
Point 1: § § §4300* §7000* Point 1: § ∇ § §4300* §7000*
Point 2: Point 2: ∇ Allowance Range Check
Point 3: Point 3: ∇
Point 4: Point 4: ∇ ο Reagent Blank
Point 5: Point 5: ∇ ο Calibration
Point 6: Point 6: ∇
Point 7: Point 7 ∇ Advanced Calibration
1-Point Cal. Point: ο with CONC-0 Slope Check: None ∇ Advanced Calibration: # ∇ Point 8 ∇ Operation # ∇
Point 9 ∇
MB Type Factor: # Calibration Stability Period: ‡
Point 10 ∇ Interval (RB/ACAL) # ∇
<Point Cal. For No. of Correction Points ∇ Use Master Curve ∇ ο Lot Calibration
# User defined Master Curve> OD Range
* Values set for working in U/L. To work in SI units (µkat/L) divide by 60 Calibrator OD Conc Low High Stability
§ For use in AB mode only, refer to leaflet for further instruction Point-1 ∇ Reagent Blank ‡ Day ‡ Hour
‡ Depends on usage pattern in the Laboratory Point-2 ∇ Calibration Day Hour
ф AU680 MB Type Factor: # 1-Point Calibration Point ∇ ο with Conc-0

Enzyme BSOSR6x03.01
2009-08
ALP, AU400/AU640 Paediatric Application ALP, AU600 Paediatric Application
System Reagent: OSR6103 Reagent ID: 003 System Reagent: OSR6103 Reagent ID: 003

Specific Test Parameters Specific test parameters


General LIH ISE Range
Test No # Test name ALP3P ∇ Sample type Ser ∇ Page 1/2
Test Name: ALP3P ∇ < > Type: Serum ∇ Operation: Yes ∇
Sample vol. 3 Dil. vol 10 μL Min. OD Max. OD
Sample: Volume 3 μL Dilution 10 μL Pre-Dilution Rate: 1 Reagent 1 vol 60 Dil. vol 60 μL L -0.1 H 2.5
Reagents: R1 Volume 60 μL Dilution 60 μL Min OD Max OD Reagent 2 vol 60 Dil. vol 50 μL Reagent OD limit
R2 Volume 60 μL Dilution 50 μL L -0.1 H 2.5 Fst. L -0.1 Fst. H 0.8
Reagent OD limit: Wave Main 410 Sub 480 ∇ Lst. L -0.1 Lst. H 0.8
Wavelength: Pri. 410 ∇ Sec. 480 ∇ First L -0.1 First H 0.8 Method RATE ∇ Dynamic range
Method: RATE ∇ Last L -0.1 Last H 0.8 Reaction + ∇ L 5* H 1500*
Reaction slope: + ∇ Dynamic Range: Point 1 Fst 13 Lst 19 Correlation factor A 1
Measuring Point 1: First 13 Last 19 L 5* H 1500* Point 2 Fst Lst B 0
Measuring Point 2: First Last Correlation Factor: Sample Pre-dil. Rate ¤
Linearity : 15 % A 1 B 0
No Lag Time: NO ∇ On-board stability period: 14 Linearity Fst 15 % Sec %
No lag time NO ∇ On-board stability period 14
Select using Space key, or select from list displayed by Guide key
Specific Test Parameters
General LIH ISE Range Test No # Test name ALP3P ∇ Sample type Ser ∇ Page 2/2
Test Name: ALP3P ∇ < > Type: Serum ∇ Level L Level H
Value/flag # ∇ # #
Value/Flag: # ∇ Level L: # Level H: # Normal range
Normal Ranges: Age L Age H Sex Age L Age H L H
Sex Year Month Year Month L H 1 # ∇ # Y # M # Y # M→ # #
ο 1. # ∇ # # # # # # 2 # ∇ # Y # M # Y # M→ # #
ο 2. # ∇ # # # # # # 3 # ∇ # Y # M # Y # M→ # #
ο 3. # ∇ # # # # # # 4 # ∇ # Y # M # Y # M→ # #
ο 4. # ∇ # # # # # # 5 # ∇ # Y # M # Y # M→ # #
ο 5. # ∇ # # # # # # 6 # ∇ # Y # M # Y # M→ # #
ο 6. # ∇ # # # # # # 7 Non select # #
7. None Selected # # 8 Out of range # #
8. Out of Range L H # # L H
Panic Value: # # Unit: U/L* Decimal places: # Panic value # #
Select the function using the Function key or the Mouse
Calibration specific
Calibration Specific
General ISE Test No # Test name ALP3P

PAEDIATRIC APPLICATION

Test Name: ALP3P ∇ < > Type Serum ∇ Cal type 1 MB ∇ Count #
Formula 1 Y=AX+B ∇ Process ∇
Calibration Type: MB ∇ Formula: Y=AX+B ∇ Counts: # Process: ∇
Selection calibrator
Cal No OD Conc Factor/OD-L Factor/OD-H
Cal. No. OD CONC Factor/OD-L Factor/OD-H
Point 1 § ∇ § §4300* §7000*
Point 1: § § §4300* §7000*
Point 2 ∇
Point 2:
Point 3: Point 3 ∇
Point 4: Point 4 ∇
Point 5: Point 5 ∇
Point 6: Point 6 ∇
Point 7: Point 7 ∇
1-Point Cal. Point: ο with CONC-0 Slope Check: None ∇ Advanced Calibration: # ∇ 1-point cal. point
MB type factor #
MB Type Factor: # Calibration Stability Period: ‡ ‡
Calibrator stability period
Select the function using the Function key or the Mouse
# User defined
# User defined ¤ Analyser default value
* Values set for working in U/L. To work in SI units (µkat/L) divide by 60
* Values set for working in U/L. To work in SI units (µkat/L) divide by 60
§ For use in AB mode only, refer to leaflet for further instruction
§ For use in AB mode only, refer to leaflet for further instruction
‡ Depends on usage pattern in the Laboratory
‡ Depends on usage pattern in the Laboratory

Enzyme BSOSR6x03.01
2009-08
ALP, AU2700/AU5400 Paediatric Application ALP, AU680/AU480 Paediatric Application
System Reagent: OSR6103 Reagent ID: 003 System Reagent: OSR6103 Reagent ID: 003
Parameters Specific Test Parameters
Specific Test Parameters General LIH ISE HbA1c Calculated Test Range
General LIH ISE Range
Test Name: ALP3P ∇ < > Type: Serum ∇ Operation Yes ∇
Test Name: ALP3P ∇ < > Type: Serum ∇ Operation: Yes ∇

Sample: Volume 1.8 Dilution 10 Pre-Dilution Rate: 1 Sample Volume 1.8 μL Dilution 10 μL OD Limit
μL μL
Pre-Dilution Rate 1 ∇ Min.OD -0.1 Max.OD 2.5
Reagents: R1 Volume 36 μL Dilution 36 μL Min OD Max OD
Rgt. Volume R1(R1-1) 36 μL Dilution 36 μL Reagent OD Limit
R2 Volume 36 μL Dilution 26 μL L -0.1 H 2.5
First Low -0.1 High 0.8
Reagent OD limit:
Last Low -0.1 High 0.8
Wavelength: Pri. 410 ∇ Sec. 480 ∇ First L -0.1 First H 0.8
R2(R2-1) 36 μL Dilution 26 μL
Method: RATE ∇ Last L -0.1 Last H 0.8
Dynamic Range Low 5* High 1500*
Reaction slope: + ∇ Dynamic Range: Common Rgt. Type Noneф Name ф Correlation Factor A 1 B 0
Measuring Point 1: First 16 Last 27 L 5* H 1500* Wavelength Pri 410 ∇nm Sec. 480 ∇nm Factor for Maker A 1 B 0
Measuring Point 2: First Last Correlation Factor: Method RATE ∇
Linearity : 15 % A 1 B 0 Reaction Slope + ∇ Onboard Stability Period 14 Day 0 Hour
No Lag Time: NO ∇ On-board stability period: 14 Measuring Point1 First 16 Last 27 LIH Influence Check # ∇
Measuring Point2 First Last Lipemia +++++ ∇
Linearity Limit 15 % Icterus +++++ ∇
Specific Test Parameters Lag Time Check NO ∇ Hemolysis ++ ∇
General LIH ISE Range
Parameters Specific Test Parameters
Test Name: ALP3P ∇ < > Type: Serum ∇ General LIH ISE HbA1c Calculated Test Range

Value/Flag: # ∇ Level L: # Level H: # Test Name: ALP3P ∇ < > Type: Serum ∇
Normal Ranges: Age L Age H
Sex Year Month Year Month L H Value/Flag: # ∇ Low High
ο 1. # ∇ # # # # # # Level # # Panic Value
Specificl Ranges: From To Low High
ο 2. # ∇ # # # # # #
Sex Year Month Year Month Low High # #
ο 3. # ∇ # # # # # # # # # # # # #
ο 1. ∇
ο 4. # ∇ # # # # # # ο 2. # ∇ # # # # # #
ο 5. # ∇ # # # # # # ο 3. # ∇ # # # # # #
ο 6. # ∇ # # # # # # ο 4. # ∇ # # # # # #
7. None Selected # # ο 5. # ∇ # # # # # #
8. Out of Range L H # # ο 6. # ∇ # # # # # #
Panic Value: # # Unit: U/L* Decimal places: # 7. No demographics # #
8. Not within expected values # #
Unit U/L* Decimal Places #
Parameters Calibration Parameters
Calibration Specific
Calibrators Calibration Specific STAT Table Calibration

PAEDIATRIC APPLICATION
General ISE
General ISE
Test Name: ALP3P ∇ < > Type Serum ∇ Test Name: ALP3P ∇ < > Type Serum ∇ ο Use Serum Cal.
Calibration Type: MB ∇ Formula: Y=AX+B ∇ Counts: # Process: ∇ Calibration Type: MB ∇ Formula: Y=AX+B ∇ Counts: # ∇
<Calibrator Parameters> Range
Cal. No. OD CONC Factor/OD-L Factor/OD-H Calibrator OD Conc Low High Slope Check None ∇
Point 1: § § §4300* §7000* Point 1: § ∇ § §4300* §7000*
Point 2: Point 2: ∇ Allowance Range Check
Point 3: Point 3: ∇
Point 4: Point 4: ∇ ο Reagent Blank
Point 5: Point 5: ∇ ο Calibration
Point 6: Point 6: ∇
Point 7: Point 7 ∇ Advanced Calibration
1-Point Cal. Point: ο with CONC-0 Slope Check: None ∇ Advanced Calibration: # ∇ Point 8 ∇ Operation # ∇
Point 9 ∇
MB Type Factor: # Calibration Stability Period: ‡
Point 10 ∇ Interval (RB/ACAL) # ∇
<Point Cal. For No. of Correction Points ∇ Use Master Curve ∇ ο Lot Calibration
# User defined Master Curve> OD Range
* Values set for working in U/L. To work in SI units (µkat/L) divide by 60 Calibrator OD Conc Low High Stability
§ For use in AB mode only, refer to leaflet for further instruction Point-1 ∇ Reagent Blank ‡ Day ‡ Hour
‡ Depends on usage pattern in the Laboratory Point-2 ∇ Calibration Day Hour
ф AU680 MB Type Factor: # 1-Point Calibration Point ∇ ο with Conc-0

Enzyme BSOSR6x03.01
2009-08
ALP
OSR6004 4 x 12 mL R1
4 x 12 mL R2
OSR6104 4 x 30 mL R1
4 x 30 mL R2
OSR6204 4 x 53 mL R1
4 x 53 mL R2
Intended Use
Kinetic colour test for the quantitative determination of alkaline phosphatase, EC 3.1.3.1 (ALP), in human serum and plasma on Beckman Coulter
analysers. For in vitro diagnostic use only.
Summary1,2
Alkaline phosphatase (ALP) is present in almost all body tissues, located at or in cell membranes. It occurs at particularly high levels in interstitial
epithelium, kidney tubules, bone (osteoblasts), liver and placenta. The precise metabolic function of ALP has not yet been fully elucidated, however the
enzyme is associated with intestinal lipid transport and bone calcification. ALP originates in approximately equal proportions from the liver and the skeletal
system. Approximately 25% of healthy individuals also have intestinal ALP which accounts for approximately 10% of the total ALP in a fasting sample.
Increases in total ALP are either due to physiological causes, or are caused by diseases of the liver or bone. Physiological increases in ALP are found in
pregnancy from the 2nd trimester onwards due to placental ALP, in growing children due to bone ALP and postprandially in individuals with blood groups B
and O, who are secretors of blood group substance H (intestinal ALP). The most common cause of elevated ALP is hepatobiliary disease, with
pathological ALP levels found in approximately 60% of patients with disease of the liver or biliary tract. ALP levels may also be elevated in primary bone
diseases, such as osteomalacia, osteogenesis imperfecta, vitamin D intoxication and primary bone tumours. ALP levels may also be increased in
secondary bone diseases, such as skeletal metastases, and in diseases such as multiple myeloma, acromegaly, renal insufficiency, hyperthyroidism,
ectopic ossification, sarcoidosis, bone tuberculosis and healing fractures. In bone diseases such as Paget’s disease, vitamin D deficiency rickets and
metastatic bone disease, ALP activity is a good indicator of bone activity, in the absence of co-existing chronic liver disease. Total ALP is only occasionally
elevated in some metabolic bone diseases such as hyperparathyroidism, osteopenia or osteoporosis. Reduced levels of ALP are found in familial
hypophosphatasia, hypoparathyroidism, achondroplasia, adynamic bone disease in dialysis patients, pituitary dwarfism, chronic radiation sickness and
malnutrition.
Test Principle3
Method based on the recommendations of the “International Federation for Clinical Chemistry” (IFCC).
Alkaline phosphatase activity is determined by measuring the rate of conversion of p-nitro-phenylphosphate (pNPP) to p-nitrophenol (pNP) in the presence
of magnesium and zinc ions and of 2-amino-2-methyl-1-propanol (AMP) as phosphate acceptor at
pH 10.4. The rate of change in absorbance due to the formation of pNP is measured bichromatically at 410/480 nm and is directly proportional to the ALP
activity in the sample.
Reaction Principle
ALP
pNPP + AMP pNP + AMP-PO4
2+
Mg
Contents, Reagent Composition in the Test
Final concentration of reactive ingredients:
2-Amino-2-Methyl-1-Propanol (AMP) pH 10.4 0.35 mol/L
p-Nitrophenyl phosphate 16 mmol/L
HEDTA 2 mmol/L
Zinc Sulphate 1 mmol/L
Magnesium Acetate 2 mmol/L
Preservative
Precautions and Warnings
Hazard Warnings and Risk Phrases:
R1 reagent: Irritant. R36/38. Irritating to eyes and skin.
R2 reagent: Irritant, contains a mixture of 5-chloro-2-methyl-4-isothiazolin-3-one [EC No 247-500-7] and 2-methyl-4-isothiazolin-3-one [EC No
220-239-6] (3:1). R43; May cause sensitisation by skin contact.
Safety Phrases:
S24,S26,S37,S60. Avoid contact with skin. In case of contact with eyes, rinse immediately with plenty of water and seek medical advice. Wear suitable
gloves. This material and its container must be disposed of as hazardous waste.
Exercise the normal precautions required for handling all laboratory reagents.
To avoid the possible build-up of azide compounds, flush waste-pipes with water after the disposal of undiluted reagent.
During the reaction p-nitrophenol is produced. This is harmful when inhaled, swallowed or absorbed through skin.
Dispose of all waste material in accordance with local guidelines.
Refer to Safety Data Sheets for further information.
Reagent Preparation
The reagents are ready for use and can be placed directly on board the instrument.
Storage and Stability
The reagents are stable, unopened, up to the stated expiry date when stored at 2…8 °C.
Absorption of atmospheric CO2 by the reagent on board the analyser can impair its stability. This effect will vary depending upon the rate of use. Bottle
replacement is recommended when one of the following conditions are encountered:
14 days have elapsed on board the analyser;
Significant shift in control values (>7 %) following local QC procedures;
Major preventative maintenance was performed on the analyser or a critical part was replaced.
Specimen
4
Serum and heparinised plasma. Complexing anticoagulants such as citrate, oxalate and EDTA should be avoided.
Strongly haemolysed samples should be avoided.
5
Stable in serum and plasma for 7 days when stored at 2…25 °C.
EN.01 BLOSR6x04.01 Enzyme
2009-08
Test Procedure
Refer to the appropriate User Guide and Setting Sheet for analyser-specific assay instructions for the sample type as listed in the Intended Use
statement. The paediatric application is suitable for use with small volume serum/plasma samples.
Calibration
The test is run in MB-mode. To provide a robust approach to generate the analyser specific MB factor, it is recommended that 5 separate calibration events
should be used. A fresh vial of calibrator, utilising System Calibrator Cat No. 66300 in the AB calibration mode, should be used for each of these runs.
When calculating the mean factor from the separate runs the data should be examined for obvious outliers which should be repeated and replaced. For the
AU2700/AU5400 this procedure needs to be performed for each ring. Quality control procedures should be undertaken immediately following calibration in
accordance with good laboratory practice.
The calibrator value is traceable to a Beckman Coulter Master Calibrator.
Re-establishment of the analyser specific MB factor is recommended when a critical part of the analyser is replaced;
Reagent blank measurement is recommended when changing to a new lot of reagent.
Quality Control
Controls Cat. No. ODC0003 and ODC0004 or other control materials with values determined by this Beckman Coulter system may be used.
Each laboratory should establish its own control frequency.
Good laboratory practice suggests that controls be tested each day patient samples are tested and each time calibration is performed. Values obtained for
the controls should fall within specified limits as defined by the user. If any trends or sudden shifts in values are detected, review all operating parameters.
Each laboratory should establish guidelines for corrective action to be taken if controls do not recover within the specified limits.
Calculation
The Beckman Coulter analysers automatically compute the alkaline phosphatase activity of each sample.
Reference Intervals2
Adults > 17y : 30 – 120 U/L

Children Male (U/L) Female (U/L)


1 – 30d 75 – 316 48 – 406
30d – 1y 82 – 383 124 – 341
1 – 3y 104 – 345 108 – 317
4 – 6y 93 – 309 96 – 297
7 – 9y 86 – 315 69 – 325
10 – 12y 42 – 362 51 – 332
13 – 15y 74 – 390 50 – 162
16 – 18y 52 – 171 47 – 119
Expected values may vary with age, sex, sample type, diet and geographical location. Each laboratory should verify the transferability of the expected
values to its own population, and if necessary determine its own reference interval according to good laboratory practice. For diagnostic purposes, results
should always be assessed in conjunction with the patient's medical history, clinical examinations and other findings.
Specific performance characteristics
Data contained within this section is representative of performance on Beckman Coulter systems. Data obtained in your laboratory may differ from these
values.
Linearity
The test is linear within an enzyme activity range of 5 – 1500 U/L (0.1 – 25.0 µkat/L).
Precision
The following data was obtained on an AU600 using 3 serum pools analysed over 10 days.
n = 60 Within Run Total
Mean U/L SD CV% SD CV%
21 0.40 1.94 0.93 4.48
260 1.45 0.56 2.57 0.99
856 7.67 0.90 11.09 1.30
Sensitivity
The lowest detectable level on an AU600 analyser was estimated at 1 U/L.
The lowest detectable level represents the lowest measurable level of ALP that can be distinguished from zero. It is calculated as the absolute mean plus
three standard deviations of 20 replicates of an analyte free sample.
Method Comparison
Patient serum samples were used to compare this ALP OSR6104 assay on the AU600 against another commercially available ALP assay. Results of
linear regression analysis were as follows:
y = 0.934x + 8 r = 0.999 n = 124 Sample range = 17 – 1500 U/L
Interfering Substances
Results of studies conducted to evaluate the susceptibility of the method to interference were as follows:
Icterus: Interference less than 10% up to 28 mg/dL or 479 µmol/L bilirubin
Haemolysis: Interference less than 10% up to 4.5 g/L haemoglobin
®
Lipemia: Interference less than 3% up to 1000 mg/dL Intralipid
6
Refer to Young for further information on interfering substances.
Setting Sheet Footnotes
# User defined ¤ Analyser default value
* Values set for working in U/L. To work in SI units (µkat/L) divide by 60.
§ For use in AB mode only, refer to leaflet for further instruction.
‡ Depends on usage pattern in the laboratory.
BIBLIOGRAPHY
1. Moss DW, Henderson RA. Clinical Enzymology. In: Burtis CA, Ashwood ER, eds. Tietz textbook of clinical chemistry. Philadelphia:WB Saunders Company, 1999; 676-684.
2. Thomas L. Alkaline phosphatase (ALP). In: Thomas L, ed. Clinical laboratory diagnostics. Use and assessment of clinical laboratory results. Frankfurt/Main:
TH-Books Verlagsgesellschaft, 1998:36-46.
3. Tietz NW, Rinker D, Shaw LM. IFCC methods for the measurement of catalytic concentration of enzymes Part 5. IFCC method for alkaline phosphatase.
J Clin Chem Clin Biochem 1983;21:731-48.
4. Moss DW, Henderson RA, Kachmar JF. Enzymes In: Tietz NW, ed. Fundamentals of clinical chemistry. Philadelphia:WB Saunders Company, 1987:387pp.
5. Ehret W, Heil W, Schmitt Y, Töpfer G, Wisser H, Zawta B, et al. Use of Anticoagulants in Diagnostic Laboratory Investigations and Stability of Blood, Plasma and Serum
Samples. WHO/DIL/LAB/99.1 Rev.2:21pp.
6. Young DS. Effects of drugs on clinical laboratory tests, 5th ed. AACC Press, 2000.

Enzyme BLOSR6x04.01 EN.01


2009-08
ALP (IFCC), AU400/AU640 Serum/Plasma Application ALP (IFCC), AU600 Serum/Plasma Application
System Reagent: OSR6004, OSR6104, OSR6204 Reagent ID: 004 System Reagent: OSR6004, OSR6104, OSR6204 Reagent ID: 004
Specific Test Parameters Specific test parameters
General LIH ISE Range
Test No # Test name ALP04 ∇ Sample type Ser ∇ Page ½
Test Name: ALP04 ∇ < > Type: Serum ∇ Operation: Yes ∇
Sample vol. 3 Dil. Vol 0 μL Min. OD Max. OD
Sample: Volume 3 μL Dilution 0 μL Pre-Dilution Rate: 1 Reagent 1 vol 60 Dil. Vol 60 μL L 0.0 H 2.5
Reagents: R1 Volume 60 μL Dilution 60 μL Min OD Max OD Reagent 2 vol 60 Dil. Vol 60 μL Reagent OD limit
R2 Volume 60 μL Dilution 60 μL L 0.0 H 2.5 Fst. L 0.0 Fst. H 1.2
Reagent OD limit: Wave Main 410 Sub 480 ∇ Lst. L 0.0 Lst. H 1.2
Wavelength: Pri. 410 ∇ Sec. 480 ∇ First L 0.0 First H 1.2 Method RATE ∇ Dynamic range
Method: RATE ∇ Last L 0.0 Last H 1.2 Reaction + ∇ L 5* H 1500*
Reaction slope: + ∇ Dynamic Range: Point 1 Fst 13 Lst 19 Correlation factor A 1
Measuring Point 1: First 13 Last 19 L 5* H 1500* Point 2 Fst Lst B 0
Measuring Point 2: First Last Correlation Factor: Sample Pre-dil Rate ¤
Linearity : 15 % A 1 B 0
No Lag Time: NO ∇ On-board stability period: 14 Linearity Fst 15 % Sec %
No lag time NO ∇ On-board stability period 14
Select using Space key, or select from list displayed by Guide key
Specific Test Parameters
General LIH ISE Range Test No # Test name ALP04 ∇ Sample type Ser ∇ Page 2/2
Test Name: ALP04 ∇ < > Type: Serum ∇ Level L Level H
Value/flag # ∇ # #
Value/Flag: # ∇ Level L: # Level H: # Normal range
Normal Ranges: Age L Age H Sex Age L Age H L H
Sex Year Month Year Month L H 1 # ∇ # Y # M # Y # M→ # #
ο 1. # ∇ # # # # # # 2 # ∇ # Y # M # Y # M→ # #
ο 2. # ∇ # # # # # # 3 # ∇ # Y # M # Y # M→ # #
ο 3. # ∇ # # # # # # 4 # ∇ # Y # M # Y # M→ # #
ο 4. # ∇ # # # # # # 5 # ∇ # Y # M # Y # M→ # #
ο 5. # ∇ # # # # # # 6 # ∇ # Y # M # Y # M→ # #
ο 6. # ∇ # # # # # # 7 Non select # #
7. None Selected # # 8 Out of range # #
8. Out of Range L H # # L H
Panic Value: # # Unit: U/L* Decimal places: # Panic value # #
Select the function using the Function key or the Mouse
Calibration specific
Calibration Specific
General ISE Test No # Test name ALP04 ∇
Test Name: ALP04 Type Serum

SERUM/PLASMA APPLICATION
∇ < > ∇ Cal type 1 MB ∇ Count #
Formula 1 Y=AX+B ∇ Process Conc ∇
Calibration Type: MB ∇ Formula: Y=AX+B ∇ Counts: # Process: CONC ∇
Selection calibrator
Cal No OD Conc Factor/OD-L Factor/OD-H
Cal. No. OD CONC Factor/OD-L Factor/OD-H
Point 1 § ∇ § § 4000* § 6500*
Point 1: § § § 4000* § 6500*
Point 2 ∇
Point 2:
Point 3: Point 3 ∇
Point 4: Point 4 ∇
Point 5: Point 5 ∇
Point 6: Point 6 ∇
Point 7: Point 7 ∇
1-Point Cal. Point: ο with CONC-0 Slope Check: None ∇ Advanced Calibration # ∇ 1-point cal. Point
MB type factor #
MB Type Factor: # Calibration Stability Period: ‡ ‡
Calibrator stability period
Select the function using the Function key or the Mouse
# User defined
# User defined ¤ Analyser default value
§ For use in the AB mode only, refer to IFU for further instruction
§ For use in the AB mode only, refer to IFU for further instruction
* Values set for working in U/L. To work in SI units (µkat/L) divide by 60
* Values set for working in U/L. To work in SI units (µkat/L) divide by 60
‡ Depends on usage pattern in the Laboratory
‡ Depends on usage pattern in the Laboratory

Enzyme BSOSR6x04.01
2009-08
ALP (IFCC), AU2700/AU5400 Serum/Plasma Application ALP (IFCC), AU680/AU480 Serum/Plasma Application
System Reagent: OSR6004, OSR6104, OSR6204 Reagent ID: 004 System Reagent: OSR6004, OSR6104, OSR6204 Reagent ID: 004
Parameters Specific Test Parameters
Specific Test Parameters General LIH ISE HbA1c Calculated Test Range
General LIH ISE Range
Test Name: ALP04 ∇ < > Type: Serum ∇ Operation Yes ∇
Test Name: ALP04 < > Type: Serum ∇ Operation: Yes ∇

Sample: Volume 1.8 Dilution 0 Pre-Dilution Rate: 1 Sample Volume 1.8 μL Dilution 0 μL OD Limit
μL μL
Pre-Dilution Rate 1 ∇ Min.OD -0.1 Max.OD 2.5
Reagents: R1 Volume 36 μL Dilution 36 μL Min OD Max OD
Rgt. Volume R1(R1-1) 36 μL Dilution 36 μL Reagent OD Limit
R2 Volume 36 μL Dilution 36 μL L -0.1 H 2.5
First Low -0.1 High 1.2
Reagent OD limit: 0.3
Last Low -0.1 High 1.2
Wavelength: Pri. 410 ∇ Sec. 480 ∇ First L -0.1 First H 1.2
R2(R2-1) 36 μL Dilution 36 μL
Method: RATE ∇ Last L -0.1 Last H 1.2
Dynamic Range Low 5* High 1500*
Reaction slope: + ∇ Dynamic Range: Common Rgt. Type Noneф Name ф Correlation Factor A 1 B 0
Measuring Point 1: First 16 Last 27 L 5* H 1500* Wavelength Pri 410 ∇nm Sec. 480 ∇nm Factor for Maker A 1 B 0
Measuring Point 2: First Last Correlation Factor: Method RATE ∇
Linearity : 15 % A 1 B 0 Reaction Slope + ∇ Onboard Stability Period 14 Day 0 Hour
No Lag Time: NO ∇ On-board stability period: 14 Measuring Point1 First 16 Last 27 LIH Influence Check # ∇
Measuring Point2 First Last Lipemia +++++ ∇
Linearity Limit 15 % Icterus ++++ ∇
Specific Test Parameters Lag Time Check NO ∇ Hemolysis +++++ ∇
General LIH ISE Range
Parameters Specific Test Parameters
Test Name: ALP04 ∇ < > Type: Serum ∇ General LIH ISE HbA1c Calculated Test Range

Value/Flag: # ∇ Level L: # Level H: # Test Name: ALP04 ∇ < > Type: Serum ∇
Normal Ranges: Age L Age H
Sex Year Month Year Month L H Value/Flag: # ∇ Low High
ο 1. # ∇ # # # # # # Level # # Panic Value
Specificl Ranges: From To Low High
ο 2. # ∇ # # # # # #
Sex Year Month Year Month Low High # #

SERUM/PLASMA
ο 3. # ∇ # # # # # # # # # # # # #
ο 1. ∇
ο 4. # ∇ # # # # # # ο 2. # ∇ # # # # # #
ο 5. # ∇ # # # # # # ο 3. # ∇ # # # # # #
ο 6. # ∇ # # # # # # ο 4. # ∇ # # # # # #
7. None Selected # # ο 5. # ∇ # # # # # #
8. Out of Range L H # # ο 6. # ∇ # # # # # #
Panic Value: # # Unit: U/L* Decimal places: # 7. No demographics # #
8. Not within expected values # #
Unit U/L* Decimal Places #
Parameters Calibration Parameters
Calibration Specific
Calibrators Calibration Specific STAT Table Calibration
General ISE
General ISE
Test Name: ALP04 ∇ < > Type Serum ∇

SERUM/PLASMA APPLICATION
Test Name: ALP04 Type Serum Use Serum Cal.

APPLICATION
∇ < > ∇ ο
Calibration Type: MB ∇ Formula: Y=AX+B ∇ Counts: # Process: CONC ∇ Calibration Type: MB ∇ Formula: Y=AX+B ∇ Counts: # ∇
<Calibrator Parameters> Range
Cal. No. OD CONC Factor/OD-L Factor/OD-H Calibrator OD Conc Low High Slope Check None ∇
Point 1: § § § 4000* § 6500* Point 1: § ∇ § §4000* §6500*
Point 2: Point 2: ∇ Allowance Range Check
Point 3: Point 3: ∇
Point 4: Point 4: ∇ ο Reagent Blank
Point 5: Point 5: ∇ ο Calibration
Point 6: Point 6: ∇
Point 7: Point 7 ∇ Advanced Calibration
1-Point Cal. Point: ο with CONC-0 Slope Check: None ∇ Advanced Calibration: # ∇ Point 8 ∇ Operation # ∇
Point 9 ∇
MB Type Factor: # Calibration Stability Period: ‡
Point 10 ∇ Interval (RB/ACAL) # ∇
<Point Cal. For No. of Correction Points ∇ Use Master Curve ∇ ο Lot Calibration
# User defined Master Curve> OD Range
§ For use in the AB mode only, refer to IFU for further instruction Calibrator OD Conc Low High Stability
* Values set for working in U/L To work in SI units (µkat/L) divide by 60 Point-1 ∇ Reagent Blank ‡ Day ‡ Hour
‡ Depends on usage pattern in the Laboratory Point-2 ∇ Calibration Day Hour
ф AU680
MB Type Factor: # 1-Point Calibration Point ∇ ο with Conc-0

Enzyme BSOSR6x04.01
2009-08
ALP (IFCC), AU400/AU640 Paediatric Application ALP (IFCC), AU600 Paediatric Application
System Reagent: OSR6004, OSR6104, OSR6204 Reagent ID: 004 System Reagent: OSR6004, OSR6104, OSR6204 Reagent ID: 004

Specific Test Parameters Specific test parameters


General LIH ISE Range
Test No # Test name ALP4P ∇ Sample type Ser ∇ Page 1/2
Test Name: ALP4P ∇ < > Type: Serum ∇ Operation: Yes ∇
Sample vol. 3 Dil. vol 10 μL Min. OD Max. OD
Sample: Volume 3 μL Dilution 10 μL Pre-Dilution Rate: 1 Reagent 1 vol 60 Dil. vol 60 μL L 0.0 H 2.5
Reagents: R1 Volume 60 μL Dilution 60 μL Min OD Max OD Reagent 2 vol 60 Dil. vol 50 μL Reagent OD limit
R2 Volume 60 μL Dilution 50 μL L 0.0 H 2.5 Fst. L 0.0 Fst. H 1.2
Reagent OD limit: Wave Main 410 Sub 480 ∇ Lst. L 0.0 Lst. H 1.2
Wavelength: Pri. 410 ∇ Sec. 480 ∇ First L 0.0 First H 1.2 Method RATE ∇ Dynamic range
Method: RATE ∇ Last L 0.0 Last H 1.2 Reaction + ∇ L 5* H 1500*
Reaction slope: + ∇ Dynamic Range: Point 1 Fst 13 Lst 19 Correlation factor A 1
Measuring Point 1: First 13 Last 19 L 5* H 1500* Point 2 Fst Lst B 0
Measuring Point 2: First Last Correlation Factor: Sample Pre-dil Rate ¤
Linearity : 15 % A 1 B 0
No Lag Time: NO ∇ On-board stability period: 14 Linearity Fst 15 % Sec %
No lag time NO ∇ On-board stability period 14
Select using Space key, or select from list displayed by Guide key
Specific Test Parameters
General LIH ISE Range Test No # Test name ALP4P ∇ Sample type Ser ∇ Page 2/2
Test Name: ALP4P ∇ < > Type: Serum ∇ Level L Level H
Value/flag # ∇ # #
Value/Flag: # ∇ Level L: # Level H: # Normal range
Normal Ranges: Age L Age H Sex Age L Age H L H
Sex Year Month Year Month L H 1 # ∇ # Y # M # Y # M→ # #
ο 1. # ∇ # # # # # # 2 # ∇ # Y # M # Y # M→ # #
ο 2. # ∇ # # # # # # 3 # ∇ # Y # M # Y # M→ # #
ο 3. # ∇ # # # # # # 4 # ∇ # Y # M # Y # M→ # #
ο 4. # ∇ # # # # # # 5 # ∇ # Y # M # Y # M→ # #
ο 5. # ∇ # # # # # # 6 # ∇ # Y # M # Y # M→ # #
ο 6. # ∇ # # # # # # 7 Non select # #
7. None Selected # # 8 Out of range # #
8. Out of Range L H # # L H
Panic Value: # # Unit: U/L* Decimal places: # Panic value # #
Select the function using the Function key or the Mouse
Calibration specific
Calibration Specific
General ISE Test No # Test name ALP4P ∇

PAEDIATRIC APPLICATION
Test Name: ALP4P ∇ < > Type Serum ∇ Cal type 1 MB ∇ Count #
Formula 1 Y=AX+B ∇ Process Conc ∇
Calibration Type: MB ∇ Formula: Y=AX+B ∇ Counts: # Process: CONC ∇
Selection calibrator
Cal No OD Conc Factor/OD-L Factor/OD-H
Cal. No. OD CONC Factor/OD-L Factor/OD-H
Point 1 § ∇ § § 4000* § 6500*
Point 1: § § § 4000* § 6500*
Point 2 ∇
Point 2:
Point 3: Point 3 ∇
Point 4: Point 4 ∇
Point 5: Point 5 ∇
Point 6: Point 6 ∇
Point 7: Point 7 ∇
1-Point Cal. Point: ο with CONC-0 Slope Check: None ∇ Advanced Calibration: # ∇ 1-point cal. point
MB type factor #
MB Type Factor: # Calibration Stability Period: ‡ ‡
Calibrator stability period
Select the function using the Function key or the Mouse
# User defined
# User defined ¤ Analyser default value
§ For use in the AB mode only, refer to IFU for further instruction
§ For use in the AB mode only, refer to IFU for further instruction
* Values set for working in U/L. To work in SI units (µkat/L) divide by 60
* Values set for working in U/L. To work in SI units (µkat/L) divide by 60
‡ Depends on usage pattern in the Laboratory
‡ Depends on usage pattern in the Laboratory

Enzyme BSOSR6x04.01
2009-08
ALP (IFCC), AU2700/AU5400 Paediatric Application ALP (IFCC), AU680/AU480 Paediatric Application
System Reagent: OSR6004, OSR6104, OSR6204 Reagent ID: 004 System Reagent: OSR6004, OSR6104, OSR6204 Reagent ID: 004
Parameters Specific Test Parameters
General LIH ISE HbA1c Calculated Test Range
Specific Test Parameters
Test Name: ALP4P ∇ < > Type: Serum ∇ Operation Yes ∇
General LIH ISE Range

Test Name: ALP4P < > Type: Serum ∇ Operation: Yes ∇


Sample Volume 1.8 μL Dilution 10 μL OD Limit
Sample: Volume 1.8 μL Dilution 10 μL Pre-Dilution Rate: 1 Pre-Dilution Rate 1 ∇ Min.OD -0.1 Max.OD 2.5
Reagents: R1 Volume 36 μL Dilution 36 μL Min OD Max OD Rgt. Volume R1(R1-1) 36 μL Dilution 36 μL Reagent OD Limit
R2 Volume 36 Dilution 26 L -0.1 H 2.5 First Low -0.1 High 1.2
μL μL
Last Low -0.1 High 1.2
Reagent OD limit: 0.3
R2(R2-1) 36 μL Dilution 26 μL
Wavelength: Pri. 410 ∇ Sec. 480 ∇ First L -0.1 First H 1.2
Dynamic Range Low 5* High 1500*
Method: RATE ∇ Last L -0.1 Last H 1.2
Common Rgt. Type Noneф Name ф Correlation Factor A 1 B 0
Reaction slope: + ∇ Dynamic Range:
Wavelength Pri 410 ∇nm Sec. 480 ∇nm Factor for Maker A 1 B 0
Measuring Point 1: First 16 Last 27 L 5* H 1500*
Method RATE ∇
Measuring Point 2: First Last Correlation Factor:
Reaction Slope + ∇ Onboard Stability Period 14 Day 0 Hour
Linearity : 15 % A 1 B 0
Measuring Point1 First 16 Last 27 LIH Influence Check # ∇
No Lag Time: NO ∇ On-board stability period: 14
Measuring Point2 First Last Lipemia +++++ ∇
Linearity Limit 15 % Icterus ++++ ∇
Lag Time Check NO ∇ Hemolysis +++++ ∇
Specific Test Parameters
General LIH ISE Range Parameters Specific Test Parameters
General LIH ISE HbA1c Calculated Test Range
Test Name: ALP4P ∇ < > Type: Serum ∇
Test Name: ALP4P ∇ < > Type: Serum ∇
Value/Flag: # ∇ Level L: # Level H: #
Normal Ranges: Age L Age H Value/Flag: # ∇ Low High
Sex Year Month Year Month L H Level # # Panic Value
ο 1. # ∇ # # # # # # Specificl Ranges: From To Low High
Sex Year Month Year Month Low High # #
ο 2. # ∇ # # # # # #
ο 1. # ∇ # # # # # #
ο 3. # ∇ # # # # # #
ο 2. # ∇ # # # # # #
ο 4. # ∇ # # # # # # # # # # # # #
ο 3. ∇
ο 5. # ∇ # # # # # # ο 4. # ∇ # # # # # #
ο 6. # ∇ # # # # # # ο 5. # ∇ # # # # # #
7. None Selected # # ο 6. # ∇ # # # # # #
8. Out of Range L H # # 7. No demographics # #
Panic Value: # # Unit: U/L* Decimal places: # 8. Not within expected values # #
Unit U/L* Decimal Places #
Parameters Calibration Parameters
Calibration Specific Calibrators Calibration Specific STAT Table Calibration
General ISE
General ISE

PAEDIATRIC APPLICATION
Test Name: ALP4P ∇ < > Type Serum ∇ ο Use Serum Cal.
Test Name: ALP4P ∇ < > Type Serum ∇
Calibration Type: MB ∇ Formula: Y=AX+B ∇ Counts: # ∇
Calibration Type: MB ∇ Formula: Y=AX+B ∇ Counts: # Process: CONC ∇
<Calibrator Parameters> Range
Calibrator OD Conc Low High Slope Check None ∇
Cal. No. OD CONC Factor/OD-L Factor/OD-H
Point 1: § ∇ § §4000* §6500*
Point 1: § § § 4000* § 6500*
Point 2: ∇ Allowance Range Check
Point 2: Point 3: ∇
Point 3: Point 4: ∇ ο Reagent Blank
Point 4: Point 5: ∇ ο Calibration
Point 5: Point 6: ∇
Point 6: Point 7 ∇ Advanced Calibration
Point 7: Point 8 ∇ Operation # ∇
1-Point Cal. Point: ο with CONC-0 Slope Check: None ∇ Advanced Calibration: # ∇ Point 9 ∇
MB Type Factor: # Calibration Stability Period: ‡ Point 10 ∇ Interval (RB/ACAL) # ∇
<Point Cal. For No. of Correction Points ∇ Use Master Curve ∇ ο Lot Calibration
Master Curve> OD Range
# User defined Calibrator OD Conc Low High Stability
§ For use in the AB mode only, refer to IFU for further instruction Point-1 ∇ Reagent Blank ‡ Day ‡ Hour
* Values set for working in U/L. To work in SI units (µkat/L) divide by 60 Point-2 ∇ Calibration Day Hour
‡ Depends on usage pattern in the Laboratory MB Type Factor: # 1-Point Calibration Point ∇ ο with Conc-0
ф AU680

Enzyme BSOSR6x04.01
2009-08
ALT
OSR6007 4 x 12 mL R1
4 x 6 mL R2
OSR6107 4 x 50 mL R1
4 x 25 mL R2
OSR6507 4 x 102 mL R1
4 x 52 mL R2
OSR60180 4 x 17 mL Liquid P-5-P R1-2
60106 6 x 4 mL Liquid Pyridoxal Phosphate
60100 4 x 25 Tablets Pyridoxal Phosphate
Intended Use
Kinetic UV test for the quantitative determination of alanine aminotransferase, EC 2.6.1.2 (ALT), in human serum and plasma on Beckman Coulter
analysers. For in vitro diagnostic use only.
OSR6507 for use on the AU2700 and AU5400 systems only. OSR60180 for use with 3-part-reagent enabled systems only.

Summary1,2,3
ALT is an aminotransferase, a group of enzymes which catalyse the reversible transformation of α-keto acids into amino acids by transfer of amino groups.
Since the specific activity of ALT in the liver is approximately 10 times that of heart and skeletal muscle, elevated serum ALT activity is mainly regarded as
an indicator of parenchymal liver disease. ALT is present in the cytosol of hepatocytes, and increased serum levels indicate deterioration in the integrity of
the hepatocyte plasma membrane. ALT has greater diagnostic sensitivity for hepatobiliary disease than AST. Activities >50 times the upper reference limit
are mainly associated with acute viral hepatitis, acute disorders of liver perfusion and acute liver necrosis due to ingestion of toxins including paracetamol
and carbon tetrachloride. Markedly elevated serum ALT levels may be found in a variety of diseases involving the liver, including hepatitis, mononucleosis
and cirrhosis. Elevated ALT levels may be detected in viral hepatitis and other forms of liver disease prior to development of overt clinical symptoms such
as jaundice. Levels greater than 15 times the upper reference limit are always indicative of acute hepatocellular necrosis of viral, toxic or circulatory origin.
Increased ALT levels may also be detected in cirrhosis and extrahepatic cholestasis. Slight or moderate increases in ALT levels may also be observed
after ingestion of alcohol, or administration of drugs including penicillin, salicylates or opiates.

Test Principle4
Method based on the recommendations of the “International Federation for Clinical Chemistry” (IFCC).
ALT transfers the amino group from alanine to 2-oxoglutarate to form pyruvate and glutamate. The addition of pyridoxal phosphate to the reaction mixture
ensures maximum catalytic activity of ALT. The pyruvate enters a lactate dehydrogenase (LDH) catalysed reaction with NADH to produce lactate and
+
NAD . The decrease in absorbance due to the consumption of NADH is measured at 340 nm and is proportional to the ALT activity in the sample.
Endogenous pyruvate is removed during the incubation period.

Reaction Principle
ALT
2-Oxoglutarate + L-Alanine L-Glutamate + Pyruvate

+ LDH +
Pyruvate + NADH + H L-lactate + NAD

Contents, Reagent Composition in the Test


Final concentration of reactive ingredients:
Tris buffer, pH: 7.15 (37°C) 100 mmol/L
L- Alanine 500 mmol/L
2-Oxoglutarate 12 mmol/L
LDH ≥ 1.8 kU/L
NADH 0.20 mmol/L
Pyridoxal Phosphate (P-5-P) 0.1 mmol/L (when Cat. No. 60106, 60100 or OSR60180 is used)
Preservative

Precautions and Warnings


Exercise the normal precautions required for handling all laboratory reagents.
To avoid the possible build-up of azide compounds, flush waste-pipes with water after the disposal of undiluted reagent.
Dispose of all waste material in accordance with local guidelines.
Safety data sheet available for professional user on request.

Reagent Preparation
With manual pyridoxal phosphate addition
Pyridoxal Phosphate Liquid (Cat. No. 60106) and Pyridoxal Phosphate tablets (Cat No. 60100) are supplied separately for pyridoxal phosphate activation.
Pipette Pyridoxal Phosphate Liquid 60106 into the R1 bottle according to the table below, and mix by gentle inversion. Alternatively, dissolve tablets
completely in the R1 bottle according to the table below, by mixing gently several times. R2 is ready for use and can be placed directly on board the
instrument. This method can also be used for 3-part-reagent enabled systems.
Cat. No. Volume Pyridoxal Phosphate Liquid No. of Pyridoxal Phosphate tablets
OSR6007 0.25 mL 1 tablet
OSR6107 1 mL 4 tablets
OSR6507 2 mL 8 tablets

Without pyridoxal phosphate activation


The reagents are ready for use and can be placed directly on board the instrument.

EN.01 BLOSR6x07.01 Enzyme


2009-08
Storage and Stability
Reagents are stable, unopened, up to the stated expiry date when stored at 2…8°C.
Without pyridoxal phosphate activation
Once open, reagents stored on board the instrument are stable for 30 days.
With pyridoxal phosphate activation
After addition of pyridoxal phosphate, R1 stored on board the instrument is stable for 7 days.
R2 stored on board the instrument is stable for 30 days.
Pyridoxal Phosphate Liquid reagent (Cat. No 60106)
Once open, Pyridoxal Phosphate Liquid reagent is stable until the expiry date printed on the label, provided that contamination is avoided through
adherence to GLP, the cap is replaced immediately after use and the reagent is stored at 2…8°C.
3-part-reagent enabled systems with P-5-P activation
Reagent Preparation
P-5-P Liquid (Cat. No. OSR60180) is specifically for use on board 3-part-reagent enabled systems. The reagent is ready for use and can be placed directly
on board the instrument in the R1 carousel. R1 and R2 are also ready for use and can be placed directly on board the instrument.
Storage and stability
Once open, P-5-P (Cat no OSR60180) is stable on board the instrument for 60 days. R1 and R2 reagent stored on board the instrument are stable for
30 days.
Specimen
Serum, and EDTA or heparinised plasma.
5
Stable in serum and plasma for 7 days when stored at 2…8°C and 3 days when stored at 15…25°C.

Test Procedure
Refer to the appropriate User Guide and Setting Sheet for analyser-specific assay instructions for the sample type as listed in the Intended Use statement.
The paediatric application is suitable for use with small volume serum/plasma samples.

Calibration
The test is run in MB-mode. To provide a robust approach to generate the analyser specific MB factor it is recommended that 5 separate calibration events
should be used. A fresh vial of calibrator, utilising System Calibrator Cat No. 66300 in the AB calibration mode, should be used for each of these runs.
When calculating the mean factor from the separate runs the data should be examined for obvious outliers which should be repeated and replaced. For the
AU2700/AU5400 this procedure needs to be performed for each ring. Quality control procedures should be undertaken immediately following calibration in
accordance with good laboratory practice.
With pyridoxal phosphate activation, the calibrator value is traceable to the IFCC reference method and IRMM/IFCC-454. Without pyridoxal phosphate
activation, the calibrator is traceable to a Beckman Coulter Master Calibrator.
Re-establishment of the analyser specific MB factor is recommended when a critical part of the analyser is replaced.
Reagent blank measurement is recommended when changing to a new lot of reagent.

Quality Control
Controls Cat. No. ODC0003 and ODC0004 or other control materials with values determined by this Beckman Coulter system may be used.
Each laboratory should establish its own control frequency however good laboratory practice suggests that controls be tested each day patient samples
are tested and each time calibration/blanking is performed.
The results obtained by any individual laboratory may vary from the given mean value. It is therefore recommended that each laboratory generates analyte
specific control target values and intervals based on multiple runs according to their requirements. These target values should fall within the corresponding
acceptable ranges given in the relevant product literature.
If any trends or sudden shifts in values are detected, review all operating parameters.
Each laboratory should establish guidelines for corrective action to be taken if controls do not recover within the specified limits.

Calculation
The Beckman Coulter analysers automatically compute the ALT activity of each sample.

Reference Intervals6,7
Male (Adult) < 50 U/L (0.85 µkat/L)
Female (Adult) < 35 U/L (0.60 µkat/L)
Newborn/Infant 13 − 45 U/L (0.22 − 0.75 µkat/L)
Expected values may vary with age, sex, sample type, diet and geographical location. Each laboratory should verify the transferability of the expected
values to its own population, and if necessary determine its own reference interval according to good laboratory practice. For diagnostic purposes, results
should always be assessed in conjunction with the patient's medical history, clinical examinations and other findings.

Specific performance characteristics


Data contained within this section is representative of performance on Beckman Coulter systems. Data obtained in your laboratory may differ from these
values.

Linearity
The test is linear within an enzyme activity range of 3 – 500 U/L (0.05 – 8.33 µkat/L).

Precision
The following data was obtained on an AU600 using 3 serum pools analysed over 10 days.
n = 60 Within Run Total
Mean U/L SD CV% SD CV%
18 0.53 2.95 0.57 3.16
56 0.80 1.43 1.00 1.78
459 2.88 0.63 3.67 0.80

Sensitivity
The lowest detectable level on an AU600 analyser was estimated at 1 U/L.
The lowest detectable level represents the lowest measurable level of ALT that can be distinguished from zero. It is calculated as the absolute mean plus
three standard deviations of 20 replicates of an analyte free sample.

Method Comparison with pyridoxal phosphate activation


Patient serum samples were used to compare ALT OSR6107 assay on the AU640 against the IFCC reference method. Results of linear regression
analysis were as follows:
y = 1.010x - 0.1 r = 0.999 n = 117 Sample range = 3 – 264 U/L
Enzyme BLOSR6x07.01 EN.01
2009-08
Interfering Substances
Results of studies conducted to evaluate the susceptibility of the method to interference were as follows:
Icterus: Interference less than 5% up to 40 mg/dL or 684 µmol/L bilirubin
Haemolysis: Interference less than 10% up to 5 g/L haemoglobin
®
Lipemia: Interference less than 3% up to 300 mg/dL Intralipid
Pyruvate: Interference less than 5% up to 1 mmol/L pyruvate
8
Refer to Young for further information on interfering substances.

Limitations
Highly lipemic samples may exceed the reaction absorbance and will be flagged with a “@”. Such samples should be diluted and re-run.

Setting Sheet Footnotes


# User defined ¤ Analyser default value
* Values set for working in U/L. To work in SI units (µkat/L) divide by 60.
§ For use in AB mode only, refer to leaflet for further instruction

BIBLIOGRAPHY
1. Thomas L. Alanine aminotransferase (ALT), Aspartate aminotransferase (AST). In:Thomas L, ed. Clinical laboratory diagnostics. Use and assessment of clinical
laboratory results. Frankfurt/Main: TH-Books Verlagsgesellschaft, 1998:55-65.
2. Moss DW, Henderson RA, Kachmar JF. Enzymes. In: Tietz NW, ed. Fundamentals of clinical chemistry. Philadelphia:WB Saunders Company, 1987:369-373.
3. Schmidt E, Schmidt FW. Diagnosis of icteric diseases. Dtsch Med Wschr 1984; 109: 139-146.
4. Schumann G, Bonora R, Ceriotti F et al. IFCC Primary Reference Procedures for the Measurement of Catalytic Activity Concentrations of Enzymes at 37°C.
Part 4. Reference Procedure for the Measurement of Catalytic Concentration of Alanine Aminotransferase. Clin Chem Lab Med 2002;40:718–24.
5. Ehret W, Heil W, Schmitt Y, Töpfer G, Wisser H, Zawta B, et al. Use of anticoagulants in diagnostic laboratory investigations and stability of blood, plasma and
serum samples. WHO/DIL/LAB/99.1 Rev.2:21pp.
6. Thomas L, Müller M, Schumann G et al. Consensus of DGKL and VDGH for interim reference intervals on enzymes in serum. J Lab Med 2005;29:301-08.
7. Painter PC, Cope JY, Smith JL. Reference information for the clinical laboratory. In: Burtis CA, Ashwood ER, eds. Tietz textbook of clinical chemistry.
Philadelphia:WB Saunders Company, 1999; 1800pp.
8. Young DS. Effects of drugs on clinical laboratory tests, 5th ed. AACC Press, 2000.

EN.01 BLOSR6x07.01 Enzyme


2009-08
ALT (IFCC, without Pyridoxal phosphate activation), AU400/AU640 ALT (IFCC, without Pyridoxal phosphate activation), AU600
Serum/Plasma Application Serum/Plasma Application
System Reagent: OSR6007, OSR6107 Reagent ID: 007 System Reagent: OSR6007, OSR6107 Reagent ID: 007
Specific Test Parameters Specific test parameters
General LIH ISE Range
Test No # Test name ALT ∇ Sample type Ser ∇ Page ½
Test Name: ALT ∇ < > Type: Serum ∇ Operation: Yes ∇
Sample vol. 10 Dil. Vol 0 µL Min. OD Max. OD
Sample: Volume 10 µL Dilution 0 µL Pre-Dilution Rate: 1 Reagent 1 vol 100 Dil. Vol 50 µL L 0.60 H 2.5
Reagents: R1 Volume 100 µL Dilution 50 µL Min OD Max OD Reagent 2 vol 50 Dil. Vol 50 µL Reagent OD limit
R2 Volume 50 µL Dilution 50 µL L 0.60 H 2.5 Fst. L 1.1 Fst. H 2.5
Reagent OD limit: Wave Main 340 Sub 660 ∇ Lst. L 1.1 Lst. H 2.5
Wavelength: Pri. 340 ∇ Sec. 660 ∇ First L 1.1 First H 2.5 Method RATE ∇ Dynamic range
Method: RATE ∇ Last L 1.1 Last H 2.5 Reaction - ∇ L 3* H 500*
Reaction slope: - ∇ Dynamic Range: Point 1 Fst 14 Lst 27 Correlation factor A 1
Measuring Point 1: First 14 Last 27 L 3* H 500* Point 2 Fst Lst B 0
Measuring Point 2: First Last Correlation Factor: Sample Pre-dil. Rate ¤
Linearity : 15 % A 1 B 0
No Lag Time: YES ∇ On-board stability period: 30 Linearity Fst 15 % Sec %
No lag time NO ∇ On-board stability period 30
Select using Space key, or select from list displayed by Guide key
Specific Test Parameters
General LIH ISE Range Test No # Test name ALT ∇ Sample type Ser ∇ Page 2/2
Test Name: ALT ∇ < > Type: Serum ∇ Level L Level H
Value/flag # ∇ # #
Value/Flag: # ∇ Level L: # Level H: # Normal range
Normal Ranges: Age L Age H Sex Age L Age H L H
Sex Year Month Year Month L H 1 # ∇ # Y # M # Y # M→ # #
R 1. # ∇ # # # # # # 2 # ∇ # Y # M # Y # M→ # #
R 2. # ∇ # # # # # # 3 # ∇ # Y # M # Y # M→ # #
R 3. # ∇ # # # # # # 4 # ∇ # Y # M # Y # M→ # #
R 4. # ∇ # # # # # # 5 # ∇ # Y # M # Y # M→ # #
R 5. # ∇ # # # # # # 6 # ∇ # Y # M # Y # M→ # #
R 6. # ∇ # # # # # # 7 Non select # #
7. None Selected # # 8 Out of range # #
8. Out of Range L H # # L H
Panic Value: # # Unit: U/L* Decimal places: # Panic value # #
Select the function using the Function key or the Mouse
Calibration specific
Calibration Specific
General ISE Test No # Test name ALT ∇
Test Name: ALT ∇ < > Type Serum ∇ Cal type 1 MB ∇ Count #

SERUM/PLASMA APPLICATION
Formula 1 Y=AX+B ∇ Process ∇
Calibration Type: MB ∇ Formula: Y=AX+B ∇ Counts: # Process: ∇
Selection calibrator
Cal No OD Conc Factor/OD-L Factor/OD-H
Cal. No. OD CONC Factor/OD-L Factor/OD-H
Point 1 § ∇ § §3650* §5470*
Point 1: § § §3650* §5470*
Point 2: Point 2 ∇
Point 3: Point 3 ∇
Point 4: Point 4 ∇
Point 5: Point 5 ∇
Point 6: Point 6 ∇
Point 7: Point 7 ∇
1-Point Cal. Point: R with CONC-0 Slope Check: None ∇ Advanced Calibration: # ∇ 1-point cal. Point
MB type factor #
MB Type Factor: # Calibration Stability Period:
Calibrator stability period
Select the function using the Function key or the Mouse
# User defined # User defined ¤ Analyser default value
* Values set for working in U/L. To work in SI units (µkat/L) divide by 60 * Values set for working in U/L. To work in SI units (µkat/L) divide by 60
§ For use in AB mode only, refer to IFU for further instruction. § For use in AB mode only, refer to IFU for further instruction.

Enzyme BSOSR6x07.02
2010-05
ALT (IFCC, without Pyridoxal phosphate activation), AU2700/AU5400 ALT (IFCC, without Pyridoxal phosphate activation), AU680/AU480
Serum/Plasma Application Serum/Plasma Application
System Reagent: OSR6007, OSR6107, OSR6507 Reagent ID: 007 System Reagent: OSR6007, OSR6107, OSR6507 Reagent ID: 007
Parameters Specific Test Parameters
Specific Test Parameters General LIH ISE HbA1c Calculated Test Range
General LIH ISE Range
Test Name: ALT ∇ < > Type: Serum ∇ Operation Yes ∇
Test Name: ALT ∇ < > Type: Serum ∇ Operation: Yes ∇

Sample: Volume 5 Dilution 0 Pre-Dilution Rate: 1 Sample Volume 5 µL Dilution 0 µL OD Limit


µL µL
Pre-Dilution Rate 1 ∇ Min.OD 0.60 Max.OD 2.5
Reagents: R1 Volume 50 µL Dilution 25 µL Min OD Max OD
Rgt. Volume R1(R1-1) 50 µL Dilution 25 µL Reagent OD Limit
R2 Volume 25 µL Dilution 25 µL L 0.60 H 2.5
First Low 1.1 High 2.5
Reagent OD limit:
Last Low 1.1 High 2.5
Wavelength: Pri. 340 ∇ Sec. 660 ∇ First L 1.1 First H 2.5
R2(R2-1) 25 µL Dilution 25 µL
Method: RATE ∇ Last L 1.1 Last H 2.5
Dynamic Range Low 3* High 500*
Reaction slope: - ∇ Dynamic Range: Common Rgt. Type Noneф Name ф Correlation Factor A 1 B 0
Measuring Point 1: First 14 Last 27 L 3* H 500* Wavelength Pri 340 ∇nm Sec. 660 ∇nm Factor for Maker A 1 B 0
Measuring Point 2: First Last Correlation Factor: Method RATE ∇
Linearity : 15 % A 1 B 0 Reaction Slope - ∇ Onboard Stability Period 30 Day 0 Hour
No Lag Time: YES ∇ On-board stability period: 30 Measuring Point1 First 14 Last 27 LIH Influence Check # ∇
Measuring Point2 First Last Lipemia +++++ ∇
Linearity Limit 15 % Icterus +++++ ∇
Specific Test Parameters Lag Time Check YES ∇ Hemolysis +++++ ∇
General LIH ISE Range
Parameters Specific Test Parameters
Test Name: ALT ∇ < > Type: Serum ∇ General LIH ISE HbA1c Calculated Test Range

Value/Flag: # ∇ Level L: # Level H: # Test Name: ALT ∇ < > Type: Serum ∇
Normal Ranges: Age L Age H
Sex Year Month Year Month L H Value/Flag: # ∇ Low High
R 1. # ∇ # # # # # # Level # # Panic Value
R 2. Specificl Ranges: From To Low High
# ∇ # # # # # #
Sex Year Month Year Month Low High # #
R 3. # ∇ # # # # # # R 1. # ∇ # # # # # #
R 4. # ∇ # # # # # # R 2. # # # # # # #

R 5. # ∇ # # # # # # R 3. # ∇ # # # # # #
R 6. # ∇ # # # # # # R 4. # ∇ # # # # # #
7. None Selected # # R 5. # ∇ # # # # # #
8. Out of Range L H # # R 6. # ∇ # # # # # #
Panic Value: # # Unit: U/L* Decimal places: # 7. No demographics # #
8. Not within expected values # #
Unit U/L* Decimal Places #
Parameters Calibration Parameters
Calibration Specific
Calibrators Calibration Specific STAT Table Calibration
General ISE
General ISE
Test Name: ALT ∇ < > Type Serum ∇

SERUM/PLASMA APPLICATION
Test Name: ALT ∇ < > Type Serum ∇ R Use Serum Cal.
Calibration Type: MB ∇ Formula: Y=AX+B ∇ Counts: # Process: ∇ Calibration Type: MB ∇ Formula: Y=AX+B ∇ Counts: # ∇
<Calibrator Parameters> Range
Cal. No. OD CONC Factor/OD-L Factor/OD-H Calibrator OD Conc Low High Slope Check None ∇
Point 1: § § §3650* §5470* Point 1: § ∇ § §3650* §5470*
Point 2: Point 2: ∇ Allowance Range Check
Point 3: Point 3: ∇
Point 4: Point 4: ∇ R Reagent Blank
Point 5: Point 5: ∇ R Calibration
Point 6: Point 6: ∇
Point 7: Point 7 ∇ Advanced Calibration
1-Point Cal. Point: R with CONC-0 Slope Check: None ∇ Advanced Calibration: # ∇ Point 8 ∇ Operation # ∇
Point 9 ∇
MB Type Factor: # Calibration Stability Period:
Point 10 ∇ Interval (RB/ACAL) # ∇
<Point Cal. For No. of Correction Points ∇ Use Master Curve ∇ R Lot Calibration
# User defined Master Curve> OD Range
* Values set for working in U/L. To work in SI units (µkat/L) divide by 60 Calibrator OD Conc Low High Stability
§ For use in AB mode only, refer to IFU for further instruction. Point-1 ∇ Reagent Blank Day Hour
Ф AU680 Point-2 ∇ Calibration Day Hour
MB Type Factor: # 1-Point Calibration Point ∇ R with Conc-0

Enzyme BSOSR6x07.02
2010-05
ALT (IFCC, with OE60106 activation), AU400/AU640 Serum/Plasma ALT (IFCC, with OE60106 activation), AU600
Application Serum/Plasma Application
System Reagent: OSR6007, OSR6107 Reagent ID: 007 System Reagent: OSR6007, OSR6107 Reagent ID: 007

Specific Test Parameters Specific test parameters


General LIH ISE Range
Test No # Test name ALT ∇ Sample type Ser ∇ Page ½
Test Name: ALT ∇ < > Type: Serum ∇ Operation: Yes ∇
Sample vol. 10 Dil. Vol 0 µL Min. OD Max. OD
Sample: Volume 10 µL Dilution 0 µL Pre-Dilution Rate: 1 Reagent 1 vol 100 Dil. Vol 50 µL L 1.0 H 2.5
Reagents: R1 Volume 100 µL Dilution 50 µL Min OD Max OD Reagent 2 vol 50 Dil. Vol 50 µL Reagent OD limit
R2 Volume 50 µL Dilution 50 µL L 1.0 H 2.5 Fst. L 1.3 Fst. H 2.5
Reagent OD limit: Wave Main 340 Sub 660 ∇ Lst. L 1.3 Lst. H 2.5
Wavelength: Pri. 340 ∇ Sec. 660 ∇ First L 1.3 First H 2.5 Method RATE ∇ Dynamic range
Method: RATE ∇ Last L 1.3 Last H 2.5 Reaction - ∇ L 3* H 500*
Reaction slope: - ∇ Dynamic Range: Point 1 Fst 14 Lst 27 Correlation factor A 1
Measuring Point 1: First 14 Last 27 L 3* H 500* Point 2 Fst Lst B 0
Measuring Point 2: First Last Correlation Factor: Sample Pre-dil. Rate ¤
Linearity : 15 % A 1 B 0
No Lag Time: YES ∇ On-board stability period: 7 Linearity Fst 15 % Sec %
No lag time NO ∇ On-board stability period 7
Select using Space key, or select from list displayed by Guide key
Specific Test Parameters
General LIH ISE Range Test No # Test name ALT ∇ Sample type Ser ∇ Page 2/2
Level L Level H
Test Name: ALT ∇ < > Type: Serum ∇
Value/flag # ∇ # #
Value/Flag: # ∇ Level L: # Level H: # Normal range
Normal Ranges: Age L Age H Sex Age L Age H L H
Sex Year Month Year Month L H 1 # ∇ # Y # M # Y # M→ # #
R 1. # ∇ # # # # # # 2 # ∇ # Y # M # Y # M→ # #
R 2. # ∇ # # # # # # 3 # ∇ # Y # M # Y # M→ # #
R 3. # ∇ # # # # # # 4 # ∇ # Y # M # Y # M→ # #
R 4. # ∇ # # # # # # 5 # ∇ # Y # M # Y # M→ # #
R 5. # ∇ # # # # # # 6 # ∇ # Y # M # Y # M→ # #
R 6. # ∇ # # # # # # 7 Non select # #
7. None Selected # # 8 Out of range # #
8. Out of Range L H # # L H
Panic value # #
Panic Value: # # Unit: U/L* Decimal places: #
Select the function using the Function key or the Mouse
Calibration specific
Calibration Specific
General ISE Test No # Test name ALT ∇

Test Name: ALT ∇ < > Type Serum ∇ Cal type 1 MB ∇ Count #

Calibration Type: MB ∇ Formula: Y=AX+B ∇ Counts: # Process: ∇


SERUM/PLASMA APPLICATION Formula 1
Selection calibrator
Y=AX+B ∇ Process ∇

Cal No OD Conc Factor/OD-L Factor/OD-H


Cal. No. OD CONC Factor/OD-L Factor/OD-H Point 1 § ∇ § §3650* §5470*
Point 1: § § §3650* §5470* Point 2 ∇
Point 2:
Point 3 ∇
Point 3:
Point 4 ∇
Point 4:
Point 5 ∇
Point 5:
Point 6 ∇
Point 6:
Point 7: Point 7 ∇
1-point cal. Point
1-Point Cal. Point: R with CONC-0 Slope Check: None ∇ Advanced Calibration: # ∇
MB type factor #
MB Type Factor: # Calibration Stability Period: Calibrator stability period
Select the function using the Function key or the Mouse
# User defined # User defined ¤ Analyser default value
* Values set for working in U/L. To work in SI units (µkat/L) divide by 60 * Values set for working in U/L. To work in SI units (µkat/L) divide by 60
§ For use in AB mode only, refer to IFU for further instruction. § For use in AB mode only, refer to IFU for further instruction.
Enzyme BSOSR6x07.02
2010-05
ALT (IFCC, with OE60106 activation), AU2700/AU5400
Serum/Plasma Application
System Reagent: OSR6007, OSR6107, OSR6507 Reagent ID: 007

Specific Test Parameters


General LIH ISE Range

Test Name: ALT ∇ < > Type: Serum ∇ Operation: Yes ∇

Sample: Volume 5 µL Dilution 0 µL Pre-Dilution Rate: 1


Reagents: R1 Volume 50 µL Dilution 25 µL Min OD Max OD
R2 Volume 25 µL Dilution 25 µL L 1.0 H 2.5
Reagent OD limit:
Wavelength: Pri. 340 ∇ Sec. 660 ∇ First L 1.3 First H 2.5
Method: RATE ∇ Last L 1.3 Last H 2.5
Reaction slope: - ∇ Dynamic Range:
Measuring Point 1: First 14 Last 27 L 3* H 500*
Measuring Point 2: First Last Correlation Factor:
Linearity : 15 % A 1 B 0
No Lag Time: YES ∇ On-board stability period: 7

Specific Test Parameters


General LIH ISE Range

Test Name: ALT ∇ < > Type: Serum ∇

Value/Flag: # ∇ Level L: # Level H: #


Normal Ranges: Age L Age H
Sex Year Month Year Month L H
R 1. # ∇ # # # # # #
R 2. # ∇ # # # # # #
R 3. # ∇ # # # # # #
R 4. # ∇ # # # # # #
R 5. # ∇ # # # # # #
R 6. # ∇ # # # # # #
7. None Selected # #
8. Out of Range L H # #
Panic Value: # # Unit: U/L* Decimal places: #

Calibration Specific
General ISE

Test Name: ALT ∇ < > Type Serum ∇

SERUM/PLASMA APPLICATION
Calibration Type: MB ∇ Formula: Y=AX+B ∇ Counts: # Process: ∇

Cal. No. OD CONC Factor/OD-L Factor/OD-H


Point 1: § § §3650* §5470*
Point 2:
Point 3:
Point 4:
Point 5:
Point 6:
Point 7:
1-Point Cal. Point: R with CONC-0 Slope Check: None ∇ Advanced Calibration: # ∇
MB Type Factor: # Calibration Stability Period:

# User defined
* Values set for working in U/L. To work in SI units (µkat/L) divide by 60
§ For use in AB mode only, refer to IFU for further instruction.
Ф AU680

Enzyme BSOSR6x07.02
2010-05
ALT (IFCC, with OE60106 activation), AU680/AU480 Serum/Plasma Application ALT (IFCC, with OSR60180 activation), AU680 Serum/Plasma Application
System Reagent: OSR6007, OSR6107, OSR6507 Reagent ID: 007 System Reagent: OSR6007, OSR6107, OSR6507 Reagent ID: 007
Parameters Specific Test Parameters Parameters Specific Test Parameters
General LIH ISE HbA1c Calculated Test Range General LIH ISE HbA1c Calculated Test Range
Test Name: ALT ∇ < > Type: Serum ∇ Operation Yes ∇ Test Name: ALT ∇ < > Type: Serum ∇ Operation Yes ∇

Sample Volume 5 µL Dilution 0 µL OD Limit Sample Volume 5 µL Dilution 0 µL OD Limit


Pre-Dilution Rate 1 ∇ Min.OD 1.0 Max.OD 2.5 Pre-Dilution Rate 1 ∇ Min.OD 1.0 Max.OD 2.5
Rgt. Volume R1(R1-1) 50 µL Dilution 25 µL Reagent OD Limit Rgt. Volume R1(R1-1) 50 µL Dilution 10 µL Reagent OD Limit
First Low 1.3 High 2.5 R1-2 5 µL Dilution 10 µL First Low 1.3 High 2.5
Last Low 1.3 High 2.5 Last Low 1.3 High 2.5
R2(R2-1) 25 µL Dilution 25 µL R2(R2-1) 25 µL Dilution 25 µL
Dynamic Range Low 3* High 500* Dynamic Range Low 3* High 500*
Common Rgt. Type Noneф Name ф Correlation Factor A 1 B 0 Common Rgt. Type R1-2 Name # Correlation Factor A 1 B 0
Wavelength Pri 340 ∇nm Sec. 660 ∇nm Factor for Maker A 1 B 0 Wavelength Pri 340 ∇nm Sec. 660 ∇nm Factor for Maker A 1 B 0
Method RATE ∇ Method RATE ∇
Reaction Slope - ∇ Onboard Stability Period 7 Day 0 Hour Reaction Slope - ∇ Onboard Stability Period 30 Day 0 Hour
Measuring Point1 First 14 Last 27 LIH Influence Check # ∇ Measuring Point1 First 14 Last 27 LIH Influence Check # ∇
Measuring Point2 First Last Lipemia +++++ ∇ Measuring Point2 First Last Lipemia +++++ ∇
Linearity Limit 15 % Icterus +++++ ∇ Linearity Limit 15 % Icterus +++++ ∇
Lag Time Check YES ∇ Hemolysis +++++ ∇ Lag Time Check YES Hemolysis +++++
∇ ∇

Parameters Specific Test Parameters Parameters Specific Test Parameters


General LIH ISE HbA1c Calculated Test Range General LIH ISE HbA1c Calculated Test Range
Test Name: ALT ∇ < > Type: Serum ∇ Test Name: ALT ∇ < > Type: Serum ∇
Value/Flag: # ∇ Low High Value/Flag: # Low High

Level # # Panic Value Level # # Panic Value
Specificl Ranges: From To Low High Specificl Ranges: From To Low High
Sex Year Month Year Month Low High # # Sex Year Month Year Month Low High # #
R 1. # ∇ # # # # # # R 1. # ∇ # # # # # #
R 2. # ∇ # # # # # # R 2. # ∇ # # # # # #
R 3. # ∇ # # # # # # R 3. # ∇ # # # # # #
R 4. # ∇ # # # # # # R 4. # ∇ # # # # # #
R 5. # ∇ # # # # # # R 5. # ∇ # # # # # #
R 6. # ∇ # # # # # # R 6. # ∇ # # # # # #
7. No demographics # # 7. No demographics # #
8. Not within expected values # # 8. Not within expected values # #
Unit U/L* Decimal Places # Unit U/L* Decimal Places #
Parameters Calibration Parameters Parameters Calibration Parameters
Calibrators Calibration Specific STAT Table Calibration Calibrators Calibration Specific STAT Table Calibration
General ISE General ISE

SERUM/PLASMA APPLICATION
Test Name: ALT ∇ < > Type Serum ∇ R Use Serum Cal. R
Test Name: ALT ∇ < > Type Serum ∇ Use Serum Cal.
Calibration Type: MB ∇ Formula: Y=AX+B ∇ Counts: # ∇ Calibration Type: MB ∇ Formula: Y=AX+B ∇ Counts: # ∇
<Calibrator Parameters> Range <Calibrator Parameters> Range
Calibrator OD Conc Low High Slope Check None ∇ Calibrator OD Conc Low High Slope Check None ∇
Point 1: § ∇ § §3650* §5470* Point 1: § § §3650* §5470*

Point 2: ∇ Allowance Range Check Point 2: Allowance Range Check

Point 3: ∇ Point 3: ∇
Point 4: ∇ R Reagent Blank Point 4: ∇ R Reagent Blank
Point 5: ∇ R Calibration Point 5: ∇ R Calibration
Point 6: ∇ Point 6: ∇
Point 7 ∇ Advanced Calibration Point 7 Advanced Calibration

Point 8 ∇ Operation # ∇ Point 8 ∇ Operation # ∇
Point 9 ∇ Point 9 ∇
Point 10 ∇ Interval (RB/ACAL) # ∇ Point 10 ∇ Interval (RB/ACAL) # ∇
<Point Cal. For No. of Correction Points ∇ Use Master Curve ∇ R Lot Calibration
<Point Cal. For No. of Correction Points ∇ Use Master Curve ∇ R Lot Calibration
Master Curve> OD Range Master Curve> OD Range
Calibrator OD Conc Low High Stability Calibrator OD Conc Low High Stability
Point-1 ∇ Reagent Blank Day Hour Point-1 ∇ Reagent Blank Day Hour
Point-2 ∇ Calibration Day Hour Point-2 Calibration Day Hour

MB Type Factor: # 1-Point Calibration Point ∇ R with Conc-0 MB Type Factor: # 1-Point Calibration Point R with Conc-0

Enzyme BSOSR6x07.02
2010-05
ALT (IFCC, without Pyridoxal phosphate activation), AU400/AU640 ALT (IFCC, without Pyridoxal phosphate activation), AU600
Paediatric Application Paediatric Application
System Reagent: OSR6007, OSR6107 Reagent ID: 007 System Reagent: OSR6007, OSR6107 Reagent ID: 007
Specific Test Parameters Specific test parameters
General LIH ISE Range
Test No # Test name ALTP ∇ Sample type Ser ∇ Page 1/2
Test Name: ALTP ∇ < > Type: Serum ∇ Operation: Yes ∇
Sample vol. 10 Dil. vol 10 µL Min. OD Max. OD
Sample: Volume 10 µL Dilution 10 µL Pre-Dilution Rate: 1 Reagent 1 vol 100 Dil. vol 50 µL L 0.60 H 2.5
Reagents: R1 Volume 100 µL Dilution 50 µL Min OD Max OD Reagent 2 vol 50 Dil. vol 40 µL Reagent OD limit
R2 Volume 50 µL Dilution 40 µL L 0.60 H 2.5 Fst. L 1.1 Fst. H 2.5
Reagent OD limit: Wave Main 340 Sub 660 ∇ Lst. L 1.1 Lst. H 2.5
Wavelength: Pri. 340 ∇ Sec. 660 ∇ First L 1.1 First H 2.5 Method RATE ∇ Dynamic range
Method: RATE ∇ Last L 1.1 Last H 2.5 Reaction - ∇ L 3* H 500*
Reaction slope: - ∇ Dynamic Range: Point 1 Fst 14 Lst 27 Correlation factor A 1
Measuring Point 1: First 14 Last 27 L 3* H 500* Point 2 Fst Lst B 0
Measuring Point 2: First Last Correlation Factor: Sample Pre-dil. Rate ¤
Linearity : 15 % A 1 B 0
No Lag Time: YES ∇ On-board stability period: 30 Linearity Fst 15 % Sec %
No lag time NO ∇ On-board stability period 30
Select using Space key, or select from list displayed by Guide key
Specific Test Parameters
General LIH ISE Range Test No # Test name ALTP ∇ Sample type Ser ∇ Page 2/2
Test Name: ALTP ∇ < > Type: Serum ∇ Level L Level H
Value/flag # ∇ # #
Value/Flag: # ∇ Level L: # Level H: # Normal range
Normal Ranges: Age L Age H Sex Age L Age H L H
Sex Year Month Year Month L H 1 # ∇ # Y # M # Y # M→ # #
R 1. # ∇ # # # # # # 2 # ∇ # Y # M # Y # M→ # #
R 2. # ∇ # # # # # # 3 # ∇ # Y # M # Y # M→ # #
R 3. # ∇ # # # # # # 4 # ∇ # Y # M # Y # M→ # #
R 4. # ∇ # # # # # # 5 # ∇ # Y # M # Y # M→ # #
R 5. # ∇ # # # # # # 6 # ∇ # Y # M # Y # M→ # #
R 6. # ∇ # # # # # # 7 Non select # #
7. None Selected # # 8 Out of range # #
8. Out of Range L H # # L H
Panic Value: # # Unit: U/L* Decimal places: # Panic value # #
Select the function using the Function key or the Mouse
Calibration specific
Calibration Specific
General ISE Test No # Test name ALTP ∇

PAEDIATRIC APPLICATION
Test Name: ALTP ∇ < > Type Serum ∇ Cal type 1 MB ∇ Count #
Formula 1 Y=AX+B ∇ Process ∇
Calibration Type: MB ∇ Formula: Y=AX+B ∇ Counts: # Process: ∇
Selection calibrator
Cal No OD Conc Factor/OD-L Factor/OD-H
Cal. No. OD CONC Factor/OD-L Factor/OD-H
Point 1 § ∇ § §3650* §5470*
Point 1: § § §3650* §5470*
Point 2: Point 2 ∇
Point 3: Point 3 ∇
Point 4: Point 4 ∇
Point 5: Point 5 ∇
Point 6: Point 6 ∇
Point 7: Point 7 ∇
1-Point Cal. Point: R With CONC-0 Slope Check: None ∇ Advanced Calibration: # ∇ 1-point cal. point
MB type factor #
MB Type Factor: # Calibration Stability Period:
Calibrator stability period
Select the function using the Function key or the Mouse
# User defined
# User defined ¤ Analyser default value
* Values set for working in U/L. To work in SI units (µkat/L) divide by 60
* Values set for working in U/L. To work in SI units (µkat/L) divide by 60
§ For use in AB mode only, refer to IFU for further instruction.
§ For use in AB mode only, refer to IFU for further instruction.

Enzyme BSOSR6x07.02
2010-05
ALT (IFCC, without Pyridoxal phosphate activation), AU2700/AU5400 ALT (IFCC, without Pyrioxal phosphate activation), AU680/AU480
Paediatric Application Paediatric Application
System Reagent: OSR6007, OSR6107, OSR6507 Reagent ID: 007 System Reagent: OSR6007, OSR6107, OSR6507 Reagent ID: 007
Parameters Specific Test Parameters
Specific Test Parameters General LIH ISE HbA1c Calculated Test Range
General LIH ISE Range
Test Name: ALTP ∇ < > Type: Serum ∇ Operation Yes ∇
Test Name: ALTP ∇ < > Type: Serum ∇ Operation: Yes ∇

Sample: Volume 5 µL Dilution 10 µL Pre-Dilution Rate: 1 Sample Volume 5 µL Dilution 10 µL OD Limit


Reagents: R1 Volume 50 µL Dilution 25 µL Min OD Max OD Pre-Dilution Rate 1 ∇ Min.OD 0.60 Max.OD 2.5
R2 Volume 25 µL Dilution 15 µL L 0.60 H 2.5 Rgt. Volume R1(R1-1) 50 µL Dilution 25 µL Reagent OD Limit
Reagent OD limit: First Low 1.1 High 2.5
Wavelength: Pri. 340 ∇ Sec. 660 ∇ First L 1.1 First H 2.5 Last Low 1.1 High 2.5
Method: RATE ∇ Last L 1.1 Last H 2.5 R2(R2-1) 25 µL Dilution 15 µL
Reaction slope: - ∇ Dynamic Range: Dynamic Range Low 3* High 500*
Measuring Point 1: First 14 Last 27 L 3* H 500* Common Rgt. Type Noneф Name ф Correlation Factor A 1 B 0
Measuring Point 2: First Last Correlation Factor: Wavelength Pri 340 ∇nm Sec. 660 ∇nm Factor for Maker A 1 B 0
Linearity : 15 % A 1 B 0 Method RATE ∇
No Lag Time: YES ∇ On-board stability period: 30 Reaction Slope - ∇ Onboard Stability Period 30 Day 0 Hour
Measuring Point1 First 14 Last 27 LIH Influence Check # ∇
Measuring Point2 First Last Lipemia +++++ ∇
Linearity Limit 15 % Icterus +++++ ∇
Specific Test Parameters Lag Time Check YES ∇ Hemolysis +++++ ∇
General LIH ISE Range

Test Name: ALTP ∇ < > Type: Serum ∇ Parameters Specific Test Parameters
General LIH ISE HbA1c Calculated Test Range
Value/Flag: # ∇ Level L: # Level H: # Test Name: ALTP Type: Serum
∇ < > ∇
Normal Ranges: Age L Age H
Sex Year Month Year Month L H Value/Flag: # ∇ Low High
R 1. # ∇ # # # # # # Level # # Panic Value
R 2. # ∇ # # # # # # Specificl Ranges: From To Low High
R 3. # ∇ # # # # # # Sex Year Month Year Month Low High # #
R 4. # # # # # # # R 1. # ∇ # # # # # #

R 5. R 2. # ∇ # # # # # #
# ∇ # # # # # #
R 3. # ∇ # # # # # #
R 6. # ∇ # # # # # #
R 4. # ∇ # # # # # #
7. None Selected # #
R 5. # ∇ # # # # # #
8. Out of Range L H # # R 6. # ∇ # # # # # #
Panic Value: # # Unit: U/L* Decimal places: # 7. No demographics # #
8. Not within expected values # #
Unit U/L* Decimal Places #
Calibration Specific Parameters Calibration Parameters

PAEDIATRIC APPLICATION
General ISE Calibrators Calibration Specific STAT Table Calibration
General ISE
Test Name: ALTP ∇ < > Type Serum ∇
Test Name: ALTP ∇ < > Type Serum ∇ R Use Serum Cal.
Calibration Type: MB ∇ Formula: Y=AX+B ∇ Counts: # Process: ∇
Calibration Type: MB ∇ Formula: Y=AX+B ∇ Counts: # ∇
Cal. No. OD CONC Factor/OD-L Factor/OD-H <Calibrator Parameters> Range
Point 1: § § §3650* §5470* Calibrator OD Conc Low High Slope Check None ∇
Point 2: Point 1: § ∇ § §3650* §5470*
Point 3: Point 2: ∇ Allowance Range Check
Point 4: Point 3: ∇
Point 5: Point 4: ∇ R Reagent Blank
Point 6: Point 5: ∇ R Calibration
Point 7: Point 6: ∇
Point 7 ∇ Advanced Calibration
1-Point Cal. Point: R with CONC-0 Slope Check: None ∇ Advanced Calibration: # ∇ Point 8 ∇ Operation # ∇
MB Type Factor: # Calibration Stability Period: Point 9 ∇
Point 10 ∇ Interval (RB/ACAL) # ∇
<Point Cal. For No. of Correction Points ∇ Use Master Curve ∇ R Lot Calibration
# User defined Master Curve> OD Range
* Values set for working in U/L. To work in SI units (µkat/L) divide by 60 Calibrator OD Conc Low High Stability
§ For use in AB mode only, refer to IFU for further instruction. Point-1 ∇ Reagent Blank Day Hour
Ф AU680 Point-2 ∇ Calibration Day Hour
MB Type Factor: # 1-Point Calibration Point ∇ R with Conc-0

Enzyme BSOSR6x07.02
2010-05
ALT (IFCC, with OE60106 activation), AU400/AU640 ALT (IFCC, with OE60106 activation), AU600
Paediatric Application Paediatric Application
System Reagent: OSR6007, OSR6107 Reagent ID: 007 System Reagent: OSR6007, OSR6107 Reagent ID: 007

Specific Test Parameters Specific test parameters


General LIH ISE Range
Test No # Test name ALTP ∇ Sample type Ser ∇ Page 1/2
Test Name: ALTP ∇ < > Type: Serum ∇ Operation: Yes ∇
Sample vol. 10 Dil. vol 10 µL Min. OD Max. OD
Sample: Volume 10 µL Dilution 10 µL Pre-Dilution Rate: 1 Reagent 1 vol 100 Dil. vol 50 µL L 1.0 H 2.5
Reagents: R1 Volume 100 µL Dilution 50 µL Min OD Max OD Reagent 2 vol 50 Dil. vol 40 µL Reagent OD limit
R2 Volume 50 µL Dilution 40 µL L 1.0 H 2.5 Fst. L 1.3 Fst. H 2.5
Reagent OD limit: Wave Main 340 Sub 660 ∇ Lst. L 1.3 Lst. H 2.5
Wavelength: Pri. 340 ∇ Sec. 660 ∇ First L 1.3 First H 2.5 Method RATE ∇ Dynamic range
Method: RATE ∇ Last L 1.3 Last H 2.5 Reaction - ∇ L 3* H 500*
Reaction slope: - ∇ Dynamic Range: Point 1 Fst 14 Lst 27 Correlation factor A 1
Measuring Point 1: First 14 Last 27 L 3* H 500* Point 2 Fst Lst B 0
Measuring Point 2: First Last Correlation Factor: Sample Pre-dil. Rate ¤
Linearity : 15 % A 1 B 0
No Lag Time: YES ∇ On-board stability period: 7 Linearity Fst 15 % Sec %
No lag time NO ∇ On-board stability period 7
Select using Space key, or select from list displayed by Guide key
Specific Test Parameters
General LIH ISE Range Test No # Test name ALTP ∇ Sample type Ser ∇ Page 2/2
Test Name: ALTP ∇ < > Type: Serum ∇ Level L Level H
Value/flag # ∇ # #
Value/Flag: # ∇ Level L: # Level H: # Normal range
Normal Ranges: Age L Age H Sex Age L Age H L H
Sex Year Month Year Month L H 1 # ∇ # Y # M # Y # M→ # #
R 1. # ∇ # # # # # # 2 # ∇ # Y # M # Y # M→ # #
R 2. # ∇ # # # # # # 3 # ∇ # Y # M # Y # M→ # #
R 3. # ∇ # # # # # # 4 # ∇ # Y # M # Y # M→ # #
R 4. # ∇ # # # # # # 5 # ∇ # Y # M # Y # M→ # #
R 5. # ∇ # # # # # # 6 # ∇ # Y # M # Y # M→ # #
R 6. # ∇ # # # # # # 7 Non select # #
7. None Selected # # 8 Out of range # #
8. Out of Range L H # # L H
Panic Value: # # Unit: U/L* Decimal places: # Panic value # #
Select the function using the Function key or the Mouse
Calibration specific
Calibration Specific
General ISE Test No # Test name ALTP ∇

PAEDIATRIC APPLICATION
Test Name: ALTP ∇ < > Type Serum ∇ Cal type 1 MB ∇ Count #
Formula 1 Y=AX+B ∇ Process ∇
Calibration Type: MB ∇ Formula: Y=AX+B ∇ Counts: # Process: ∇
Selection calibrator
Cal No OD Conc Factor/OD-L Factor/OD-H
Cal. No. OD CONC Factor/OD-L Factor/OD-H
Point 1 § ∇ § §3650* §5470*
Point 1: § § §3650* §5470*
Point 2: Point 2 ∇
Point 3: Point 3 ∇
Point 4: Point 4 ∇
Point 5: Point 5 ∇
Point 6: Point 6 ∇
Point 7: Point 7 ∇
1-Point Cal. Point: R with CONC-0 Slope Check: None ∇ Advanced Calibration: # ∇ 1-point cal. point
MB type factor #
MB Type Factor: # Calibration Stability Period:
Calibrator stability period
Select the function using the Function key or the Mouse
# User defined
* Values set for working in U/L. To work in SI units (µkat/L) divide by 60 # User defined ¤ Analyser default value
§ For use in AB mode only, refer to IFU for further instruction. * Values set for working in U/L. To work in SI units (µkat/L) divide by 60
§ For use in AB mode only, refer to IFU for further instruction.
Enzyme BSOSR6x07.02
2010-05
ALT (IFCC, with OE60106 activation), AU2700/AU5400
Paediatric Application
System Reagent: OSR6007, OSR6107, OSR6507 Reagent ID: 007

Specific Test Parameters


General LIH ISE Range

Test Name: ALTP ∇ < > Type: Serum ∇ Operation: Yes ∇

Sample: Volume 5 µL Dilution 10 µL Pre-Dilution Rate: 1


Reagents: R1 Volume 50 µL Dilution 25 µL Min OD Max OD
R2 Volume 25 µL Dilution 15 µL L 1.0 H 2.5
Reagent OD limit:
Wavelength: Pri. 340 ∇ Sec. 660 ∇ First L 1.3 First H 2.5
Method: RATE ∇ Last L 1.3 Last H 2.5
Reaction slope: - ∇ Dynamic Range:
Measuring Point 1: First 14 Last 27 L 3* H 500*
Measuring Point 2: First Last Correlation Factor:
Linearity : 15 % A 1 B 0
No Lag Time: YES ∇ On-board stability period: 7

Specific Test Parameters


General LIH ISE Range

Test Name: ALTP ∇ < > Type: Serum ∇

Value/Flag: # ∇ Level L: # Level H: #


Normal Ranges: Age L Age H
Sex Year Month Year Month L H
R 1. # ∇ # # # # # #
R 2. # ∇ # # # # # #
R 3. # ∇ # # # # # #
R 4. # ∇ # # # # # #
R 5. # ∇ # # # # # #
R 6. # ∇ # # # # # #
7. None Selected # #
8. Out of Range L H # #
Panic Value: # # Unit: U/L* Decimal places: #

Calibration Specific
General ISE

PAEDIATRIC APPLICATION
Test Name: ALTP ∇ < > Type Serum ∇

Calibration Type: MB ∇ Formula: Y=AX+B ∇ Counts: # Process: ∇

Cal. No. OD CONC Factor/OD-L Factor/OD-H


Point 1: § § §3650* §5470*
Point 2:
Point 3:
Point 4:
Point 5:
Point 6:
Point 7:
1-Point Cal. Point: R with CONC-0 Slope Check: None ∇ Advanced Calibration: # ∇
MB Type Factor: # Calibration Stability Period:

# User defined
* Values set for working in U/L. To work in SI units (µkat/L) divide by 60
§ For use in AB mode only, refer to IFU for further instruction.
Ф AU680

Enzyme BSOSR6x07.02
2010-05
ALT (IFCC, with OE60106 activation), AU680/AU480 Paediatric Application ALT (IFCC, with OSR60180 activation), AU680 Paediatric Application
System Reagent: OSR6007, OSR6107, OSR6507 Reagent ID: 007 System Reagent: OSR6007, OSR6107, OSR6507 Reagent ID: 007
Parameters Specific Test Parameters Parameters Specific Test Parameters
General LIH ISE HbA1c Calculated Test Range General LIH ISE HbA1c Calculated Test Range
Test Name: ALTP ∇ < > Type: Serum ∇ Operation Yes ∇ Test Name: ALTP ∇ < > Type: Serum ∇ Operation Yes ∇

Sample Volume 5 µL Dilution 10 µL OD Limit Sample Volume 5 µL Dilution 10 µL OD Limit


Pre-Dilution Rate 1 ∇ Min.OD 1.0 Max.OD 2.5 Pre-Dilution Rate 1 ∇ Min.OD 1.0 Max.OD 2.5
Rgt. Volume R1(R1-1) 50 µL Dilution 25 µL Reagent OD Limit Rgt. Volume R1(R1-1) 50 µL Dilution 10 µL Reagent OD Limit
First Low 1.3 High 2.5 R1-2 5 µL Dilution 10 µL First Low 1.3 High 2.5
Last Low 1.3 High 2.5 Last Low 1.3 High 2.5
R2(R2-1) 25 µL Dilution 15 µL R2(R2-1) 25 µL Dilution 15 µL
Dynamic Range Low 3* High 500* Dynamic Range Low 3* High 500*
Common Rgt. Type Noneф Name ф Correlation Factor A 1 B 0 Common Rgt. Type R1-2 Name # Correlation Factor A 1 B 0
Wavelength Pri 340 ∇nm Sec. 660 ∇nm Factor for Maker A 1 B 0 Wavelength Pri 340 ∇nm Sec. 660 ∇nm Factor for Maker A 1 B 0
Method RATE ∇ Method RATE ∇
Reaction Slope - ∇ Onboard Stability Period 7 Day 0 Hour Reaction Slope - ∇ Onboard Stability Period 30 Day 0 Hour
Measuring Point1 First 14 Last 27 LIH Influence Check # ∇ Measuring Point1 First 14 Last 27 LIH Influence Check # ∇
Measuring Point2 First Last Lipemia +++++ ∇ Measuring Point2 First Last Lipemia +++++ ∇
Linearity Limit 15 % Icterus +++++ ∇ Linearity Limit 15 % Icterus +++++ ∇
Lag Time Check YES ∇ Hemolysis +++++ ∇ Lag Time Check YES Hemolysis +++++
∇ ∇

Parameters Specific Test Parameters Parameters Specific Test Parameters


General LIH ISE HbA1c Calculated Test Range General LIH ISE HbA1c Calculated Test Range
Test Name: ALTP ∇ < > Type: Serum ∇ Test Name: ALTP ∇ < > Type: Serum ∇
Value/Flag: # ∇ Low High Value/Flag: # Low High

Level # # Panic Value Level # # Panic Value
Specificl Ranges: From To Low High Specificl Ranges: From To Low High
Sex Year Month Year Month Low High # # Sex Year Month Year Month Low High # #
R 1. # ∇ # # # # # # R 1. # ∇ # # # # # #
R 2. # ∇ # # # # # # R 2. # ∇ # # # # # #
R 3. # ∇ # # # # # # R 3. # ∇ # # # # # #
R 4. # ∇ # # # # # # R 4. # ∇ # # # # # #
R 5. # ∇ # # # # # # R 5. # ∇ # # # # # #
R 6. # ∇ # # # # # # R 6. # ∇ # # # # # #
7. No demographics # # 7. No demographics # #
8. Not within expected values # # 8. Not within expected values # #
Unit U/L* Decimal Places # Unit U/L* Decimal Places #
Parameters Calibration Parameters Parameters Calibration Parameters
Calibrators Calibration Specific STAT Table Calibration Calibrators Calibration Specific STAT Table Calibration
General ISE General ISE
Test Name: ALTP ∇ < > Type Serum ∇ R Use Serum Cal. R
Test Name: ALTP ∇ < > Type Serum ∇ Use Serum Cal.

PAEDIATRIC APPLICATION
Calibration Type: MB ∇ Formula: Y=AX+B ∇ Counts: # ∇ Calibration Type: MB ∇ Formula: Y=AX+B ∇ Counts: # ∇
<Calibrator Parameters> Range <Calibrator Parameters> Range
Calibrator OD Conc Low High Slope Check None ∇ Calibrator OD Conc Low High Slope Check None ∇
Point 1: § ∇ § §3650* §5470* Point 1: § § §3650* §5470*

Point 2: ∇ Allowance Range Check Point 2: Allowance Range Check

Point 3: ∇ Point 3: ∇
Point 4: ∇ R Reagent Blank Point 4: ∇ R Reagent Blank
Point 5: ∇ R Calibration Point 5: ∇ R Calibration
Point 6: ∇ Point 6: ∇
Point 7 ∇ Advanced Calibration Point 7 Advanced Calibration

Point 8 ∇ Operation # ∇ Point 8 ∇ Operation # ∇
Point 9 ∇ Point 9 ∇
Point 10 ∇ Interval (RB/ACAL) # ∇ Point 10 ∇ Interval (RB/ACAL) # ∇
<Point Cal. For No. of Correction Points ∇ Use Master Curve ∇ R Lot Calibration
<Point Cal. For No. of Correction Points ∇ Use Master Curve ∇ R Lot Calibration
Master Curve> OD Range Master Curve> OD Range
Calibrator OD Conc Low High Stability Calibrator OD Conc Low High Stability
Point-1 ∇ Reagent Blank Day Hour Point-1 ∇ Reagent Blank Day Hour
Point-2 ∇ Calibration Day Hour Point-2 Calibration Day Hour

MB Type Factor: # 1-Point Calibration Point ∇ R with Conc-0 MB Type Factor: # 1-Point Calibration Point R with Conc-0

Enzyme BSOSR6x07.02
2010-05
α-AMYLASE
OSR6006 4 x 10 mL R1
OSR6106 4 x 40 mL R1
Intended Use
Kinetic colour test for the quantitative determination of α-amylase, [1,4-α-D-glucan 4-glucanohydrolase, EC 3.2.1.1], in human serum, plasma and urine on
Beckman Coulter analysers. For in vitro diagnostic use only.
Summary1,2,3
Amylases are a group of hydrolases that split complex carbohydrates composed of alpha-D-glucose units linked through carbon atoms 1 and 4 located on
adjacent glucose residues. In the body, amylase is present in a number of organs and tissues. The greatest concentration is present in the pancreas,
where the enzyme is synthesised by the acinar cells and then secreted into the intestinal tract by way of the pancreatic duct system. The salivary glands
also secrete a potent amylase to initiate hydrolysis of starches while the food is still in the mouth and oesophagus.
Diseases resulting in elevation of plasma alpha-amylase include: acute pancreatitis, parotitis, alcoholism, renal insufficiency and diseases such as viral
hepatitis, AIDS, abdominal typhoid, sarcoidosis and trauma to the upper abdomen. There is also a detectable increase in amylase after an ERCP
procedure.
In acute pancreatitis, amylase increases 5-6 hours after the onset of symptoms and remains elevated for 2-5 days. The increase in plasma activity does
not reflect disease severity and conversely, extensive destruction of the pancreas may not cause a significant increase in the plasma concentration of
pancreatic alpha-amylase.
Alpha amylase is excreted by glomerular filtration and then 50% of it is reabsorbed by the tubules. This reabsorption is significantly reduced in transient
tubular damage, after burns, in the presence of diabetic ketoacidosis and acute pancreatitis as well as proteinuria resulting in an increase of alpha-amylase
clearance. The measurement of alpha amylase in urine is indicated in the investigation of hyperamylasemia associated with macroamylasemia or renal
insufficiency. Hypoamylasemia has been observed in advanced cystic fibrosis, severe liver disease and pancreatectomy. Also due to the decreased
concentration of the salivary fraction hypoamylasemia has been found in obese subjects.
Test Principle4
The α-amylase colour test employs 2-chloro-4-nitrophenyl-α-D-maltotrioside (CNPG3) as substrate. This substrate reacts directly with α-amylase and does
not require the presence of ancillary enzymes. The release of 2-chloro-4-nitrophenol (CNP) from the substrate and the resulting absorbance increase at
410 nm is directly proportional to the α-amylase activity in the sample.
Reaction Principle
CNPG3 + H2O α-amylase CNP + Maltotriose

Contents, Reagent Composition in the Test


Final concentration of reactive ingredients:
MES (pH 6.05) 36.1 mmol/L
Calcium acetate 3.60 mmol/L
NaCl 37.2 mmol/L
Potassium thiocyanate 253 mmol/L
CNPG3 1.63 mmol/L
Preservative
Precautions and Warnings
Exercise the normal precautions required for handling all laboratory reagents.
To avoid the possible build-up of azide compounds, flush waste-pipes with water after the disposal of undiluted reagent.
Dispose of all waste material in accordance with local guidelines.
Safety data sheet available for professional user on request.
Reagent Preparation
The reagent is ready for use and can be placed directly on board the instrument.
Care should be taken when handling this reagent to avoid contamination with skin and body fluids.
Storage and Stability
The reagent is stable, unopened, up to the stated expiry date when stored at 2...8°C. Once open the reagent stored on board the instrument is stable for
30 days.
Discard reagents if any discolouration is observed.
Specimen
Serum, heparinised plasma. Haemolysed and strongly icteric samples should be avoided, separate from blood cells as soon as possible.
5
Stable in serum and plasma for 7 days when stored at 2…25°C.
1,6
Plasma using EDTA, oxalate and citrate should be avoided.
1 6
Urine: Timed or random sample. Adjust pH to approximately 7.0 before storage.
5
Stable in urine for 10 days when stored at 2…8°C and 2 days when stored at 15…25°C.
Test Procedure
Refer to the appropriate User Guide and Setting Sheet for analyser-specific assay instructions for the sample type as listed in the Intended Use statement.
Calibration
The test is run in MB-mode. To provide a robust approach to generate the analyser specific MB factor, it is recommended that 5 separate calibration events
should be used. A fresh vial of calibrator, utilising System Calibrator Cat No. 66300 for serum application and Urine calibrator Cat no. ODC0025 for urine
application, in the AB calibration mode, should be used for each of these runs. When calculating the mean factor from the separate runs the data should be
examined for obvious outliers which should be repeated and replaced. For the AU2700/AU5400 this procedure needs to be performed for each ring.
Quality control procedures should be undertaken immediately following calibration in accordance with good laboratory practice.
The calibrator values are traceable to a Beckman Coulter master calibrator.
Re-establishment of the analyser specific MB factor is recommended when a critical part of the analyser is replaced.
Reagent blank measurement is recommended when changing to a new lot of reagent.

EN.01 BLOSR6x06.01 Enzyme


2009-08
Quality Control
Controls Cat. No. ODC0003 and ODC0004 or other control materials with values determined by this Beckman Coulter system may be used for the
serum/plasma application.
Biorad Liquichek Urine Chemistry Controls Cat. No. 397 and 398 or other control materials with values determined by this Beckman Coulter system may
be used for the urine application.
Each laboratory should establish its own control frequency.
Good laboratory practice suggests that controls be tested each day patient samples are tested and each time calibration is performed. Values obtained for
the controls should fall within specified limits as defined by the user. If any trends or sudden shifts in values are detected, review all operating parameters.
Each laboratory should establish guidelines for corrective action to be taken if controls do not recover within the specified limits.
Calculation
The Beckman Coulter analysers automatically compute the α-amylase activity of each sample.
Reference Intervals
3
Serum/Plasma 22 – 80 U/L (0.36 – 1.33 µkat/L)
7
Urine 42 – 321 U/L (0.7 – 5.35 µkat/L)
Expected values may vary with age, sex, sample type, diet and geographical location. Each laboratory should verify the transferability of the expected
values to its own population, and if necessary determine its own reference interval according to good laboratory practice. For diagnostic purposes, results
should always be assessed in conjunction with the patient's medical history, clinical examinations and other findings.
Specific Performance Characteristics
Data contained within this section is representative of performance on Beckman Coulter systems. Data obtained in your laboratory may differ from these
values.
Linearity
The test is linear within an enzyme activity range of 10 – 2000 U/L (0.2 – 33.3 µkat/L) for serum and plasma. The test is linear within an enzyme activity
range of 5 – 4800 U/L (0.1 – 80 µkat/L) for urine.
Precision
The following data was obtained on an AU600 using 3 serum pools analysed over 10 days.
n = 60 Within Run Total
Mean U/L SD CV% SD CV%
45 0.88 1.97 1.17 2.63
140 1.12 0.80 4.39 3.14
1562 12.62 0.81 28.82 1.84
The following data was obtained on an AU640 using 3 urine pools analysed over 20 days.
n = 80 Within Run Total
Mean U/L SD CV% SD CV%
45.97 0.69 1.51 2.13 4.64
494.94 3.99 0.81 14.1 2.85
3207.84 50.73 1.58 101.63 3.17
Sensitivity
The lowest detectable level using serum settings on an AU600 analyser was calculated as 2 U/L.
The lowest detectable level using urine settings on an AU2700 analyser was calculated as 2 U/L.
The lowest detectable level represents the lowest measurable level of α-amylase that can be distinguished from zero. It is calculated as the absolute mean
plus three standard deviations of 20 replicates of an analyte free sample.
Method Comparison
Patient serum samples were used to compare this α-Amylase OSR6106 assay on the AU600 against another commercially available α-amylase assay.
Results of linear regression analysis were as follows:
y = 0.879x + 6.313 r = 0.999 n = 112 Sample range = 11 – 1530 U/L
Patient urine samples were used to compare this α-Amylase OSR6106 assay on the AU2700 against another commercially available α-amylase assay.
Results of linear regression analysis were as follows:
y = 1.067x – 4.492 r = 0.995 n = 125 Sample range = 18 – 339 U/L
Interfering Substances
Results of serum studies conducted to evaluate the susceptibility of the method to interference were as follows:
Icterus: Interference less than 10% up to 20 mg/dL or 342 µmol/L bilirubin
Haemolysis: Interference less than 10% up to 2.5 g/L haemoglobin
®
Lipemia: Interference less than 5% up to 1000 mg/dL Intralipid
Results of urine studies conducted to evaluate the susceptibility of the method to interference were as follows:
Ascorbate: Interference less than 10% up to 50 mg/dL ascorbate
Icterus: Interference less than 10% up to 40 mg/dL or 684 µmol/L bilirubin
Haemolysis: Interference less than 5% up to 5 g/L haemoglobin
8
Refer to Young for further information on interfering substances.
Setting Sheet Footnotes
# User defined ¤ Analyser default value
* Values set for working in U/L. To work in SI units (µkat/L) divide by 60.
§ For use in AB mode only, refer to leaflet for further instruction
BIBLIOGRAPHY
1. Lorentz K. α-Amylase. In: Thomas L, hrsg. Labor und Diagnose. Indikation und Bewertung von Laborbefunden für die Medizinische Diagnostik. Frankfurt/Main:
TH-Books Verlagsgesellschaft, 2005:51-56.
2. Tietz NW, ed. Clinical guide to laboratory tests, 3rd ed. Philadelphia: WB Saunders Company,1995:47pp.
3. Moss DW, Henderson RA. Clinical Enzymology. In: Burtis CA, Ashwood ER, eds. Tietz textbook of clinical chemistry. Philadelphia:WB Saunders Company,
1999;689-698.
4. IFCC methods for measurement of catalytic concentration of enzymes. Part 9. IFCC method for alpha-amylase [1,4-α-D-glucan 4-glucanohydrolase, EC 3.2.1.1].
International Federation of Clinical Chemistry. Clin Chim Acta. 1999;281(1-2):S5-39.
5. Ehret W, Heil W, Schmitt Y, Töpfer G, Wisser H, Zawta B, et al. Use of anticoagulants in diagnostic laboratory investigations and stability of blood, plasma and
serum samples. WHO/DIL/LAB/99.1 Rev.2:22,46pp.
6. Moss DW, Henderson RA, Kachmar JF. Enzymes. In: Tietz NW, ed. Fundamentals of clinical chemistry. Philadelphia:WB Saunders Company, 1987:393-396.
7. Ballsells D, Gella FJ, Gubern G, Canalias F, Reference values for α-amylase in human serum and urine using 2-chloro-4-nitrophenyl-α-D-maltotrioside as
substrate. Clin Chim Acta 1998;274:213–217.
8. Young DS. Effects of drugs on clinical laboratory tests, 5th ed. AACC Press, 2000.

Enzyme BLOSR6x06.01 EN.01


2009-08
AMYLASE, AU400/AU640 Serum/Plasma Application AMYLASE, AU600 Serum/Plasma Application
System Reagent: OSR6006, OSR6106 Reagent ID: 006 System Reagent: OSR6006, OSR6106 Reagent ID: 006

Specific Test Parameters Specific test parameters


General LIH ISE Range
Test No # Test name AMY ∇ Sample type Ser ∇ Page ½
Test Name: AMY ∇ < > Type: Serum ∇ Operation: Yes ∇
Sample vol. 2 Dil. Vol 0 μL Min. OD Max. OD
Sample: Volume 2 μL Dilution 0 μL Pre-Dilution Rate: 1 Reagent 1 vol 200 Dil. Vol 0 μL L -0.1 H 1.5
Reagents: R1 Volume 200 μL Dilution 0 μL Min OD Max OD Reagent 2 vol 0 Dil. Vol 0 μL Reagent OD limit
R2 Volume 0 μL Dilution 0 μL L -0.1 H 1.5 Fst. L -0.1 Fst. H 0.2
Reagent OD limit: Wave Main 410 Sub 480 ∇ Lst. L -0.1 Lst. H 0.2
Wavelength: Pri. 410 ∇ Sec. 480 ∇ First L -0.1 First H 0.2 Method RATE ∇ Dynamic range
Method: RATE ∇ Last L -0.1 Last H 0.2 Reaction + ∇ L 10* H 2000*
Reaction slope: + ∇ Dynamic Range: Point 1 Fst 5 Lst 10 Correlation factor A 1
Measuring Point 1: First 5 Last 10 L 10* H 2000* Point 2 Fst Lst B 0
Measuring Point 2: First Last Correlation Factor: Sample Pre-dil. Rate ¤
Linearity : 15 % A 1 B 0
No Lag Time: NO ∇ On-board stability period: 30 Linearity Fst 15 % Sec %
No lag time NO ∇ On-board stability period 30
Select using Space key, or select from list displayed by Guide key
Specific Test Parameters
General LIH ISE Range Test No # Test name AMY ∇ Sample type Ser ∇ Page 2/2
Test Name: AMY ∇ < > Type: Serum ∇ Level L Level H
Value/flag # ∇ # #
Value/Flag: # ∇ Level L: # Level H: # Normal range
Normal Ranges: Age L Age H Sex Age L Age H L H
Sex Year Month Year Month L H 1 # ∇ # Y # M # Y # M→ # #
ο 1. # ∇ # # # # # # 2 # ∇ # Y # M # Y # M→ # #
ο 2. # ∇ # # # # # # 3 # ∇ # Y # M # Y # M→ # #
ο 3. # ∇ # # # # # # 4 # ∇ # Y # M # Y # M→ # #
ο 4. # ∇ # # # # # # 5 # ∇ # Y # M # Y # M→ # #
ο 5. # ∇ # # # # # # 6 # ∇ # Y # M # Y # M→ # #
ο 6. # ∇ # # # # # # 7 Non select # #
7. None Selected # # 8 Out of range # #
8. Out of Range L H # # L H
Panic Value: # # Unit: U/L* Decimal places: # Panic value # #
Select the function using the Function key or the Mouse
Calibration specific
Calibration Specific
General ISE Test No # Test name AMY ∇
Test Name: AMY ∇ < > Type Serum ∇ Cal type 1 MB Count #

SERUM/PLASMA APPLICATION

Formula 1 Y=AX+B ∇ Process ∇
Calibration Type: MB ∇ Formula: Y=AX+B ∇ Counts: # Process: ∇
Selection calibrator
Cal No OD Conc Factor/OD-L Factor/OD-H
Cal. No. OD CONC Factor/OD-L Factor/OD-H
Point 1 § ∇ § §7120* §10680*
Point 1: § § §7120* §10680* Point 2 ∇
Point 2:
Point 3 ∇
Point 3:
Point 4 ∇
Point 4:
Point 5 ∇
Point 5:
Point 6: Point 6 ∇
Point 7: Point 7 ∇
1-point cal. Point
1-Point Cal. Point: ο with CONC-0 Slope Check: None ∇ Advanced Calibration: # ∇
MB type factor #
MB Type Factor: # Calibration Stability Period: Calibrator stability period
Select the function using the Function key or the Mouse
# User defined # User defined ¤ Analyser default value
* Values set for working in U/L. To work in SI units (µkat/L) divide by 60 * Values set for working in U/L. To work in SI units (µkat/L) divide by 60
§ For use in AB mode only, refer to leaflet for further instruction. § For use in AB mode only, refer to leaflet for further instruction.

Enzyme BSOSR6x06.01
2009-08
AMYLASE, AU2700/AU5400 Serum/Plasma Application AMYLASE, AU680/AU480 Serum/Plasma Application
System Reagent: OSR6006, OSR6106 Reagent ID: 006 System Reagent: OSR6006, OSR6106 Reagent ID: 006
Parameters Specific Test Parameters
Specific Test Parameters General LIH ISE HbA1c Calculated Test Range
General LIH ISE Range
Test Name: AMY ∇ < > Type: Serum ∇ Operation Yes ∇
Test Name: AMY ∇ < > Type: Serum ∇ Operation: Yes ∇

Sample: Volume 1.6 Dilution 0 Pre-Dilution Rate: 1 Sample Volume 1.6 μL Dilution 0 μL OD Limit
μL μL
Pre-Dilution Rate 1 ∇ Min.OD -0.1 Max.OD 1.5
Reagents: R1 Volume 160 μL Dilution 0 μL Min OD Max OD
Rgt. Volume R1(R1-1) 160 μL Dilution 0 μL Reagent OD Limit
R2 Volume 0 μL Dilution 0 μL L -0.1 H 1.5
First Low -0.1 High 0.2
Reagent OD limit:
Last Low -0.1 High 0.2
Wavelength: Pri. 410 ∇ Sec. 480 ∇ First L -0.1 First H 0.2
R2(R2-1) 0 μL Dilution 0 μL
Method: RATE ∇ Last L -0.1 Last H 0.2
Dynamic Range Low 10* High 2000*
Reaction slope: + ∇ Dynamic Range: Common Rgt. Type Noneф Name ф Correlation Factor A 1 B 0
Measuring Point 1: First 5 Last 10 L 10* H 2000* Wavelength Pri 410 ∇nm Sec. 480 ∇nm Factor for Maker A 1 B 0
Measuring Point 2: First Last Correlation Factor: Method RATE ∇
Linearity : 15 % A 1 B 0 Reaction Slope + ∇ Onboard Stability Period 30 Day 0 Hour
No Lag Time: NO ∇ On-board stability period: 30 Measuring Point1 First 5 Last 10 LIH Influence Check # ∇
Measuring Point2 First Last Lipemia +++++ ∇
Linearity Limit 15 % Icterus ++++ ∇
Specific Test Parameters Lag Time Check NO ∇ Hemolysis +++ ∇
General LIH ISE Range
Parameters Specific Test Parameters
Test Name: AMY ∇ < > Type: Serum ∇ General LIH ISE HbA1c Calculated Test Range

Value/Flag: # ∇ Level L: # Level H: # Test Name: AMY ∇ < > Type: Serum ∇
Normal Ranges: Age L Age H
Sex Year Month Year Month L H Value/Flag: # ∇ Low High
ο 1. # ∇ # # # # # # Level # # Panic Value
Specificl Ranges: From To Low High
ο 2. # ∇ # # # # # #
Sex Year Month Year Month Low High # #
ο 3. # ∇ # # # # # # # # # # # # #
ο 1. ∇
ο 4. # ∇ # # # # # # ο 2. # ∇ # # # # # #
ο 5. # ∇ # # # # # # ο 3. # ∇ # # # # # #
ο 6. # ∇ # # # # # # ο 4. # ∇ # # # # # #
7. None Selected # # ο 5. # ∇ # # # # # #
8. Out of Range L H # # ο 6. # ∇ # # # # # #
Panic Value: # # Unit: U/L* Decimal places: # 7. No demographics # #
8. Not within expected values # #
Unit U/L* Decimal Places #
Parameters Calibration Parameters
Calibration Specific
Calibrators Calibration Specific STAT Table Calibration
General ISE
General ISE
Test Name: AMY ∇ < > Type Serum ∇

SERUM/PLASMA APPLICATION
Test Name: AMY ∇ < > Type Serum ∇ ο Use Serum Cal.
Calibration Type: MB ∇ Formula: Y=AX+B ∇ Counts: # Process: ∇ Calibration Type: MB ∇ Formula: Y=AX+B ∇ Counts: # ∇
<Calibrator Parameters> Range
Cal. No. OD CONC Factor/OD-L Factor/OD-H Calibrator OD Conc Low High Slope Check None ∇
Point 1: § § §7120* §10680* Point 1: § ∇ § §7120* §10680*
Point 2: Point 2: ∇ Allowance Range Check
Point 3: Point 3: ∇
Point 4: Point 4: ∇ ο Reagent Blank
Point 5: Point 5: ∇ ο Calibration
Point 6: Point 6: ∇
Point 7: Point 7 ∇ Advanced Calibration
1-Point Cal. Point: ο with CONC-0 Slope Check: None ∇ Advanced Calibration: # ∇ Point 8 ∇ Operation # ∇
Point 9 ∇
MB Type Factor: # Calibration Stability Period:
Point 10 ∇ Interval (RB/ACAL) ∇
<Point Cal. For No. of Correction Points ∇ Use Master Curve ∇ ο Lot Calibration
Master Curve> OD Range
# User defined
Calibrator OD Conc Low High Stability
* Values set for working in U/L. To work in SI units (µkat/L) divide by 60
Point-1 ∇ Reagent Blank Day Hour
§ For use in AB mode only, refer to leaflet for further instruction.
Point-2 ∇ Calibration Day Hour
ф AU680
MB Type Factor: # 1-Point Calibration Point ∇ ο with Conc-0

Enzyme BSOSR6x06.01
2009-08
AMYLASE, AU400/AU640 Urine Application AMYLASE, AU600 Urine Application
System Reagent: OSR6006, OSR6106 Reagent ID: 006 Reagent: OSR6006, OSR6106 Reagent ID: 006

Specific Test Parameters Specific test parameters


General LIH ISE Range
Test No # Test name AMY ∇ Sample type Uri ∇ Page 1/2
Test Name: AMY ∇ < > Type: Urine ∇ Operation: Yes ∇
Sample vol. 1 Dil. vol 0 μL Min. OD Max. OD
Sample: Volume 1 μL Dilution 0 μL Pre-Dilution Rate: 1 Reagent 1 vol 150 Dil. vol 0 μL L -0.1 H 1.5
Reagents: R1 Volume 150 μL Dilution 0 μL Min OD Max OD Reagent 2 vol 0 Dil. vol 0 μL Reagent OD limit
R2 Volume 0 μL Dilution 0 μL L -0.1 H 1.5 Fst. L -0.1 Fst. H 0.2
Reagent OD limit: Wave Main 410 Sub 480 ∇ Lst. L -0.1 Lst. H 0.2
Wavelength: Pri. 410 ∇ Sec. 480 ∇ First L -0.1 First H 0.2 Method RATE ∇ Dynamic range
Method: RATE ∇ Last L -0.1 Last H 0.2 Reaction + ∇ L 5* H 4800*
Reaction slope: + ∇ Dynamic Range: Point 1 Fst 5 Lst 10 Correlation factor A 1
Measuring Point 1: First 5 Last 10 L 5* H 4800* Point 2 Fst Lst B 0
Measuring Point 2: First Last Correlation Factor: Sample Pre-dil. Rate ¤
Linearity : 15 % A 1 B 0
No Lag Time: NO ∇ On-board stability period: 30 Linearity Fst 15 % Sec %
No lag time NO ∇ On-board stability period 30
Select using Space key, or select from list displayed by Guide key
Specific Test Parameters
General LIH ISE Range Test No # Test name AMY ∇ Sample type Uri ∇ Page 2/2
Test Name: AMY ∇ < > Type: Urine ∇ Level L Level H
Value/flag # ∇ # #
Value/Flag: # ∇ Level L: # Level H: # Normal range
Normal Ranges: Age L Age H Sex Age L Age H L H
Sex Year Month Year Month L H 1 # ∇ # Y # M # Y # M→ # #
ο 1. # ∇ # # # # # # 2 # ∇ # Y # M # Y # M→ # #
ο 2. # ∇ # # # # # # 3 # ∇ # Y # M # Y # M→ # #
ο 3. # ∇ # # # # # # 4 # ∇ # Y # M # Y # M→ # #
ο 4. # ∇ # # # # # # 5 # ∇ # Y # M # Y # M→ # #
ο 5. # ∇ # # # # # # 6 # ∇ # Y # M # Y # M→ # #
ο 6. # ∇ # # # # # # 7 Non select # #
7. None Selected # # 8 Out of range # #
8. Out of Range L H # # L H
Panic Value: # # Unit: U/L* Decimal places: # Panic value # #
Select the function using the Function key or the Mouse

URINE APPLICATION
Calibration specific
Calibration Specific
General ISE Test No # Test name AMY ∇
Test Name: AMY ∇ < > Type Urine ∇ Cal type 1 MB ∇ Count #
Formula 1 Y=AX+B ∇ Process ∇
Calibration Type: MB ∇ Formula: Y=AX+B ∇ Counts: # Process: ∇
Selection calibrator
Cal No OD Conc Factor/OD-L Factor/OD-H
Cal. No. OD CONC Factor/OD-L Factor/OD-H
Point 1 § ∇ § §9300* §17300*
Point 1: § § §9300* §17300*
Point 2 ∇
Point 2:
Point 3 ∇
Point 3:
Point 4: Point 4 ∇
Point 5: Point 5 ∇
Point 6: Point 6 ∇
Point 7: Point 7 ∇
1-Point Cal. Point: With CONC-0 Slope Check: None # 1-point cal. point
ο ∇ Advanced Calibration: ∇
MB type factor #
MB Type Factor: # Calibration Stability Period: Calibrator stability period
Select the function using the Function key or the Mouse
# User defined # User defined ¤ Analyser default value
* Values set for working in U/L. To work in SI units (µkat/L) divide by 60 * Values set for working in U/L. To work in SI units (µkat/L) divide by 60
§ For use in AB mode only, refer to leaflet for further instruction. § For use in AB mode only, refer to leaflet for further instruction.

Enzyme BSOSR6x06.01
2009-08
AMYLASE, AU2700/AU5400 Urine Application AMYLASE, AU680/AU480 Urine Application
System Reagent: OSR6006, OSR6106 Reagent ID: 006 System Reagent: OSR6006, OSR6106 Reagent ID: 006
Parameters Specific Test Parameters
Specific Test Parameters General LIH ISE HbA1c Calculated Test Range
General LIH ISE Range
Test Name: AMY ∇ < > Type: Urine ∇ Operation Yes ∇
Test Name: AMY ∇ < > Type: Urine ∇ Operation: Yes ∇

Sample: Volume 1.0 Dilution 0 Pre-Dilution Rate: 1 Sample Volume 1.0 μL Dilution 0 μL OD Limit
μL μL
Pre-Dilution Rate 1 ∇ Min.OD -0.1 Max.OD 1.5
Reagents: R1 Volume 150 μL Dilution 0 μL Min OD Max OD
Rgt. Volume R1(R1-1) 150 μL Dilution 0 μL Reagent OD Limit
R2 Volume 0 μL Dilution 0 μL L -0.1 H 1.5
First Low -0.1 High 0.2
Reagent OD limit:
Last Low -0.1 High 0.2
Wavelength: Pri. 410 ∇ Sec. 480 ∇ First L -0.1 First H 0.2
R2(R2-1) 0 μL Dilution 0 μL
Method: RATE ∇ Last L -0.1 Last H 0.2
Dynamic Range Low 5* High 4800*
Reaction slope: + ∇ Dynamic Range: Common Rgt. Type Noneф Name ф Correlation Factor A 1 B 0
Measuring Point 1: First 5 Last 10 L 5* H 4800* Wavelength Pri 410 ∇nm Sec. 480 ∇nm Factor for Maker A 1 B 0
Measuring Point 2: First Last Correlation Factor: Method RATE ∇
Linearity : 15 % A 1 B 0 Reaction Slope + ∇ Onboard Stability Period 30 Day 0 Hour
No Lag Time: NO ∇ On-board stability period: 30 Measuring Point1 First 5 Last 10
Measuring Point2 First Last
Linearity Limit 15 %
Specific Test Parameters Lag Time Check NO ∇
General LIH ISE Range
Parameters Specific Test Parameters
Test Name: AMY ∇ < > Type: Urine ∇ General LIH ISE HbA1c Calculated Test Range

Value/Flag: # ∇ Level L: # Level H: # Test Name: AMY ∇ < > Type: Urine ∇
Normal Ranges: Age L Age H
Sex Year Month Year Month L H Value/Flag: # ∇ Low High
ο 1. # ∇ # # # # # # Level # # Panic Value
Specificl Ranges: From To Low High
ο 2. # ∇ # # # # # #
Sex Year Month Year Month Low High # #
ο 3. # ∇ # # # # # # # # # # # # #
ο 1. ∇
ο 4. # ∇ # # # # # # ο 2. # ∇ # # # # # #
ο 5. # ∇ # # # # # # ο 3. # ∇ # # # # # #
ο 6. # ∇ # # # # # # ο 4. # ∇ # # # # # #
7. None Selected # # ο 5. # ∇ # # # # # #
8. Out of Range L H # # ο 6. # ∇ # # # # # #
Panic Value: # # Unit: U/L* Decimal places: # 7. No demographics # #
8. Not within expected values # #

URINE APPLICATION
Unit U/L* Decimal Places #
Parameters Calibration Parameters
Calibration Specific
Calibrators Calibration Specific STAT Table Calibration
General ISE
General ISE
Test Name: AMY ∇ < > Type Urine ∇ Test Name: AMY ∇ < > Type Urine ∇ ο Use Serum Cal.
Calibration Type: MB ∇ Formula: Y=AX+B ∇ Counts: # Process: ∇ Calibration Type: MB ∇ Formula: Y=AX+B ∇ Counts: # ∇
<Calibrator Parameters> Range
Cal. No. OD CONC Factor/OD-L Factor/OD-H Calibrator OD Conc Low High Slope Check None ∇
Point 1: § § §9300* §17300* Point 1: § ∇ § §9300* §17300*
Point 2: Point 2: ∇ Allowance Range Check
Point 3: Point 3: ∇
Point 4: Point 4: ∇ ο Reagent Blank
Point 5: Point 5: ∇ ο Calibration
Point 6: Point 6: ∇
Point 7: Point 7 ∇ Advanced Calibration
1-Point Cal. Point: ο with CONC-0 Slope Check: None ∇ Advanced Calibration: # ∇ Point 8 ∇ Operation # ∇
Point 9 ∇
MB Type Factor: # Calibration Stability Period:
Point 10 ∇ Interval (RB/ACAL) # ∇
<Point Cal. For No. of Correction Points ∇ Use Master Curve ∇ ο Lot Calibration
Master Curve> OD Range
# User defined
Calibrator OD Conc Low High Stability
* Values set for working in U/L. To work in SI units (µkat/L) divide by 60
Point-1 ∇ Reagent Blank Day Hour
§ For use in AB mode only, refer to leaflet for further instruction.
Point-2 ∇ Calibration Day Hour
ф AU680
MB Type Factor: # 1-Point Calibration Point ∇ ο with Conc-0

Enzyme BSOSR6x06.01
2009-08
α-AMYLASE
OSR6182 4 x 40 mL R1
4 x 10 mL R2
Intended Use
Kinetic colour test for the quantitative determination of α-amylase, [1,4-α-D-glucan 4-glucanohydrolase, EC 3.2.1.1], in human serum, plasma
and urine on Beckman Coulter analysers. For in vitro diagnostic use only.
Summary1,2,3
Amylases are a group of hydrolases that split complex carbohydrates composed of alpha-D-glucose units linked through carbon atoms 1 and 4
located on adjacent glucose residues. In the body, amylase is present in a number of organs and tissues. The greatest concentration is present
in the pancreas, where the enzyme is synthesised by the acinar cells and then secreted into the intestinal tract by way of the pancreatic duct
system. The salivary glands also secrete a potent amylase to initiate hydrolysis of starches while the food is still in the mouth and oesophagus.
Diseases resulting in elevation of plasma alpha-amylase include: acute pancreatitis, parotitis, alcoholism, renal insufficiency and diseases such
as viral hepatitis, AIDS, abdominal typhoid, sarcoidosis and trauma to the upper abdomen. There is also a detectable increase in amylase after
an ERCP procedure.
In acute pancreatitis, amylase increases 5-6h after the onset of symptoms and remains elevated for 2-5 days. The increase in plasma activity
does not reflect disease severity and conversely, extensive destruction of the pancreas may not cause a significant increase in the plasma
concentration of pancreatic alpha-amylase.
Alpha amylase is excreted by glomerular filtration and then 50% of it is reabsorbed by the tubules. This reabsorption is significantly reduced in
transient tubular damage, after burns, in the presence of diabetic ketoacidosis and acute pancreatitis as well as proteinuria resulting in an
increase of alpha-amylase clearance. The measurement of alpha amylase in urine is indicated in the investigation of hyperamylasemia
associated with macroamylasemia or renal insufficiency.
Hypoamylasemia has been observed in advanced cystic fibrosis, severe liver disease and pancreatectomy. Also due to the decreased
concentration of the salivary fraction hypoamylasemia has been found in obese subjects.
Test Principle4
Method based on the recommendations of the “International Federation of Clinical Chemistry” (IFCC).
The α-amylase colour test employs 4,6-ethylidene(G7)-p-nitrophenyl(G1)-α-D-maltoheptaoside (ethylidene-G7PNP) as substrate. This substrate
reacts with α-amylase and the fragments with α-glucosidase to give a 100% release of p-Nitrophenol (PNP). The increase of absorbance at
410 nm is directly proportional to the α-amylase activity in the sample.
Reaction Principle (Simplified)4
α-amylase
5 ethylidene-G7PNP + 5H20 2 ethylidene-G5 + 2 G2PNP + 2 ethylidene-G4
+ 2 G3PNP + ethylidene-G3 + G4PNP
α-glucosidase
2 G2 PNP + 2 G3PNP + G4PNP + 14 H20 5PNP + 14G
(PNP = p-Nitrophenol; G = Glucose)
Contents, Reagent Composition in the Test
Final concentration of reactive ingredients:
HEPES buffer (pH 7.15) 50 mmol/L
Sodium chloride 70 mmol/L
Calcium chloride 1 mmol/L
4,6-ethylidene-G7PNP > 1.4 mmol/L
α-Glucosidase ≥ 4.8 kU/L
Preservatives
Precautions and Warnings
R43. May cause sensitisation by skin contact.
S24,S37,S60. Avoid contact with skin. Wear suitable gloves. This material and its container must be disposed of as hazardous waste.
Exercise the normal precautions required for handling all laboratory reagents.
Dispose of all waste material in accordance with local guidelines.
Refer to Safety Data Sheets for further information.
Reagent Preparation
The reagents are ready for use and can be placed directly on board the instrument.
Care should be taken when handling this reagent to avoid contamination with skin and body fluids.
Storage and Stability
The reagents are stable, unopened, up to the stated expiry date when stored at 2...8°C. Once open reagents stored on board the instrument are
stable for 90 days.
Discard reagents if any discolouration is observed.
Specimen
Serum or heparinised plasma.
5
Stable in serum and plasma for 7 days when stored at 2…25°C.
1,6
Plasma using EDTA, oxalate and citrate should be avoided.
1 6
Urine: Timed or random sample. Adjust pH to approximately 7.0 before storage.
5
Stable in urine for 10 days when stored at 2…8°C and 2 days when stored at 15…25°C.
Test Procedure
Refer to the appropriate User Guide and Setting Sheet for analyser-specific assay instructions for the sample type as listed in the Intended Use statement.

EN.01 BLOSR6x82.01 Enzyme


2009-08
Calibration
The test is run in MB-mode. To provide a robust approach to generate the analyser specific MB factor, it is recommended that 5 separate
calibration events should be used. A fresh vial of calibrator, utilising System Calibrator Cat No. 66300 for serum application and Urine calibrator
Cat no. ODC0025 for urine application, in the AB calibration mode, should be used for each of these runs. When calculating the mean factor
from the separate runs the data should be examined for obvious outliers which should be repeated and replaced. For the AU2700/AU5400 this
procedure needs to be performed for each ring. Quality control procedures should be undertaken immediately following calibration in
accordance with good laboratory practice.
The calibrator values are traceable to a manual IFCC reference method and IRMM/IFCC-456.
Re-establishment of the analyser specific MB factor is recommended when a critical part of the analyser is replaced or as deemed necessary by the user.
Reagent blank measurement is recommended when changing to a new lot of reagent.
Quality Control
Controls Cat. No. ODC0003 and ODC0004 or other control materials with values determined by this Beckman Coulter system may be used for
the serum application.
Biorad Liquichek Urine Chemistry Controls Cat. No. 397 and 398 or other control materials with values determined by this Beckman Coulter
system may be used for the urine application.
Each laboratory should establish its own control frequency however good laboratory practice suggests that controls be tested each day patient
samples are tested and each time calibration/blanking is performed.
The results obtained by any individual laboratory may vary from the given mean value. It is therefore recommended that each laboratory
generates analyte specific control target values and intervals based on multiple runs according to their requirements. These target values should
fall within the corresponding acceptable ranges given in the relevant product literature.
If any trends or sudden shifts in values are detected, review all operating parameters.
Each laboratory should establish guidelines for corrective action to be taken if controls do not recover within the specified limits.
Calculation
The Beckman Coulter analysers automatically compute the α-amylase activity of each sample.
Reference Intervals7
Serum/Plasma 28 - 100 U/L (0.46 – 1.66 µkat/L)
Urine
Male ≤ 490 U/L (8.16 µkat/L) or 280 U/g creatinine
Female ≤ 450 U/L (7.50 µkat/L) or 380 U/g creatinine
Expected values may vary with age, sex, sample type, diet and geographical location. Each laboratory should verify the transferability of the
expected values to its own population, and if necessary determine its own reference interval according to good laboratory practice. For
diagnostic purposes, results should always be assessed in conjunction with the patient's medical history, clinical examinations and other
findings.
Specific Performance Characteristics
Data contained within this section is representative of performance on Beckman Coulter systems. Data obtained in your laboratory may differ
from these values.
Linearity
The test is linear within an enzyme activity range of 10 – 1500 U/L (0.2 – 25.0 µkat/L) for serum and plasma.
The test is linear within an enzyme activity range of 10 – 4800 U/L (0 – 80.0 µkat/L) for urine.
Precision
The following data was obtained on an AU640 using 3 serum pools analysed over 20 days.
n = 80 Within Run Total
Mean U/L SD CV% SD CV%
51 0.36 0.71 0.59 1.16
132 0.73 0.56 1.82 1.38
1267 6.98 0.55 12.94 0.99
The following data was obtained on an AU640 using 3 urine pools analysed over 20 days
n = 80 Within Run Total
Mean U/L SD CV% SD CV%
46 0.62 1.35 1.53 3.34
585 5.04 0.86 7.41 1.27
3732 21.11 0.57 44.65 1.20
Sensitivity
The lowest detectable level using serum settings on an AU640 analyser was calculated as 1 U/L.
The lowest detectable level using urine settings on an AU640 analyser was calculated as 2 U/L.
The lowest detectable level represents the lowest measurable level of α-amylase that can be distinguished from zero. It is calculated as the
absolute mean plus three standard deviations of 20 replicates of an analyte free sample.
Method Comparison
Patient serum samples were used to compare this α-Amylase OSR6182 assay on the AU640 against another commercially available amylase
assay. Results of linear regression analysis were as follows:
y = 0.985x + 0.346 r = 1.000 n = 116 Sample range = 30 – 1482 U/L
Patient urine samples were used to compare this α-Amylase OSR6182 assay on the AU640 against another commercially available amylase
assay. Results of linear regression analysis were as follows:
y = 1.003x – 5.213 r = 1.000 n = 98 Sample range = 18 – 4470 U/L

Interfering Substances
Results of serum studies conducted to evaluate the susceptibility of the method to interference were as follows:
Icterus: Interference less than 3% up to 40 mg/dL or 684 µmol/L bilirubin
Haemolysis: Interference less than 10% up to 5 g/L haemoglobin
®
Lipemia: Interference less than 3% up to 1000 mg/dL Intralipid
Results of urine studies conducted to evaluate the susceptibility of the method to interference were as follows:
Ascorbate: Interference less than 5% up to 50 mg/dL ascorbate
Icterus: Interference less than 10% up to 40 mg/dL or 684 µmol/L bilirubin
Haemolysis: Interference less than 5% up to 5 g/L haemoglobin
8
Refer to Young for further information on interfering substances.

Enzyme BLOSR6x82.01 EN.01


2009-08
Setting Sheet Footnotes
# User defined ¤ Analyser default value
* Values set for working in U/L. To work in SI units (µkat/L) divide by 60.
§ For use in AB mode only, refer to leaflet for further instruction
BIBLIOGRAPHY
1. Lorentz K. α-Amylase. In: Thomas L, hrsg. Labor und Diagnose.. Indikation und Bewertung von Laborbefunden für die Medizinische Diagnostik. Frankfurt/Main:
TH-Books Verlagsgellschaft, 2005:51-56.
2. Tietz NW, ed. Clinical guide to laboratory tests, 3rd ed. Philadelphia: WB Saunders Company,1995:47pp.
3. Moss DW, Henderson RA. Clinical Enzymology. In: Burtis CA, Ashwood ER, eds. Tietz textbook of clinical chemistry. Philadelphia:WB Saunders Company,
1999; 689-692.
4. Lorentz K. Approved recommendation on IFCC methods for the measurement of catalytic concentration of enzymes. Part 9. IFCC method for α-amylase (1,4-α-
D-glucan 4-glucanohydrolase, EC 3.2.1.1). International Federation of Clinical Chemistry and Laboratory Medicine (IFCC). Committee on enzymes. Clin Chem
Lab Med 1998;36(3):185-203.
5. Ehret W, Heil W, Schmitt Y, Töpfer G, Wisser H, Zawta B, et al. Use of anticoagulants in diagnostic laboratory investigations and stability of blood, plasma and
serum samples. WHO/DIL/LAB/99.1 Rev.2:22, 46pp.
6. Moss DW, Henderson RA, Kachmar JF. Enzymes. In: Tietz NW, ed. Fundamentals of clinical chemistry. Philadelphia: WB Saunders Company, 1987:393-396.
7. Junge W, Wortmann W, Wilke B, Waldenstrom J, Weittenhiller A, Finke J, Klein G, Development and evaluation of assays for the determination of total and
pancreatic amylase at 37°C according to the principle recommended by the IFCC. Clin Biochem 2001;34:607–615.
8. Young DS. Effects of drugs on clinical laboratory tests, 5th ed. AACC Press, 2000.

EN.01 BLOSR6x82.01 Enzyme


2009-08
AMYLASE (EPS Liquid), AU400/AU640 Serum/Plasma Application AMYLASE (EPS Liquid), AU600 Serum/Plasma Application
System Reagent: OSR6182 Reagent ID: 082 System Reagent: OSR6182 Reagent ID: 082

Specific Test Parameters Specific test parameters


General LIH ISE Range
Test No # Test name AMY ∇ Sample type Ser ∇ Page ½
Test Name: AMY ∇ < > Type: Serum ∇ Operation: Yes ∇
Sample vol. 4 Dil. Vol 0 μL Min. OD Max. OD
Sample: Volume 4 μL Dilution 0 μL Pre-Dilution Rate: 1 Reagent 1 vol 200 Dil. Vol 0 μL L -0.1 H 2.0
Reagents: R1 Volume 200 μL Dilution 0 μL Min OD Max OD Reagent 2 vol 50 Dil. Vol 0 μL Reagent OD limit
R2 Volume 50 μL Dilution 0 μL L -0.1 H 2.0 Fst. L -0.1 Fst. H 0.2
Reagent OD limit: Wave Main 410 Sub 480 ∇ Lst. L -0.1 Lst. H 0.2
Wavelength: Pri. 410 ∇ Sec. 480 ∇ First L -0.1 First H 0.2 Method RATE ∇ Dynamic range
Method: RATE ∇ Last L -0.1 Last H 0.2 Reaction + ∇ L 10* H 1500*
Reaction slope: + ∇ Dynamic Range: Point 1 Fst 20 Lst 27 Correlation factor A 1
Measuring Point 1: First 20 Last 27 L 10* H 1500* Point 2 Fst Lst B 0
Measuring Point 2: First Last Correlation Factor: Sample Pre-dil. Rate ¤
Linearity : 15 % A 1 B 0
No Lag Time: NO ∇ On-board stability period: 90 Linearity Fst 15 % Sec %
No lag time NO ∇ On-board stability period 90
Select using Space key, or select from list displayed by Guide key
Specific Test Parameters
General LIH ISE Range Test No # Test name AMY ∇ Sample type Ser ∇ Page 2/2
Test Name: AMY ∇ < > Type: Serum ∇ Level L Level H
Value/flag # ∇ # #
Value/Flag: # ∇ Level L: # Level H: # Normal range
Normal Ranges: Age L Age H Sex Age L Age H L H
Sex Year Month Year Month L H 1 # ∇ # Y # M # Y # M→ # #
ο 1. # ∇ # # # # # # 2 # ∇ # Y # M # Y # M→ # #
ο 2. # ∇ # # # # # # 3 # ∇ # Y # M # Y # M→ # #
ο 3. # ∇ # # # # # # 4 # ∇ # Y # M # Y # M→ # #
ο 4. # ∇ # # # # # # 5 # ∇ # Y # M # Y # M→ # #
ο 5. # ∇ # # # # # # 6 # ∇ # Y # M # Y # M→ # #
ο 6. # ∇ # # # # # # 7 Non select # #
7. None Selected # # 8 Out of range # #
8. Out of Range L H # # L H
Panic Value: # # Unit: U/L* Decimal places: # Panic value # #
Select the function using the Function key or the Mouse
Calibration specific
Calibration Specific
General ISE Test No # Test name AMY ∇
Test Name: AMY ∇ < > Type Serum ∇ Cal type 1 MB Count #

SERUM/PLASMA APPLICATION

Formula 1 Y=AX+B ∇ Process ∇
Calibration Type: MB ∇ Formula: Y=AX+B ∇ Counts: # Process: ∇
Selection calibrator
Cal No OD Conc Factor/OD-L Factor/OD-H
Cal. No. OD CONC Factor/OD-L Factor/OD-H
Point 1 § ∇ § §5570* §8360*
Point 1: § § §5570* §8360*
Point 2 ∇
Point 2:
Point 3: Point 3 ∇
Point 4: Point 4 ∇
Point 5: Point 5 ∇
Point 6: Point 6 ∇
Point 7: Point 7 ∇
1-Point Cal. Point: Slope Check: None # 1-point cal. Point
ο with CONC-0 ∇ Advanced Calibration: ∇
MB type factor #
MB Type Factor: # Calibration Stability Period: 999 Calibrator stability period 999
Select the function using the Function key or the Mouse
# User defined # User defined ¤ Analyser default value
* Values set for working in U/L. To work in SI units (µkat/L) divide by 60 * Values set for working in U/L. To work in SI units (µkat/L) divide by 60
§ For use in AB mode only, refer to IFU for further instruction § For use in AB mode only, refer to IFU for further instruction

Enzyme BSOSR6x82.01
2009-08
AMYLASE (EPS liquid), AU2700/AU5400 Serum/Plasma Application AMYLASE (EPS liquid), AU680/AU480 Serum/Plasma Application
System Reagent: OSR6182 Reagent ID: 082 System Reagent: OSR6182 Reagent ID: 082
Parameters Specific Test Parameters
Specific Test Parameters General LIH ISE HbA1c Calculated Test Range
General LIH ISE Range
Test Name: AMY ∇ < > Type: Serum ∇ Operation Yes ∇
Test Name: AMY ∇ < > Type: Serum ∇ Operation: Yes ∇

Sample: Volume 2 Dilution 0 Pre-Dilution Rate: 1 Sample Volume 2 μL Dilution 0 μL OD Limit


μL μL
Pre-Dilution Rate 1 ∇ Min.OD -0.1 Max.OD 2.0
Reagents: R1 Volume 100 μL Dilution 10 μL Min OD Max OD
Rgt. Volume R1(R1-1) 100 μL Dilution 10 μL Reagent OD Limit
R2 Volume 25 μL Dilution 10 μL L -0.1 H 2.0
First Low -0.1 High 0.2
Reagent OD limit:
Last Low -0.1 High 0.2
Wavelength: Pri. 410 ∇ Sec. 480 ∇ First L -0.1 First H 0.2
R2(R2-1) 25 μL Dilution 10 μL
Method: RATE ∇ Last L -0.1 Last H 0.2
Dynamic Range Low 10* High 1500*
Reaction slope: + ∇ Dynamic Range: Common Rgt. Type Noneф Name ф Correlation Factor A 1 B 0
Measuring Point 1: First 20 Last 27 L 10* H 1500* Wavelength Pri 410 ∇nm Sec. 480 ∇nm Factor for Maker A 1 B 0
Measuring Point 2: First Last Correlation Factor: Method RATE ∇
Linearity : 15 % A 1 B 0 Reaction Slope + ∇ Onboard Stability Period 90 Day 0 Hour
No Lag Time: NO ∇ On-board stability period: 90 Measuring Point1 First 20 Last 27 LIH Influence Check # ∇
Measuring Point2 First Last Lipemia +++++ ∇
Linearity Limit 15 % Icterus +++++ ∇
Specific Test Parameters Lag Time Check NO ∇ Hemolysis +++++ ∇
General LIH ISE Range
Parameters Specific Test Parameters
Test Name: AMY ∇ < > Type: Serum ∇ General LIH ISE HbA1c Calculated Test Range

Value/Flag: # ∇ Level L: # Level H: # Test Name: AMY ∇ < > Type: Serum ∇
Normal Ranges: Age L Age H
Sex Year Month Year Month L H Value/Flag: # ∇ Low High
ο 1. # ∇ # # # # # # Level # # Panic Value
Specificl Ranges: From To Low High
ο 2. # ∇ # # # # # #
Sex Year Month Year Month Low High # #
ο 3. # ∇ # # # # # # # # # # # # #
ο 1. ∇
ο 4. # ∇ # # # # # # ο 2. # ∇ # # # # # #
ο 5. # ∇ # # # # # # ο 3. # ∇ # # # # # #
ο 6. # ∇ # # # # # # ο 4. # ∇ # # # # # #
7. None Selected # # ο 5. # ∇ # # # # # #
8. Out of Range L H # # ο 6. # ∇ # # # # # #
Panic Value: # # Unit: U/L* Decimal places: # 7. No demographics # #
8. Not within expected values # #
Unit U/L* Decimal Places #
Parameters Calibration Parameters
Calibration Specific
Calibrators Calibration Specific STAT Table Calibration
General ISE
General ISE

SERUM/PLASMA APPLICATION
Test Name: AMY ∇ < > Type Serum ∇ Test Name: AMY ∇ < > Type Serum ∇ ο Use Serum Cal.
Calibration Type: MB ∇ Formula: Y=AX+B ∇ Counts: # Process: ∇ Calibration Type: MB ∇ Formula: Y=AX+B ∇ Counts: # ∇
<Calibrator Parameters> Factor Range
Cal. No. OD CONC Factor/OD-L Factor/OD-H Calibrator OD Conc Low High Slope Check None ∇
Point 1: § § §6450* §9680* Point 1: § ∇ § §6450* §9680*
Point 2: Point 2: ∇ Allowance Range Check
Point 3: Point 3: ∇
Point 4: Point 4: ∇ ο Reagent Blank
Point 5: Point 5: ∇ ο Calibration
Point 6: Point 6: ∇
Point 7: Point 7 ∇ Advanced Calibration
1-Point Cal. Point: ο with CONC-0 Slope Check: None ∇ Advanced Calibration: # ∇ Point 8 ∇ Operation # ∇
Point 9 ∇
MB Type Factor: # Calibration Stability Period: 999
Point 10 ∇ Interval (RB/ACAL) # ∇
<Point Cal. For No. of Correction Points ∇ Use Master Curve ∇ ο Lot Calibration
# User defined Master Curve> OD Range
Calibrator OD Conc Low High Stability
* Values set for working in U/L. To work in SI units (µkat/L) divide by 60
§ For use in AB mode only, refer to IFU for further instruction Point-1 ∇ Reagent Blank 999 Day 0 Hour
ф AU680 Point-2 ∇ Calibration Day Hour
MB Type Factor: # 1-Point Calibration Point ∇ ο with Conc-0

Enzyme BSOSR6x82.01
2009-08
AMYLASE (EPS Liquid), AU400/AU640 Urine Application AMYLASE (EPS Liquid), AU600 Urine Application
System Reagent: OSR6182 Reagent ID: 082 System Reagent: OSR6182 Reagent ID: 082

Specific Test Parameters Specific test parameters


General LIH ISE Range
Test No # Test name AMY ∇ Sample type Uri ∇ Page 1/2
Test Name: AMY ∇ < > Type: Urine ∇ Operation: Yes ∇
Sample vol. 1.5 Dil. vol 0 μL Min. OD Max. OD
Sample: Volume 1.5 μL Dilution 0 μL Pre-Dilution Rate: 1 Reagent 1 vol 150 Dil. vol 0 μL L -0.1 H 2.0
Reagents: R1 Volume 150 μL Dilution 0 μL Min OD Max OD Reagent 2 vol 38 Dil. vol 0 μL Reagent OD limit
R2 Volume 38 μL Dilution 0 μL L -0.1 H 2.0 Fst. L -0.1 Fst. H 0.2
Reagent OD limit: Wave Main 410 Sub 480 ∇ Lst. L -0.1 Lst. H 0.2
Wavelength: Pri. 410 ∇ Sec. 480 ∇ First L -0.1 First H 0.2 Method RATE ∇ Dynamic range
Method: RATE ∇ Last L -0.1 Last H 0.2 Reaction + ∇ L 10* H 4800*
Reaction slope: + ∇ Dynamic Range: Point 1 Fst 20 Lst 27 Correlation factor A 1
Measuring Point 1: First 20 Last 27 L 10* H 4800* Point 2 Fst Lst B 0
Measuring Point 2: First Last Correlation Factor: Sample Pre-dil. Rate ¤
Linearity : 15 % A 1 B 0
No Lag Time: NO ∇ On-board stability period: 90 Linearity Fst 15 % Sec %
No lag time NO ∇ On-board stability period 90
Select using Space key, or select from list displayed by Guide key
Specific Test Parameters
General LIH ISE Range Test No # Test name AMY ∇ Sample type Uri ∇ Page 2/2
Test Name: AMY ∇ < > Type: Urine ∇ Level L Level H
Value/flag # ∇ # #
Value/Flag: # ∇ Level L: # Level H: # Normal range
Normal Ranges: Age L Age H Sex Age L Age H L H
Sex Year Month Year Month L H 1 # ∇ # Y # M # Y # M→ # #
ο 1. # ∇ # # # # # # 2 # ∇ # Y # M # Y # M→ # #
ο 2. # ∇ # # # # # # 3 # ∇ # Y # M # Y # M→ # #
ο 3. # ∇ # # # # # # 4 # ∇ # Y # M # Y # M→ # #
ο 4. # ∇ # # # # # # 5 # ∇ # Y # M # Y # M→ # #
ο 5. # ∇ # # # # # # 6 # ∇ # Y # M # Y # M→ # #
ο 6. # ∇ # # # # # # 7 Non select # #
7. None Selected # # 8 Out of range # #
8. Out of Range L H # # L H
Panic Value: # # Unit: U/L* Decimal places: # Panic value # #
Select the function using the Function key or the Mouse

URINE APPLICATION
Calibration specific
Calibration Specific
General ISE Test No # Test name AMY ∇
Test Name: AMY ∇ < > Type Urine ∇ Cal type 1 MB ∇ Count #
Formula 1 Y=AX+B ∇ Process ∇
Calibration Type: MB ∇ Formula: Y=AX+B ∇ Counts: # Process: ∇
Selection calibrator
Cal No OD Conc Factor/OD-L Factor/OD-H
Cal. No. OD CONC Factor/OD-L Factor/OD-H
Point 1 § ∇ § §10500* §19500*
Point 1: § § §10500* §19500*
Point 2 ∇
Point 2:
Point 3: Point 3 ∇
Point 4: Point 4 ∇
Point 5: Point 5 ∇
Point 6: Point 6 ∇
Point 7: Point 7 ∇
1-Point Cal. Point: with CONC-0 Slope Check: None # 1-point cal. point
ο ∇ Advanced Calibration: ∇
MB type factor #
MB Type Factor: # Calibration Stability Period: 999 Calibrator stability period 999
Select the function using the Function key or the Mouse
# User defined # User defined ¤ Analyser default value
* Values set for working in U/L. To work in SI units (µkat/L) divide by 60 * Values set for working in U/L. To work in SI units (µkat/L) divide by 60
§ For use in AB mode only, refer to IFU for further instruction § For use in AB mode only, refer to IFU for further instruction

Enzyme BSOSR6x82.01
2009-08
AMYLASE (EPS liquid), AU2700/AU5400 Urine Application AMYLASE (EPS liquid), AU680/AU480 Urine Application
System Reagent: OSR6182 Reagent ID: 082 System Reagent: OSR6182 Reagent ID: 082
Parameters Specific Test Parameters
Specific Test Parameters General LIH ISE HbA1c Calculated Test Range
General LIH ISE Range
Test Name: AMY ∇ < > Type: Urine ∇ Operation Yes ∇
Test Name: AMY ∇ < > Type: Urine ∇ Operation: Yes ∇

Sample: Volume 1.0 Dilution 0 Pre-Dilution Rate: 1 Sample Volume 1.0 μL Dilution 0 μL OD Limit
μL μL
Pre-Dilution Rate 1 ∇ Min.OD -0.1 Max.OD 2.0
Reagents: R1 Volume 100 μL Dilution 0 μL Min OD Max OD
Rgt. Volume R1(R1-1) 100 μL Dilution 0 μL Reagent OD Limit
R2 Volume 25 μL Dilution 0 μL L -0.1 H 2.0
First Low -0.1 High 0.2
Reagent OD limit:
Last Low -0.1 High 0.2
Wavelength: Pri. 410 ∇ Sec. 480 ∇ First L -0.1 First H 0.2
R2(R2-1) 25 μL Dilution 0 μL
Method: RATE ∇ Last L -0.1 Last H 0.2
Dynamic Range Low 10* High 4800*
Reaction slope: + ∇ Dynamic Range: Common Rgt. Type Noneф Name ф Correlation Factor A 1 B 0
Measuring Point 1: First 20 Last 27 L 10* H 4800* Wavelength Pri 410 ∇nm Sec. 480 ∇nm Factor for Maker A 1 B 0
Measuring Point 2: First Last Correlation Factor: Method RATE ∇
Linearity : 15 % A 1 B 0 Reaction Slope + ∇ Onboard Stability Period 90 Day 0 Hour
No Lag Time: NO ∇ On-board stability period: 90 Measuring Point1 First 20 Last 27
Measuring Point2 First Last
Linearity Limit 15 %
Specific Test Parameters Lag Time Check NO ∇
General LIH ISE Range
Parameters Specific Test Parameters
Test Name: AMY ∇ < > Type: Urine ∇ General LIH ISE HbA1c Calculated Test Range

Value/Flag: # ∇ Level L: # Level H: # Test Name: AMY ∇ < > Type: Urine ∇
Normal Ranges: Age L Age H
Sex Year Month Year Month L H Value/Flag: # ∇ Low High
ο 1. # ∇ # # # # # # Level # # Panic Value
Specificl Ranges: From To Low High
ο 2. # ∇ # # # # # #
Sex Year Month Year Month Low High # #
ο 3. # ∇ # # # # # # # # # # # # #
ο 1. ∇
ο 4. # ∇ # # # # # # ο 2. # ∇ # # # # # #
ο 5. # ∇ # # # # # # ο 3. # ∇ # # # # # #
ο 6. # ∇ # # # # # # ο 4. # ∇ # # # # # #
7. None Selected # # ο 5. # ∇ # # # # # #
8. Out of Range L H # # ο 6. # ∇ # # # # # #
Panic Value: # # Unit: U/L* Decimal places: # 7. No demographics # #
8. Not within expected values # #

URINE APPLICATION
Unit U/L* Decimal Places #
Parameters Calibration Parameters
Calibration Specific
Calibrators Calibration Specific STAT Table Calibration
General ISE
General ISE
Test Name: AMY ∇ < > Type Urine ∇ Test Name: AMY ∇ < > Type Urine ∇ ο Use Serum Cal.
Calibration Type: MB ∇ Formula: Y=AX+B ∇ Counts: # Process: ∇ Calibration Type: MB ∇ Formula: Y=AX+B ∇ Counts: # ∇
<Calibrator Parameters> Factor Range
Cal. No. OD CONC Factor/OD-L Factor/OD-H Calibrator OD Conc Low High Slope Check None ∇
Point 1: § § §10500* §19500* Point 1: § ∇ § §10500* §19500*
Point 2: Point 2: ∇ Allowance Range Check
Point 3: Point 3: ∇
Point 4: Point 4: ∇ ο Reagent Blank
Point 5: Point 5: ∇ ο Calibration
Point 6: Point 6: ∇
Point 7: Point 7 ∇ Advanced Calibration
1-Point Cal. Point: ο with CONC-0 Slope Check None ∇ Advanced Calibration: # ∇ Point 8 ∇ Operation # ∇
Point 9 ∇
MB Type Factor: # Calibration Stability Period: 999
Point 10 ∇ Interval (RB/ACAL) # ∇
<Point Cal. For No. of Correction Points ∇ Use Master Curve ∇ ο Lot Calibration
# User defined Master Curve> OD Range
Calibrator OD Conc Low High Stability
* Values set for working in U/L. To work in SI units (µkat/L) divide by 60
§ For use in AB mode only, refer to IFU for further instruction Point-1 ∇ Reagent Blank 999 Day 0 Hour
ф AU680 Point-2 ∇ Calibration Day Hour
MB Type Factor: # 1-Point Calibration Point ∇ ο with Conc-0

Enzyme BSOSR6x82.01
2009-08
AST
OSR6009 4x 6 mL R1
4x 6 mL R2
OSR6109 4x 25 mL R1
4x 25 mL R2
OSR6209 4x 50 mL R1
4x 50 mL R2
OSR6509 4x 104 mL R1
4x 104 mL R2
OSR60180 4x 17 mL Liquid P-5-P R1-2
60106 6x 4 mL Liquid Pyridoxal Phosphate
60100 4x 25 Tablets Pyridoxal Phosphate
Intended Use
Kinetic UV test for the quantitative determination of aspartate aminotransferase, EC 2.6.1.1 (AST), in human serum and plasma on Beckman Coulter
analysers. For in vitro diagnostic use only.
OSR6509 for the use on the AU2700 and AU5400 systems only. OSR60180 for use with 3-part-reagent enabled systems only.

Summary1,2,3
AST occurs in a wide variety of tissues including liver, cardiac muscle, skeletal muscle, brain, kidneys, lungs, pancreas, erythrocytes and leucocytes, with
highest activities found in liver and skeletal muscle. Measurement of AST is indicated in the diagnosis, differentiation and monitoring of hepatobiliary
disease, myocardial infarction and skeletal muscle damage. AST measurement may also be performed as part of medical screening examinations. In
some cases, AST may be useful in monitoring the course of myocardial infarction. Where recent myocardial infarction is suspected, AST has a diagnostic
sensitivity of 96%, with a diagnostic sensitivity of 86% at 12 hours after onset of chest pain. AST levels may be increased in viral hepatitis and liver disease
associated with hepatic necrosis, with 20 to 50 fold elevations frequently encountered. The evaluation of AST activity in relation to ALT (De Ritis ratio;
AST/ALT) is a useful indicator of liver damage. Ratios <1.0 are indicative of mild liver damage, and are particularly associated with diseases of an
inflammatory nature. Ratios >1.0 are indicative of severe liver disease, usually involving necrosis. Increased AST levels may be detected in cirrhosis,
extrahepatic cholestasis, progressive muscular dystrophy, dermatomyositis, acute pancreatitis, haemolytic disease, gangrene, crushed muscle injuries and
pulmonary emboli. Slight or moderate increases in AST levels may also be observed after ingestion of alcohol, or administration of drugs including
penicillin, salicylates or opiates.

Test Principle4
Method based on the recommendations of the “International Federation for Clinical Chemistry” (IFCC).
In this method, aspartate aminotransferase (AST) catalyses the transamination of aspartate and 2-oxoglutarate, forming L-glutamate and oxalacetate. The
addition of pyridoxal phosphate to the reaction mixture ensures maximum catalytic activity of AST. The oxalacetate is reduced to L-malate by malate
dehydrogenase (MDH), while NADH is simultaneously converted to NAD+. The decrease in absorbance due to the consumption of NADH is measured at
340 nm and is proportional to the AST activity in the sample. Endogenous pyruvate is removed by the LDH-reaction during the incubation period.

Reaction Principle
AST
2-Oxoglutarate + L-Aspartate L-Glutamate + Oxalacetate
+ MDH +
Oxalacetate + NADH + H L-Malate + NAD

Contents, Reagent Composition in the Test


Final concentration of reactive ingredients:
Tris buffer, pH 7.65 (37°C) 80 mmol/L
L-aspartate 240 mmol/L
2-Oxoglutarate 12 mmol/L
LDH ≥ 0.9 kU/L
MDH ≥ 0.6 kU/L
NADH 0.20 mmol/L
Pyridoxal phosphate (P-5-P) 0.1 mmol/L (when Cat. No. 60106, 60100 or OSR60180 is used)
Preservative
Precautions and Warnings
Exercise the normal precautions required for handling all laboratory reagents.
To avoid the possible build-up of azide compounds, flush waste-pipes with water after the disposal of undiluted reagent.
Dispose of all waste material in accordance with local guidelines.
Safety data sheet available for professional user on request.

Reagent Preparation
With manual pyridoxal phosphate addition
Pyridoxal Phosphate Liquid (Cat. No. 60106), and Pyridoxal Phosphate tablets (Cat. No. 60100) are supplied separately for pyridoxal phosphate activation.
Pipette Pyridoxal Phosphate Liquid 60106 into the R1 bottle according to the table below, and mix by gentle inversion. Alternatively, dissolve tablets
completely in the R1 bottle according to the table below, by mixing gently several times. R2 is ready for use and can be placed directly on board the
instrument. This method can also be used for 3-part-reagent enabled systems.
Cat No. Volume Pyridoxal Phosphate Liquid No. of tablets
OSR6009 0.25 mL 1 tablet
OSR6109 1 mL 4 tablets
OSR6209 2 mL 8 tablets
OSR6509 4 mL 17 tablets
EN.01 BLOSR6x09.01 Enzyme
2009-08
Without pyridoxal phosphate activation
The reagents are ready for use and can be placed directly on board the instrument.

Storage and Stability


Reagents are stable, unopened, up to the stated expiry date when stored at 2…8°C.
Without pyridoxal phosphate activation
Once open, reagents stored on board the instrument are stable for 30 days.
With pyridoxal phosphate activation
After addition of pyridoxal phosphate, R1 stored on board the instrument is stable for 7 days.
R2 stored on board the instrument is stable for 30 days.
Pyridoxal Phosphate Liquid reagent (Cat no 60106)
Once open, Pyridoxal Phosphate Liquid reagent is stable until the expiry date printed on the label, provided that contamination is avoided through
adherence to GLP, the cap is replaced immediately after use and the reagent is stored at 2…8°C.
3-part-reagent enabled systems with P-5-P activation
Reagent Preparation
P-5-P Liquid (Cat. No. OSR60180) is specifically for use on board 3-part-reagent enabled systems. The reagent is ready for use and can be placed directly
on board the instrument in the R1 carousel. R1 and R2 are also ready for use and can be placed directly on board the instrument.
Storage and stability
Once open, P-5-P (Cat no OSR60180) is stable on board the instrument for 60 days. R1 and R2 reagent stored on board the instrument are stable for
30 days.
Specimen
5
Serum and heparinised plasma: Stable in serum for 7 days when stored at 2…8°C and 4 days when stored at 15...25°C.
Haemolysed samples should be avoided as the concentration of AST in erythrocytes is approximately 15 times higher than that of normal serum.

Test Procedure
Refer to the appropriate User Guide and Setting Sheet for analyser-specific assay instructions for the sample type as listed in the Intended Use statement.
The paediatric application is suitable for use with small volume serum/plasma samples.
Calibration
The test is run in MB-mode. To provide a robust approach to generate the analyser specific MB factor it is recommended that 5 separate calibration events
should be used. A fresh vial of calibrator, utilising System Calibrator Cat No. 66300 in the AB calibration mode, should be used for each of these runs.
When calculating the mean factor from the separate runs the data should be examined for obvious outliers which should be repeated and replaced. For the
AU2700/AU5400 this procedure needs to be performed for each ring. Quality control procedures should be undertaken immediately following calibration in
accordance with good laboratory practice.
With pyridoxal phosphate activation the calibrator value is traceable to the IFCC reference method. Without pyridoxal phosphate activation,the calibrator
value is traceable to a Beckman Coulter Master Calibrator.
Re-establishment of the analyser specific MB factor is recommended when a critical part of the analyser is replaced.
Reagent blank measurement is recommended when changing to a new lot of reagent.
Quality Control
Controls Cat. No. ODC0003 and ODC0004 or other control materials with values determined by this Beckman Coulter system may be used.
Each laboratory should establish its own control frequency however good laboratory practice suggests that controls be tested each day patient samples
are tested and each time calibration/blanking is performed.
The results obtained by any individual laboratory may vary from the given mean value. It is therefore recommended that each laboratory generates analyte
specific control target values and intervals based on multiple runs according to their requirements. These target values should fall within the corresponding
acceptable ranges given in the relevant product literature.
If any trends or sudden shifts in values are detected, review all operating parameters.
Each laboratory should establish guidelines for corrective action to be taken if controls do not recover within the specified limits.
Calculation
The Beckman Coulter analysers automatically compute the AST activity of each sample.
Reference Intervals6,7
Male (Adult) < 50 U/L (0.85 µkat/L)
Female (Adult) < 35 U/L (0.60 µkat/L)
Newborn 25 − 75 U/L (0.42 − 1.25 µkat/L)
Infant 15 − 60 U/L (0.25 − 1 µkat/L)
Expected values may vary with age, sex, sample type, diet and geographical location. Each laboratory should verify the transferability of the expected
values to its own population, and if necessary determine its own reference interval according to good laboratory practice. For diagnostic purposes, results
should always be assessed in conjunction with the patient's medical history, clinical examinations and other findings.
Specific performance characteristics
Data contained within this section is representative of performance on Beckman Coulter systems. Data obtained in your laboratory may differ from these
values.
Linearity
The test is linear within an enzyme activity range of 3 – 1000 U/L (0.05 – 16.7 µkat/L).
Precision
The following data was obtained on an AU600 using 3 serum pools analysed over 10 days.
n = 60 Within Run Total
Mean U/L SD CV% SD CV%
27 0.53 1.93 1.16 4.23
71 0.59 0.84 0.95 1.35
415 2.42 0.58 5.05 1.22
Sensitivity
The lowest detectable level on an AU600 analyser was estimated at 1 U/L.
The lowest detectable level represents the lowest measurable level of AST that can be distinguished from zero. It is calculated as the absolute mean plus
three standard deviations of 20 replicates of an analyte free sample.

Enzyme BLOSR6x09.01 EN.01


2009-08
Method Comparison with pyridoxal phosphate activation
Patient serum samples were used to compare this AST OSR6109 assay on the AU640 against the IFCC reference method. Results of linear regression
analysis were as follows:
y = 1.032x - 0.2 r = 0.997 n = 112 Sample range = 9 – 250 U/L

Interfering Substances
Results of studies conducted to evaluate the susceptibility of the method to interference were as follows:
Icterus: Interference less than 10% up to 40 mg/dL or 684 µmol/L bilirubin
®
Lipemia: Interference less than 5% up to 300 mg/dL Intralipid
Pyruvate: Interference less than 10% up to 1 mmol/L pyruvate
8
Refer to Young for further information on interfering substances.
Limitations
Highly lipemic samples may exceed the reaction absorbance and will be flagged with a “@”. Such samples should be diluted and re-run.
Setting Sheet Footnotes
# User defined ¤ Analyser default value
* Values set for working in U/L. To work in SI units (µkat/L) divide by 60.
§ For use in AB mode only, refer to leaflet for further instruction

BIBLIOGRAPHY
1. Thomas L. Alanine aminotransferase (ALT), Aspartate aminotransferase (AST). In:Thomas L, ed. Clinical laboratory diagnostics. Use and assessment of clinical
laboratory results. Frankfurt/Main: TH-Books Verlagsgesellschaft, 1998:55-65.
2. Moss DW, Henderson RA, Kachmar JF. Enzymes. In: Tietz NW, ed. Fundamentals of clinical chemistry. Philadelphia:WB Saunders Company, 1987:369-373.
3. Schmidt E, Schmidt FW. Diagnosis of icteric diseases. Dtsch Med Wschr 1984;109:139-146.
4. Schumann G, Bonora R, Ceriotti F et al. IFCC Primary Reference Procedures for the Measurement of Catalytic Activity Concentrations of Enzymes at 37°C.
Part 5. Reference Procedure for the Measurement of Catalytic Concentration of Aspartate Aminotransferase. Clin Chem Lab Med 2002;40:725–733.
5. Ehret W, Heil W, Schmitt Y, Töpfer G, Wisser H, Zawta B, et al. Use of Anticoagulants in Diagnostic Laboratory Investigations and Stability of Blood, Plasma
and Serum Samples. WHO/DIL/LAB/99.1 Rev.2:23pp.
6. Thomas L, Müller M, Schumann G et al. Consensus of DGKL and VDGH for interim reference intervals on enzymes in serum. J Lab Med 2005;29:301-08.
7. Painter PC, Cope JY, Smith JL. Reference information for the clinical laboratory. In: Burtis CA, Ashwood ER, eds. Tietz textbook of clinical chemistry.
Philadelphia:WB Saunders Company, 1999;1802pp.
8. Young DS. Effects of drugs on clinical laboratory tests, 5th ed. AACC Press, 2000.

EN.01 BLOSR6x09.01 Enzyme


2009-08
AST (IFCC, without Pyridoxal phosphate activation), AU400/AU640 AST (IFCC, without Pyridoxal phosphate activation), AU600
Serum/Plasma Application Serum/Plasma Application
System Reagent: OSR6009, OSR6109, OSR6209 Reagent ID: 009 System Reagent: OSR6009, OSR6109, OSR6209 Reagent ID: 009

Specific Test Parameters Specific test parameters


General LIH ISE Range
Test No # Test name AST ∇ Sample type Ser ∇ Page ½
Test Name: AST ∇ < > Type: Serum ∇ Operation: Yes ∇
Sample vol. 10 Dil. Vol 0 µL Min. OD Max. OD
Sample: Volume 10 µL Dilution 0 µL Pre-Dilution Rate: 1 Reagent 1 vol 50 Dil. Vol 50 µL L 0.60 H 2.5
Reagents: R1 Volume 50 µL Dilution 50 µL Min OD Max OD Reagent 2 vol 50 Dil. Vol 100 µL Reagent OD limit
R2 Volume 50 µL Dilution 100 µL L 0.60 H 2.5 Fst. L 1.1 Fst. H 2.5
Reagent OD limit: Wave Main 340 Sub 660 ∇ Lst. L 1.1 Lst. H 2.5
Wavelength: Pri. 340 ∇ Sec. 660 ∇ First L 1.1 First H 2.5 Method RATE ∇ Dynamic range
Method: RATE ∇ Last L 1.1 Last H 2.5 Reaction - ∇ L 3* H 1000*
Reaction slope: - ∇ Dynamic Range: Point 1 Fst 14 Lst 27 Correlation factor A 1
Measuring Point 1: First 14 Last 27 L 3* H 1000* Point 2 Fst Lst B 0
Measuring Point 2: First Last Correlation Factor: Sample Pre-dil. Rate ¤
Linearity : 15 % A 1 B 0
No Lag Time: YES ∇ On-board stability period: 30 Linearity Fst 15 % Sec %
No lag time NO ∇ On-board stability period 30
Select using Space key, or select from list displayed by Guide key
Specific Test Parameters
General LIH ISE Range Test No # Test name AST ∇ Sample type Ser ∇ Page 2/2
Test Name: AST ∇ < > Type: Serum ∇ Level L Level H
Value/flag # ∇ # #
Value/Flag: # ∇ Level L: # Level H: # Normal range
Normal Ranges: Age L Age H Sex Age L Age H L H
Sex Year Month Year Month L H 1 # ∇ # Y # M # Y # M→ # #
R 1. # ∇ # # # # # # 2 # ∇ # Y # M # Y # M→ # #
R 2. # ∇ # # # # # # 3 # ∇ # Y # M # Y # M→ # #
R 3. # ∇ # # # # # # 4 # ∇ # Y # M # Y # M→ # #
R 4. # ∇ # # # # # # 5 # ∇ # Y # M # Y # M→ # #
R 5. # ∇ # # # # # # 6 # ∇ # Y # M # Y # M→ # #
R 6. # ∇ # # # # # # 7 Non select # #
7. None Selected # # 8 Out of range # #
8. Out of Range L H # # L H
Panic Value: # # Unit: U/L* Decimal places: # Panic value # #
Select the function using the Function key or the Mouse
Calibration specific
Calibration Specific
General ISE Test No # Test name AST ∇

SERUM/PLASMA APPLICATION
Test Name: AST ∇ < > Type Serum ∇ Cal type 1 MB ∇ Count #
Formula 1 Y=AX+B ∇ Process ∇
Calibration Type: MB ∇ Formula: Y=AX+B ∇ Counts: # Process: ∇
Selection calibrator
Cal No OD Conc Factor/OD-L Factor/OD-H
Cal. No. OD CONC Factor/OD-L Factor/OD-H
Point 1 § ∇ § §3650* §5470*
Point 1: § § §3650* §5470*
Point 2 ∇
Point 2:
Point 3 ∇
Point 3:
Point 4: Point 4 ∇
Point 5: Point 5 ∇
Point 6: Point 6 ∇
Point 7: Point 7 ∇
1-Point Cal. Point: R With CONC-0 Slope Check: None # 1-point cal. Point
∇ Advanced Calibration: ∇
MB type factor #
MB Type Factor: # Calibration Stability Period: Calibrator stability period
Select the function using the Function key or the Mouse
# User defined # User defined ¤ Analyser default value
* Values set for working in U/L. To work in SI units (µkat/L) divide by 60 * Values set for working in U/L. To work in SI units (µkat/L) divide by 60
§ For use in AB mode only, refer to leaflet for further instruction. § For use in AB mode only, refer to IFU for further instruction.
Enzyme BSOSR6x09.02
2010-05
AST (IFCC, without Pyridoxal phosphate activation), AU2700/AU5400 AST (IFCC, without Pyridoxal phosphate activation), AU680/AU480
Serum/Plasma Application
Serum/Plasma Application System Reagent: OSR6009, OSR6109, OSR6209, OSR6509 Reagent ID: 009
System Reagent: OSR6009, OSR6109, OSR6209, OSR6509 Reagent ID: 009 Parameters Specific Test Parameters
General LIH ISE HbA1c Calculated Test Range
Specific Test Parameters
General LIH ISE Range Test Name: AST ∇ < > Type: Serum ∇ Operation Yes ∇

Test Name: AST ∇ < > Type: Serum ∇ Operation: Yes ∇


Sample Volume 5 µL Dilution 0 µL OD Limit
Sample: Volume 5 µL Dilution 0 µL Pre-Dilution Rate: 1 Pre-Dilution Rate 1 ∇ Min.OD 0.60 Max.OD 2.5
Reagents: R1 Volume 25 µL Dilution 25 µL Min OD Max OD Rgt. Volume R1(R1-1) 25 µL Dilution 25 µL Reagent OD Limit
R2 Volume 25 µL Dilution 50 µL L 0.60 H 2.5 First Low 1.1 High 2.5
Reagent OD limit: Last Low 1.1 High 2.5
Wavelength: Pri. 340 ∇ Sec. 660 ∇ First L 1.1 First H 2.5 R2(R2-1) 25 µL Dilution 50 µL
Method: RATE ∇ Last L 1.1 Last H 2.5 Dynamic Range Low 3* High 1000*
Reaction slope: - ∇ Dynamic Range: Common Rgt. Type Noneф Name ф Correlation Factor A 1 B 0
Measuring Point 1: First 14 Last 27 L 3* H 1000* Wavelength Pri 340 ∇nm Sec. 660 ∇nm Factor for Maker A 1 B 0
Measuring Point 2: First Last Correlation Factor: Method RATE ∇
Linearity : 15 % A 1 B 0 Reaction Slope - ∇ Onboard Stability Period 30 Day 0 Hour
No Lag Time: YES ∇ On-board stability period: 30 Measuring Point1 First 14 Last 27 LIH Influence Check # ∇
Measuring Point2 First Last Lipemia +++++ ∇
Linearity Limit 15 % Icterus +++++ ∇
Lag Time Check YES ∇ Hemolysis + ∇
Specific Test Parameters
General LIH ISE Range Parameters Specific Test Parameters
General LIH ISE HbA1c Calculated Test Range
Test Name: AST ∇ < > Type: Serum ∇
Test Name: AST ∇ < > Type: Serum ∇
Value/Flag: # ∇ Level L: # Level H: #
Normal Ranges: Age L Age H Value/Flag: # ∇ Low High
Sex Year Month Year Month L H Level # # Panic Value
R 1. # ∇ # # # # # # Specificl Ranges: From To Low High
R 2. # ∇ # # # # # # Sex Year Month Year Month Low High # #
R 3. # # # # # # # R 1. # ∇ # # # # # #

R 4. R 2. # ∇ # # # # # #
# ∇ # # # # # #
R 3. # ∇ # # # # # #
R 5. # ∇ # # # # # #
R 4. # ∇ # # # # # #
R 6. # ∇ # # # # # #
R 5. # ∇ # # # # # #
7. None Selected # # R 6. # ∇ # # # # # #
8. Out of Range L H # #
7. No demographics # #
Panic Value: # # Unit: U/L* Decimal places: # 8. Not within expected values # #
Unit U/L* Decimal Places #
Parameters Calibration Parameters
Calibration Specific Calibrators Calibration Specific STAT Table Calibration
General ISE General ISE

Test Name: AST ∇ < > Type Serum ∇ Test Name: AST ∇ < > Type Serum ∇ R Use Serum Cal.

SERUM/PLASMA APPLICATION
Calibration Type: MB ∇ Formula: Y=AX+B ∇ Counts: # Process: ∇ Calibration Type: MB ∇ Formula: Y=AX+B ∇ Counts: # ∇
<Calibrator Parameters> Range
Cal. No. OD CONC Factor/OD-L Factor/OD-H Calibrator OD Conc Low High Slope Check None ∇
Point 1: § § §3650* §5470* Point 1: § ∇ § §3650* §5470*
Point 2: Point 2: ∇ Allowance Range Check
Point 3: Point 3: ∇
Point 4: Point 4: ∇ R Reagent Blank
Point 5: Point 5: ∇ R Calibration
Point 6: ∇
Point 6:
Point 7 ∇ Advanced Calibration
Point 7:
Point 8 ∇ Operation # ∇
1-Point Cal. Point: R with CONC-0 Slope Check: None ∇ Advanced Calibration: # ∇ Point 9 ∇
MB Type Factor: # Calibration Stability Period: Point 10 ∇ Interval (RB/ACAL) # ∇
<Point Cal. For No. of Correction Points ∇ Use Master Curve ∇ R Lot Calibration
Master Curve> OD Range
# User defined Calibrator OD Conc Low High Stability
* Values set for working in U/L. To work in SI units (µkat/L) divide by 60 Point-1 ∇ Reagent Blank Day Hour
§ For use in AB mode only. Refer to IFU for further instruction Point-2 ∇ Calibration Day Hour
ф AU680
MB Type Factor: # 1-Point Calibration Point ∇ R with Conc-0

Enzyme BSOSR6x09.02
2010-05
AST (IFCC, with OE60106 activation), AU400/AU640 Serum/Plasma AST (IFCC, with OE60106 activation), AU600
Application Serum/Plasma Application
System Reagent: OSR6009, OSR6109, OSR6209 Reagent ID: 009 System Reagent: OSR6009, OSR6109, OSR6209 Reagent ID: 009

Specific Test Parameters Specific test parameters


General LIH ISE Range
Test No # Test name AST ∇ Sample type Ser ∇ Page 1/2
Test Name: AST ∇ < > Type: Serum ∇ Operation: Yes ∇
Sample vol. 10 Dil. vol 0 µL Min. OD Max. OD
Sample: Volume 10 µL Dilution 0 µL Pre-Dilution Rate: 1 Reagent 1 vol 50 Dil. vol 50 µL L 1.0 H 2.5
Reagents: R1 Volume 50 µL Dilution 50 µL Min OD Max OD Reagent 2 vol 50 Dil. vol 100 µL Reagent OD limit
R2 Volume 50 µL Dilution 100 µL L 1.0 H 2.5 Fst. L 1.3 Fst. H 2.5
Reagent OD limit: Wave Main 340 Sub 660 ∇ Lst. L 1.3 Lst. H 2.5
Wavelength: Pri. 340 ∇ Sec. 660 ∇ First L 1.3 First H 2.5 Method RATE ∇ Dynamic range
Method: RATE ∇ Last L 1.3 Last H 2.5 Reaction - ∇ L 3* H 1000*
Reaction slope: - ∇ Dynamic Range: Point 1 Fst 14 Lst 27 Correlation factor A 1
Measuring Point 1: First 14 Last 27 L 3* H 1000* Point 2 Fst Lst B 0
Measuring Point 2: First Last Correlation Factor: Sample Pre-dil. Rate ¤
Linearity : 15 % A 1 B 0
No Lag Time: YES ∇ On-board stability period: 7 Linearity Fst 15 % Sec %
No lag time NO ∇ On-board stability period 7
Select using Space key, or select from list displayed by Guide key
Specific Test Parameters
General LIH ISE Range Test No # Test name AST ∇ Sample type Ser ∇ Page 2/2
Level L Level H
Test Name: AST ∇ < > Type: Serum ∇
Value/flag # ∇ # #
Value/Flag: # ∇ Level L: # Level H: # Normal range
Normal Ranges: Age L Age H Sex Age L Age H L H
Sex Year Month Year Month L H 1 # ∇ # Y # M # Y # M→ # #
R 1. # ∇ # # # # # # 2 # ∇ # Y # M # Y # M→ # #
R 2. # ∇ # # # # # # 3 # ∇ # Y # M # Y # M→ # #
R 3. # ∇ # # # # # # 4 # ∇ # Y # M # Y # M→ # #
R 4. # ∇ # # # # # # 5 # ∇ # Y # M # Y # M→ # #
R 5. # ∇ # # # # # # 6 # ∇ # Y # M # Y # M→ # #
R 6. # ∇ # # # # # # 7 Non select # #
7. None Selected # # 8 Out of range # #
8. Out of Range L H # # L H
Panic value # #
Panic Value: # # Unit: U/L* Decimal places: #
Select the function using the Function key or the Mouse
Calibration specific
Calibration Specific
General ISE Test No # Test name AST ∇

Test Name: AST ∇ < > Type Serum ∇ Cal type 1 MB ∇ Count #

Calibration Type: MB ∇ Formula: Y=AX+B ∇ Counts: # Process: ∇


SERUM/PLASMA APPLICATION Formula 1
Selection calibrator
Y=AX+B ∇ Process ∇

Cal No OD Conc Factor/OD-L Factor/OD-H


Cal. No. OD CONC Factor/OD-L Factor/OD-H Point 1 § ∇ § §3650* §5470*
Point 1: § § §3650* §5470* Point 2 ∇
Point 2: Point 3 ∇
Point 3: Point 4 ∇
Point 4:
Point 5 ∇
Point 5:
Point 6 ∇
Point 6:
Point 7 ∇
Point 7:
1-point cal. point
1-Point Cal. Point: R with CONC-0 Slope Check: None ∇ Advanced Calibration: # ∇ MB type factor #
MB Type Factor: # Calibration Stability Period: Calibrator stability period
Select the function using the Function key or the Mouse
# User defined # User defined ¤ Analyser default value
* Values set for working in U/L. To work in SI units (µkat/L) divide by 60 * Values set for working in U/L. To work in SI units (µkat/L) divide by 60
§ For use in AB mode only, refer to IFU for further instruction. § For use in AB mode only, refer to IFU for further instruction.

Enzyme BSOSR6x09.02
2010-05
AST (IFCC, with OE60106 activation), AU2700/AU5400 Serum/Plasma
Application
System Reagent: OSR6009, OSR6109, OSR6209, OSR6509 Reagent ID: 009

Specific Test Parameters


General LIH ISE Range

Test Name: AST ∇ < > Type: Serum ∇ Operation: Yes ∇

Sample: Volume 5 µL Dilution 0 µL Pre-Dilution Rate: 1


Reagents: R1 Volume 25 µL Dilution 25 µL Min OD Max OD
R2 Volume 25 µL Dilution 50 µL L 1.0 H 2.5
Reagent OD limit:
Wavelength: Pri. 340 ∇ Sec. 660 ∇ First L 1.3 First H 2.5
Method: RATE ∇ Last L 1.3 Last H 2.5
Reaction slope: - ∇ Dynamic Range:
Measuring Point 1: First 14 Last 27 L 3* H 1000*
Measuring Point 2: First Last Correlation Factor:
Linearity : 15 % A 1 B 0
No Lag Time: YES ∇ On-board stability period: 7

Specific Test Parameters


General LIH ISE Range

Test Name: AST ∇ < > Type: Serum ∇

Value/Flag: # ∇ Level L: # Level H: #


Normal Ranges: Age L Age H
Sex Year Month Year Month L H
R 1. # ∇ # # # # # #
R 2. # ∇ # # # # # #
R 3. # ∇ # # # # # #
R 4. # ∇ # # # # # #
R 5. # ∇ # # # # # #
R 6. # ∇ # # # # # #
7. None Selected # #
8. Out of Range L H # #
Panic Value: # # Unit: U/L* Decimal places: #

Calibration Specific
General ISE

Test Name: AST ∇ < > Type Serum ∇

SERUM/PLASMA APPLICATION
Calibration Type: MB ∇ Formula: Y=AX+B ∇ Counts: # Process: ∇

Cal. No. OD CONC Factor/OD-L Factor/OD-H


Point 1: § § §3650* §5470*
Point 2:
Point 3:
Point 4:
Point 5:
Point 6:
Point 7:
1-Point Cal. Point: R with CONC-0 Slope Check: None ∇ Advanced Calibration: # ∇
MB Type Factor: # Calibration Stability Period:

# User defined
* Values set for working in U/L. To work in SI units (µkat/L) divide by 60
§ For use in AB mode only. Refer to IFU for further instruction
ф AU680

Enzyme BSOSR6x09.02
2010-05
AST (IFCC, with OE60106 activation ), AU680/AU480 AST (IFCC, with OSR60180 activation ), AU680
Serum/Plasma Application Serum/Plasma Application
System Reagent: OSR6009, OSR6109, OSR6209, OSR6509 Reagent ID: 009 System Reagent: OSR6009, OSR6109, OSR6209, OSR6509 Reagent ID: 009
Parameters Specific Test Parameters Parameters Specific Test Parameters
General LIH ISE HbA1c Calculated Test Range General LIH ISE HbA1c Calculated Test Range

Test Name: AST ∇ < > Type: Serum ∇ Operation Yes ∇ Test Name: AST ∇ < > Type: Serum ∇ Operation Yes ∇

Sample Volume 5 µL Dilution 0 µL OD Limit Sample Volume 5 µL Dilution 0 µL OD Limit


Pre-Dilution Rate 1 ∇ Min.OD 1.0 Max.OD 2.5 Pre-Dilution Rate 1 ∇ Min.OD 1.0 Max.OD 2.5
Rgt. Volume R1(R1-1) 25 µL Dilution 25 µL Reagent OD Limit Rgt. Volume R1(R1-1) 25 µL Dilution 10 µL Reagent OD Limit
First Low 1.3 High 2.5 R1-2 5 µL Dilution 10 µL First Low 1.3 High 2.5
Last Low 1.3 High 2.5 Last Low 1.3 High 2.5
R2(R2-1) 25 µL Dilution 50 µL R2(R2-1) 25 µL Dilution 50 µL
Dynamic Range Low 3* High 1000* Dynamic Range Low 3* High 1000*
Common Rgt. Type Noneф Name ф Correlation Factor A 1 B 0 Common Rgt. Type R1-2 Name # Correlation Factor A 1 B 0
Wavelength Pri 340 ∇nm Sec. 660 ∇nm Factor for Maker A 1 B 0 Wavelength Pri 340 ∇nm Sec. 660 ∇nm Factor for Maker A 1 B 0
Method RATE ∇ Method RATE ∇
Reaction Slope - ∇ Onboard Stability Period 7 Day 0 Hour Reaction Slope - ∇ Onboard Stability Period 30 Day 0 Hour
Measuring Point1 First 14 Last 27 LIH Influence Check # ∇ Measuring Point1 First 14 Last 27 LIH Influence Check # ∇
Measuring Point2 First Last Lipemia +++++ ∇ Measuring Point2 First Last Lipemia +++++ ∇
Linearity Limit 15 % Icterus +++++ ∇ Linearity Limit 15 % Icterus +++++ ∇
Lag Time Check YES ∇ Hemolysis + ∇ Lag Time Check YES ∇ Hemolysis + ∇

Parameters Specific Test Parameters Parameters Specific Test Parameters


General LIH ISE HbA1c Calculated Test Range General LIH ISE HbA1c Calculated Test Range

Test Name: AST ∇ < > Type: Serum ∇ Test Name: AST ∇ < > Type: Serum ∇

Value/Flag: # ∇ Low High Value/Flag: # ∇ Low High


Level # # Panic Value Level # # Panic Value
Specificl Ranges: From To Low High Specificl Ranges: From To Low High
Sex Year Month Year Month Low High # # Sex Year Month Year Month Low High # #
R 1. # ∇ # # # # # # R 1. # ∇ # # # # # #
R 2. # ∇ # # # # # # R 2. # ∇ # # # # # #
R 3. # ∇ # # # # # # R 3. # ∇ # # # # # #
R 4. # ∇ # # # # # # R 4. # ∇ # # # # # #
R 5. # ∇ # # # # # # R 5. # ∇ # # # # # #
R 6. # ∇ # # # # # # R 6. # ∇ # # # # # #
7. No demographics # # 7. No demographics # #
8. Not within expected values # # 8. Not within expected values # #
Unit U/L* Decimal Places # Unit U/L* Decimal Places #
Parameters Calibration Parameters Parameters Calibration Parameters
Calibrators Calibration Specific STAT Table Calibration Calibrators Calibration Specific STAT Table Calibration
General ISE General ISE

Test Name: AST ∇ < > Type Serum ∇ R Use Serum Cal. Test Name: AST ∇ < > Type Serum ∇ R Use Serum Cal.

Calibration Type: MB ∇ Formula: Y=AX+B ∇ Counts: # ∇ Calibration Type: MB ∇ Formula: Y=AX+B ∇ Counts: # ∇

SERUM/PLASMA APPLICATION
<Calibrator Parameters> Range <Calibrator Parameters> Range
Calibrator OD Conc Low High Slope Check None ∇ Calibrator OD Conc Low High Slope Check None ∇
Point 1: § ∇ § §3650* §5470* Point 1: § ∇ § §3650* §5470*
Point 2: ∇ Allowance Range Check Point 2: ∇ Allowance Range Check
Point 3: ∇ Point 3: ∇
Point 4: ∇ R Reagent Blank Point 4: ∇ R Reagent Blank
Point 5: ∇ R Calibration Point 5: ∇ R Calibration
Point 6: ∇ Point 6: ∇
Point 7 ∇ Advanced Calibration Point 7 ∇ Advanced Calibration
Point 8 ∇ Operation # ∇ Point 8 ∇ Operation # ∇
Point 9 ∇ Point 9 ∇
Point 10 ∇ Interval (RB/ACAL) # ∇ Point 10 ∇ Interval (RB/ACAL) # ∇
<Point Cal. For No. of Correction Points ∇ Use Master Curve ∇ R Lot Calibration <Point Cal. For No. of Correction Points ∇ Use Master Curve ∇ R Lot Calibration
Master Curve> OD Range Master Curve> OD Range
Calibrator OD Conc Low High Stability Calibrator OD Conc Low High Stability
Point-1 ∇ Reagent Blank Day Hour Point-1 ∇ Reagent Blank Day Hour
Point-2 ∇ Calibration Day Hour Point-2 ∇ Calibration Day Hour
MB Type Factor: # 1-Point Calibration Point ∇ R with Conc-0 MB Type Factor: # 1-Point Calibration Point ∇ R with Conc-0

Enzyme BSOSR6x09.02
2010-05
AST (IFCC, without Pyridoxal phosphate activation), AU400/AU640 AST (IFCC, without Pyridoxal phosphate activation), AU600
Paediatric Application Paediatric Application
System Reagent: OSR6009, OSR6109, OSR6209 Reagent ID: 009 System Reagent: OSR6009, OSR6109, OSR6209 Reagent ID: 009
Specific Test Parameters Specific test parameters
General LIH ISE Range
Test No # Test name ASTP ∇ Sample type Ser ∇ Page ½
Test Name: ASTP ∇ < > Type: Serum ∇ Operation: Yes ∇
Sample vol. 10 Dil. Vol 10 µL Min. OD Max. OD
Sample: Volume 10 µL Dilution 10 µL Pre-Dilution Rate: 1 Reagent 1 vol 50 Dil. Vol 50 µL L 0.60 H 2.5
Reagents: R1 Volume 50 µL Dilution 50 µL Min OD Max OD Reagent 2 vol 50 Dil. Vol 90 µL Reagent OD limit
R2 Volume 50 µL Dilution 90 µL L 0.60 H 2.5 Fst. L 1.1 Fst. H 2.5
Reagent OD limit: Wave Main 340 Sub 660 ∇ Lst. L 1.1 Lst. H 2.5
Wavelength: Pri. 340 ∇ Sec. 660 ∇ First L 1.1 First H 2.5 Method RATE ∇ Dynamic range
Method: RATE ∇ Last L 1.1 Last H 2.5 Reaction - ∇ L 3* H 1000*
Reaction slope: - ∇ Dynamic Range: Point 1 Fst 14 Lst 27 Correlation factor A 1
Measuring Point 1: First 14 Last 27 L 3* H 1000* Point 2 Fst Lst B 0
Measuring Point 2: First Last Correlation Factor: Sample Pre-dil. Rate ¤
Linearity : 15 % A 1 B 0
No Lag Time: YES ∇ On-board stability period: 30 Linearity Fst 15 % Sec %
No lag time NO ∇ On-board stability period 30
Select using Space key, or select from list displayed by Guide key
Specific Test Parameters
General LIH ISE Range Test No # Test name ASTP ∇ Sample type Ser ∇ Page 2/2
Test Name: ASTP ∇ < > Type: Serum ∇ Level L Level H
Value/flag # ∇ # #
Value/Flag: # ∇ Level L: # Level H: # Normal range
Normal Ranges: Age L Age H Sex Age L Age H L H
Sex Year Month Year Month L H 1 # ∇ # Y # M # Y # M→ # #
R 1. # ∇ # # # # # # 2 # ∇ # Y # M # Y # M→ # #
R 2. # ∇ # # # # # # 3 # ∇ # Y # M # Y # M→ # #
R 3. # ∇ # # # # # # 4 # ∇ # Y # M # Y # M→ # #
R 4. # ∇ # # # # # # 5 # ∇ # Y # M # Y # M→ # #
R 5. # ∇ # # # # # # 6 # ∇ # Y # M # Y # M→ # #
R 6. # ∇ # # # # # # 7 Non select # #
7. None Selected # # 8 Out of range # #
8. Out of Range L H # # L H
Panic Value: # # Unit: U/L* Decimal places: # Panic value # #
Select the function using the Function key or the Mouse
Calibration specific
Calibration Specific
General ISE Test No # Test name ASTP ∇

PAEDIATRIC APPLICATION
Test Name: ASTP ∇ < > Type Serum ∇ Cal type 1 MB ∇ Count #
Formula 1 Y=AX+B ∇ Process ∇
Calibration Type: MB ∇ Formula: Y=AX+B ∇ Counts: # Process: ∇
Selection calibrator
Cal No OD Conc Factor/OD-L Factor/OD-H
Cal. No. OD CONC Factor/OD-L Factor/OD-H
Point 1 § ∇ § §3650* §5470*
Point 1: § § §3650* §5470*
Point 2 ∇
Point 2:
Point 3 ∇
Point 3:
Point 4: Point 4 ∇
Point 5: Point 5 ∇
Point 6: Point 6 ∇
Point 7: Point 7 ∇
1-Point Cal. Point: R with CONC-0 Slope Check: None # 1-point cal. Point
∇ Advanced Calibration: ∇
MB type factor #
MB Type Factor: # Calibration Stability Period: Calibrator stability period
Select the function using the Function key or the Mouse
# User defined # User defined ¤ Analyser default value
* Values set for working in U/L. To work in SI units (µkat/L) divide by 60 * Values set for working in U/L. To work in SI units (µkat/L) divide by 60
§ For use in AB mode only, refer to IFU for further instruction. § For use in AB mode only, refer to IFU for further instruction.
Enzyme BSOSR6x09.02
2010-05
AST (IFCC, without Pyridoxal phosphate activation), AU2700/AU5400 AST (IFCC, without Pyridoxal phosphate activation), AU680/AU480
Paediatric Application
Paediatric Application System Reagent: OSR6009, OSR6109, OSR6209, OSR6509 Reagent ID: 009
System Reagent: OSR6009, OSR6109, OSR6209, OSR6509 Reagent ID: 009 Parameters Specific Test Parameters
Specific Test Parameters General LIH ISE HbA1c Calculated Test Range
General LIH ISE Range Test Name: ASTP Type: Serum
∇ < > ∇ Operation Yes ∇
Test Name: ASTP ∇ < > Type: Serum ∇ Operation: Yes ∇
Sample Volume 5 µL Dilution 10 µL OD Limit
Sample: Volume 5 µL Dilution 10 µL Pre-Dilution Rate: 1