BXC0431A BXC0431B BXC0431C BXC0431D BXC0431E BXC0431F

For In Vitro Diagnostics Use Only

R1 3x10ml R1 6x30ml R1 4x70ml Lot Number

R1 3x10ml R1 6x30ml R1 4x70ml
R2 1x10ml R2 3x20ml R2 2x50ml
R2 1x10ml R2 3x20ml R2 2x50ml
R4 1x1ml R4 1x1ml R4 1x1ml Catalogue Number
Storage Temperature
LDL CHOLESTEROL STORE AT 2-8°C
Expiry Date (Year / Month)
PRODUCT CODE: BXC0431
INSTRUCTIONS FOR USE Warning, Read Enclosed Documents
Instructions For Use

FOR IN-VITRO DIAGNOSTIC USE ONLY Manufactured By

H202+4Amino Antipyrine+HDOAS Blue colour Additional Reagents required:

LDL CHOLESTEROL complex +3H20 Calibrator :BXC0315B/C 1x3 ml/5x3ml
(DIRECT) – A25/A15 APPLICATION Summary: Calibrator:
Kit Contents: BXC0431A BXC0431B BXC0431C
Low Density Lipoproteins (LDL) plays a key role in causing and Dissolve the Calibrator with 3 ml of distilled water and mix gently to
R1 Enzyme Colour influencing the progression of atherosclerosis and coronary sclerosis in dissolve the contents. Make sure that the solid material is dislodged
3 x 10 ml 6 x 30 ml 4 x 70 ml particular. The LDLs are derived from VLDLs (Very Low Density from the bottom of the vial. Return the vial to its upright position and
Reagent
1 x 10ml 3 x 20 ml 2 x 50 ml Lipoproteins) rich in triglycerides by the action of various lipolytic allow to stand for at least 10 minutes before use.
R2 Reacting solution
enzymes and are synthesized in the liver. The elimination of LDL from After dissolving R4 is stable for 7 days when aliquotted and stored
BXC0431D BXC0431E BXC0431F plasma takes place mainly by liver parenchymal cells via specific LDL at -20oC..
receptors.
R1 Enzyme Colour 3 x 10 ml 6 x 30 ml 4 x 70ml Elevated LDL concentrations in blood and an increase in their Specimen:
Reagent 1 x 10 ml 3 x 20 ml 2 x 50 ml residence time coupled with an increase in the biological modification Collect serum using standard sampling tubes
R2 Reacting Solution 1 x 1 ml 1 x 1 ml 1 x 1 ml rate results in the destruction of the endothelial function and a higher Li-heparin
R4 Calibrator LDL-cholesterol uptake in the monocyte / macrophage system as well Lithium Heparin plasma recovers 3% lower than the serum recoveries.
as by smooth muscle cells in vessel walls. The majority of cholesterol EDTA plasma recovers 10% lower than serum recoveries.
Intended Use: stored in atherosclerotic plaques originates from LDL. The LDL-
Homogenous Enzymatic Selective Protection Method for the cholesterol value is the most powerful clinical predictor among all of 7 days at +2°C - +8°C
quantitative determination of LDL Cholesterol in Serum and Plasma. Stability:
the single parameters with respect to coronary atherosclerosis. 30 days at - 70°C
Therefore, therapies focusing on lipid reduction primarily target the
Test Principle: reduction of LDL-cholesterol which is then expressed in an Testing Procedure:
LDL in the Sample is protected from reaction by the addition of the improvement of the endothelial function, prevention of atherosclerosis Materials Provided:
Enzyme Colour Reagent, Polyanion and Amphoteric Surfactant. and reducing its progression as well as preventing plaque rupture.  Working Solutions as described above
Cholesterol Esterase and Cholesterol Oxidase react with non LDL
Additional Materials Required:
lipoproteins, VLDL, HDL and the Hydrogen Peroxide liberated during
 Calibrators and controls as indicated below
the reaction is decomposed by catalase in the R1: Reagent Concentration:  0.9% NaCI
Good’s Buffer pH 6.8 25 mmol/l
CM / VLDL / HDL Cholesterol + H202+02 Cholesterol Estarase Cholistenone Cholesterol Oxidase 5000 U/l Manual Procedure: Differential Measurement:
+ Fatty acids+H202 R1Enzyme Colour Cholesterol Esterase 5000 U/l Wavelength Temperature Cuvette Measurement
Reagent Catalase 1000000 U/l Hg 600 nm Reagent Blank
2H2O2 CATALASE O2+2H2O Ascorbate Oxidase 5 U/ml (Side (one reagent
37oC 1 cm light path
Peroxidase 4 KU/l wavelength blank per
In the 2nd Step of the reaction, the reacting solution removes the Aminoantipyrine 4 mmol/l 700 nm) series only)
protecting reagent from LDL and catalase is inactivated by Sodium R2 Reacting Solution
POD 20000 U/l
Azide. In this step Cholesterol Esterase and Cholesterol Oxidase react Pipette into test tubes as follows:
only with LDLC. Hydrogen Peroxide produced by the enzyme reaction Reagent Handling and Preparation: Reagent Blank Sample / Calibrator
with LDL C yields a colour complex upon oxidative condensation with R1: Ready for use. Sample / Calibrator --- 5 µl
HDOAS and $-Amino antipyrine in the presence of Peroxidase. By R2: Ready for use. R1 450 µl 450 µl
measuring the absorbance of the blue colour complex produced at Unopened kit components: Up to the expiration date at 2 - 8°C Mix well and incubate at: 37°C for 5 minutes. Measure A1
approximately 600 nm, LDL-C concentration in the sample is On board stability R1: 28 days R2 150 µl 150 µl
calculated. R2: 28 days Incubate at 37°C for 5 minutes. Measure A2. Calculate Delta
Chol Esterase / Oxidase Prolonged exposure to the atmosphere could cause darkening of the Absorbance by subtracting A2 and A1.
LDL-Cholesterol +H202 Cholestenone +fatty R2. After each use it is suggested that the reagent is kept tightly
acids + H202. capped and stored at 2-8oC.
POD

Biorex Diagnostics Limited, Unit 2C Antrim Technology Park, Antrim, BT41 1QS (United Kingdom)
Tel: +44 (0) 2894 468786 | Fax: +44 (0) 2894 469933 | Website: www.biorexdiagnostics.com BXC0431 – LDL CHOLESTEROL| Revision No.10 AUG/11 | Page 1 of 2
Calculation: Limitations - Interference: Determination of LDL-Cholesterol in Serum Based on Precipitation
A sample x Calibrator conc. = LDL conc. Criterion: Recovery within 10% of initial value. of LDL with Dextran Sulfate. Ärztl. Lab. 1985;31 :325-330.
A Calibrator Icterus: No significant interference up to an index I of 50 (approximate 2. Bablok W. et al. A General Regression Procedure for Method
50 mg/dl bilirubin) Transformation. J Clin Chem Clin Biochem 1988;26:783-790.
Manual Procedure: End Point: Haemolysis: No significant interference up to an index H of 500 3. Bachorik P.S., Ross J.W. National cholesterol education program
Wavelength Temperature Cuvette Measurement (approximate haemoglobin concentration: 500 mg/dl). recommendations for measurement of low-density lipoprotein
Hg 600 nm Reagent Blank Lipemia (Intralipid): No significant interference up to an index L of 750. cholesterol: executive summary. Clin Chem 1995;41 :1414-1420.
(Side (one reagent No significant interference from native triglycerides up to 1500 mg/dl. 4. Cohn J.S., Mc Namara J.R., Schaefer E.J.. Lipoprotein Cholesterol
37oC 1 cm light path No significant interference from HDL, VLD, or chylomicrons. Concentrations in the Plasma of Human Subjects as Measured in
wavelength blank per
700 nm) series only) In rare cases, elevated immunoglobulin concentrations can lead to the Fed and Fasted States. Clin Chem 1988;34:2456-2459.
falsely elevated LDL-cholesterol results. 5. Cremer F., Nagel D., Mann H. et al. Ten-year follow-up results from
Pipette into test tubes as follows: Abnormal liver function does affect lipid metabolism; consequently the Goettingen Risk, Incidence and Prevalence Study (GR(PS). 1.
Reagent Blank Sample / Calibrator HDL and LDL results are of limited diagnostic value Risk factors for myocardial infarction in a cohort of 5790 men.
Sample / Calibrator --- 5 µl Atherosclerosis 1997;129:221-230.
R1 450 µl 450 µl Normal Values:
Mix well and incubate at: 37°C for 5 minutes.
Levels in terms of risk for coronary heart disease: BIOSYSTEMS A25 / A15 APPLICATION:
R2 150 µl 150 µl Adult Levels
Incubate at 37°C for 5 minutes. Measure the absorbance of the Recommended < 130 mg/dl
Calibrator and the sample against the reagent blank. (desirable) (<3.37 mmol/l) A 25 A 15
130-159 mg/dl TEST NAME LDL LDL
Moderate Risk
LDL Calculation: (3.37-4.12 mmol/l) ANALYSIS MODE BIR DIFF BIR DIFF
Absorbance sample x Calibrator conc. = LDL conc. ≥ 160 mg/dl SAMPLE TYPE SERUM SERUM
Absorbance Calibrator Risk
(≥ 4.14 mmol/l) UNITS mg/dl mg/dl
Recommended values according to the GRIPS study: GENERAL
REACTION TYPE INCREASING INCREASING
Measuring/reportable range: mg/dl mmol/l
1 - 400 mg/dl (0.02 – 10.4 mmol/l) DECIMALS 1 1
For patients with
Determine samples with LDL-cholesterol concentration> 400 mg/dl via 145 3,8 manifested coronary
NO.OF REPLICATES 1 1
the rerun function. On instruments without rerun function, manually heart disease TEST NAME IN PT REP
dilute the samples with 0.9% NaCI or distilled/deionized water (e.g. 1 + For patients having PROCEDURE READING MONOCHR MONOCHR
9). Multiply the result by the appropriate dilution factor (e.g. factor 10). 170 4,4 one or more risk VOLUMES SAMPLE 3 3
factors REAGENT 1 300 300
Automated Analysis:
For persons exhibiting REAGENT 2 100 100
no risk factors and WASHING 1.2 1.2
This reagent is suitable for all Automated systems. Applications for
200 5,2 without manifested
automated systems can be requested from our technical department. PREDIL FACTOR
coronary heart
POST DIL FACTOR 2 2
disease
Imprecision: FILTERS MAIN 600 600
Each laboratory should investigate the transferability of the expected
Reproducibility was determined using controls. The following results REFERENCE
values to its own patient population and if necessary determine its
were obtained: TIMES READING 1 285s 168s
own reference range.
Intra Assay – Within run
For diagnostic purposes the LDL-cholesterol results should always be READING 2 600s 600s
MW SD CV assayed in conjunction with the patient's medical history, clinical
Sample REAGENT 2 300s 192s
mg/dl mg/dl % examinations and other findings. CALIBRATION TYPE MULTIPLE MULTIPLE
Control serum 1 89.6 0.39 0.43
Control serum 2 120.2 0.63 0.54 CALIBRATOR REPLICATES 3 3
Quality Control: CALIBRATION
Control serum 3 152.9 0.81 0.53 BLANK REPLICATES 3 3
Lipid Controls BXC0330A / BXC0316 A
The control intervals and limits must be adapted to the individual CALIBRATION CURVE
Inter Assay – Between Run laboratory and country-specific requirements. Values obtained should BLANK ABS LIMIT 0.15 0.15
MW SD CV fall within established limits. Each laboratory should establish corrective OPTIONS KINETIC BLANK LIMIT
Sample
mg/dl mg/dl % measures to be taken if values fall outside the limits. LINEARITY LIMIT 400 400
Control serum 1 89.6 0.45 0.51
Control serum 2 120.2 0.65 0.54 Health & Safety:
Control serum 3 152.9 0.51 0.33 This kit is designed for use by suitably qualified laboratory personnel
only. Exercise the normal precautions required for the handling of
Method comparison: laboratory reagents. Do not ingest the material. Dispose of material
A comparison of the Biorex LDL-D (y) with a commercial obtainable according to local guidelines.
assay (x) gave following result (mg/dl):
y = 0.97 x + 5.12; r = 0.983
References:
1. Armstrong V., Seidel D. Evaluation of a Commercial Kit for the
Biorex Diagnostics Limited, Unit 2C Antrim Technology Park, Antrim, BT41 1QS (United Kingdom)
Tel: +44 (0) 2894 468786 | Fax: +44 (0) 2894 469933 | Website: www.biorexdiagnostics.com BXC0431-LDL CHOLESTEROL | Revision No.10 AUG/11 | Page 2 of 2