0120767131322COINV4.

GLUC2
Glucose HK - Hemolysate Application Gen.2 Substrates
Order information
Analyzer(s) on which cobasc pack(s) can be used
COBASINTEGRA 400 plus
20767131 322 Glucose HK (200 tests) System-ID 0767131
COBAS INTEGRA 800
10759350 190 Calibrator f.a.s. (12 3 mL) System-ID 0737186
12149435 122 Precinorm U plus (10 3 mL) System-ID 0779997
12149443 122 Precipath U plus (10 3 mL) System-ID 0780006
10171743 122 Precinorm U (20 5 mL) System-ID 0779970
10171735 122 Precinorm U (4 5 mL) System-ID 0779970
10171778 122 Precipath U (20 5 mL) System-ID 0779989
10171760 122 Precipath U (4 5 mL) System-ID 0779989
05117003 190 PreciControl ClinChem Multi 1 (20 5 mL) System-ID 0774693
05947626 190 PreciControl ClinChem Multi 1 (4 5 mL) System-ID 0774693
05117216 190 PreciControl ClinChem Multi 2 (20 5 mL) System-ID 0774707
05947774 190 PreciControl ClinChem Multi 2 (4 5 mL) System-ID 0774707
05067235 191 Glucose Hemolyzing Reagent Gen.2 (1000 mL)

English SR HEPES buffer: 30mmol/L, pH7.0; Mg2+: 4mmol/L; HK (yeast):

System information 130kat/L; G6PDH (microbial): 250kat/L
Test GLUH2, test ID 0613 (HemolysateApplicationGen.2); testGLU2P, R1 is in positionB and SR is in positionC.
testID 0001 (HemolysateapplicationGen.2 plasma-level)
Precautions and warnings
Intended use Pay attention to all precautions and warnings listed in
In vitro test for the quantitative determination of the glucose concentration in Section1/Introduction of this Method Manual.
hemolysate.
Reagent handling
Summary1,2,3 Ready for use
Glucose is the major carbohydrate present in the peripheral blood.
Oxidation of glucose is the major source of cellular energy in the body. Storage and stability
Glucose derived from dietary sources is converted to glycogen for storage Glucose HK
in the liver or to fatty acids for storage in adipose tissue. The concentration
of glucose in blood is controlled within narrow limits by many hormones, the Shelf life at 28C See expiration date on
most important of which are produced by the pancreas. cobasc pack label
The most frequent cause of hyperglycemia is diabetes mellitus resulting
from a deficiency in insulin secretion or action. A number of secondary COBASINTEGRA400 plus system
factors also contribute to elevated blood glucose levels. These include On-board in use at 1015C 8weeks
pancreatitis, thyroid dysfunction, renal failure, and liver disease.
COBASINTEGRA800 system
Hypoglycemia is less frequently observed. A variety of conditions may
cause low blood glucose levels such as insulinoma, hypopituitarism, or On-board in use at 8C 8weeks
insulin induced hypoglycemia.
Glucose Hemolyzing Reagent Gen.2
Glucose measurement in hemolysate is a convenient method for routine
diabetes monitoring and in cases where only small sample volumes are Shelf life at 1525C See expiration date on
available. reagent label
Test principle Stability after opening 6weeks
Enzymatic reference method with hexokinase.4,5
Storage after opening 1525C
Hexokinase(HK) catalyzes the phosphorylation of glucose by ATP to form
glucose6phosphate and ADP. To follow the reaction, a second enzyme, Specimen collection and preparation
glucose6phosphate dehydrogenase (G6PDH) is used to catalyze For specimen collection and preparation only use suitable tubes or
oxidation of glucose6phosphate by NADP+ to form NADPH. collection containers.
HK
Only the specimens listed below were tested and found acceptable:
Whole blood: The samples should be hemolyzed immediately after
D-glucose + ATP D-glucose-6-phosphate + ADP collection. Collect 20L of capillary blood. Take venous blood or blood from
the earlobe or from the fingertip. Earlobe or fingertip should be well supplied
G6PDH with blood at the time of collection.
D-glucose-6-phosphate + NADP+ Hemolysate preparation
D-6-phosphogluconate + NADPH + H+ 1. Place 0.5mL of GlucoseHemolyzingReagentGen.2 in a test tube.
2. Add the filled 20L capillary and close the test tube.
The concentration of the NADPH formed is directly proportional to the
glucose concentration. It is determined by measuring the increase in 3. Mix gently, avoiding the formation of foam.
absorbance at 340nm. 4. Allow to stand for at least 5minutes at room temperature prior to
Reagents - working solutions glucose determination. Do not centrifuge.
R1 TRIS buffer: 100mmol/L, pH7.8; Mg2+: 4mmol/L; Stability in hemolysate6 8days at 1525C
ATP:1.7mmol/L; NADP+: 1mmol/L 2weeks at 28C

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GLUC2
Glucose HK - Hemolysate Application Gen.2 Substrates

Materials provided Note

See Reagents working solutions section for reagents. Do not pretreat calibrator.
Materials required (but not provided) Calibrator f.a.s. is automatically diluted 1:26 (1+25) with water by the
GlucoseHemolyzingReagentGen.2, 1000mL, Cat.No.05067235191, for instrument. A conversion factor (laboratory correlation factor) of26 is
hemolysate preparation. See above for sample pretreatment procedure. applied in order to achieve correct patient results. Enter the assigned lot-
specific glucose value of the undiluted calibrator, indicated in the package
Assay insert of the Calibrator f.a.s.
For optimum performance of the assay follow the directions given in this a) Isotope Dilution Mass Spectrometry
document for the analyzer concerned. Refer to the appropriate operators
manual for analyzerspecific assay instructions. Quality control

Application for hemolysate Reference range PrecinormU, PrecinormUplus or

PreciControlClinChemMulti1
COBASINTEGRA400plus test definition Dilute control serum 1:26 (1+25) in
Measuring mode Absorbance distilled/deionized water. Add control
Abs. calculation mode Endpoint manually.
Reaction mode R1SSR Pathological range PrecipathU, PrecipathUplus or
PreciControlClinChemMulti2
Reaction direction Increase
Dilute control serum 1:26 (1+25) in
Wavelength A/B 340/378nm distilled/deionized water. Add control
Calc. first/last 33/53 manually.
Unit mmol/L Control interval 24hours recommended
Laboratory correlation factor 26 Control sequence User defined
Control after calibration Recommended
Pipetting parameters
For quality control, use control materials as listed in the Order information
Diluent (H2O) section. In addition, other suitable control material can be used.
R1 150L The control intervals and limits should be adapted to each laboratorys
individual requirements. Values obtained should fall within the defined
Sample 20L 12L limits. Each laboratory should establish corrective measures to be taken if
SR 30L values fall outside the defined limits.
Total volume 212L Follow the applicable government regulations and local guidelines for
quality control.
COBASINTEGRA800 test definition Calculation
Measuring mode Absorbance COBASINTEGRAanalyzers automatically calculate the analyte
concentration of each sample. For more details, please refer to Data
Abs. calculation mode Endpoint Analysis in the Online Help (COBASINTEGRA400plus/800 analyzers).
Reaction mode R1SSR
Conversion factors: mmol/L18.02=mg/dL
Reaction direction Increase
mg/dL0.0555=mmol/L
Wavelength A/B 340/378nm
Limitations - interference
Calc. first/last 44/77
Criterion: Recovery within 10% of initial value.
Unit mmol/L Icterus:7 No significant interference up to an Iindex of 60 for conjugated
Laboratory correlation factor 26 and unconjugated bilirubin (approximate conjugated and unconjugated
bilirubin concentration: 1026mol/L or 60mg/dL).
Pipetting parameters Lipemia (Intralipid):7 No significant interference up to an Lindex of 1000.
There is poor correlation between the Lindex (corresponds to turbidity) and
Diluent (H2O) triglycerides concentration.
R1 150L Drugs: No interference was found at therapeutic concentrations using
Sample 20L 12L common drug panels.8,9
SR 30L In very rare cases, gammopathy, in particular type IgM (Waldenstrms
macroglobulinemia), may cause unreliable results.10
Total volume 212L For diagnostic purposes, the results should always be assessed in
conjunction with the patients medical history, clinical examination and other
Calibration findings.
Calibrator Calibrator f.a.s. ACTION REQUIRED
Special Wash Programming: The use of special wash steps is mandatory
Use deionized water as zero when certain test combinations are run together on COBASINTEGRA
calibrator. analyzers. Refer to the CLEAN Method Sheet for further instructions and for
Calibration mode Linear regression the latest version of the Extra wash cycle list.
Where required, special wash/carry-over evasion programming must
Calibration replicate Duplicate recommended be implemented prior to reporting results with this test.
Calibration interval Each lot and as required following
quality control procedures.
Traceability: This method has been standardized against IDMSa).

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GLUC2
Glucose HK - Hemolysate Application Gen.2 Substrates

Limits and ranges Glucose values for samples hemolyzed with the GlucoseHemolyzing
Measuring range Reagent Gen.2 obtained on a COBASINTEGRA800 analyzer (GLUH2,
Hemolysate (test ID 0613): testID 0613) using the COBASINTEGRA GlucoseHK reagent
1.545mmol/L (27811mg/dL) (GLUC2)(y) were compared with samples hemolyzed with the Hemolyzing
Reagent Fluid determined on a Roche/Hitachi917 analyzer (ACN548)(x).
Hemolysate plasma-level (test ID 0001): Single measurement was done.
1.750mmol/L (31901mg/dL)
Determine samples having higher concentrations via the rerun function. Roche/Hitachi917 analyzer
Dilution of samples via the rerun function is a 1:10 dilution. Results from Pretreatment HemolyzingReagentFluid (1+50)
samples diluted using the rerun function are automatically multiplied by a
factor of10. Sample size (n) 58
Lower limits of measurement Corr. coefficient (r) 0.999
Lower detection limit of the tests (test IDs 0613 and 0001): Linear regression y=1.018x+0.120mmol/L
0.4mmol/L (7.2mg/dL)
The lower detection limit represents the lowest measurable analyte level Passing/Bablok14 y=1.023x+0.050mmol/L
that can be distinguished from zero. It is calculated as the value lying The sample concentrations were between 1.69 and 43.9mmol/L (30.5 and
3standard deviations above that of the lowest standard (standard1+3SD, 791mg/dL).
repeatability, n=21).
References
Expected values 1 Sacks DB. Carbohydrates. In: Tietz NW, ed. Fundamentals of Clinical
Whole blood: 3.65.3mmol/L (6595mg/dL)11 Chemistry. 5th ed. Philadelphia: WB Saunders 2001;427-461.
2 Khan MI, Weinstock RS. Carbohydrates. In: Henry JB, ed. Clinical
Whole blood plasma-level: 4.05.9mmol/L (72106mg/dL)* Diagnosis and Management by Laboratory Methods. 21st ed.
*calculated by a conversion factor of 1.1112 Philadelphia: WB Saunders 2007:185-199.
Hematocrit level may influence the difference between plasma and whole 3 Sacks DB. Carbohydrates. In: Burtis CA, Ashwood ER, eds. Tietz
blood glucose levels due to lower glucose values in erythrocytes compared Textbook of Clinical Chemistry. 4th ed. Philadelphia: WB Saunders
with plasma concentration. Higher hematocrit levels lead to an increased 2006:837-901.
plasma glucose level compared to whole blood.11,13 4 Neeley WE. Simple automated determination of serum or plasma
Each laboratory should investigate the transferability of the expected values glucose by a hexokinase/glucose-6-phosphate dehydrogenase method.
to its own patient population and if necessary determine its own reference Clin Chem 1972;18:509-515.
ranges.
5 Bondar RJ, Mead DC. Evaluation of glucose-6-phosphate
Specific performance data dehydrogenase from Leuconostoc mesenteroides in the hexokinase
Representative performance data on the COBASINTEGRA analyzers are method for determining glucose in serum. Clin Chem 1974;20:586-590.
given below. Results obtained in individual laboratories may differ. 6 Data on file at Roche Diagnostics.
Precision 7 Glick MR, Ryder KW, Jackson SA. Graphical Comparisons of
Precision was determined using human samples and controls in an internal Interferences in Clinical Chemistry Instrumentation.
protocol with repeatability (n=21) and intermediate precision (1aliquot per ClinChem1986;32:470-475.
run, 1run per day, 21days). The following results were obtained: 8 Breuer J. Report on the Symposium Drug effects in Clinical Chemistry
Methods. Eur J Clin Chem Clin Biochem 1996;34:385-386.
Repeatability Level 1 Level 2
9 Sonntag O, Scholer A. Drug interference in clinical chemistry:
Mean 3.43mmol/L 7.60mmol/L recommendation of drugs and their concentrations to be used in drug
(61.8mg/dL) (137mg/dL) interference studies. Ann Clin Biochem 2001;38:376-385.
CV 0.7% 0.4% 10 Bakker AJ, Mcke M. Gammopathy interference in clinical chemistry
assays: mechanisms, detection and prevention.
Intermediate precision Level 1 Level 2 ClinChemLabMed2007;45(9):1240-1243.
Mean 3.44mmol/L 7.60mmol/L 11 Tietz NW, ed. Clinical Guide to Laboratory Tests, 4th ed. Philadelphia.
WB Saunders 2006;444-455.
(62.0mg/dL) (137mg/dL)
12 D'Orazio P, Burnett RW, Fogh-Andersen N, et al. Approved IFCC
CV 1.0% 1.1% Recommendation on Reporting Results for Blood Glucose
Method comparison (Abbreviated). Clin Chem 2005;51:1573-1576.
Glucose values for samples hemolyzed with the GlucoseHemolyzing 13 Kruse-Jarres JD, Schttler A, Witt I. Kohlenhydratstoffwechsel. In:
Reagent Gen.2 obtained on a COBASINTEGRA400 analyzer (GLUH2, Greiling H, Gressner AM, eds. Lehrbuch der Klinischen Chemie und
test ID 0613) using the COBASINTEGRA GlucoseHK reagent Pathobiochemie. Stuttgart: Schattauer 1987:186-222.
(GLUC2)(y) were compared with those determined using the 14 Bablok W, Passing H, Bender R, et al. A general regression procedure
corresponding reagent on a COBASINTEGRA800 analyzer (GLUH2, test for method transformation. Application of linear regression procedures
ID 0613)(x). Single measurement was done. for method comparison studies in clinical chemistry, Part III.
JClinChemClinBiochem 1988 Nov;26(11):783-790.
COBASINTEGRA800 analyzer
A point (period/stop) is always used in this Method Sheet as the decimal
Pretreatment GlucoseHemolyzingReagent Gen.2 (1+25) separator to mark the border between the integral and the fractional parts of
Sample size (n) 57 a decimal numeral. Separators for thousands are not used.
Corr. coefficient (r) 0.999 Symbols
Roche Diagnostics uses the following symbols and signs in addition to
Linear regression y=0.992x+0.013mmol/L those listed in the ISO 152231 standard:
Passing/Bablok14 y=0.993x+0.002mmol/L
Contents of kit
The sample concentrations were between 1.79 and 44.9mmol/L (32.3 and
809mg/dL). Volume after reconstitution or mixing
GTIN Global Trade Item Number

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GLUC2
Glucose HK - Hemolysate Application Gen.2 Substrates

COBAS, COBASC, COBASINTEGRA, PRECICONTROL, PRECINORM and PRECIPATH are trademarks of

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Additions, deletions or changes are indicated by a change bar in the margin.
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GLUH2 4/4 2016-04, V 4.0 English