GLUCOSE OXIDASE

(LIQUID) REAGENT
 1. The reagent should be stored refrigerated at 2-8°C.
2. The reagent is stable until the expiration date
when stored at 2-8°C.
INTENDED USE
For the in vitro quantitative determination of Glucose in REAGENT DETERIORATION
serum. Do not use if:
1. The reagent fails to recover stated control
METHODOLOGY values or meet stated linearity.
Early enzymatic methods for glucose determination 2. The reagent develops turbidity, or other
used Glucose Oxidase to catalyze the oxidation of evidence of microbial growth.
glucose to hydrogen peroxide and gluconic acid.' The SPECIMEN COLLECTION AND STORAGE
hydrogen peroxide that is formed is measured by the 1. Non-hemolyzed serum or heparinized plasma is
oxidation of a chromagens2 Many chromagens were recommended.
investigated but many were discarded because of 2. Serum must be separated from the clot promptly
possible carcinogenicity, toxicity, instability or because since the rate of glucose decrease is approximately
they were affected by many interfering substances. 7% per hour in whole bloods
Trinder modified Emerson 4to develop an efficient 3. Glucose in serum or plasma is stable for twenty-
peroxidase phenolaminophenazone system for the four hours when stored refrigerated (2-8°C)
quantitation of hydrogen peroxide by formulation of a
red quinoneimine dye. This method is less influenced INTERFERENCES
by interfering substances and does not suffer from the 1. Grossly lipemic or icteric samples will cause false
many drawbacks of earlier methods. The present glucose values, consequently a patient blank should
procedure is based on the above principle but utilizes a be run.
non-corrosive phenol substitute for added safety and 2. For a comprehensive list of interfering substances
convenience. see Young, et al6
3. Bilirubin to a level of 20 mg/dL and Hemoglobin to a
Principle: level of 500 mg/dL have both been found to exhibit
Glucose Oxidase negligible interference in this assay.
D-Glucose + H2O + 02 H202 + D-Gluconate
ADDITIONAL EQUIPMENT REQUIRED BUT NOT
PROVIDED
POD
1. A clinical chemistry analyzer capable maintaining
H202 + 4-Aminoantipydne + Phenol
constant temperature (37°C), and measuring
Quinoneimine dye + H2O
absorbance at 500nm.
2. Deionized water and related equipment, e.g.:
REAGENT COMPOSITION
pipettes
Active Ingredients Concentration
3. Analyzer specific consumables, e.g.: sample cups
Glucose Oxidase (microbial) >15,000 U/L 4. Control, and Calibrator materials such as those
Peroxidase (horseradish) >1,000 U/L provided by JAS Diagnostics.
4-Aminoantipyrine 0.3 mM
Phenol 0.5 mM
Phosphate Buffer ASSAY PROCEDURE
Non-reactive Stabilizers and fillers These instructions are to be used as a general
Sodium Azide 0.02%. guideline for adapting to select automated instruments.
pH 7.3 ± 0.15 Refer to your specific JAS instrument application
instructions available upon request.
Precautions
1. This reagent is for in vitro diagnostic use only.
System Parameters
2. The reagent should not be used if it has developed
Glucose Oxidase
turbidity or other evidence of microbial growth.
TEMPERATURE: 37°C
3. The reagent should not be used if it fails to meet
WAVELENGTH: 500 nm
linearity claims or fails to recover control values n the
ASSAY TYPE: Endpoint
DIRECTION: Increase
SAMPLE/ RGT RATIO: 1 : 100
e.g.Sample Vol. 0.003 mL (3 µL)
stated range.
Reagent Vol. 0.300 mL (300 µL)
4. All specimens and controls should be handled as
INCUBATION TIME: 10 Min
potentially infectious, using safe laboratory procedures.
Procedure Note:
REAGENT PREPARATION
The reagent and sample volumes may be altered
Reagent comes in a ready to use form.
proportionally to accommodate various instrument
requirements.
REAGENT STORAGE
Calculations: S.D.(mg/dL) 1.1 1.3 3.9
(A = Absorbance) C.V. (%) 1.1 0.7 1.3

A (patient) x Concentration of standard = Glucose Run to Run:

A (standard) (mg/dL) (mg/dL) Mean (mg/dL) 86 198 283
S.D.(mg/dL) 2.1 6.3 9.2
Example: C.V. (%) 2.5 3.2 3.3
A (patient) =0.10
A (standard) = 0.300 Sensitivity:
Concentration of standard = 150 (mg/dL) The sensitivity for the reagent was investigated by reading the
change in absorbance at 500nm for a saline sample, and a
0.10 x 150 = 50 mg/dL Glucose (mg/dL) serum with a known concentration. Ten replicates of each
0.30 sample were performed. The results of this investigation
indicated that, on the analyzer used, the reagent showed little
Limitations: or no reagent drift on a zero sample. Under the reaction
1. Samples with values exceeding 500 mg/dL should conditions described, 1mg/dl of glucose gives an absorbance
be diluted 1:1 with saline and re-run. The final of 0.002.
answer should be multiplied by two.
1. Keston, AS., Abstr., 129 th Meeting Amer. Chem. Soc.,
CALIBRATION p.31 (1956).
Use an aqueous Glucose standard or an appropriate
2. Teller, J.D., Abstr., 130 th Meeting Amer. Chem. Soc.,
serum calibrator. Atlantic City, NJ., p69c (1956).

QUALITY CONTROL 3. Trinder, P., Ann. Clin. Biochem., 6:24 (1969).

The integrity of the reaction should be monitored by use
of a two level control with known Glucose values 4. Emerson, E.J., et al, J. Org. Chem., 3:153 (1938)and
8:417 (1943).
EXPECTED VALUES 7
5. Tietz N.W. Fundamentals of Clinical Chemistry,
Normal range is reported to be: 70 -105 mg/dL. Philadelphia, W.B. Saunders, p243 (1976).
It is strongly recommended that each laboratory
establish its own normal range. 6. Young, D.S., et al, Clin. Chem. 21:1D (1975).

PERFORMANCE 7. Tietz N.W. Fundamentals of Clinical Chemistry,

Linearity: Philadelphia, W.B. Saunders, p155 (1970).
When run as recommended the assay is linear to 500
mg/dL
JAS Diagnostics Inc. Glucose Oxidase (Liquid) Reagent
7220 NW 68 Street Catalog No: _ Size
Method Comparison: Miami Florida 33166 GL02-125 2 x 125 mL
Studies performed between this procedure and a Tel. (305) 418-2320 GL02-1L 1 x 1000 mL
similar methodology yielded the following results: Fax. (305) 418-2321
Correlation Coefficient: 0.999
Slope: 1.024
Intercept: -1.134 (mg/dL)
PI:GLO2.04 REV: 10/19/09

Precision:
Within Run Level 1 Level 2 Level 3
Mean (mg/dL) 101 172 293