Biochemistry GMC Bhavnagar

Glucose estimation from serum or plasma by Hexokinase/G-6-PDH Method.

Calibration: Technician Quality Control: Technician Routine operation: Technician Overall Monitoring: Quality Manager

Type of Sample: Plasma, Urine, CSF

Type of container and additives: Sodium fluoride/ Potassium oxalate vaccutte,glacial acetic acid container

Patient Preparation: As per Primary Sample Collection Manual sample_collection_manual

Stability: At Room temperature 18°–28°C (64°–82°F) stability ≤ 24 hours

Handling and transport: As per Primary Sample collection manual

Storage: 24 hours at 2-8° C

Centrifuge, Auto-Pipette, Disposable Tips, Disposable sample cups, FullyAuto Chemistry Analyzer

R1.ATP .2Na 9.0 mg/mL

NAD 5.0 mg/mL

G-6-PDH 3,000 U/L

Hexokinase 15,000 U/L

Remove any air bubbles present in the reagents with a new applicator stick or allow the reagents to settle at the appropriate storage temperature to allow the bubbles to dissipate. To minimize volume depletion, do not use a transfer pipette to remove bubbles

Unopened reagent stable at 2-8°C until expiration date.

On board System temperature reagent is stable for 30 days.

Instability or deterioration should be suspected if there are precipitates, visible signs of leakage or contamination, turbidity, or if calibration or controls do not meet the appropriate criteria.

1. Calibrator Material- Multicostitute(MCC)

2. Frequency: Reagent lot change QC out of range After service or maintenance Replacement in any parts of Instrument

3. Procedure:

1. Start the equipment.
2. Calibrators are ready to use.
3. Put calibrator 15-20 minutes at room temperature
4. Prior to use, mix the contents by inverting the vial. Do not shake the vial to prevent foam formation.
5. Take a 150 µl both level of calibrator solution in to separate aliquot.
6. Go to the calibration and give the calibration order.
7. Verify calibration with at least two levels of controls
8. If control results fall outside acceptable ranges,root cause analysis or recalibration may be necessary.

1. Name: BioRad Level 1 & 2

2. Frequency: As per Quality Control Procedure

3. Procedure for Reconstitution of IQC

1. Reconstitution of QC material with 5 ml Distilled water by using calibrated fixed volume pipette.
2. Leave to stand for 30 min in the dark place.
3. Swirl gently several times during the reconstitution period to ensure that the contents are completely dissolved.
4. Prior to use, mix the contents by inverting the vial. Do not shake the vial as the information of foam should be avoided.
5. Ensure that no lyophilized material remains un-reconstituted.
6. Prepare aliquots of 150 µl from the reconstituted QC material.
7. Store these aliquots at -15° C to -20° C.
8. Prior to use, make sure that aliquots should be at room temperature for at least 15 min.

4. Procedure to run IQC

1. Press Control order
2. Select Assay /Panel, to be run.
3. Select the control/s and its level/s
4. Give Carrier Number and Position number
5. Press F3 / Add order
6. Put respected carrier in RSH rack
7. Check IQC results, in case outliers call residents.

Glucose is phosphorylated by hexokinase (HK) in the presence of adenosine triphosphate (ATP) and magnesium ions to produce glucose‐6-phosphate (G‐6-P) and adenosine diphosphate (ADP). Glucose-6-phosphate dehydrogenase (G-6-PDH) specifically oxidizes G-6-P to 6-phosphogluconate with the concurrent reduction of nicotinamide adenine dinucleotide (NAD) to nicotinamide adenine dinucleotide reduced (NADH). One micromole of NADH is produced foreach micromole of glucose consumed. The NADH produced absorbs light at 340 nm and can be detected spectrophotometrically as an increased absorbance.

Required Sample Volume: 150 µl of the sample

Temperature: 37°

Take 150-200µl of the sample from Primary tube to examination. Write the sample ID on the aliquot.

1. Press Patient order
2. Select Assay /Panel, to be run
3. Give Carrier Number and Position number
4. Press F3 / Add order
5. Put respected carrier in RSH rack

1. Linearity: 05 to 800 mg/dL

If values exceed this linearity limit 800 mg/dl, dilute the sample by Manual Dilution Procedure, or the Automatic Dilution Protocol provided in the assay parameters.

Automated Dilution Protocol:

When using the Automated Dilution Protocol, the system performs a dilution of the specimen and automatically corrects the concentration by multiplying the result by the appropriate dilution factor.

Manual Dilution Procedure:

Dilute the specimen with saline (0.85% to 0.90% NaCl). Enter the dilution factor in the Patient or Control order screen

2. The limit of detection (LOD): 2.5 mg/dl

3. The limit of quantification (LOQ): 5.0 mg/dL

4. Unit: mg/dl

The common causes of High Glucose Level are as follows:

Diabetes Mellitus, pancreatic tumor, hyperthyroidism, adrenal cortical hyperfunction

The common causes of Low Glucose Level are as follows:

Jaundice, Alcoholism, Insulin Therapy, Hypoglycemic Drugs

The Following analytes were tested up to the levels indicated at Sugar concentrations of 1 mg/dl and 5.03 found not to interfere:

No interference from Hemoglobin up to 1000 mg/dL

No interference from Intralipid up to 1000 mg/dL

No interference from bilirubin up to30 mg/dl

Routine: 6.0 hours

Urgent: 2.0 hours

Software backup

Machine raw data

Follow storage & discard procedure

Follow the universal work precautions. Reagent R1 contains sodium azide 0.1% as a preservative. Avoid contact with skin & mucosa. Contact with acid liberates very toxic gas.

1. Burtis CA,Ashwood ER,editors,Tietz Textbook of clinical chemistry
2. Kaplan LA,Pesce AJ editors. Clinical Chemistry Theory Analysis, and Correlation, 3rd ed.
3. ARCHITECT Glucose 3L82-21 AND 3L82-41 B3L8X0