Albumin Examination Procedure

1.Purpose of examination: Albumin estimation from serum or plasma by Colorimetric (Bromocresol Green)

2.Responsibility and Authority:

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

3.Sample Details:

• Type of Sample: Serum, Plasma
• Type of container and additives: Plain without any additives
• 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

4.Required Equipment: Centrifuge, Auto-Pipette, Disposable Tips, Disposable sample cups, FullyAuto Chemistry Analyzer

5.Required reagents: R1. Bromocresol green 0.320 g/L.

6.Reagent Handling

1. 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
2. Reagent Storage and stability
3. Unopened reagent stable at 2-8°C until expiration date.
4. On board System temperature reagent is stable for 42 days
5. Instability or deterioration should be suspected if there are precipitates, visible signs of leakage or
6. contamination, turbidity, or if calibration or controls do not meet the appropriate criteria.

7.Calibration Procedure:

• Consolidated Chemistry Calibrator
• Frequency:
• Reagent lot change
• QC out of range
• After service or maintenance
• Replacement in any parts of Instrument

Procedure:

1. Start the equipment.WDI abbotte fully.docx
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 calibrator solution in separate aliquots.
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.

8.Quality control Procedure: Name: Bio Rad Level 1 & 2 Frequency: As per Quality Control Procedure

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. Ensure that no lyophilized material remains un-reconstituted.
5. Prepare aliquots of 150 µl from the reconstituted QC material.
6. Store these aliquots at -15° C to -20° C.
7. Prior to use, make sure that aliquots should be at room temperature for at least 15 min.

Procedure to run IQC

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

9.Principle of the procedure used for examinations: bromocresol green in the assay reagent specifically with albumin in the patient sample to produce a colored complex. The absorbance of the complex at 604 nm is directly proportional to the albumin concentration in the sample.

10.Sample Preparation:

• Required SampleVolume: 150 µl of the sample
• Temperature: 37°
• Take 150-200µl of the sample from Primary tube to the secondary aliquot. Write the sample ID on the aliquot.

Procedure to run Patient sample

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

11.Performance Characteristics:

• Linearity: 0.3 to 9.4 g/dL
• The limit of detection (LOD): 0.3 g/dL
• The limit of quantification(LOQ): 0.3 g/dL
• Unit: g/dL

Normal and critical ranges:

Albumin

12.Laboratory Clinical interpretation:

• Albumin is the major serum protein in normal individuals. Elevated serum albumin levels are usually the result of dehydration.Decreased albumin levels are found in a wide variety of conditions, including kidney disease, liver disease, malabsorption, malnutrition,severe burns, infections, and cancer.

13.Interference and cross reaction: The Following analytes were tested up to the levels indicated at Albumin concentrations of 0.3mg/dl and 9.4 mg/dl, and found not to interfere:

• No interference from Bilirubin up to 30.0 mg/dL
• No interference from Hemoglobin up to750 mg/dL
• No interference from Intralipid up to1000 mg/dL

14.Potential source of variation: Turn around time (TAT): Routine: 6.0 hours Urgent: 2.0 hours

15.Recording of observation:

• Software backup
• Machine raw data

16.Storage & Disposal of waste: Follow storage & discard procedure

17.Environmental & Safety control: Follow the universal work precautions. Reagent R1 contains sodium azide 0.1% as a preservative. Avoid contact with skin & mucosa. contact with acid liberate very toxic gas.

18.References:

1. osafety manual,3rd edUS department of labor,Occupational safety and health administration.
2. US department of health and human services. Biosafety in MIcrobiological and biomedical laboratories.
3. World health organization. Biition