IRON Examination Procedure

1.Purpose of examination:

• Iron estimation from serum or plasma by Ferene method.

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:guanidine hydrochloride (382.120 g/L). R2 :ferene-S (4.944 g/L) L-ascorbic acid (96.866 g/L).

6.Reagent Handling

• 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
• Reagent Storage and stability
• Unopened reagent stable at 2-8°C until expiration date.
• The onboard 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.

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:

• Start the equipment.WDI abbotte fully.docx
• Calibrators are ready to use.
• Put calibrator 15-20 minutes at room temperature
• Prior to use, mix the contents by inverting the vial. Do not shake the vial to prevent foam formation.
• Take a 150 µl calibrator solution in separate aliquots.
• Go to the calibration and give the calibration order.
• Verify calibration with at least two levels of controls.
• 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: At an acidic pH, iron is released from transferrin to which it is bound, and then quantitatively reduced to a ferrous state. The iron forms with ferene-S (3-(2-pyridyl)-5,6-bis-[2-(5-furylsulfonic acid)]-1,2,4- triazine), a stable colored complex of which the color intensity is proportional to the amount of iron 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:7 to 1143 μg/dL
• The limit of detection (LOD):4 μg/dL
• The limit of quantitation (LOQ):7 μg/dL
• Unit: μg/dL

Normal and critical ranges:

12.Laboratory Clinical interpretation: Increased iron concentrations are seen in hemolytic anemias, hemochromatosis, and acute liver disease. Decreased iron concentrations are seen in iron deficiency and anemia of chronic disease such as in chronic renal disease.1 Major causes of iron deficiency include gastrointestinal and menstrual bleeding.

13.Interference and cross reaction:

• No interference from lipemia up to 1500 mg/dl.
• No interference from hemoglobin up to 200mg/dl.
• No interference from conjugated bilirubin up to 60mg/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: Prevention:

• Do not handle until all safety precautions have been read and understood.
• Avoid breathing mist / vapors / spray.
• Wash hands thoroughly after handling.
• Use only outdoors or in a well-ventilated area.
• Avoid release to the environment.
• Contaminated work clothing should not be allowed out of the workplace.
• Wear protective gloves / protective clothing / eye protection.

Response:

• IF SWALLOWED: Rinse mouth.
• IF ON SKIN: Wash with plenty of water.
• IF IN EYES: Rinse cautiously with water for several minutes. Remove contact lenses, if present and easy to do. Continue rinsing.
• IF INHALED: Remove person to fresh air and keep comfortable for breathing.
• IF exposed or concerned: Get medical advice /attention.
• If skin irritation or rash occurs: Get medical advice / attention.
• If eye irritation persists: Get medical advice /attention.
• Take off contaminated clothing and wash it before reuse.

Disposal: Dispose of contents / container in accordance with local regulations.

18.References:

1. World Health Organization. Laboratory Biosafety Manual. 3rd ed.Geneva: World Health Organization; 2004.
2. Burtis CA, Bruns DE, editors. Tietz Fundamentals of Clinical Chemistry and Molecular Diagnostics. 7th ed. St. Louis, MO:Saunders Elsevier; 2015.Burtis CA, Bruns DE, editors. Tietz Fundamentals of Clinical Chemistry and Molecular Diagnostics. 7th ed. St. Louis, MO: Saunders Elsevier; 2015.