• Reaction type : End Point (2 Step)
• Reaction time : 5 + 5 mins.
• Wavelength: 578 nm.
• Flowcell temperature: 37 degree C
• Zero setting with:  Distilled water
• Sample volume: 0.01 ml (10 NI)
• Reagent volume: 0.750 ml + 0.250 ml
• Linearity : 185 mg/dl

Perform the assay as given below:

• Mix and incubate for 5 minutes at 37 degree C. 
• Measure the absorbance Al at 578 nm.

• Men : 30 - 70 mg/di
• Women : 30 - 85 mg/di

Expected range varies from population to population. It is therefore recommended that each laboratory should establish its own normal range.

SPECIMEN COLLECTION & PRESERVATION

Blood should be collected in a clean and dry container. Although serum is preferred, anticoagulants such as heparin, citrate, oxalate, EDTA and sodium fluoride do not have significant influence on the assay when they are used in their usual amounts. Specimen analysis should be done immediately after collection. Do not use the specimens that are repeatedly subjected to freezing and thawing.

PROCEDURE LIMITATIONS

If the HDL-C value exceeds the linearity limit of 185 mg/dl, then dilute the specimen suitably with normal saline and repeat the assay. In such case the assay value should be multiplied with the dilution factor to obtain the correct HDL-C value of the specimen.

QUALITY CONTROL

To ensure adequate quality control, it is recommended that each batch should include a normal and an abnormal commercial reference control serum. It should be realized that the use of quality control material checks both instrument and reagent functions together. Factors which might affect the performance of this test include instrument function, temperature control, cleanliness of glassware and accuracy of pipetting.