Name: Ozioko Joshua

Reg no: 2020/BS/13891

Date: 12/01/2024

Week 19th report

Title: HDL-Cholesterol test

Aim: to assess cardiovascular health, including the risk for heart disease.

Introduction: A HDL cholesterol test measures the amount of cholesterol found inside high-density
lipoproteins (HDL) in a sample of blood. High density lipoproteins are one of the major classes of plasma
lipoproteins. They are composed of a number of a heterogeneous particles, including cholesterol and
vary with respect to size and content of lipid and apolipoprotein. HDL is known as the good cholesterol
because it carries LDL, triglycerides, and harmful fats and returns them to the liver for processing. When
HDL reaches the liver, the liver breaks down the LDL, turns it into bile, and removes it from the body.

Principle: Low density lipoproteins (LDL and VLDL) and chylomicron fractions are precipitated
quantitatively by the addition of phosphotungstic acid in the presence of magnesium ions. After
centrifugation, the cholesterol concentration in the HDL fraction, which remains in the supernatant, is
determined.

Materials: plasma sample, Test tubes, Centrifuge, Micropipette, Spectrophotometer, Water-bath,

Randox HDL- cholesterol test pack.

Procedures:

1st phase

Pipette into plain sample bottle for centrifugation.

Standard Sample

Standard 500ul

Sample 500ul

Precipitant 1000ul 1000ul

Mix and allow to sit for 10 minutes at room temperature. Then centrifuge for 10 minutes at 4000rpm.
After centrifugation separate off the clear supernatant.

2nd phase

Pipette into test tubes

Reagent blank Standard Sample

Distilled water 100ul ---------- ---------

Standard supernatant --------- 100ul -----------

Sample supernatant --------- ---------- 100ul

Reagent 1000ul 1000ul 1000ul

Mix and incubate for 5 minutes at 37°c .

Measure the Absorbance of standard and sample against reagent blank at 500nm.

Result

Abs Concentration

Standard 0.713 5.12

Sample 0.431 2.96

Calculations

Conc of sample = Abs of sample/ Abs of standard × conc of standard

0.431/0.713×5.12 = 2.96mmol/L

Conclusion: accurate measurement of HDL- Cholesterol is of vital important when assessing patients risk
from coronary heart disease (CHD).

Precautions:

1) I made sure I wore my lab coat, shoes and hand gloves.

2) I carefully picked the supernatant after centrifugation to avoid error.