Cholesterol-1.pptx curudyjfujcubdjcgjguvhh

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1. Clinical lab practice 2 Javeed Ahmed BS MLT ( Shifa Tameer Milat University Islamabad Lecturer Allied College Of Health Sciences Multan Lecture No# 11 Topic # Estimation of Serum cholesterol Date 02/07/21

2. ESTIMATION OF SERUM TOTAL CHOLESTROL

3. LIPID PROFILE Total Cholesterol Levels LDL levels –Low density lipoprotein cholesterol HDL levels -High density lipoprotein cholesterol Triglyceride levels Done in Fasting state

4. CHOLESTEROL: Cholesterol is a important sterol synthesized in tissues liver and intestinal cells Approx. three fourth is synthesize in the body and and one fourth originates from dietary intake In plasma, both forms are transported in lipoproteins.

5. TYPES OF CHOLESTEROL: LDL Carries cholesterol to the Peripheral Tissues It can be deposited and causes Peripheral vascular diseases, atherosclerosis. The Bad Cholesterol HDL Transports cholesterol from peripheral tissues to the liver for excretion. Good Cholesterol

6. FUNCTIONS OF CHOLESTROL: Synthesizes: Vitamin D. Steroid hormones: Testosterone, estrogen, progesterone, aldosterone, cortisol. Bile

7. Membrane component – Cholesterol plays has a role in membrane fluidity but it's most important function is in reducing the permeability of the cell membrane . Nerve signal conduction

8. SOURCES: Food of animal origin Synthesized by Liver

9. REAGENT–Enzymatic Reagent Cholesterase Cholesterol oxidase Peroxidase Phenol 4-aminophenazone Conc. of Standard Solution: 200mg/dl Sample : Serum

10. PRINCIPLE: Cholesterol esters are hydrolyzed to free cholesterol by cholesterase . The free cholesterol produced is oxidized by cholesterol oxidase to produce H2O2. H2O2couples with 4-aminophenazone and phenol to yield Quinoamine ( chromogen ) by peroxidase.

11. EQUIPMENT Spectrophotometer Micropipette.

12. PROCEDURE: Take 3 cuvettes, label them as Blank (B), Standard (S), Test (T) S T B

13. PROCEDURE: Label three cuvettes as B -Blank, S -Standard, T-test. Add 1ml of enzyme reagent in all three of them Add 10 l of Standard Solution in S cuvette Add 10 l of Serum (sample) in T cuvette Allow them to stand for 10 min at 25˚C

14. Place them in spectrophotometer and adjust the wavelength to 540nm Observe the absorbance. Conc. of Sample = Absorbance of Unknown Absorbance of Known Conc. of Standard Solution

15. PROCEDURE BLANK 'B' STANDARD 'S‘ TEST 'T' ENZYMATICREAGENT 1ml 1ml 1ml STANDARD SOLUTION ------ 10 Microliter -------- SAMPLE SOLUTION ---- -------- 10 Microliter

16. INTERPRETATION: Desirable: < 200mg/dl (adults) Borderline: 200 -239 mg/dl High Risk: > 240 mg/dl

17. CAUSES OF HYPERCHOLESTEREMIA: Diabetes mellitus Hypothyroidism Nephrotic syndrome Obstructive Jaundice Familial hypercholesterolemia Also depends on age, race, geographical distribution, food consumption and lifestyle.