3. Outline of Today’s lecture 1. Introduction of Triglyceride 2. Clinical significance of Triglyceride 3. Principle of Triglyceride estimation

4. Objectives  To determine serum Triglyceride level  Master how to determine the conc. of Triglyceride in serum.  Understand the clinical significance of Triglyceride

5. What are triglycerides? A triglyceride is an ester formed from glycerol and three fatty acid groups. Triglycerides are the main constituents of natural fats and oils.

6. The major lipids present in the plasma are fatty acids, triglycerides, cholesterol and phospholipids. Triglycerides consist of glycerol esterified with three long-chain fatty acids, such as stearic acid (18 carbon atoms) or palmitic (16 carbon atoms) acids. Triglyceride is present in dietary fat, and can be synthesized in the liver and adipose tissue to provide a source of energy, which can be mobilized when required, for example, during starvation.

7. Triglycerides containing both saturated and unsaturated fatty acids which are important components of cell membrane. Triglycerides are the main constituents of vvvv eeee gggg eeee tttt aaaa bbbb llll eeee o o o o iiii llll and aaaa nnnn iiii mmmm aaaa llll f f f f fats. They are a major component of hhhh uuuu mmmm aaaa nnnn s s s s kkkk iiii nnnn o o o o iiii llll ssss.

8. Lipoprotein Structure and Composition

9. © 2006 Thomson-Wadsworth Roles of Triglycerides Fat stores –Energy –Protection- fat layer protect organs –Insulation- fat does not conduct heat

10. Causes of High Triglycerides High carbohydrate diets Several diseases (type 2 diabetes, chronic renal failure, nephrotic syndrome) Certain drugs (corticosteroids, estrogens, retinoids, higher doses of beta-blockers) Various genetic dyslipidemias Physical inactivity Cigarette smoking

11. Increased Triglycerides Occur With The Following Conditions: Hyperlipoproteinemia type II, IIb, III, IV, and I. Liver disease, alcoholism. Liver disease, alcoholism. Nephritic syndrome, renal disease. Myocardial infarction. Hypothyroidism.

12. Decreased Triglyceride Levels Occur With The Following Conditions: Congenital α-β-lipoproteinemia. Malnutrition. Hyperthyroidism. Recent weight loss. Chronic obstructive lung disease.

13. How can you low high triglycerides? You can make dddd iiii eeee tttt and lifestyle changes to help lower your levels. Stay at a hhhh eeee aaaa llll tttt hhhh yyyy w w w w eeee iiii gggg hhhh tttt. Limit fats and sugars in your diet. Be more active. Quit ssss mmmm oooo kkkk iiii nnnn gggg.

14. Determination of serum Triglyceride Types of Samples: Plasma Water + solids (e.g. glucose, urea, albumin, fibrinogen) No cells Serum Serum = plasma – clotting factors

15. Requirements Requirements:  Automatic pipettes  Tips  Cuvettes  Spectrophotometer  Reagents Specimen: Specimen: Serum or plasma Serum or plasma

16. Preparation of Cuvettes Procedure Procedure Sample (Reagent+Serum) Standard (Reagent+ St. sample) Blank (only Reagent)

17. Triglyceride Lipase Glycerol + Fatty acids Glycerol + ATP Gk Glycerol-3-phosphate + ADP Glycerol-3-Phosphate + O 2 GPO Dihydroxyacetone-phosphate + H 2 O 2 Dihydroxyacetone-phosphate + H 2 O 2 2H 2 O 2 + 4-aminoantipyrine + 4-chlorophenol POD 2H 2 O 2 + 4-aminoantipyrine + 4-chlorophenol POD Quinoneimine + HCL + 4H 2 O Quinoneimine + HCL + 4H 2 O

18. Procedure Kit Method ReagentsBlankSampleStandard Sample ( Sample (  l)- 10 10- Standard ( Standard (  l)--10 Working reagent (ml) 111

19. Procedure Add the reagents and sample as directed Mix and let stand at room temperature for 10 min. Read the absorbance at 505 nm (495-550). Zero the spectrophotometer with the blank reagent. Read the absorbance's of the standard and samples within 1 hr Enter the absorbance's readings and calculate the concentration of the sample.

20. Calculations: Concentration of the sample (s) = Absorbance of the sample X Conc. of the Std. Absorbance of the standard

21. 50 – 150 mg/dl

22. Abnormal ??? Clinical Interpretation Interpretation Level mmol/LmmolLLevel mg/dLmgdL Normal range, low risk < 1.7 < 150 Some risk 2.26–5.65200-499 Very high, high risk >5.65> 500