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LIPID PROFILE
INTRODUCTION <ul><li>DEFINITION : Lipids are group of organic substances of fatty nature which are insoluble in water, sol...
IMPORTANCE of LIPIDS <ul><li>Reservoir of energy </li></ul><ul><li>Constituents of cell membrane </li></ul><ul><li>Constit...
CLASSIFICATION OF LIPIDS LIPIDS SIMPLE LIPIDS DERIVED LIPIDS FATS WAXES COMPOUND LIPIDS PHOSPHO LIPIDS CEREBROSIDES SULFO ...
 
LIPOPROTEINS <ul><li>Spherical complexes of lipids & proteins. </li></ul><ul><li>Transport various lipids & fat soluble vi...
LIPOPROTEINS-CLASSIFICATION LIPOPROTEINS MAJOR LIPOPROTEINS MINOR/ABNORMAL LIPOPROTEINS CHYLOMICRONS V.L.D.L L.D.L H.D.L I...
PLASMA LIPOPROTEINS 1.063-1.125 HDL 2 5 Beta 0-12 1.019-1.063 180-300 A LDL 4 Beta OR Pre-Beta 12-20 1.006-1.019 IDL 3 PRE...
PLASMA LIPOPROTEINS PRE-Beta 1.045-1.080 Lp (a) 7 ORIGIN >400 1.125-1.210 50-20 A HDL 3 6 MOBILITY Sf DENSITY(Kg/L) DIAMET...
PLASMA LIPOPROTEINS 26-32 2-7 15-20 3-5 45-55 HDL 18-24 4-8 45-50 6-8 18-22 LDL 15-20 45-65 16-22 4-8 6-10 VLDL 3-6 80-95 ...
APOLIPOPROTEINS <ul><li>Required for assembly & structure of lipoproteins. </li></ul><ul><li>Serve to activate enzymes imp...
MAJOR APOLIPOPROTEINS Structural protein for chylomicrons Chylomicrons Intestine Apo-B48 Unknown VLDL Liver Apo-AV Unknown...
MAJOR APOLIPOPROTEINS Unknown HDL Spleen,brain, Testes, adrenal Apo-D Inhibits lipoprotein binding to receptiors Chylomicr...
MAJOR APOLIPOPROTEINS Unknown Lp(a) Liver Apo-(a) Unknown Chylomicrons,VLDL,HDL Unknown Apo-L Unknown Chylomicrons,VLDL,HD...
LIPID PROFILE TESTS <ul><li>TOTAL LIPIDS </li></ul><ul><li>SERUM TOTAL CHOLESTEROL </li></ul><ul><li>SERUM H.D.L – C </li>...
INDICATIONS <ul><li>Screening for primary & secondary hyperlipidemias  </li></ul><ul><li>Monitoring for risk of atheroscle...
BLOOD SAMPLING & STORAGE <ul><li>Variation & error can be introduced before or during venepuncture or when samples are han...
BLOOD SAMPLING & STORAGE <ul><li>Biological variation: </li></ul><ul><li>Age = cholesterol levels increase with age. </li>...
BLOOD SAMPLING & STORAGE <ul><li>Biological variation: </li></ul><ul><li>5. Medical disorders = thyroid , hepatic & kidney...
BLOOD SAMPLING & STORAGE <ul><li>Biological variation: </li></ul><ul><li>It is recommended that lipoproteins measurement s...
BLOOD SAMPLING & STORAGE <ul><li>FASTING : </li></ul><ul><li>Ideally patient should fast for 12 hours before sampling.  </...
BLOOD SAMPLING & STORAGE <ul><li>POSTURE: </li></ul><ul><li>Decreases of as much 10 % in conc. of TC,LDL-C,HDL-C & apo-a I...
BLOOD SAMPLING & STORAGE <ul><li>POSTURE: </li></ul><ul><li>4. If recumbent position is used, this position should be used...
BLOOD SAMPLING & STORAGE <ul><li>EXERCISE:  </li></ul><ul><li>Mild exercise produces a slight decrease in conc. Of cholest...
BLOOD SAMPLING & STORAGE <ul><li>MENSTRUAL CYCLE: </li></ul><ul><li>The plasma cholesterol & triglyceride conc. Tend to be...
BLOOD SAMPLING & STORAGE <ul><li>DIET:  </li></ul><ul><li>A high fat diet increases serum triglycerides. </li></ul><ul><li...
BLOOD SAMPLING & STORAGE <ul><li>DIET:  </li></ul><ul><li>A high carbohydrate diet decreases the serum conc. of VLDL-C , T...
BLOOD SAMPLING & STORAGE <ul><li>DIET: </li></ul><ul><li>Large protein meals at lunch or in the evening also increase the ...
BLOOD SAMPLING & STORAGE <ul><li>SMOKING: </li></ul><ul><li>The plasma cholesterol , triglyceride & LDL cholesterol conc. ...
BLOOD SAMPLING & STORAGE <ul><li>ALCOHOL INGESTION: </li></ul><ul><li>When moderate amount of alcohol is ingested for 1 we...
BLOOD SAMPLING & STORAGE <ul><li>VENOUS Vs CAPILLARY SAMPLES: </li></ul><ul><li>Measurements in the capillary samples seem...
BLOOD SAMPLING & STORAGE <ul><li>PLASMA Vs SERUM: </li></ul><ul><li>Either can be used when TC,TG & HDL-C are measured. </...
BLOOD SAMPLING & STORAGE <ul><li>ANTICOAGULANTS: </li></ul><ul><li>Some anticoagulants such as citrate exert large osmotic...
BLOOD SAMPLING & STORAGE <ul><li>ANTICOAGULANTS: </li></ul><ul><li>EDTA is preferred anticoagulant even though TC & TG con...
BLOOD SAMPLING & STORAGE <ul><li>STORAGE: </li></ul><ul><li>TC,TG,HDL-C can be satisfactorily analyzed in frozen samples. ...
ESTIMATION OF PLASMA LIPIDS <ul><li>Cholesterol  & triglycerides are plasma lipids of most interest in diagnosis & managem...
CHOLESTEROL – chemical method <ul><li>Standard reference method. </li></ul><ul><li>Modified ABELL KENDALL method. </li></u...
CHOLESTEROL –  enzymatic method <ul><li>PRINCIPLE : These measure total cholesterol directly in plasma/serum through a ser...
CHOLESTEROL –  enzymatic method <ul><li>Cholesteryl esters + h2o ----------  cholesterol + free fatty acids </li></ul><ul>...
CHOLESTEROL –  enzymatic method <ul><li>ADVANTAGES : </li></ul><ul><li>Less subject to interference by non-sterol substanc...
CHOLESTEROL –  enzymatic method <ul><li>DISADVANTAGES : </li></ul><ul><li>They are not absolute specific for cholesterol <...
CHOLESTEROL –  enzymatic method <ul><li>DISADVANTAGES : </li></ul><ul><li>4. Interference of bilirubin is complex & can pr...
SERUM CHOLESTEROL: INCREASED IN  <ul><li>PRIMARY HYPER </li></ul><ul><li>LIPOPROTEINEMIA </li></ul><ul><li>SECONDARY HYPER...
SERUM CHOLESTEROL: DECREASED IN <ul><li>Severe liver damage. </li></ul><ul><li>Hyperthyroidism </li></ul><ul><li>Malnutrit...
TRIGLYCERIDE-ENZYMATIC METHOD <ul><li>PRINCIPLE :  </li></ul><ul><li>Serum TG are   hydrolysed to glycerol & free fatty ac...
TRIGLYCERIDE: INCREASED IN  <ul><li>Familiar hypertriglyceridemia </li></ul><ul><li>Von gierke disease </li></ul><ul><li>D...
TRIGLYCERIDE: DECREASED IN <ul><li>A beta lipoproteinemia </li></ul><ul><li>Malnutrition </li></ul><ul><li>Recent weight l...
TRIGLYCERIDE <ul><li>Eruptive xanthoma </li></ul><ul><li>Lipemia retinalis </li></ul>> 5000 mg/dl 1)Hyper lipidemia type I...
HDL-CHOLESTEROL: HOMOGENOUS ASSAY <ul><li>PRINCIPLE :   The method   depends   on the   properties of detergent which solu...
HDL-CHOLESTEROL: INCREASED IN <ul><li>Moderate consumption of alcohol, insulin & estrogen. </li></ul><ul><li>Hyper alpha l...
HDL-CHOLESTEROL: DECREASED IN <ul><li>Stress </li></ul><ul><li>Acute M.I </li></ul><ul><li>Stroke </li></ul><ul><li>Surger...
LDL-CHOLESTEROL <ul><li>FRIDEWALD CALCULATION :  </li></ul><ul><li>LDL-C  (MMOL/L)  = (TC – HDL CHOLESTEROL) – PLASMA TG /...
LDL-CHOLESTEROL <ul><li>FRIDEWALD CALCULATION  :  </li></ul><ul><li>DISADVANTAGES :  </li></ul><ul><li>Underestimate LDL-C...
LDL-CHOLESTEROL: INCREASED IN <ul><li>Familiar hypercholesterolemia </li></ul><ul><li>Familiar combined hyperlipidemia </l...
LDL-CHOLESTEROL: DECREASED IN <ul><li>Severe illness </li></ul><ul><li>Certain drugs </li></ul><ul><li>A beta lipoproteine...
VLDL-C/ PLASMA TG ratio <ul><li>VLDL-C/PLASMA TG ratio may be useful in evaluation of type III hyperlipoproteinemia </li><...
NORMAL LEVELS 8.0-11.0 MG/DL PHOSPHOLIPID PHOSPHORUS 6 9.0-15.0 MMOL/L FREE FATTY ACIDS 5 150-380 MG/DL PHOSPHOLIPIDS 4 10...
SERUM CHOLESTEROL & TG RISK CATEGORIES > or = 500 > or = 190 > 6.0 > 11.0 > or = 240 < 40 HIGH RISK 200-499 > or = 160 3.0...
CARDIAC RISK RATIO <ul><li>Cardiac risk ratio = LDL-C/HDL-C </li></ul><ul><li>Normal = 0.5 – 3.0 </li></ul><ul><li>Moderat...
MAJOR RISK FACTORS THAT MODIFY LDL GOALS <ul><li>Cigarette smoking </li></ul><ul><li>Hypertension </li></ul><ul><li>Low HD...
ATHEROGENIC INDEX <ul><li>ATHEROGENIC INDEX = combination of ratio of LDL-C to HDL-C X apo-B with ratio of apo-B to apo-AI...
ESTIMATION OF LIPOPROTEINS  <ul><li>ULTRACENTRIFUGAL METHODS </li></ul><ul><li>ELECTROPHORETIC METHOD </li></ul><ul><li>PO...
LIPOPROTEIN ELECTROPHORESIS <ul><li>USE  : identify rare familiar disorders ( e.g TYPE I, III, V HYPERLIPIDEMIA ) </li></u...
COMPLICATIONS OF HYPERLIPIDEMIA <ul><li>ACUTE MYOCARDIAL INFARCTION </li></ul><ul><li>STROKE </li></ul><ul><li>DIABETES ME...
REFERENCES <ul><li>HENRY’S clinical diagnosis & management by lab. methods 21/e </li></ul><ul><li>TIETZ’S fundamentals of ...
<ul><li>THANKS </li></ul>
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Dr ravi lipid profile

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Lipid profile is an important group of tests used to diagnose hyperlipidemias. it is also used in Investigating Myocardial infarction , Diabetes mellitus & nephrotic syndrome

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Dr ravi lipid profile

  1. 1. LIPID PROFILE
  2. 2. INTRODUCTION <ul><li>DEFINITION : Lipids are group of organic substances of fatty nature which are insoluble in water, soluble in fat solvents & which are related to fatty acids. </li></ul>
  3. 3. IMPORTANCE of LIPIDS <ul><li>Reservoir of energy </li></ul><ul><li>Constituents of cell membrane </li></ul><ul><li>Constituents of neural tissue </li></ul><ul><li>As a insulator & protective coating </li></ul><ul><li>As vitamins </li></ul>
  4. 4. CLASSIFICATION OF LIPIDS LIPIDS SIMPLE LIPIDS DERIVED LIPIDS FATS WAXES COMPOUND LIPIDS PHOSPHO LIPIDS CEREBROSIDES SULFO LIPIDS FATTY ACIDS STEROIDS
  5. 6. LIPOPROTEINS <ul><li>Spherical complexes of lipids & proteins. </li></ul><ul><li>Transport various lipids & fat soluble vitamins to & from tissues. </li></ul><ul><li>Contain core of hydrophobic lipids surrounded by hydrophilic lipids & proteins. </li></ul>
  6. 7. LIPOPROTEINS-CLASSIFICATION LIPOPROTEINS MAJOR LIPOPROTEINS MINOR/ABNORMAL LIPOPROTEINS CHYLOMICRONS V.L.D.L L.D.L H.D.L I.D.L Lp (a) Lp X Beta – V.L.D.L
  7. 8. PLASMA LIPOPROTEINS 1.063-1.125 HDL 2 5 Beta 0-12 1.019-1.063 180-300 A LDL 4 Beta OR Pre-Beta 12-20 1.006-1.019 IDL 3 PRE-Beta 20-400 0.95-1.006 300-700 A VLDL 2 ORIGIN >400 < 0.95 750-12,000 A CHYLOMICRONS 1 MOBILITY Sf DENSITY(Kg/L) DIAMETER TYPE
  8. 9. PLASMA LIPOPROTEINS PRE-Beta 1.045-1.080 Lp (a) 7 ORIGIN >400 1.125-1.210 50-20 A HDL 3 6 MOBILITY Sf DENSITY(Kg/L) DIAMETER TYPE
  9. 10. PLASMA LIPOPROTEINS 26-32 2-7 15-20 3-5 45-55 HDL 18-24 4-8 45-50 6-8 18-22 LDL 15-20 45-65 16-22 4-8 6-10 VLDL 3-6 80-95 2-4 1-3 1-2 CHYLOMICRON PL % TG % Ch. ESTERS % CHOLESTEROL % PROTEIN % LIPOPROTEIN
  10. 11. APOLIPOPROTEINS <ul><li>Required for assembly & structure of lipoproteins. </li></ul><ul><li>Serve to activate enzymes important in lipoprotein metabolism. </li></ul><ul><li>Mediate binding of lipoprotein to cell surface receptor. </li></ul>
  11. 12. MAJOR APOLIPOPROTEINS Structural protein for chylomicrons Chylomicrons Intestine Apo-B48 Unknown VLDL Liver Apo-AV Unknown HDL,Chylomicrons Intestine, Apo-AIV Structural protein for hdl HDL,Chylomicrons Liver Apo-AII Structural protein for hdl, activates LCAT HDL,Chylomicrons Intestine, Liver Apo-AI Function Lipoprotein association Source Apolipo- protein
  12. 13. MAJOR APOLIPOPROTEINS Unknown HDL Spleen,brain, Testes, adrenal Apo-D Inhibits lipoprotein binding to receptiors Chylomicrons,VLDL,HDL Liver Apo-CIII Cofactor for LPL Chylomicrons,VLDL,HDL Liver Apo-CII Unknown Chylomicrons,VLDL,HDL Liver Apo-CI Structural protein for VLDL,LDL,IDL,Lp(a) VLDL,IDL,LDL,Lp(a) Liver Apo-B100 Function Lipoprotein association Source Apolipo- protein
  13. 14. MAJOR APOLIPOPROTEINS Unknown Lp(a) Liver Apo-(a) Unknown Chylomicrons,VLDL,HDL Unknown Apo-L Unknown Chylomicrons,VLDL,HDL Liver Apo-J B2 glycoprotein Chylomicrons,VLDL,LDL,HDL Liver Apo-H Ligand for binding to LDL receptor Chylomicrons remnants,IDL,HDL Liver Apo-E Function Lipoprotein association Source Apolipo- protein
  14. 15. LIPID PROFILE TESTS <ul><li>TOTAL LIPIDS </li></ul><ul><li>SERUM TOTAL CHOLESTEROL </li></ul><ul><li>SERUM H.D.L – C </li></ul><ul><li>TC / H.D.L – C </li></ul><ul><li>SERUM TRIGLYCERIDES </li></ul><ul><li>SERUM PHOSPHOLIPIDS </li></ul><ul><li>ELECTROPHORESIS </li></ul>
  15. 16. INDICATIONS <ul><li>Screening for primary & secondary hyperlipidemias </li></ul><ul><li>Monitoring for risk of atherosclerosis </li></ul><ul><li>Monitoring treatment of hyperlipidemias. </li></ul>
  16. 17. BLOOD SAMPLING & STORAGE <ul><li>Variation & error can be introduced before or during venepuncture or when samples are handled & stored before analysis. </li></ul><ul><li>It is important to standardize conditions under which blood specimen are drawn & prepared for analysis. </li></ul>
  17. 18. BLOOD SAMPLING & STORAGE <ul><li>Biological variation: </li></ul><ul><li>Age = cholesterol levels increase with age. </li></ul><ul><li>Sex = women have lower level than men except in childhood & after early 50’s. </li></ul><ul><li>Season = cholesterol levels are slightly higher in winter. </li></ul><ul><li>Food intake = daily intake of fat increases cholesterol levels. Therefore it is important that patient should be on their usual diet for 2 wks & are neither gaining nor losing weight. </li></ul>
  18. 19. BLOOD SAMPLING & STORAGE <ul><li>Biological variation: </li></ul><ul><li>5. Medical disorders = thyroid , hepatic & kidney disease. </li></ul><ul><li>6. Life style factors = </li></ul><ul><li>Fasting </li></ul><ul><li>Posture </li></ul><ul><li>Venous occlusion </li></ul><ul><li>Anticoagulants </li></ul><ul><li>Recent M.I </li></ul><ul><li>Stroke </li></ul><ul><li>Cardiac catheterization </li></ul><ul><li>Trauma </li></ul><ul><li>Acute infection </li></ul><ul><li>pregnancy </li></ul>
  19. 20. BLOOD SAMPLING & STORAGE <ul><li>Biological variation: </li></ul><ul><li>It is recommended that lipoproteins measurement should be made no sooner than 8 weeks after any form of trauma or acute bacterial / viral infection & 3 – 4 months after child birth. </li></ul>
  20. 21. BLOOD SAMPLING & STORAGE <ul><li>FASTING : </li></ul><ul><li>Ideally patient should fast for 12 hours before sampling. </li></ul><ul><li>The concentration of LDL-C/HDL-C declines after eating </li></ul><ul><li>Chylomicrons are cleared with in 6 – 9 hours & their presence after 12 hours fast is abnormal. </li></ul><ul><li>TC/HDL-C levels can be measured in non fasting individuals. </li></ul><ul><li>NCEP recommended that patient fast for at least 9 hour before blood specimen are taken for lipid/lipoprotein analysis. </li></ul>
  21. 22. BLOOD SAMPLING & STORAGE <ul><li>POSTURE: </li></ul><ul><li>Decreases of as much 10 % in conc. of TC,LDL-C,HDL-C & apo-a I & B, have been observed after 20 minutes recumbence. </li></ul><ul><li>Postural changes are reversible when patient resumes original position. </li></ul><ul><li>NCEP guidelines = patient to be seated for 5 min. prior to sampling to prevent hemo concentration. </li></ul>
  22. 23. BLOOD SAMPLING & STORAGE <ul><li>POSTURE: </li></ul><ul><li>4. If recumbent position is used, this position should be used each time patient is sampled. </li></ul><ul><li>5. Prolonged venous occlusion leads to increase in cholesterol conc. By 10 – 15 %. </li></ul><ul><li>6. Torniquet should not be for more than minutes or two. </li></ul>
  23. 24. BLOOD SAMPLING & STORAGE <ul><li>EXERCISE: </li></ul><ul><li>Mild exercise produces a slight decrease in conc. Of cholesterol & triglyceride that may persists for several days. </li></ul><ul><li>Those who walk for about 4 hours each week have an average cholesterol conc. 5 % lower & HDL-C conc. </li></ul><ul><li>3.4 % higher than inactive persons. </li></ul>
  24. 25. BLOOD SAMPLING & STORAGE <ul><li>MENSTRUAL CYCLE: </li></ul><ul><li>The plasma cholesterol & triglyceride conc. Tend to be highest at midcycle, the time of maximum estrogen secretion </li></ul><ul><li>The cyclical variation in cholesterol is not observed with anovulatory cycles. </li></ul>
  25. 26. BLOOD SAMPLING & STORAGE <ul><li>DIET: </li></ul><ul><li>A high fat diet increases serum triglycerides. </li></ul><ul><li>Ingestion of monounsaturated fat reduces cholesterol & LDL cholesterol. </li></ul><ul><li>Plasma triglyceride conc. Is reduced when sucrose intake is reduced. </li></ul>
  26. 27. BLOOD SAMPLING & STORAGE <ul><li>DIET: </li></ul><ul><li>A high carbohydrate diet decreases the serum conc. of VLDL-C , TG, cholesterol & protein. </li></ul><ul><li>Individuals who eat many small meals throughout the day tend to have conc. of total LDL & HDL-C that are lower than when same type & amount of food is eaten in three meals. </li></ul>
  27. 28. BLOOD SAMPLING & STORAGE <ul><li>DIET: </li></ul><ul><li>Large protein meals at lunch or in the evening also increase the serum cholesterol for atleast 1 hour after a meal. </li></ul><ul><li>In vegeterian individuals, conc. Of LDL & VLDL-C are reduced by 37 % & 12%. </li></ul>
  28. 29. BLOOD SAMPLING & STORAGE <ul><li>SMOKING: </li></ul><ul><li>The plasma cholesterol , triglyceride & LDL cholesterol conc. are higher by about 3 % </li></ul><ul><li>9.1 % & 1.7 % respectively in smokers than in non smokers. </li></ul><ul><li>HDL cholesterol is lower in smokers than in non smokers.s </li></ul>
  29. 30. BLOOD SAMPLING & STORAGE <ul><li>ALCOHOL INGESTION: </li></ul><ul><li>When moderate amount of alcohol is ingested for 1 week, the serum TG conc. Is increased by more than 20 mg/dl. </li></ul><ul><li>Prolonged moderate ingestion may increase HDL-C conc., which is assoc . with reduced plasma conc. Of cholesterol ester transfer protein.( CETP ) </li></ul>
  30. 31. BLOOD SAMPLING & STORAGE <ul><li>VENOUS Vs CAPILLARY SAMPLES: </li></ul><ul><li>Measurements in the capillary samples seem to be little lower than venous samples. </li></ul><ul><li>2. Measurements in the fingerpick samples tends to be more variable than in venous samples. </li></ul>
  31. 32. BLOOD SAMPLING & STORAGE <ul><li>PLASMA Vs SERUM: </li></ul><ul><li>Either can be used when TC,TG & HDL-C are measured. </li></ul><ul><li>Plasma is preferred when lipoprotein are measured by ultra centrifugal / electrophoretic methods. </li></ul><ul><li>Serum can be used when it is necessary to store samples for weeks or months. </li></ul>
  32. 33. BLOOD SAMPLING & STORAGE <ul><li>ANTICOAGULANTS: </li></ul><ul><li>Some anticoagulants such as citrate exert large osmotic effect. </li></ul><ul><li>This results in falsely low plasma lipid & lipoprotein concentration. </li></ul><ul><li>Heparin because of high M.W can alter electrophoretic mobilities of lipoprotein. </li></ul>
  33. 34. BLOOD SAMPLING & STORAGE <ul><li>ANTICOAGULANTS: </li></ul><ul><li>EDTA is preferred anticoagulant even though TC & TG conc. In EDTA plasma are 3 % lower than in serum. </li></ul><ul><li>EDTA retards certain oxidative & enzymatic alterations that occur in lipoproteins during storage. </li></ul><ul><li>TC values of EDTA plasma should be multiplied by 1.03 to make it comparable to serum values. </li></ul>
  34. 35. BLOOD SAMPLING & STORAGE <ul><li>STORAGE: </li></ul><ul><li>TC,TG,HDL-C can be satisfactorily analyzed in frozen samples. </li></ul><ul><li>Apolipoproteins can also be measured in frozen samples. </li></ul><ul><li>Serum / plasma must be stored at – 70 deg if stored for long time. </li></ul><ul><li>For short time storage ( upto a month or two ) the sample can be kept at – 20 deg. </li></ul>
  35. 36. ESTIMATION OF PLASMA LIPIDS <ul><li>Cholesterol & triglycerides are plasma lipids of most interest in diagnosis & management of lipoprotein disorders </li></ul>
  36. 37. CHOLESTEROL – chemical method <ul><li>Standard reference method. </li></ul><ul><li>Modified ABELL KENDALL method. </li></ul><ul><li>PRINCIPLE : </li></ul><ul><li>1)Cholesteryl esters hydrolyzed with alcoholic KOH </li></ul><ul><li>2) Unesterfied cholesterols are extracted with petroleum ether. </li></ul><ul><li>3) Then they are measured with LIEBERMANN BURCHARD REAGENT. </li></ul>
  37. 38. CHOLESTEROL – enzymatic method <ul><li>PRINCIPLE : These measure total cholesterol directly in plasma/serum through a series of reaction in which cholesterol esters are hydrolyzed , the 3- OH group of cholesterol is oxidized & H2O2 is liberated which is measured . </li></ul>
  38. 39. CHOLESTEROL – enzymatic method <ul><li>Cholesteryl esters + h2o ---------- cholesterol + free fatty acids </li></ul><ul><li>Cholesterol + o2 --------------------------------------- </li></ul><ul><li>---cholesten-4 en-3one + h2o2 </li></ul><ul><li>H2o2 + phenol + 4-aminoantipyrine -------- quinoenimine +2h2o </li></ul><ul><li>The absorbance of quinoenimine produced is measured at 500 nm. </li></ul>
  39. 40. CHOLESTEROL – enzymatic method <ul><li>ADVANTAGES : </li></ul><ul><li>Less subject to interference by non-sterol substances that react in chemical method. </li></ul><ul><li>Consume only micro liters quantity of sample. </li></ul><ul><li>Do not require preliminary extraction step. </li></ul><ul><li>Rapid method. </li></ul><ul><li>If cholesterol ester hydrolase step is omitted, they can be used to measure Unesterfied cholesterol. </li></ul>
  40. 41. CHOLESTEROL – enzymatic method <ul><li>DISADVANTAGES : </li></ul><ul><li>They are not absolute specific for cholesterol </li></ul><ul><li>Cholesterol oxidase reacts with sterols other than cholesterol & also with plant sterols. </li></ul><ul><li>Ascorbic acid & bilirubin interfere with enzymatic methods by consuming h2o2. </li></ul>
  41. 42. CHOLESTEROL – enzymatic method <ul><li>DISADVANTAGES : </li></ul><ul><li>4. Interference of bilirubin is complex & can produce falsely high or low values. </li></ul><ul><li>5. Interference by bilirubin seems to be significant only at conc. > 5 mg/dl. </li></ul><ul><li>6. At this level, it has been reported to decrease cholesterol values by 5-15 %. </li></ul>
  42. 43. SERUM CHOLESTEROL: INCREASED IN <ul><li>PRIMARY HYPER </li></ul><ul><li>LIPOPROTEINEMIA </li></ul><ul><li>SECONDARY HYPER </li></ul><ul><li>LIPOPROTEINEMIA </li></ul><ul><li>Diabetes mellitus </li></ul><ul><li>Hypothyroidism </li></ul><ul><li>Nephrotic syndrome </li></ul><ul><li>Chronic uremia </li></ul><ul><li>Hemodialysis </li></ul><ul><li>Hepatic glycogenoses. </li></ul><ul><li>Obstru. Liver disease. </li></ul><ul><li>Chronic alcoholism </li></ul>
  43. 44. SERUM CHOLESTEROL: DECREASED IN <ul><li>Severe liver damage. </li></ul><ul><li>Hyperthyroidism </li></ul><ul><li>Malnutrition </li></ul><ul><li>Myelo proliferative disorders </li></ul><ul><li>Chronic anemia </li></ul><ul><li>Infection </li></ul><ul><li>Drugs </li></ul><ul><li>Hypo beta lipoproteinemia </li></ul><ul><li>A beta lipoproteinemia </li></ul><ul><li>Tangier’s disease </li></ul>
  44. 45. TRIGLYCERIDE-ENZYMATIC METHOD <ul><li>PRINCIPLE : </li></ul><ul><li>Serum TG are hydrolysed to glycerol & free fatty acids by lipase. </li></ul><ul><li>In presence of ATP & glycerokinase , glycerol is converted to Gly – PO4 which is then oxidised to yield h2o2. </li></ul><ul><li>H2o2 reacts with ESPAS to form colored complex, the intensity of which is measured at 546 nm </li></ul><ul><li>( 530-570 nm ) </li></ul>
  45. 46. TRIGLYCERIDE: INCREASED IN <ul><li>Familiar hypertriglyceridemia </li></ul><ul><li>Von gierke disease </li></ul><ul><li>Diabetes mellitus </li></ul><ul><li>Hypothyroidism </li></ul><ul><li>Nephrosis </li></ul><ul><li>Chronic renal failure </li></ul><ul><li>Diet </li></ul><ul><li>Drugs </li></ul>
  46. 47. TRIGLYCERIDE: DECREASED IN <ul><li>A beta lipoproteinemia </li></ul><ul><li>Malnutrition </li></ul><ul><li>Recent weight loss </li></ul><ul><li>Vigorous exercise </li></ul><ul><li>Drugs </li></ul><ul><li>NOTE: TG levels are inversely related to </li></ul><ul><li>HDL-C levels. </li></ul>
  47. 48. TRIGLYCERIDE <ul><li>Eruptive xanthoma </li></ul><ul><li>Lipemia retinalis </li></ul>> 5000 mg/dl 1)Hyper lipidemia type I or IV 2)Substantial risk of pancreatitis > 1000 mg/dl High risk of pancreatitis > 500 mg/dl Peri vascular disease 250-500 mg/dl No disease < 150 mg/dl Condition Concentration of TG
  48. 49. HDL-CHOLESTEROL: HOMOGENOUS ASSAY <ul><li>PRINCIPLE : The method depends on the properties of detergent which solubilizes only the HDL so that HDL-C is released to react with the cholesterol esterase & cholesterol oxidase & chromogen to give color. </li></ul><ul><li>The intensity of color is formed proportional to conc. Of HDL in sample, the absorbance of which is measured at 600 nm( 600- 670 nm ) </li></ul>
  49. 50. HDL-CHOLESTEROL: INCREASED IN <ul><li>Moderate consumption of alcohol, insulin & estrogen. </li></ul><ul><li>Hyper alpha lipoproteinemia </li></ul><ul><li>Hypo beta lipoproteinemia </li></ul>
  50. 51. HDL-CHOLESTEROL: DECREASED IN <ul><li>Stress </li></ul><ul><li>Acute M.I </li></ul><ul><li>Stroke </li></ul><ul><li>Surgery </li></ul><ul><li>Starvation </li></ul><ul><li>Diabetes mellitus </li></ul><ul><li>Hypothyroidism </li></ul><ul><li>Liver disease </li></ul><ul><li>Nephrosis </li></ul><ul><li>Uremia </li></ul><ul><li>Chronic anemia </li></ul>
  51. 52. LDL-CHOLESTEROL <ul><li>FRIDEWALD CALCULATION : </li></ul><ul><li>LDL-C (MMOL/L) = (TC – HDL CHOLESTEROL) – PLASMA TG / 2.175 </li></ul><ul><li>LDL-C (MG/DL) = (TC – HDL CHOLESTEROL) – PLASMA TG / 5 </li></ul>
  52. 53. LDL-CHOLESTEROL <ul><li>FRIDEWALD CALCULATION : </li></ul><ul><li>DISADVANTAGES : </li></ul><ul><li>Underestimate LDL-C in chronic alcholics </li></ul><ul><li>Unsuitable for monitoring </li></ul><ul><li>Misclassifies 15 – 40 % of patients when TG levels are between 200 to 400 mg/dl </li></ul><ul><li>Fails if fasting TG levels > 400 mg/dl. </li></ul><ul><li>Not reliable if suspect type III dyslipidemia </li></ul>
  53. 54. LDL-CHOLESTEROL: INCREASED IN <ul><li>Familiar hypercholesterolemia </li></ul><ul><li>Familiar combined hyperlipidemia </li></ul><ul><li>Diabetes mellitus </li></ul><ul><li>Hypothyroidism </li></ul><ul><li>Nephrosis </li></ul><ul><li>Chronic renal failure </li></ul>
  54. 55. LDL-CHOLESTEROL: DECREASED IN <ul><li>Severe illness </li></ul><ul><li>Certain drugs </li></ul><ul><li>A beta lipoproteinemia </li></ul>
  55. 56. VLDL-C/ PLASMA TG ratio <ul><li>VLDL-C/PLASMA TG ratio may be useful in evaluation of type III hyperlipoproteinemia </li></ul><ul><li>Expressed in mol/mol or mass/mass </li></ul><ul><li>Ranges 0.230-0.575 in samples without </li></ul><ul><li>beta VLDL </li></ul><ul><li>Type III subjects have ratio > 0.689, usually in range of 0.689 – 0.0919 </li></ul>
  56. 57. NORMAL LEVELS 8.0-11.0 MG/DL PHOSPHOLIPID PHOSPHORUS 6 9.0-15.0 MMOL/L FREE FATTY ACIDS 5 150-380 MG/DL PHOSPHOLIPIDS 4 10-90 MG/DL TRIGLYCERIDES 3 150-250 MG/DL TOTAL CHOLESTEROL 2 400-800 MG/DL TOTAL LIPIDS 1 level lipid S.No
  57. 58. SERUM CHOLESTEROL & TG RISK CATEGORIES > or = 500 > or = 190 > 6.0 > 11.0 > or = 240 < 40 HIGH RISK 200-499 > or = 160 3.0-6.0 7.1-11.0 200-239 MODERATE RISK 150-199 < or = 130 4.4-7.1 AVERAGE RISK < 150 < 30 < 100 0.5-3.0 3.3-4.4 150- 250 > Or = 50 NORMAL SERUM-TG VLDL-C LDL-C LDL/HDL TC/HDL-C TC HDL-C RISK CAT.
  58. 59. CARDIAC RISK RATIO <ul><li>Cardiac risk ratio = LDL-C/HDL-C </li></ul><ul><li>Normal = 0.5 – 3.0 </li></ul><ul><li>Moderate risk = 3.0 – 6.0 </li></ul><ul><li>High risk = > 6.0 </li></ul>
  59. 60. MAJOR RISK FACTORS THAT MODIFY LDL GOALS <ul><li>Cigarette smoking </li></ul><ul><li>Hypertension </li></ul><ul><li>Low HDL cholesterol ( < 40 mg/dl ) </li></ul><ul><li>Family history of premature CHD </li></ul><ul><li>Age ( men > 45 ; women > 55 ) </li></ul><ul><li>Diabetes mellitus </li></ul><ul><li>Pre existing CHD </li></ul>
  60. 61. ATHEROGENIC INDEX <ul><li>ATHEROGENIC INDEX = combination of ratio of LDL-C to HDL-C X apo-B with ratio of apo-B to apo-AI </li></ul><ul><li>Atherogenic index = </li></ul><ul><li>(TC-HDL-C) x apo-B/ (apo AI X HDL-C) </li></ul>
  61. 62. ESTIMATION OF LIPOPROTEINS <ul><li>ULTRACENTRIFUGAL METHODS </li></ul><ul><li>ELECTROPHORETIC METHOD </li></ul><ul><li>POLY ANION PRECIPITATION METHOD </li></ul><ul><li>HOMOGENOUS ASSAY </li></ul>
  62. 63. LIPOPROTEIN ELECTROPHORESIS <ul><li>USE : identify rare familiar disorders ( e.g TYPE I, III, V HYPERLIPIDEMIA ) </li></ul><ul><li>INDICATION : </li></ul><ul><li>serum TG > 300 mg/dl </li></ul><ul><li>fasting serum is lipemic </li></ul><ul><li>significant hyperglycemia, impaired glucose intolerance </li></ul><ul><li>serum uric acid > 8.5 mg/dl </li></ul><ul><li>clinical evidence of CHD or atherosclerosis in patient < 40 years of age. </li></ul>
  63. 64. COMPLICATIONS OF HYPERLIPIDEMIA <ul><li>ACUTE MYOCARDIAL INFARCTION </li></ul><ul><li>STROKE </li></ul><ul><li>DIABETES MELLITUS </li></ul><ul><li>GALL STONES </li></ul><ul><li>METABOLIC SYNDROME </li></ul><ul><li>PANCREATITIS </li></ul>
  64. 65. REFERENCES <ul><li>HENRY’S clinical diagnosis & management by lab. methods 21/e </li></ul><ul><li>TIETZ’S fundamentals of clinical chemistry 6/e </li></ul>
  65. 66. <ul><li>THANKS </li></ul>

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