CLINCHEM LIPIDS LAB
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CLINCHEM LIPIDS LAB - Marcador
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| 🇬🇧 | 🇬🇧 |
| Cholesterol Assay Based on Agappe INTENDED USE | This reagent is intended for in vitro quantitative determination of Cholesterol in serum or plasma |
| Cholesterol assay Methodology | • CHOD PAP |
| What is in vitro testing | - testing within the tube |
| Cholesterol assay linearity | 600 mg/ dL |
| Importance of LCF | If the patient has an increase lipid contents the serum/plasma will be blurry and the clarity will be decrease because of the increase lipids. For the machine to read the linearity we need a clearing factor. |
| Clinical Significance Cholesterol Assay | • It is the main lipid found in the blood, bile & brain tissues . It is also one of the most important steroids of the body & is a precursor of many steroid hormones. • Two thirds of cholesterol present in the blood is esterified. The liver metabolizes the cholesterol & it is transported in the blood stream by lipoproteins • Increased levels are found in hypercholesterolaemia , hyperlipidaemia , hypothyroidism, uncontrolled diabetes, nephritic syndrome & cirrhosis • Decreased levels are found in malabsorption , malnutrition , hyperthyroidism, anemia , & liver diseases. |
| Principle: Cholesterol Assay | Principle |
| Normal range, sample used, preparation | Normal range |
| Laboratory procedure | Lab procedure |
| Triglycerides Assay Based on Agappe INTENDED USE | This reagent is intended for in vitro quantitative determination of triglycerides in serum or plasma. |
| TAG METHODOLOGY | GPO TOPS methodology |
| Linearity | 1000 mg/ dL |
| Clinical Significance Triglycerides Assay | • Triglycerides are simple lipids, formed in the liver by glycerol & fatty acids • They are transported by VLDL, LDL & constitute about 95% of fat, stored as source of energy in the tissue & plasma. • Increased levels are found in hyperlipidemias , diabetes , nephrotic syndrome & hypothyroidism • Increased levels are risk factor for arteriosclerotic coronary disease , peripheral vascular disease, acute pancreatitis & hyperlipoproteinemia • Decreased levels are found in malnutrition & hyperthyroidism. |
| HDL Assay Based on Agappe INTENDED USE | This reagent is intended for in vitro quantitative determination of HDL cholesterol in serum • Direct determination of HDL Cholesterol |
| HDL assay method | Selective method |
| HDL linearity | Up to 150 mg/ dL |
| Clinical Significance HDL Assay | Blood total cholesterol levels have long been known to be related to coronary heart disease ( • In recent years, in addition to total cholesterol, high density lipoprotein cholesterol (HDL C ) has become an important tool used to assess an individual risk of developing CHD since a strong negative relationship between HDL C concentration and the incidence of CHD was reported |
| Principle of HDL Assay | The reaction between cholesterol other than HDL and the enzyme for cholesterol assay is suppressed by the electrostatic interaction between polyanions & cationic substances. • Hydrogen peroxide is formed by the free cholesterol in HDL by cholesterol oxidase. Oxidative condensation of EMSE and 4 AA is caused by hydrogen peroxide in the presence of peroxidase , and the absorbance of the resulting red purple quinone is measured to obtain the cholesterol value in HDL. |
| METHODOLOGIES General Sample and Patient Consideration | Cholesterol values rise with age • 12 hrs fasting ; CMs almost completely cleared within 6 9hrs after food ingestion • Total cholesterol and HDL C generally don’t need for fasting • Plasma or serum can be used for only cholesterol , TAG , and HDL C are measured; LDL C is calculated Friedewald equation • Plasma should be separated from cells within 3 hrs • Plasma (EDTA) is preferred for lipoprotein measurement |
| Cholesterol Assay considerations | -px should be in his/her usual diet 2 weeks prior to testing -total cholesterol is measured rather than its forms -Serum is the preferred specimen AC cause water shifting from RC dilution -For Colorimetric methods , sample for cholesterol assay is incubated 20 mins and 10min at 37 degrees Celsius. |
| Cholesterol Assay CHEMICAL METHODS Definition of terms: | • Saponification hydrolysis/splitting up cholesterol ester to free cholesterol and FAs • Extraction separation of cholesterol from protein carriers • Precipitation further isolation of FAs • Colorimetry addition of color reagent as an indicato |
| EXPLAIN CHOLESTEROL ASSAY CHEMICAL METHODS | Cholesterol assay |
| Explain cholesterol assay enzymatic method | Cholesterol assay |
| Triglyceride Assay Patient Preparation and Specimen Considerations | 12 14 hrs fasting fresh serum best sample • Plasma has slightly higher TAG level • Heparinized or EDTA treated plasma can be used • Glucose and phospholipids cause interference in non enzymatic mtd addition of zeolite (magnesium aluminum silicate) removes interference |
| WHAT ARE THE QUANTITATIVE ANALYSIS OF LIPOPROTEINS | ELECTROPHORESIS AND ULTRACENTRIFUGATION |
| EXPLAIN ELECTROPHOREISIS | ELECTRO |
| Quantitative Analysis of Lipoprotein | • Polyanion Precipitation Method and standing plasma test |
| Polyanion Precipitation Method | - lipoproteins are precipitated with polyanion heparin sulfate , dextran sulphate , phosphotungstate ) in the presence of calcium , magnesium , or manganese. most commonly used for HDL assay |
| Standing plasma test | Quali |
| Lipoprotein Disorders | Familial Hypercholesterolemia(Type 2A) defective or deficient LDL receptors • Familial Dystbetalipoproteinemia ( Type 3 accumulation of plasma VLDL rich in cholesterol VLDL fraction migrates abnormally in beta region Beta VLDL Abetalipoproteinemia Bassen Kornzweig Syndrome) defective apo B synthesis VLDL, LDL, CM = absent in plasma deficient Vitamins A,E,K; Vit D is not affected • Tangier’s Disease complete absence of HDL due to mutation in the ABCA1 gene on chromosome 9 • Tay Sachs Disease neurodegenerative disorder of lipid metabolism characterized by a deficiency of the enzyme Hexosaminidase A = accumulation of sphingolipids in brain |