Bio ,, OSPE.docx

7/28/2019 Bio ,, OSPE.docx

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Lipid Profile

Lipid Profile :

1. LDL Cholesterol2. Cholesterol3. VLDL Cholesterol4. HDL Cholesterol5. Triglycerides6. Cholesterol/HDL ratio7. LDL /HDL ratio

Indications:

1. CAD Family history2. Obesity3. Hypertension4. Diabetes5. Renal diseases6. Liver diseases7. Thyroid related disorders

Estimation of cholesterol in the serum (principle)

:

Peroxidase

H2O2+ indicator -------------------> red color, measured at 505

nm.


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Estimation of HDL cholesterol

Principle

When serum is reacted with the Phosphotungstic acid and Magnesium chloride, it causes the

precipitation of LDL and VLDL proteins. HDL fraction however remains in the supernatant. The HDL

Cholesterol is then estimated by the same procedure as described for total cholesterol above and follows

the same principleTwo Steps

1. Precipitation of LDL and VLDL with phosphotungestic acid2. Estimation of HDL Cholesterol as mentioned before (3 reactions)

Estimation of triglycerides in serum

Lipase Glycerol kinase Glycerol phosphate oxidase Peroxidase

TAG ---------> FA + Glycerol ----------------> G3P -----------------------> DHAP +H2O2 --------------> red colorIndicator


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Estimation of LDL Cholesterol

Direct method: Costly and cumbersome

Indirect method: Needs no additional procedures, only calculations are requiredFriedewald formula (TG must be 400 mg/dl)

LDL C = (TC HDL C) 0.2 * TG

Where LDLC= LDL cholesterol, HDLC = HDL Cholesterol, TC =total cholesterol and TG= triglycerides

Normal Values:

HDL Male: >45 mg/dl or >0.75 mmol/l (SI units) Female: >55 mg/dl or >0.91 mmol/l (SI units)

Triglyceride Adult/elderly

Male: 40-160 mg/dl Female: 35-135 mg/dl

Cholesterol

Desirable 200 mg/dlBorderline high 200 - 240 mg/dl

High 240 mg/dl.

Hypercholesterolemia

Primary hyperlipoproteinemia

AtherosclerosisMyocardial Infaraction

Diabetes melitis

HypothyroidismObstructive jaundice

Hypocholesterolemia

Familial hypobeta l

ipoproteinemia

HyperthyroidismPernicious anemiaHemolytic jaundiceSevere malnutrition


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Primary biliary cirrhosis

The total cholesterol / HDL cholesterol ratio

The total chol /HDL is helpful in estimating the risk of developing

atherosclerosis. High ratios indicate a higher risk of heart attacks, whereas low ratios

indicate a lower risk.

An average ratio would be about 4.5. Ideally, one should strive for ratios of 2

or 3 (less than 4).

LDL/HDL ratio:Risk Level Low

risk

Average

risk

Moderate

risk

High

risk

LDL/HDL

Ratio

3.3 -

4.4

4.4 - 7.1


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Cholesterol concentration?

A patient's blood sample was processed for cholesterol estimation as

per the above protocol. Following data were observed.

Absorbance of the sample was 0.32, absorbance of the standard

was 0.4, and the conc. of standard was 200 mg/dl.

Determine the concentration of cholesterol in patients blood.

Answer:

Standard (200 mg/dl) 0.4

Sample (unknown) 0.32

Cholesterol concn = 200 X 0.32 /0.4 = 160 mg/dl


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Cardiac Markers

1. Proteins that leak out of fatally injured myocytesi. Myoglobin

ii. MB fraction of Creatine Kinase (CK-MB)iii. Troponins T and I

iv. Lactate dehydrogenase (LDH)v. Aspartate Aminotransferase (AST) or SGOT

2. The rate of appearance of these markers depends on:i. Intracellular location and molecular weight

ii. Blood flow and lymphatic drainage of area of infarctiii. Rate of elimination of marker from the blood

Quantitative estimation: CK-MB

Based on rate of formation of ATP per unit time


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Immunological Method:

Principle: Ck activity is measured in the presence of antibody (Ab) against CK-M monomer. This Ab completely inhibits the activity of CK-MM and half of CK-MB; while not

affecting the B subunit activity in CK-BB or CK-MB. We use the ordinal CK method to quantitatively determine CK-B activity.

The CK-MB activity is obtained by multiplying the CK-B activity by two

****% CK- MB activity = CK-MB activity x 100

Total Ck activity

AST activity in Serum

Quantitative determination Kinetic Method: change of rate of absorbance per unit time

Principles:

LDH activity in serum

Quantitative determination Kinetic method based on reaction:

The rate of NADH+ formation is indicated by increasing absorbance at 340nm and isdirectly proportional to serum LDH activity


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Clinical

significance

SpecificityDurationPeaksRisesMarkers

Very early DNS1 day6-8 hrs1-2 hrsMyoglobin

Early DNS4-5days12-24hrs4-6hsCPK

Early DS4-5days12-24 hrs4-6hsCK-MB

Early DNS5days48-60 hrs6hsAST, SGOT

Early DS3-5 days24-48 hrs3-6 hscTnI

Early D, FUS10-14 days24-48 hrs3-6 hscTnT

FUNS14days72-144 hs8-12hsLDH


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