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Lipid Profile

Lipid Profile :
1. 2. 3. 4. 5. 6. 7. LDL Cholesterol Cholesterol VLDL Cholesterol HDL Cholesterol Triglycerides Cholesterol/HDL ratio LDL /HDL ratio

Indications:
1. 2. 3. 4. 5. 6. 7. CAD Family history Obesity Hypertension Diabetes Renal diseases Liver diseases Thyroid related disorders

Estimation of cholesterol in the serum (principle)

: Peroxidase H2O2+ indicator -------------------> red color, measured at 505 nm.

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 principle Two Steps 1. Precipitation of LDL and VLDL with phosphotungestic acid 2. Estimation of HDL Cholesterol as mentioned before (3 reactions)

Estimation of triglycerides in serum

Lipase

Glycerol kinase

Glycerol phosphate oxidase

Peroxidase Indicator

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

Estimation of LDL Cholesterol

Direct method: Costly and cumbersome Indirect method: Needs no additional procedures, only calculations are required Friedewald 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/dl

Borderline high 200 - 240 mg/dl High 240 mg/dl.

Hypercholesterolemia Primary hyperlipoproteinemia Atherosclerosis Myocardial Infaraction Diabetes melitis Hypothyroidism Obstructive jaundice

Hypocholesterolemia Familial hypobeta l ipoproteinemia Hyperthyroidism Pernicious anemia Hemolytic jaundice Severe malnutrition

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 3.3 4.4 Average risk 4.4 - 7.1 Moderate risk <6 High risk <7

LDL/HDL Ratio

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) Sample (unknown) 0.4 0.32

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

Cardiac Markers
1. Proteins that leak out of fatally injured myocytes i. 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 infarct iii. Rate of elimination of marker from the blood

Quantitative estimation: CK-MB Based on rate of formation of ATP per unit time

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 is directly proportional to serum LDH activity

Markers

Rises

Peaks

Duration

Specificity

Clinical significance

Myoglobin

1-2 hrs

6-8 hrs

1 day

NS

Very early D

CPK

4-6hs

12-24hrs

4-5days

NS

Early D

CK-MB AST, SGOT

4-6hs 6hs

12-24 hrs 48-60 hrs

4-5days 5days

S NS

Early D Early D

cTnI

3-6 hs

24-48 hrs

3-5 days

Early D

cTnT

3-6 hs

24-48 hrs

10-14 days

Early D, FU

LDH

8-12hs

72-144 hs

14days

NS

FU

Serum Cardiac Marker

O- 4 hrs MI: Myoglobin is released 4- 48 hrs: CPK, CK-MB , cTnI, cTnT , AST > 48 hrs: LDH, cTnT