LIPID Determination Flashcards
LABORATORY TEST FOR
LIPOPROTEIN AND LIPID DISORDER
- The patient should remain on a regular diet for at least 3 weeks prior to the blood sample
being taken. - There should be no significant weight gain or loss.
- No consumption of alcohol for at least 2 days
-The sample must be collected after a fast of atleast 12 hour
Triglyceride Determination methods
Zilversmith and Van Handel
Enzymatic Method
Zilversmith and Van Handel
◦ 1st step: extract using organic
solvents like
Ether and alcohol
Reagent used 1st step in Zilversmith and Van Handel
Bloor’s reagent
What added in 2nd step in Zilversmith and Van Handel
KOH (potassium hydroxide)
In 3rd step what does it measure in Zilversmith and Van Handel
Glycerol
In 3rd step in zilversmith and van handel the Oxidize glycerol by periodate solution of
Formaldehyde
4th step in zilversmith and van handel the color reaction of formaldehyde is
Pink chromopore
Steps of zilversmith and van handel
Step 1: extracting using organic solvent
Step 2: add KOH
Step 3: measurement of glycerol
Step 4: color reaction of formaldehyde (pink chromopore)
It is a method Uses Lipase – no extraction needed
Enzymatic method
Triglyceride + 3 H2O → Glycerol + 3 F.A. What enzyme is used
Lipase
Glycerol + ATP → Glycerophosphate + ADP what enzyme used
Glycerokinase
- A) Glycerophosphate + O2 → Dihydroxyacetone + H2O2 what enzymes used
Glycerophosphate oxidase
- B) Glycerophosphate + NAD → Dihydroxyacetone + NADH + H
what are the enzymes used
Glycerophosphate dehydrogenase
NADH + Tetrazolium dye → Formazan + NAD what are the enzymes used
Diaphorase
ADP + Phosphoenol pyruvate →ATP + Pyruvate what enzymes is used
Pyvurate kinase
Pyruvate + NADH + H → Lactate + NAD
What enzymes is used
Lactate dehydrogenase
DETERMINATION
OF PLASMA
TRIGLYCERIDES
Hydrolysis of triglyceride to form glycerol and free fatty acids
Measurement of glycerol present (either as glycerol or after conversion to another product)
Measure spectrophotometrically, fluorometrically or colorimetrically
Method used fluorometry
Hantzsch method
Hantzsch method end color
Yellow
Wavelength used in hantzsch method
400-485nm
ATP III Classification for TAG Values
Normal
Borderline high
High
Very high
Normal triglycerides result/reference value
<150mg/dl
Borderline high of triglyceride result/reference value
150-199mg/dl