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cholestrol determination in blood


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#1 eman

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Posted 20 January 2012 - 11:25 AM

can any one tell me about cholestrol determiantion in blood

#2 Bluefunk

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Posted 16 February 2012 - 07:54 PM

When we measure cholesterol, we extract all the lipids in the cell using a bligh and dyer differential extraction (google it, its simple). In our lab, we use an Iatroscan TLC-FID (thin layer chromatography flame ionization detector), which separates the lipids and when compared to standards, quantifies them by detecting the ions formed when it is burnt).

#3 Inmost sun

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Posted 17 February 2012 - 07:08 AM

in vitro assays based on cholesterol oxidase reaction

#4 Zagami Francesco

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Posted 09 December 2017 - 07:17 AM

Cholesterol total (by Zagami)

Reagent 1: Hepes buffer 50 mM pH 7.0, 2.03 g/L magnesium chloride, 0.508 g/L 4-aminoantipyrine, 1500 U/L cholesterol esterase, 2000 U/L cholesterol oxidase, 10 ml/L of 0.5 g/L Tween-20, 1 g/L NaN3.

Reagent 2: Hepes buffer 50 mM pH 7.0, 1.47 g/L N-Ethyl-N-(3-sulfopropyl)-3-methoxyaniline sodium salt (Fluka cod. 04340), 3000 U/L peroxidase, 10 ml/L of 0.5 g/L Tween-20, 1 g/L NaN3.

Procedure

Sample and Reagent 1 and 2 is: 16 µl: 1000 µl: 500 μl, setting a dominant wavelength of 505 nm, sub-wavelength of 660 nm, after mixing the sample and reagent 1, 37 °C incubation 3 – 5 min, read after each tube absorbance A1, reagent 2 was added the reaction mix 5 min, read the absorbance A2, the change in absorbance per tube ΔAbssample = A2 - A1, after deduction of the reagent blank sample to calculate the change in absorbance value of the standard ΔAbsstandard = A2 - A1, then for to calculate unknown sample concentration apply the following formula

total cholesterol (mg/dl) = ΔAbssample/ΔAbsstandardX Standard cholesterol






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