Test Catageory
فئة التحليل
Clinical Chemistry
Refrence Books
كتب مراجع
1. Bishop’s Clinical chemistry
Skills Required
المهارات المطلوبة
1. Pipetting
2. Colorimetry
Video Resources
مصادر فيديو
Principle
Free cholesterol, including that originally present, is then oxidized by cholesterol oxidase (CHOD) to cholest-4-en-3-one and H2O2. 3. In presence of peroxidase (POD), the formed hydrogen peroxide formed effects the oxidative coupling of phenol and 4- aminoantipyrine (4-AAP) to form a Pink – Red colored quinoneimine dye.
Equipments
1. 10 – 100 ul + 100 – 100 ul Automatic Pipette
2. Small glss tubes
3. cholesterol mono-reagent
4. yellow and blue tips
5. Colorimeter or Spectrophotometer
Sample Type
Serum or plasma is acceptable for the Test
But make sure the Patient is fasting to get accurate results
Procedure
1. in 3 clean dry tubes add reagents and sample as mentioned in the Table below
| Blank (B) | STandard (STD) | Test (T) | |
|---|---|---|---|
| W. Reagent | 1000 μl | 1000 μl | 1000 μl |
| Standard | – | 10 μl | – |
| Sample | – | – | 10 μl |
Procedure
2. Mix Well and incubate for 10 mins at room Tempreature or 5 min @ 37c’
3. using the colorimeter or spectrophotometer read the absorbance against blank at filter wavelenght 540 nm
4. then do the calculations based on the following formula
Cholesterol = Absorbance of test / Absorbance of standard x Conc of standard
The STD Conc is 200 mg/dl or may vary depending on the reagent brand.
Normal Range
M. Sulieman
mohammad@mlsgaate.com
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