Triglycerides ( GPO / PAP METHOD )

 Clinical significance :-

                       Triglyceride are form of fatty acid esters. They are produced in the liver by binding glycerol  and other fatty acids. They are transported by VLDL and LDL and act as a storage source for energy. Increased level are found in hyperlipidemias,  diabetes , nephrotic syndrome,  hypothyroidism and hyper lipidemia . Increased level of risk factor of arterosclerotic coronary disease and peripheral vascular disease. Decreased level are found in malnutrition, hyper thyroidism, hypolipidemia.

Principle :-

               Lipoprotein lipase hydrolyses triglycerides to glycerol and free fatty acids. The glycerol formed with ATP in the presence of glycerol kinase  forms glycerol 3 phosphate which is oxidised by the enzyme glycerol 3-phosphate which is oxidised  by the enzyme glycerol phosphate oxidised to form hydrogen peroxide.  The hydrogen peroxide further react with phenolic compound and 4-aminoantipyrine by the catalytic action of peroxidase to form a red colour quinonimine dye Complex.  Intensity of the colour formed is directly proportional to the amount of triglyceride present in the sample.

 Material required :-

• Clean and dry glassware.                                     
• Laboratory glass Pipettes or  micro Pipettes and tips. 
• Colorimeter.

Normal Range :-

                    Serum (Suspicious) : 150 mg/dl and above 

                          (Elevated)   : 200 mg/dl and above 

Procedure :-

                       Pipette into clean and dry test tubes labeled as Blank ( B ) , Standard ( S ), and Test ( T ) :

Addition           Sequence	B	S	T
Reagent	1ml	1ml	1ml
Standard	-	10ul	-
Sample	-	-	10ul

 Mix well  and  incubate  at  37 ‘ C for 10 mins. Measure absorbance of the standard ( Abs . S ) and Test ( Abs . T ) against Reagent blank at 520 nm.

 

   Calculation  :-

                                              Abs . T

                  TRIGLYCERIDES ( mg/dl ) = ..............  * 200

                                              Abs . S   

Preparation and reagent and stability :-

                                        All Reagent are ready to use and are stable till the expiry date mentioned on the label , when stored at 2' to 8' C.

Quality Control :-

                  For accuracy it is necessary to run known controls with every assay.

Limitation & Precautions :-

 

1. Storage conditions as mentioned on the kit to be adhered.
2. Do not freeze or expose the reagent to high temperature and protect from direct as it may affect the performance of the kit.
3. Use clean glassware free from dust .

 

General System Parameters:-

          Reaction type           : End point 

          Wave length             : 505 nm ( 490 - 530 nm )

          Temperature             : 37'C

          Incubation              : 10 minutes 

          Reagent Volume          : 1.0 ml

          Sample Volume           : 10 ul 

          Standard concentration  : 200 mg/dl

          Zero setting            : Reagent Blank

          Light Path              : 1 cm