TRIGLYCERIDES

(GOP – PAP Method)

For the determination of Triglycerides in serum and plasma
(For In vitro Diagnostic use only)

SUMMARY:

Triglycerides are the 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.

PRINCPLE:

Lipoprotein Lipase
Triglycerides Glycerol + Free fatty acids
Glycerol Kinase
Glycerol + ATP Glycerol 3 Phosphate + ADP
Glycerol 3 PO
Glycerol 3 Phosphate + O2 Dihydroxyacetone Phosphate + H2O2
Peroxidase
H2O2 + 4 Aminoantipyrine + Phenol Red Quinoneimine dye + H2O
The intensity of the color formed is directly proportional to the amount of Triglycerides present in the sample.

CONTENTS:

PACK SIZE TGL REAGENT (A1) STANDARD (S)

1x50ml 1x50ml 2ml

2x50ml 2x50ml 2ml

4x25ml 4x25ml 2ml

STORAGE/STABILITY:

Triglycerides reagent is stable at 2 – 8˚C till the expiry mentioned on the label.
Triglycerides Standard is stable at 2-8˚C till the expiry mentioned on the label.

REAGENT PREPARATION:

Reagents are ready to use. Protect from bright light.

SAMPLE:
Serum or Plasma. Triglycerides is reported to be stable in the sample for 5 days at 2-8˚C.

GENERAL SYSTEM PARAMETER:

Reaction Mode End Point Sample Volume 10 µl

Wavelength 505 nm Reagent volume 1000µl
Blank Reagent Standard 200 mg/dl
Incubation 10 min 37˚C/15 min R.T Reaction Slope Increasing
Delay Time 5 Sec Linearity 1000 mg/dl
Read Time 5 Sec Units mg/dl

ASSAY PROCEDURE:
Wavelength/ Filter : 505 nm (Hg 546 nm)/Green
Temperature : R.T/37˚C
Light Path : 1 cm
Pipette into clean dry test tube labeled as Blank (B), Standard (S) and Test (T)

Addition Sequence B(µl) S(µl) T(µl)

Triglycerides 1000 1000 1000
Reagent(A1)
Standard(S) - 10 -
Sample - - 10
Mix well and incubate at 37˚C for 10 min or at R.T. for 15 min. Measure the absorbance of the standard
(Absorbance of Standard) and test Sample (Absorbance of Test) against reagent blank at 505 nm (546 nm)
within 60 min.

CALCULATION:
Absorbance of Test
Triglycerides in mg/dl = ------------------------ × 200
Absorbance of Standard

NORMAL REFERENCE VALUES:

Serum and Plasma (Suspicious): 150 mg/dl and above
(Elevated) : 200 mg/dl and above
It is recommended that each laboratory establish its own normal range representing its patient population.

LINEARITY:
This procedure is linear upto 1000 mg/dl. If values exceed this limit , dilute the sample with normal
saline(NaCl 0.9%) and repeat assay. Calculate the value using the proper dilution factor.

NOTE:
Fasting Samples of 12 to 14 hours are preferred. Fatty meals and alcohol may cause elevated results. Patients
should not drink alcohol for 24 hours before the test.

REFERENCES:
1.Trinder.P., (1969) Ann. Clin. Biochem . 6:24
2.Bucolo.G, David.H (1973) Clin. Chem 19:476
3.Fossati.P, Prencipe.L (1982) Clin. Chem. 28:277