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VETSPEC™REAGENTS - Triglycerides

Intended Use

For IN VITRO diagnostic use in the quantitative determination of Triglycerides in serum using manual or automated applications.

Method Principle

Serum Triglycerides are hydrolyzed by microbial Lipase to Glycerol and free fatty acids. The resultant Glycerol, in the presence of Glycerol Kinase (GK), ATP and Mg2+ ions is phosphorilated to Glycerol-1-phosphate. This latter metabolite is then oxidized by Glycerophosphate Oxidase (GPO) to produce Hydrogen Peroxide. The Hydrogen Peroxide thus produced is quantitatively determined by coupling 4-aminoantipyrine with N-ethyl-N-(2-hydroxy-3-sulphopropyl)-m-toluidine (TOOS), where a Quinonemine dye with maximum absorption at 550 nm is produced. The following reaction scheme illustrates the reactions that occur in this method:

Lipase
     Triglycerides ──────> Glycerol + FFA

GK
                     Glycerol + ATP ─────────> 1-glycero-phosphate + ADP
   Mg2+

GPO
                   1-glycerophosphate + O2 ─────> H2O2 + Dihydroxyacetonephosphate

 Peroxidase
           H2O2 + TOOS + 4AA ────────> Quinoneimine dye + 2H2O
 

 

Preparation Of Working Reagent

Catachem Triglycerides Working Reagent is prepared by adding the required volume of distilled or deionized water to the required number of vials of Triglycerides reagent.

Reagent Storage And Stability

Catachem Triglycerides reagent is stored at 2-8°C. When stored as directed, this reagent is stable until the expiration date stated on the label. When prepared and stored as directed, the Working TG Reagent is stable for 3 days at room temperature (18-26°C) and for 30 days at  2-8°C.

Method Performance Characteristics

Sensitivity: 0.0008-0012 absorbance units per mg/dL.
Linear Range: 0-1000 mg/dL.
Precision: Within-run and day-to-day precision is summarized below:

Triglycerides

Within-Run Precision

Total Precision

MEAN

SD

CV

SD

CV

mg/dL

mg/dL

%

mg/dL

%

69

1.10

1.60

1.50

2.10

386

1.12

1.30

2.90

0.70

714

3.50

0.50

7.40

1.00

 

Correlation

A comparison of Catachem Triglycerides method, using an automated analyzer and a reference method based on the GPO Trinder reaction resulted in the following regression statistics:

Correlation Data

Parameter

Data Observed

N

118

Range (mg/dL)

29-715

Regression

Y = 1.036x + 5.8

Correlation

r = 0.998

Sy,x

7.00