Lipids in serum consist of triglycerides, cholesterols, phospholipids, free fatty acids and slight amounts of fat-soluble components such as fatsoluble vitamins and carotenes. Triglycerides, as major components of very low density lipoprotein (VLDL) and chylomicrons, play an important role in metabolism as energy sources and transporters of dietary fat.

This kit is based on an enzymatic method using N-ethyl-N- (2-hydroxy-3-sulfopropyl)-3,5-dimethoxyaniline sodium salt (DAOS) as a blue pigment. It is used for the quantitative determination of triglycerides in mouse serum. It is a simultaneous multi-sample assay format using a microplate, but measurements can also be made using a test tube.

Kit Contents

Buffer (50 mmol/L Good''s (PIPES) buffer, pH 6.5) 3 vials x 105 mL

Chromogen Substrate 3 vials x for 105 mL (when reconstituted)

Lipoprotein lipase 99 units/mL

Adenosine 5''-Triphosphate Disodium Salt Trihydrate (ATP) 1.7 mmol/L

Glycerolkinase 30 units/mL

Glycerol-3-phosphate Oxidase (GPO) 1.0 unit/mL

Peroxidase (POD) 5.5 units/mL

N-Ethyl-N-(2-hydroxy-3-sulfopropyl)-3,5-dimethoxyaniline sodium salt (DAOS) 0.54 mmol/L

4-Aminoantipyrine 0.11 mmol/L

Ascorbate Oxidase 4.5 units/mL

Standard Solution 1 vial x 10 mL

Glycerol 31.2 mg/dL (Corresponding to 300 mg/dL Triolein) [Assay principle] Triglycerides in a sample are hydrolyzed to glycerol and free fatty acids in a reaction catalyzed by lipoprotein lipase (LPL). Glycerol is converted to glycerol-3-phosphate by glycerolkinase (GK) in the presence of ATP. Glycerol-3-phosphate so formed is oxidized by glycerol-3-phosphate oxidase (GPO) in a reaction that produces hydrogen peroxide. The hydrogen peroxide produced causes DAOS and 4-Aminoantipyrine to undergo a quantitative oxidative condensation catalysed by peroxidase (POD), producing a blue pigment. The amount of triglycerides contained in the sample is determined by measuring the absorbance of the blue color.

Sensitivity (assay in a test tube)

The absorbance is below 0.10, when measuring purified water as a sample.

The absorbance is 0.09 to 0.25, when measuring 300 mg/dL triglycerides as a sample.

Specificity

The triglycerides concentration is less than ±12%, when measuring the known concentration of control serum as a sample.