Free Glycerol Determination Kit

General description

Glycerol, also referred to as glycerin or glycerine, is a 3 carbon sugar alcohol that forms the backbone of fatty acids. It is a central component for synthesis of all lipids, and acts as a backbone for triglycerides and phospholipids, which play an important role in cell membrane structure. Due to its low toxicity, glycerol is widely used in pharmaceutical, food and cosmetic formulations and is the main waste by-product of biodiesel production via transesterification. In addition to kits, the individual reagents and glycerol standard are available separately when fewer reactions are needed.

Application

The Free Glycerol Determination Kit has been used to determine the level of glycerol released into the medium from mammalian cells grown in vitro .

The Free Glycerol Determination Kit measures free, endogenous glycerol using coupled enzyme reactions and does not include initial lipase hydrolysis. Glycerol is phosphorylated by adenosine-5′-triphosphate (ATP) forming glycerol-1-phosphate (G-1-P) and adenosine-5′-diphosphate (ADP) in the reaction catalyzed by glycerol kinase (GK). G-1-P is then oxidized by glycerol phosphate oxidase (GPO) to dihydroxyacetone phosphate (DAP) and hydrogen peroxide (H₂O₂). Peroxidase catalyzes the coupling of H₂O₂ with 4-aminoantipyrine (4-AAP) and sodium N-ethyl-N-(3-sulfopropyl) m-anisidine (ESPA) to produce a quinoneimine dye that shows an absorbance maximum at 540 nm. The increase in absorbance at 540 nm is directly proportional to the free glycerol concentration of the sample.

The triglyceride and free glycerol kits are for the quantitative determination of glycerol, total triglycerides or free triglycerides.

Suitability

Suitable for the quantitative enzymatic determination of glycerol in serum or plasma.

Linkage

In addition to kits, the individual reagents and glycerol standard are available separately when fewer reactions are needed.