can we determine triglycerides in blood serum through lipase enzyme. any valuable method which is directly hydrolysis of triglycerides in blood serum. Lipase directly synthesized triglycerides through lipase. which type of triglycerides synthesized through lipase enzyme .
can we use blood serum and lipase enzyme both are mixable. can lipase enzyme directly synthesized triglyceride in blood serum because lipase have hydrolysis property and having only target triglyceride. lipase have main part of the triglyceride .
The kits are available for assay of serum triglycerides. The lipase converts triglycerides to glycerol and three fatty acid molecules. The glycerol is phosphorylated and a colorimetric assay is developed.
Enzymatic hydrolysis of serum triglycerides by lipase generates free fatty acids and glycerol. The
glycerol released is phosphorylated by adenosine triphosphate (ATP) forming glycerol-1-phosphate
(G-1-P) and adenosine-5’-diphosphate (ADP) in the reaction catalyzed by glycerol kinase. G-1-P is
then oxidized by glycerol phosphate oxidase to dihydroxyacetone phosphate (DAP) and hydrogen
peroxide (H2O2). A quinoeimine dye is produced by the peroxidase catalyzed coupling of 4-
aminoantipyrine (4-AAP) and sodium N-ethyl-N-(3-sulfopropyl)m-anisidine (ESPA) with H2O2, which
shows an absorbance maximum at 540nm. The increase in absorbance at 540nm is directly
proportional to glycerol (and triglyceride) concentration of the sample.
Triglyceride Fatty Acids + Glycerol
Triglyceride determination using the enzyme can degrade many different compounds of lipids, but using the following steps in the assay and as noted in the attached equations, these steps can be determination only Triglycerid..
Simply use Roche Accutrend Plus System with its triglyceride test strip. Just small fingertip whole blood applied into the strip , and then the meter can measure the concentration of triglyceride with 600mg/dL in maximum.
https://diagnostics.roche.com/global/en/products/instruments/accutrend-plus.html#selectProfile
"Cayman's Triglyceride Colorimetric Assay uses the enzymatic hydrolysis of the triglycerides by lipase to glycerol and free fatty acids. The glycerol released is subsequently measured by a coupled enzymatic reaction system. The glycerol formed in reaction 1 is phosphorylated to glycerol- 3-phosphate in a reaction catalyzed by glycerol kinase. The glycerol-3- phosphate is oxidized by glycerol phosphate oxidase producing dihydroxyacetone phosphate and hydrogen peroxide. Peroxidase catalyzes the redox-coupled reaction of H2O2 with 4-aminoantipyrine (4-AAP) and N-Ethyl-N-(3-sulfopropyl)- m-anisidine (ESPA), producing a brilliant purple color. The absorbance is measured at 540 nm."
https://www.caymanchem.com/pdfs/10010303.pdf
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