| 摘要 |
1373106 Determination of triglyceride CALBIOCHEM 30 Nov 1971 [16 Dec 1970] 55609/71 Heading G1B A glycerol ester of a fatty acid in an aqueous liquid, for example serum or milk is assayed by hydrolyzing the ester by adding a lipase and a protease to the liquid, then determining the amount of glycerol present in the resulting solution. The lipase may be from Chromobacterium viscosum or Rhizopus delemar; the protease may be chymotrypsin (exemplified), trypsin, a streptomyces griseus protease, elastase, papain or bromelin. The assay may be carried out (a) using a reagent comprising the lipase and protease, glycerol kinase, ATP, phosphoenol pyruvate (PEP), a magnesium ion source, pyruvate kinase (PK), lactate dehydrogenase (LDH) and NADH and measuring the decrease in optical density resulting from the oxidation of the NADH; (b) using the reagent as in (a), without the NADH and LDH, and reacting the pyruvate ion formed with dinitrophenyl hydrazine; (c) using the reagent as in (a), without the PEP, PK NADH and LDH, adding glycerol phosphate dehydrogenase and NAD, and measuring increase in fluorescence produced by formation of NADH, or optionally also including in the reaction mixture dinitrophenyl hydrazine which forms a coloured product with dihydroxyacetone phosphate; or (d) using a reaction mixture which comprises the lipase, protease, NAD and glycerol dehydrogenase and measuring the increase in optical density, resulting from formation of NADH, or alternatively adding dinitrophenylhydrazine to form a coloured product with the dihydroxyacetone in the reaction mixture. In other methods the NADH formed may be determined by reaction with a tetrazolium salt in the presence of a hydrogen transfer agent such as diaphorase or phenazine methosulphate to form a coloured dye. |
