15-HYDROXY-5Z,8Z,11Z,13E-EICOZATETRAENIC ACID MULTIPLE LABELED WITH TRITIUM AT DOUBLE BONDS AND METHOD OF ITS SYNTHESIS Russian patent published in 1997 - IPC

FIELD: biochemistry. SUBSTANCE: product: 15-hydroxy-5Z,8Z,11Z,13E-[³H]-eicozatetraenic acid multiple labeled with tritium. [³H]-Arachidonic acid (substrate) multiple labeled with tritium at double bonds is incubated with enzyme 15-lipoxygenase (specific activity is 10.9 mcmole/min/mg) isolated from soybean. After 10-15 min incubation intermediate substance is reduced by addition of two-fold molar excess of NaBH₄, reaction mixture is acidified to pH = 3.5 and the end product is isolated. Substrate taken at amount 12-16 mcg is dissolved in 0.7-1.0 ml 0.1 M sodium-borate buffer, pH = 9.0, and enzyme is added at amount providing the ratio substrate : enzyme = 1: (15-20) (mcg/mcg). Incubation is carried out at 20-22 C. Synthesized product is used in medicine and veterinary science. EFFECT: improved method of synthesis. 2 cl, 4 tbl