Petrarulo M, Pellegrino S, Marangella M, Cosseddu D, Linari F
Laboratorio Calcolosi Renale, Ospedale Mauriziano Umberto I, Turin, Italy.
Clin Chim Acta. 1993 Sep 30;218(2):193-200. doi: 10.1016/0009-8981(93)90183-5.
A rapid and sensitive liquid chromatographic technique to determine L-glutamate:glyoxylate and aminotransferase (EC 2.6.1.4) activity in human liver is described. Homogenised tissue was incubated for 60 min in the presence of substrates and the 2-oxoglutarate generated was converted into the corresponding phenylhydrazone which was determined using reversed-phase high-performance liquid chromatography. The procedure allowed the detection of the enzyme activity expressed by 7.5 micrograms of liver protein, it was more sensitive and less time-consuming than the spectrophotometric procedure previously used. No significant differences were found between normal controls and patients with primary hyperoxaluria. In an 8-month-infant with primary hyperoxaluria type 1, the enzyme activity was reduced to 16% of the average control values.
描述了一种快速灵敏的液相色谱技术,用于测定人肝脏中L-谷氨酸:乙醛酸转氨酶(EC 2.6.1.4)的活性。将匀浆组织在底物存在下孵育60分钟,产生的2-氧代戊二酸转化为相应的苯腙,使用反相高效液相色谱法进行测定。该方法能够检测到7.5微克肝脏蛋白所表达的酶活性,比之前使用的分光光度法更灵敏且耗时更短。正常对照组与原发性高草酸尿症患者之间未发现显著差异。在一名1型原发性高草酸尿症的8个月婴儿中,酶活性降至平均对照值的16%。