采用高效液相色谱法测定血浆中的支链酮酸。

Measurement of branched-chain ketoacids in plasma by high-performance liquid chromatography.

作者信息

Walser M, Swain L M, Alexander V

机构信息

Department of Pharmacology and Molecular Science, Johns Hopkins University School of Medicine, Baltimore, Maryland 21205.

出版信息

Anal Biochem. 1987 Aug 1;164(2):287-91. doi: 10.1016/0003-2697(87)90494-5.

DOI:10.1016/0003-2697(87)90494-5

PMID:3674376

Abstract

Branched-chain ketoacids were isolated from plasma or serum samples by acidification, passage through a cationic exchange resin, ether extraction, and extraction of the ether layer with phosphate buffer. The recovery of 2-[1-14C]ketoisocaproate taken through these procedures averaged 95 +/- 3%. Branched-chain ketoacids were measured by high-performance liquid chromatography using a single mobile phase (sodium phosphate:acetonitrile). In normal human subjects, mean +/- SD fasting levels of 2-ketoisocaproate, 2-keto-3-methylvalerate, and 2-ketoisovalerate were 29 +/- 8, 18 +/- 4, and 12 +/- 3 microM, respectively. In normal rats, slightly different results were found: 24 +/- 10, 19 +/- 7 and 17 +/- 6 microM, respectively. In both species, levels of each ketoacid expressed as fractions of total branched-chain ketoacids were much less variable.

摘要

通过酸化、阳离子交换树脂柱处理、乙醚萃取以及用磷酸盐缓冲液萃取乙醚层，从血浆或血清样本中分离出支链酮酸。经过这些步骤，2-[1-¹⁴C]酮异己酸的回收率平均为95±3%。采用单一流动相（磷酸钠：乙腈）的高效液相色谱法测定支链酮酸。在正常人类受试者中，2-酮异己酸、2-酮-3-甲基戊酸和2-酮异戊酸的空腹水平均值±标准差分别为29±8、18±4和12±3微摩尔/升。在正常大鼠中，结果略有不同：分别为24±10、19±7和17±6微摩尔/升。在这两个物种中，以总支链酮酸的分数表示的每种酮酸水平变化较小。