Miwa H, Yamamoto M, Asano T
Faculty of Pharmaceutical Sciences, Fukuoka University, Japan.
Anal Biochem. 1990 Feb 15;185(1):17-23. doi: 10.1016/0003-2697(90)90248-8.
Both hydroxymonocarboxylic acids and dicarboxylic acids in urine were converted into their 2-nitrophenylhydrazides without lengthy and cumbersome sample workup and were separated from each other by two-step extraction with diethyl ether at different pH values. HPLC analysis of each acid group was achieved isocratically within 30 min. By the use of a visible-range detector (400 nm) the detection limits ranged from 1 to 2 pmol and from 2 to 5 pmol per injection for the hydroxymonocarboxylic acids and dicarboxylic acids, respectively. The analytical results showed good recovery and reproducibility. Analysis profiles of the two acid groups in normal and diabetic subjects could be performed with 200 microliters of urine. The present method is superior over previously published methods because of its great simplicity and its time-, cost-, and labor-saving nature.
尿液中的一元羟基羧酸和二元羧酸均无需冗长繁琐的样品预处理即可转化为它们的2-硝基苯腙，并通过在不同pH值下用乙醚进行两步萃取彼此分离。每个酸组的高效液相色谱分析在30分钟内等度完成。使用可见范围检测器（400nm）时，一元羟基羧酸和二元羧酸每次进样的检测限分别为1至2皮摩尔和2至5皮摩尔。分析结果显示出良好的回收率和重现性。正常人和糖尿病患者的两个酸组的分析谱可以用200微升尿液进行。本方法由于其极大的简便性以及省时、成本低和省力的特点，优于先前发表的方法。
