Graduate School of Pharmaceutical Sciences, University of Tokyo, Tokyo 1130033, Japan.
Graduate School of Pharmaceutical Sciences, University of Tokyo, Tokyo 1130033, Japan.
J Pharm Biomed Anal. 2021 May 10;198:114019. doi: 10.1016/j.jpba.2021.114019. Epub 2021 Mar 9.
Branched-chain α-keto acids (BCKAs, namely, α-ketoisovaleric acid (KIV), α-ketoisocaproic acid (KIC), and α-keto-β-methylvaleric acid (KMV)) are related to many diseases such as myeloid leukemia, liver cancer, and diabetes mellitus. A rapid quantitative analytical method for BCKAs using pillar array columns was developed. α-Keto acids were labeled with 1,2-diamino-4,5-methylenedioxybenzene (DMB), followed by their separation on octadecylsilane-treated pillar array columns with MeOH/HO as the mobile phase. Five DMB-labelled α-keto acids including the internal standard were separated in 160 s. The lower limits of quantification for DMB-α-keto acids were 2-5 μM. The intra- and interday precisions were 2.9-6.6 % and 5.2-10.7 %, respectively. The developed method was applied to BCKA quantification in human plasma samples; KIV, KIC, and KMV concentrations were determined to be 13.8, 24.2, and 15.2 μM, respectively. The method realized rapid, sensitive, and precise analysis of BCKAs and can be applied for clinical diagnosis.
支链α-酮酸（BCKAs，即α-酮异戊酸（KIV）、α-酮基-β-甲基戊酸（KMV）和α-酮异己酸（KIC））与多种疾病有关，如髓样白血病、肝癌和糖尿病。建立了一种使用柱阵列柱快速定量分析 BCKAs 的方法。α-酮酸用 1,2-二氨基-4,5-亚甲基二氧基苯（DMB）标记，然后用十八烷基硅烷处理的柱阵列柱，以 MeOH/HO 为流动相进行分离。5 种 DMB 标记的α-酮酸（包括内标）在 160 s 内得到分离。DMB-α-酮酸的定量下限为 2-5 μM。日内和日间精密度分别为 2.9-6.6%和 5.2-10.7%。该方法应用于人血浆样品中 BCKA 的定量分析；分别测定出 KIV、KIC 和 KMV 的浓度为 13.8、24.2 和 15.2 μM。该方法实现了 BCKAs 的快速、灵敏和精确分析，可用于临床诊断。
