Fast analysis using pillar array columns: Quantification of branched-chain α-keto acids in human plasma samples
Fast analysis using pillar array columns: Quantification of branched-chain α-keto acids in human plasma samples
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DOI：
10.1016/j.jpba.2021.114019
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发表时间：
2021-03-13
影响因子：
3.4
通讯作者：
Tsunoda, Makoto
中科院分区：
文献类型：
--
作者：
Fujiwara, Takuya;Funatsu, Takashi;Tsunoda, Makoto
支链α-酮酸（BCKA，即α-酮异戊酸（KIV）、α-酮异己酸（KIC）和α-酮-β-甲基戊酸（KMV））与骨髓性白血病、肝癌、糖尿病等多种疾病有关。开发了一种使用柱阵列柱快速定量分析 BCKA 的方法。 α-酮酸用 1,2-二氨基-4,5-亚甲二氧基苯 (DMB) 标记，然后在十八烷基硅烷处理的柱阵列柱上以 MeOH/H2O 作为流动相进行分离。五种 DMB 标记的 α-酮酸（包括内标）在 160 秒内分离。 DMB-α-酮酸的定量下限为2-5μM。日内和日间精度分别为2.9-6.6%和5.2-10.7%。所开发的方法应用于人血浆样本中BCKA的定量； KIV、KIC和KMV浓度分别测定为13.8、24.2和15.2μM。该方法实现了BCKA的快速、灵敏、精确分析，可应用于临床诊断。 (C) 2021 Elsevier B.V. 保留所有权利。
Branched-chain alpha-keto acids (BCKAs, namely, alpha-ketoisovaleric acid (KIV), alpha-ketoisocaproic acid (KIC), and alpha-keto-beta-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. alpha-Keto acids were labeled with 1,2-diamino-4,5-methylenedioxybenzene (DMB), followed by their separation on octadecylsilane-treated pillar array columns with MeOH/H2O as the mobile phase. Five DMB-labelled alpha-keto acids including the internal standard were separated in 160 s. The lower limits of quantification for DMB-alpha-keto acids were 2-5 mu M. 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 mu M, respectively. The method realized rapid, sensitive, and precise analysis of BCKAs and can be applied for clinical diagnosis. (C) 2021 Elsevier B.V. All rights reserved.