Determination of FKPase-catalyzed formation of GDP-L-fucose from L-fucose by high performance liquid chromatography

ZHANG Yiming ¹ ,LI Wenkai ² ,MA Xuan ² ,LIU Huanyi ² ,FENG Zhe ² ,ZHANG Yaling ²

（1. Xi’an Tianhong Biotechnology Co., Xi’an　710018, China； 2. Shaanxi Normal University, Xi’an　710100, China ）

Abstract: A high-performance liquid chromatography method was established to determine the formation of GDP-L-fucose from L-fucose catalyzed by FKP. Agilent C₁₈ (250 mm × 4.6 mm, 5 μm) was used as the chromatographic column, with 2.72 g/L potassium dihydrogen phosphate (containing 0.2% tetrabutylammonium bromide, pH 4.0 adjusted with phosphoric acid) and acetonitrile as the mobile phase, flow rate of 1.0 mL/min, detection wavelength of 254 nm, injection volume of 20 μL, column temperature of 30 ℃, and determined using a diode array detector. The GDP-L-fucose peak was single and free from interference from other peaks. The mass concentrations of GDP-L-fucose and adenosine diphosphate showed good linear relationships with their respective chromatographic peak areas in the ranges of 1.005-21.100 and 0.05-0.2 μg/mL, with linear correlation coefficients of 0.999 9 and detection limits of 0.016 and 0.3 μ g/mL, respectively. The average recovery rates of sample spiked were 99.6%, the relative standard deviations of the measurement results were 0.04% (n=6). This method is applicable for the determination of the conversion of L-fucose to GDP-L-fucose catalyzed by FKP enzyme.Keywords: high performance liquid chromatography；diode array；GDP-L-fucose；enzyme-catalysed

引用本文： ZHANG Yiming, LI Wenkai, MA Xuan, et al. Determination of FKPase-catalyzed formation of GDP-L-fucose from L-fucose by high performance liquid chromatography[J]. Chemical Analysis and Meterage, 2025, 34(4): 50.)

通讯作者：ZHANG Yiming, Master, Engineer, research direction in organic synthesis and chromatographic separation

中图分类号： O657.7

文章编号：1008-6145(2025)04-0050-07

文献标识码： A

收稿日期：2025-03-10

出版日期：2025-04-20

网刊发布日期：2025-04-25