FMOC-Cl柱前衍生HPLC-MS/MS测定白芨中16种氨基酸成分  

Determination of 16 Amino Acids in Bletillae Rhizoma by HPLC-MS/MS with 9-Fluorenylmethyl Chloroformate Pre-Column Derivatization

作　　者：刘诗菡 金文芳 杨玉玉 邵千山 李春晓 范宝磊[2] LIU Shihan;JIN Wenfang;YANG Yuyu;SHAO Qianshan;LI Chunxiao;FAN Baolei(College of Pharmacy,Hubei University of Science and Technology,Xianning 437000;Hubei University of Science and Technology,Xianning 437000)

机构地区：[1]湖北科技学院药学院,湖北咸宁437000 [2]湖北科技学院,湖北咸宁437000

出　　处：《分析科学学报》2024年第6期675-682,共8页Journal of Analytical Science

基　　金：湖北省重大科技专项资金(2022BCE066)。

摘　　要：基于9-芴甲氧羰酰氯(FMOC-Cl)柱前衍生,建立了白芨中16种氨基酸成分的高效液相色谱-串联质谱(HPLC-MS/MS)分析方法。样品经提取、FMOC-Cl衍生后,样品溶液经Shim-pack VP-ODS色谱柱(300 mm×2.0 mm,5μm)分离,以乙腈-水溶液为流动相进行梯度洗脱分离。采用电喷雾电离负离子多反应监测模式检测。实验结果表明,16种氨基酸在一定浓度范围内线性关系良好(相关系数R 2>0.999),实际样品加标回收率在91.5%~105.6%之间,相对标准偏差均小于5%。通过多元统计分析,产地相近的白芨中氨基酸含量具有相似性,并且确定了8种核心指标。该方法简便、准确、灵敏,适用于白芨中16种氨基酸的快速定性、定量分析,测定结果可为白芨的质量评价以及后期的开发利用提供技术支持。A high-performance liquid chromatography-tandem mass spectrometry(HPLC-MS/MS)method based on pre-column derivatization with 9-Fluorenylmethyl chloroformate(FMOC-Cl)was developed for the analysis of 16 amino acid components in Bletillae Rhizoma.The samples were extracted and derivatized with FMOC-Cl and then separated on a Shim-pack VP-ODS column(300 mm×2.0 mm,5μm)with gradient elution using acetonitrile-water solution as the mobile phase and detected by electrospray ionization MS/MS in negative ion multiple reaction monitoring mode.The results showed that the linear relationships of the 16 amino acids were good in the concentration range(R 2>0.999),with high precision,good stability and good repeatability.The relative standard deviations were all less than 5%,and the recoveries were in the range of 91.5%-105.6%.Through multivariate statistical analysis,the amino acid contents in Bletillae Rhizoma of similar origin were similar and identified eight core indicators.The method is simple,accurate,sensitive,and can be used for rapid qualitative and quantitative analysis of 16 amino acids in Bletillae Rhizoma.The results can provide technical support for quality evaluation and later development and utilization of Bletillae Rhizoma.

关 键 词：氨基酸 白芨 FMOC-Cl衍生 高效液相色谱-串联质谱 多元统计分析 

分 类 号：R917[医药卫生—药物分析学] O657.63[医药卫生—药学]