色谱

色谱 ›› 2014, Vol. 32 ›› Issue (12): 1326-1332.DOI: 10.3724/SP.J.1123.2014.08025

• 研究论文 • 上一篇    下一篇

超高效液相色谱-串联质谱法定性和高效液相色谱法定量分析天南星中氨基酸成分

王星, 池玉梅, 康安   

  1. 南京中医药大学, 江苏 南京 210023
  • 收稿日期:2014-08-26 修回日期:2014-10-05 出版日期:2014-12-08 发布日期:2014-11-28
  • 通讯作者: 池玉梅,E-mail:ymchii@njutcm.edu.cn.
  • 基金资助:

    江苏高校优势学科建设工程资助项目(PADA).

Qualitative and quantitative analysis of the amino acids in Rhizoma Arisaematis by ultra high performance liquid chromatography-tandem mass spectrometry and high performance liquid chromatography

WANG Xing, CHI Yumei, KANG An   

  1. Nanjing University of Traditional Chinese Medicine, Nanjing 210023, China
  • Received:2014-08-26 Revised:2014-10-05 Online:2014-12-08 Published:2014-11-28

PDF

265

被引次数

16

摘要：

为建立中药材中氨基类极性非紫外活性成分的定性与定量分析方法,以中药材天南星为研究对象,采用柱前衍生化技术,以异硫氰酸苯酯(PITC)为衍生化试剂,经C18色谱柱(100 mm×2.1 mm, 3.5 μm)分离和超高效液相色谱-串联质谱(UHPLC-MS/MS)分析,共解析了天南星中20个成分,包括18个氨基酸和2个胺类化合物。经优化衍生化条件,应用高效液相色谱法(HPLC),以Diamonsil C18色谱柱(250 mm×4.6 mm, 5 μm)分离,以乙腈和0.05 mol/L醋酸铵-醋酸缓冲液(pH 6.5)为流动相,梯度洗脱,在254 nm下检测,建立了同时测定15种氨基酸含量的方法,经方法学考察符合含量测定要求。谷氨酸、色氨酸在2~100 mg/L范围内、精氨酸在6~300 mg/L范围内、其余各氨基酸在0.8~40 mg/L范围内均呈良好的线性关系,相关系数均不小于0.9995;平均回收率在95%~105%之间,RSD均小于3%;并成功应用于12批中药材的测定。本方法简便、灵敏、准确,具有可操作性,可用于快速鉴定中药中的氨基类成分以及进行含量测定。

关键词: 氨基类成分, 超高效液相色谱-串联质谱法, 高效液相色谱, 天南星, 异硫氰酸苯酯, 柱前衍生化

Abstract:

A method for the identification and determination of the polar amino components without ultraviolet activity in traditional Chinese medicines was developed. With Rhizoma Arisaematis as the object of this study, using pre-column derivatization with phenyl isothiocyanate (PITC) as the derivatization reagent, compounds were separated and identified on a C18 column (100 mm×2.1 mm, 3.5 μm) by ultra high performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS). A total of 20 components, including 18 amino acids and 2 amine compounds were identified. Furthermore, after the optimization of the derivatization conditions, 15 amino acids were determined by high performance liquid chromatography (HPLC) on Diamonsil C18 column (250 mm×4.6 mm, 5 μm), detected at 254 nm and gradiently eluted by acetonitrile and 0.05 mol/L ammonium acetate-acetic acid (pH 6.5) as the mobile phases. The results of methodological study demonstrated that the method can meet the requirements of the determination. All calibration curves expressed good linearity: Glu, Try in the range of 2-100 mg/L, Arg in the range of 6-300 mg/L, others in the range of 0.8-40 mg/L, with the correlation coefficients ≥ 0.9995. The average recovery of this method was among 95%-105% and the RSD was less than 3%. The developed method was successfully applied to quantitative determination of amino compounds in 12 batches of Rhizoma Arisaematis samples. The method is simple, sensitive, accurate, and can be used for rapid identification and determination of amino components in traditional Chinese medicines.

Key words: amino compounds, high performance liquid chromatography (HPLC), phenyl isothiocyanate (PITC), pre-column derivatization, Rhizoma Arisaematis, ultra high performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS)

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