Analytical and Bioanalytical Chemistry

, Volume 411, Issue 17, pp 3857–3870 | Cite as

A novel two-dimensional liquid chromatography system for the simultaneous determination of three monoterpene indole alkaloids in biological matrices

  • Sha-Sha Liu
  • Kun Yang
  • Zhi-Liang Sun
  • Xiaofeng Zheng
  • Xia Bai
  • Zhao-Ying LiuEmail author
Research Paper


The present paper describes a novel two-dimensional liquid chromatography (2D-LC) system, which is comprised of a first-dimensional ion exchange chromatography (IEX1) column, trap column, and second-dimensional reversed-phase chromatography (RP2) column system. The biological sample is separated by the first-dimensional LC using an IEX column to remove interferences. The analytes are transferred to the trap column after heart-cutting. Then, the analytes are transferred to the second-dimensional LC using an RP2 column for further separation and ultraviolet detection. This 2D-LC system can offer a large injection volume to provide sufficient sensitivity and exhibits a strong capacity for removing interferences. Here, the determination of three monoterpene indole alkaloids (MIAs; gelsemine, koumine, and humantenmine) from Gelsemium in biological matrices (plasma, tissue, and urine) was used this 2D-LC system. After a rapid and easy sample preparation method based on protein precipitation, the sample was injected into the 2D-LC. The method was developed and validated in terms of the selectivity, LOD, LOQ, linearity, precision, accuracy, and stability. The sample preparation time for the three MIAs was 15 min. The LOD for these compounds was 10 ng/mL, which was lower than the developed HPLC methods. The results showed that this method had good quantitation performance and allowed the determination of gelsemine, koumine, and humantenmine in biological matrices. The method is rapid, exhibits high selectivity, has good sensitivity, and is low-cost, thus making it well-suited for application in the pharmaceutical and toxicological analysis of Gelsemium.

Graphical abstract


Two-dimensional liquid chromatography Gelsemium Indole alkaloid Gelsemine Koumine Humantenmine 



We thank the American Journal Expert for the language help.


This study was funded by Hunan Provincial Natural Science Foundation of China (Grant Number 2017JJ1017) and Scientific Research Fund of Hunan Provincial Education Department (Grant Number 18A088).

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interest.


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Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature 2019

Authors and Affiliations

  • Sha-Sha Liu
    • 1
    • 2
  • Kun Yang
    • 1
    • 2
  • Zhi-Liang Sun
    • 1
    • 2
  • Xiaofeng Zheng
    • 1
    • 2
  • Xia Bai
    • 1
    • 2
  • Zhao-Ying Liu
    • 1
    • 2
    Email author
  1. 1.Hunan Engineering Technology Research Center of Veterinary DrugsHunan Agricultural UniversityChangshaChina
  2. 2.College of Veterinary MedicineHunan Agricultural UniversityChangshaChina

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