2. Academic Validation
  3. Development and in-house validation of a sensitive LC-MS/MS method for simultaneous quantification of gelsemine, koumine and humantenmine in porcine plasma

Development and in-house validation of a sensitive LC-MS/MS method for simultaneous quantification of gelsemine, koumine and humantenmine in porcine plasma

  • J Chromatogr B Analyt Technol Biomed Life Sci. 2018 Feb 15;1076:54-60. doi: 10.1016/j.jchromb.2018.01.019.
Kun Yang 1,  Xue-Ming Long 2,  Yan-Chun Liu 1,  Fu-Hua Chen 2,  Xiao-Feng Liu 1,  Zhi-Liang Sun 3,  Zhao-Ying Liu 4
Affiliations

Affiliations

  • 1 Hunan Engineering Research Center of Veterinary Drug, College of Veterinary Medicine, Hunan Agricultural University, Changsha 410128, China.
  • 2 Institute of Hunan Province Veterinary Drugs and Feed Supervision, Changsha 410006, China.
  • 3 Hunan Engineering Research Center of Veterinary Drug, College of Veterinary Medicine, Hunan Agricultural University, Changsha 410128, China; Hunan Co-Innovation Center for Utilization of Botanical Functional Ingredients, College of Veterinary Medicine, Hunan Agricultural University, Changsha 410128, China. Electronic address: sunzhiliang1965@aliyun.com.
  • 4 Hunan Engineering Research Center of Veterinary Drug, College of Veterinary Medicine, Hunan Agricultural University, Changsha 410128, China; Hunan Co-Innovation Center for Utilization of Botanical Functional Ingredients, College of Veterinary Medicine, Hunan Agricultural University, Changsha 410128, China; National and Local Union Engineering Research Center for the Veterinary Herbal Medicine Resources and Initiative, Hunan Agricultural University, Changsha 410128, China. Electronic address: liu_zhaoying@hunau.edu.cn.
PMID: 29406028 DOI: 10.1016/j.jchromb.2018.01.019
Abstract

Three monomers of G. elegans Indole Alkaloids (gelsemine, koumine and humantenmine) were simultaneously detected in porcine plasma for the first time with the development and validation of a sensitive and reliable LC-ESI-MS/MS method. Using a gradient mobile phase at a constant flow rate of 0.2 mL/min via electrospray ionization (positive ion mode) in a multiple reaction monitoring (MRM) scan, gelsemine, koumine and humantenmine were eluted, separated and detected at an appropriate retention time. The porcine plasma was prepared using protein precipitation with 1% formic acid-acetonitrile: methanol (2:1, v/v). Using matrix-matched calibration curves and weighted least squares linear regression, a good linearity (r2 > 0.99) was achieved with a concentration range of 0.1-200 μg/L for gelsemine, koumine and humantenmine; estimated LOD and LOQ values were 0.10 μg/L and 0.2 μg/L, respectively. The mean of the recoveries was in the range of 82.68-100.35% of porcine plasma at four different levels, and the intra-day and inter-day precision (CV) were lower than 15% with a range of 2.46-8.76% and 2.73-10.83%, respectively. The proposed method has proved to be suitable for accurate, quantitative determination of gelsemine, koumine and humantenmine in porcine plasma.

Keywords

Gelsemine; Gelsemium elegans; Humantenmine; Indole alkaloid; Koumine; Plasma.

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