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

作者：Yang, Kun; Long, Xue-Ming; Liu, Yan-Chun; Chen, Fu-Hua; Liu, Xiao-Feng; Sun, Zhi-Liang^*; Liu, Zhao-Ying^*

来源：Journal of Chromatography B-Analytical Technologies in the Biomedical and Life Sciences, 2018, 1076: 54-60.

DOI：10.1016/j.jchromb.2018.01.019

• Indole alkaloid
• Gelsemine
• Gelsemium elegans
• Plasma
• Koumine
• Humantenmine

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 (r(2) > 0.99) was achieved with a concentration range of 0.1-200 mu g/L for gelsemine, koumine and humantenmine; estimated LOD and LOQ values were 0.10 mu g/L and 0.2 mu g/L, respectively. The mean of the recoveries was in the range of 82.68-400.35% 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.

• 出版日期2018-2-15
• 单位湖南农业大学