Page 29 - 理化检验-化学分册2025年第三期

淳泽利，等：超高效液相色谱-串联质谱法同时测定雪上一支蒿中7种生物碱类化合物的含量

Simultaneous Determination of 7 Alkaloid Compounds in Aconitum Brachypodum
Diels by Ultra-High Performance Liquid Chromatography-Tandem Mass
Spectrometry

CHUN Zeli 1,2,3 , XIAO Xin 1,2,3 , ZHANG Yong 1,2,3 , ZHOU Ming , LIU Yun , BAO Yuxin 1,2,3*
2,3
2,3
(1. Analytical and Testing Center, Zunyi Medical University, Zunyi 563000, China; 2. School of Forensic Medicine,
Zunyi Medical University, Zunyi 563000, China; 3. Center of Forensic Expertise, the Affiliated Hospital of
Zunyi Medical University, Zunyi 563000, China)
Abstract: The treated Aconitum brachypodum Diels powder sample (0. 1 g) was taken, and extracted ultrasonically in 5 mL
of 80% (volume fraction) methanol solution at 25 ℃ for 20 min. The supernatant was passed through a 0. 22 μm filter membrane to
obtain the test solution Ⅰ, which was used for determination of mesaconitine, hypaconitine, benzoylhypaconine, and benzoylmesaconine.
An appropriate amount of the test solution Ⅰ was taken and diluted 100 times with 50% (volume fraction, the same below) methanol
solution to obtain the test solution Ⅱ, which was used for determination of bullatine B. An appropriate amount of the test solution Ⅱ was
taken and diluted 10 times with 50% methanol solution to obtain the test solution Ⅲ, which was used for determination of aconitine
and benzoylaconine. Acquity UPLC BEH C 18 column was used as the stationary phase, and a mixture of 0. 1% (volume fraction)
formic acid solution and acetonitrile at different volume ratios was used as the mobile phase for gradient elution. The multiple
reaction monitoring mode was carried out by the electrospray ion source under positive ion mode in mass spectrometry analysis. As
shown by the results, linear relationships between the corresponding peak areas and mass concentrations of 7 alkaloid compounds
were found in the range of 1. 0-100. 0 μg · L −1 , with detection limits (3S/N) of 0. 002 1-0. 019 5 μg · L −1 . RSDs (n=6) of the
determined values in the instrument precision, repeatability, and stability tests were less than 5. 0%. Test for recovery was made
by the standard addition method, giving results in the range of 94. 5%-105%. This method was used for analysis of 12 batches of
Aconitum brachypodum Diels samples. The contents of aconitine, benzoylaconine, and bullatine B were higher, and the contents
of the other four alkaloid compounds were relatively lower.
Keywords: Aconitum brachypodum Diels; ultra-high performance liquid chromatography-tandem mass spectrometry
(UHPLC-MS/MS) ; alkaloid compound

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