Fan Baolei, Xu Sheng, Bi Jianli, Huang Shengtang, Zu Zengyi, Qian Chunqi
Hubei University of Science and Technology, 2 Yong'an Ave, Xian'an District, Xianning, Hubei 430081, China.
Michigan State University, 846 Service Road, East Lansing, Michigan 48864, United States.
ACS Pharmacol Transl Sci. 2020 Dec 6;4(1):118-127. doi: 10.1021/acsptsci.0c00133. eCollection 2021 Feb 12.
Simultaneous determination of the content of six alkaloids (aconitine, hypoaconitine, mesaconitine, benzoylaconine, benzoylhypaconine, and benzoylmesaconine) in rat plasma is enabled by HPLC-MS/MS combined with microsolid phase extraction (micro-SPE). To study its pharmacokinetics in rat plasma, the extracted plasma sample was passed through a C18 extraction column and eluted with acetonitrile. The six alkaloids in the extract can be completely separated as peaks with good shape. The six components in the plasma sample showed a good linear relationship within their respective linear ranges ( > 0.997). The analysis of the six alkaloids can be completed within 20 min. This method has high intraday and interday precision, and the room temperature stability and freeze-thaw stability are good. The matrix effect of the plasma samples is between 86.4 and 114%. The metabolism of the six alkaloids in plasma is analyzed using a two-compartment model, which is characterized by fast absorption, slow elimination, and good linear fit, > 0.99. The peak time ( ) for aconitine, hypaconitine, and neoaconitine ranged from 29.95 to 42.07 min, while the peak time ( ) for benzoaconitine, benzohypaconitine, and benzoxinaconitine ranged from 42.88 to 73.08 min. With the increased dosage, the bioavailability of alkaloids decreased gradually. The method for the determination of alkaloids in rat plasma by high performance liquid chromatography-tandem mass spectrometry is sensitive and accurate, which is suitable for rat plasma analysis. The results provide a scientific basis for metabolic study of alkaloids , and pave the way for clinical use of medicinal materials and extracts.
高效液相色谱-串联质谱联用结合微固相萃取（micro-SPE）可同时测定大鼠血浆中六种生物碱（乌头碱、次乌头碱、中乌头碱、苯甲酰乌头碱、苯甲酰次乌头碱和苯甲酰中乌头碱）的含量。为研究其在大鼠血浆中的药代动力学，将提取的血浆样品通过C18萃取柱并用乙腈洗脱。提取物中的六种生物碱可作为峰完全分离，峰形良好。血浆样品中的六种成分在各自线性范围内呈现良好的线性关系（>0.997）。六种生物碱的分析可在20分钟内完成。该方法具有较高的日内和日间精密度，室温稳定性和冻融稳定性良好。血浆样品的基质效应在86.4%至114%之间。采用二室模型分析血浆中六种生物碱的代谢，其特点是吸收快、消除慢、线性拟合良好（>0.99）。乌头碱、次乌头碱和新乌头碱的峰时间（）为29.95至42.07分钟，而苯甲酰乌头碱、苯甲酰次乌头碱和苯甲酰新乌头碱的峰时间（）为42.88至73.08分钟。随着剂量增加，生物碱的生物利用度逐渐降低。高效液相色谱-串联质谱法测定大鼠血浆中生物碱的方法灵敏、准确，适用于大鼠血浆分析。研究结果为生物碱的代谢研究提供了科学依据，为药材及提取物的临床应用奠定了基础。
