附子煎煮过程中13种生物碱含量的动态变化规律研究

目的: 采用超高效液相色谱-三重四极杆质谱,精确测定附子水煎液中酯型生物碱、原碱、去甲猪毛菜碱等成分的含量,探讨附子煎煮过程中13种生物碱的动态变化规律,为临床应用和炮制提供参考. 方法: 采用Agilent SB-C₁₈色谱柱(2.1 mm×20 mm,1.8 μm),以2 mmol·L^-1醋酸铵溶液-乙腈为流动相, 梯度洗脱,流速0.4 mL·min^-1,柱温35 ℃;质谱采用ESI⁺离子源,多反应监测模式(MRM),检测13种生物碱(乌头碱、新乌头碱、次乌头碱、苯甲酰乌头原碱、苯甲酰新乌头原碱、苯甲酰次乌头原碱、乌头原碱、新乌头原碱、次乌头原碱、附子灵、多根乌头碱、宋果灵、去甲猪毛菜碱)以及内标氢溴酸高乌甲素的定量离子和定性离子. 结果: 生附片煎煮过程中双酯型生物碱(乌头碱、新乌头碱、次乌头碱)含量迅速降低,4 h后趋于稳定且含量很低;单酯型生物碱(苯甲酰乌头原碱、苯甲酰新乌头原碱、苯甲酰次乌头原碱)含量先升后降,于4~6 h达到高峰;原碱中乌头原碱、新乌头原碱、次乌头原碱含量一直快速增加,而附子灵、宋果灵、多根乌头碱和去甲猪毛菜碱含量缓慢增加,约在4 h达到最高峰,后趋于稳定或略有降低.黑顺片煎煮过程中生物碱成分的动态变化趋势与生附片相似,但变化幅度显著小于生附片. 结论: 生附片煎煮2~4 h,双酯型生物碱含量已很低,能保证临床用药安全,且总生物碱等主要成分的含量显著高于黑顺片水煎液.

Objective: To employ UPLC/QQQ-MS-MS for the determination of components such as ester alkaloids,aconine alkaloids,salsolinol and so on in decoction of Aconiti Lateralis Radix Praeparata(Fuzi),and investigate content changes of 13 alkaloids during the decocting process of Fuzi,so as to provide a valuable reference for clinical application and processing. Methods: The determination was performed on an Agilent SB-C₁₈ column(2.1 mm×20 mm,1.8 μm)with the mobile phase of 2 mmol·L^-1 ammonium acetate solution-acetonitrile(gradient elution)at the flow rate of 0.4 mL·min^-1,and the column temperature was 35 ℃.MS equipped with ESI⁺ ion source and MRM mode were employed to detect qualitative and quantitation ions of the 13 alkaloids,including aconitine,mesaconitine,hypaconitine,benzoylaconine,benzoylmesaconine,benzoylhypaconine,aconine,mesaconine,hypaconine,fuziline,karacoline,songorine,salsolinol,and the internal standard lannaconitine. Results: As for crude Fuzi,contents of diester-diterpenoid alkaloids(aconitine,mesaconitine,hypaconitine)decreased rapidly,and maintained at low level after 4 h decocting.The contents of monoester-diterpenoid alkaloids(benzoylaconine,benzoylmesaconine,benzoylhypaconine)firstly increased and reached the peak in 4-6 h and then gradually decreased.The contents of aconine alkaloids(mesaconine,aconine and hypaconine)increased throughout the decoction process.The contents of fuziline,songorine,karacoline and salsolinol firstly increased and reached to the peak at about 4 h and then tended to be stable or reduced slightly.Trends of the 13 alkaloids were almost the same in crude Fuzi compared to Heishunpian,but the amplitude of changes in Heishunpian was observably smaller than that in crude Fuzi. Conclusion: After 2-4 h decocting,the contents of diester-diterpenoid alkaloids in crude Fuzi were at very low level which could ensure clinical safety and the contents of total alkaloids were significantly higher than that in Heishunpian decotion.