HPLC-QTOF-MS法测定风湿骨痛胶囊中9个乌头类生物碱的含量

目的: 建立高效液相色谱四极杆飞行时间串联质谱法(HPLC-QTOF-MS法)测定风湿骨痛胶囊中乌头碱、次乌头碱、新乌头碱、苯甲酰乌头原碱、苯甲酰次乌头原碱、苯甲酰新乌头原碱、乌头原碱、次乌头原碱、新乌头原碱9个成分含量. 方法: 使用Agilent ZORBAX Extend-C₁₈ RRHT色谱柱(2.1 mm×50 mm,1.8 μm),以甲醇(A)-水(含0.1%甲酸和2.5 mmol·L^-1醋酸铵)(B)为流动相,梯度洗脱(0~3.5 min,15%A;3.5~4.5 min,15%A→40%A;4.5~20 min,40%A;20~20.1 min,40%A→15%A;20.1~25 min,15%A),流速0.21 mL·min^-1,柱温30℃;采用电喷雾离子(ESI)源,正离子方式检测,利用一级质谱的提取离子流色谱图进行定量,同时利用二级质谱进行成分的确认. 结果: 上述9种生物碱在一定范围内具有较好的线性关系,r值均大于0.999 7,精密度、重复性和稳定性的RSD均小于5%,加样回收率均在98.1%~101.8%之间.3批胶囊中以上 9个成分的含量测定结果分别是0.069、0.058、0.063 μg·粒^-1;1.091、1.015、0.908 μg·粒^-1;0.139、0.118、0.091 μg·粒^-1;28.481、25.446、30.492 μg·粒^-1;30.990、32.985、40.105 μg·粒^-1;87.188、67.171、93.819 μg·粒^-1;0.958、1.060、1.363 μg ·粒^-1;3.116、2.805、2.621 μg·粒^-1;1.892、1.594、1.708 μg·粒^-1. 结论: 本实验建立的方法经验证,可为控制该制剂的质量提供依据.

Objective: To simultaneously determine the contents of aconitine,hypaconitine,mesaconitine,benzoylaconine,benzoylhypaconine,benzoylmesaconine,aconine,hypaconine and mesaconine in Fengshi Gutong capsules by HPLC-QTOF-MS. Methods: The determination was performed on the Agilent ZORBAX Extend-C₁₈RRHT column (2.1 mm×50 mm,1.8 μm) with the mobile phase consisting of methanol(A)-water (containing 0.1% formic acid and 2.5 mmol·L^-1 ammonium acetate)(B) in gradient elution (0-3.5 min,15%A;3.5-4.5 min,15%A→40%A;4.5-20 min,40%A;20-20.1 min,40%A→15%A;20.1-25 min,15%A).The flow rate was 0.21 mL·min^-1.The column temperature was 30℃.MS instrument was equipped with ESI⁺ ion source.The exacted ion chromatograms of the first order mass spectrums were used to determine the quantities of different compounds in samples while the second order mass spectrums of product ions were used for confirming the compounds. Results: All the 9 alkaloids showed good linearity (r >0.999 7).The RSD of the precision,repeatability and stability tests were less than 5%.The average recoveries were in the range of 98.1%-101.8%.The contents of nine alkaloids in 3 batches of tested samples were 0.069,0.058,0.063;1.091,1.015,0.908;0.139 0,0.117 8,0.091;28.481,25.446,30.492;30.990,32.985,40.105;87.188,67.171,93.819;0.958,1.060,1.363;3.116,2.805,2.621;1.892,1.594,1.708 μg per capsule respectively. Conclusion: It is proved that the method can be used for quality control of Fengshi Gutong capsules.