UPLC-MS/MS法同时测定止痒乳膏中10个成分的含量

Simultaneous determination of ten compounds in Zhiyang cream by UPLC-MS/MS method

目的:建立超高效液相色谱-串联质谱法(UPLC-MS/MS)同时测定止痒乳膏中10个主要成分(氧化苦参碱、黄柏碱、芍药苷、升麻素苷、药根碱、小檗碱、巴马汀、大黄素、大黄酚、大黄素甲醚)的含量。方法:采用ZORBAX Eclipse Plus C18色谱柱(2.1 mm×50 mm,1.8μm),以甲醇(A)-0.1%甲酸水溶液(B)为流动相,梯度洗脱(0~4 min,5%A→28%A;4~10 min,28%A→30%A;10~11 min,30%A→73%A;11~16 min,73%A→75%A),流速0.4 m L·min^(-1),柱温30℃;采用电喷雾离子源(ESI),多反应监测模式(MRM),正负离子同时检测。结果:氧化苦参碱、黄柏碱、芍药苷、升麻素苷、药根碱、小檗碱、巴马汀、大黄素、大黄酚、大黄素甲醚的线性范围分别为4.526~452.6μg·m L^(-1)(r=0.999 2)、1.625~32.50μg·m L^(-1)(r=0.999 0)、7.500~150.0μg·m L^(-1)(r=0.998 7)、3.626~181.3μg·m L^(-1)(r=0.998 9)、0.155 0~3.100μg·m L^(-1)(r=0.999 3)、3.376~168.8μg·m L^(-1)(r=0.996 4)、0.015 50~15.50μg·m L^(-1)(r=0.999 6)、0.122 5~24.50μg·m L^(-1)(r=0.999 8)、0.069 00~1.380μg·m L^(-1)(r=0.996 8)和0.016 30~0.815 0μg·m L^(-1)(r=0.998 9);平均加样回收率(n=6)均在96.53%~100.2%范围内,RSD均小于2.04%。3批样品中氧化苦参碱、黄柏碱、芍药苷、升麻素苷、药根碱、小檗碱、巴马汀、大黄素、大黄酚和大黄素甲醚含量测定结果分别为2.98~3.17、0.24~0.25、1.69~1.75、0.19、0.003 6~0.004 3、0.30~0.31、0.004 0~0.004 3、0.023~0.026、0.018~0.020和0.000 9~0.001 1 mg·g^(-1)。结论:该测定方法测定结果可为止痒乳膏的质量控制提供依据。

Objective： To establish a UPLC-MS/MS method for simultaneous determination of ten components（oxymatrine,phellodendrine,paeoniflorin, prim-O-glucosylcimifugin, jatrorrhizine,berberine,palmatine,emodin,chrysophanol,physcion） in Zhiyang cream.Methods：The chromatographic separation was performed on an Aglient ZORBAX Eclipse Plus C18 column（2.1 mm×50 mm, 1.8 μm）, with a mobile phase consisting of methanol（A）-0.1% formic acid in water（B） at the flow rate of 0.4 m L·min-（-1）（0-4 min, 5%A → 28%A; 4-（-1）0 min, 28%A → 30%A; 10-（-1）1 min, 30%A → 73%A; 11-（-1）6 min, 73%A → 75%A）.The column temperature was 30 ℃.Electrospray ionization source（ESI）, and multiple reaction monitoring（MRM）, combined with positiveand negative scanning switch were adopted.Results：The method has a good linearity in the range of 4.526-452.6 μg·m L-（-1）（r=0.999 2） for oxymatrine, 1.625-32.50μg·m L-（-1）（r=0.999 0） for phellodendrine, 7.500-（-1）50.0μg·m L-（-1）（r=0.998 7） for paeoniflorin,3.626-（-1）81.3 μg·m L-（-1）（r=0.998 9） for prim-O-glucosylcimifugin,0.155 0-3.100 μg·m L-（-1）（r=0.999 3） for jatrorrhizine,3.376-（-1）68.8 μg·m L-（-1）（r=0.996 4） for berberine,0.015 50-（-1）5.50 μg·m L-（-1）（r=0.999 6） for palmatine,0.122 5-24.50 μg·m L-（-1）（r=0.999 8） for emodin,0.069 00-（-1）.380 μg·m L-（-1）（r=0.996 8） for chrysophanol,0.016 30-0.815 0μg·m L-（-1）（r=0.998 9） for physcion.The average recoveries of ten active components were 96.53%-（-1）00.2% with RSDs（n=6） of less than 2.04%.The contents of oxymatrine, phellodendrine, paeoniflorin, prim-O-glucosylcimifugin, jatrorrhizine, berberine, palmatine, emodin, chrysophanol and physcion in 3 batches of samples were 2.98-3.17,0.24-0.25,1.69-（-1）.75,0.19,0.003 6-0.004 3,0.30-0.31,0.004 0-0.004 3,0.023-0.026,0.018-0.020 and 0.000 9-0.001 1 mg·g-（-1）, respectively.Conclusion：The determination results of the established method may provide a basis for the quality control of Zhiyang cream.