高效液相色谱与QDA质谱联用同时测定附子理中丸中6个单酯及双酯型生物碱含量
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摘要
目的:建立同时测定附子理中丸中6个单酯及双酯型生物碱含量的方法。方法:应用高效液相色谱(HPLC)与QDA质谱联用同时测定附子理中丸中苯甲酰新乌头原碱、苯甲酰次乌头原碱、苯甲酰乌头原碱、新乌头碱、次乌头碱和乌头碱6个成分的含量。使用Phenomenex Luna C_(18)色谱柱(4.6 mm×250 mm,5μm),以乙腈-0.2%乙酸水(用三乙胺调p H 6.2)为流动相,梯度洗脱,流速1.0 m L·min~(-1),分流比为1∶4,使用ESI~+离子源,选择性离子扫描(SIM)模式下对苯甲酰新乌头原碱(m/z 590.0)、苯甲酰乌头原碱(m/z 604.0)、苯甲酰次乌头原碱(m/z 574.0)、新乌头碱(m/z 632.0)、次乌头碱(m/z 616.0)和乌头碱(m/z 646.0)进行测定。结果:苯甲酰新乌头原碱、苯甲酰乌头原碱、苯甲酰次乌头原碱、次乌头碱、新乌头碱和乌头碱6个成分的质量浓度分别在0.000 2~51.75μg·m L~(-1)(r=0.999 2)、0.000 2~50.95μg·m L~(-1)(r=0.999 5)、0.000 2~50.25μg·m L~(-1)(r=0.999 8)、0.000 2~49.80μg·m L~(-1)(r=0.999 9)、0.000 2~50.60μg·m L~(-1)(r=0.999 5)和0.000 2~52.00μg·m L~(-1)(r=0.999 7)范围内线性关系良好;平均回收率(n=6)分别为101.8%、101.3%、98.2%、104.8%、99.9%和89.9%,RSD分别为6.4%、2.6%、2.7%、6.1%、3.8%和5.7%。3批附子理中丸中6个生物碱成分即苯甲酰新乌头原碱、苯甲酰次乌头原碱、苯甲酰乌头原碱、新乌头碱、次乌头碱和乌头碱的含量范围分别为7.095~7.366、1.007~1.214、0.594~0.742、1.261~1.438、1.492~1.578和0.135~0.163μg·g~(-1)。结论:该方法可测定附子理中丸中6个生物碱成分含量,为附子理中丸的质量标准提高打下了基础。
Objective:To establish the quantitative method of six diterpenoid alkaloids in Fuzilizhong pills.Methods:The contents of benzoylmesaconitine,benzoylaconitine,benzoylhypaconitine,mesaconitine,aconitine,and hypaconitine in Fuzilizhong pills were determined by HPLC coupled with QDA mass detector.The chromatographic column was Phenomenex Luna C_(18)(4.6 mm×250 mm,5μm).The mobile phase was the mixture of acetonitrile-0.2%acetic acid(adjust p H to 6.2 with triethylamine)in gradient mode,the flow rate was1.0 m L·min~(-1),and the split ratio was 1∶4.The ESI~+source of the mass spectrometer was used in selected ion monitoring(SIM)mode for the detection of benzoylmesaconitine(m/z 590.0),benzoylaconitine(m/z 604.0),benzoylhypaconitine(m/z 574.0),mesaconitine(m/z 632.0),aconitine(m/z 646.0)and hypaconitine(m/z616.0).Results:The linear ranges of benzoylmesaconitine,benzoylaconitine,benzoylhypaconitine,hypaconitine,mesaconitine,and aconitine were 0.000 2-51.75μg·m L~(-1)(r=0.999 2),0.000 2-50.95μg·m L~(-1)(r=0.999 5),0.000 2-50.25μg·m L~(-1)(r=0.999 8),0.000 2-49.80μg·m L~(-1)(r=0.999 9),0.000 2-50.60μg·m L~(-1)(r=0.999 5)and 0.000 2-52.00μg·m L~(-1)(r=0.999 7),respectively.The average recovery rates(n=6)of the above compounds were 101.8%,101.3%,98.2%,104.8%,99.9%and 89.9%with RSD of 6.4%,2.6%,2.7%,6.1%,3.8%and5.7%,respectively.The contents of benzoylmesaconitine,benzoylhypaconitine,benzoylaconitine,mesaconitine,aconitine and hypaconitine were 7.095-7.366,1.007-1.214,0.594-0.742,1.261-1.438,1.492-1.578 and 0.135-0.163μg·g~(-1),respectively.Conclusion:The method can be used to determine the content of six diterpenoid alkaloids in Fuzilizhong pills.The present study provides a scientific basis for the quality control of Fuzilizhong pills.
Objective:To establish the quantitative method of six diterpenoid alkaloids in Fuzilizhong pills.Methods:The contents of benzoylmesaconitine,benzoylaconitine,benzoylhypaconitine,mesaconitine,aconitine,and hypaconitine in Fuzilizhong pills were determined by HPLC coupled with QDA mass detector.The chromatographic column was Phenomenex Luna C_(18)(4.6 mm×250 mm,5μm).The mobile phase was the mixture of acetonitrile-0.2%acetic acid(adjust p H to 6.2 with triethylamine)in gradient mode,the flow rate was1.0 m L·min~(-1),and the split ratio was 1∶4.The ESI~+source of the mass spectrometer was used in selected ion monitoring(SIM)mode for the detection of benzoylmesaconitine(m/z 590.0),benzoylaconitine(m/z 604.0),benzoylhypaconitine(m/z 574.0),mesaconitine(m/z 632.0),aconitine(m/z 646.0)and hypaconitine(m/z616.0).Results:The linear ranges of benzoylmesaconitine,benzoylaconitine,benzoylhypaconitine,hypaconitine,mesaconitine,and aconitine were 0.000 2-51.75μg·m L~(-1)(r=0.999 2),0.000 2-50.95μg·m L~(-1)(r=0.999 5),0.000 2-50.25μg·m L~(-1)(r=0.999 8),0.000 2-49.80μg·m L~(-1)(r=0.999 9),0.000 2-50.60μg·m L~(-1)(r=0.999 5)and 0.000 2-52.00μg·m L~(-1)(r=0.999 7),respectively.The average recovery rates(n=6)of the above compounds were 101.8%,101.3%,98.2%,104.8%,99.9%and 89.9%with RSD of 6.4%,2.6%,2.7%,6.1%,3.8%and5.7%,respectively.The contents of benzoylmesaconitine,benzoylhypaconitine,benzoylaconitine,mesaconitine,aconitine and hypaconitine were 7.095-7.366,1.007-1.214,0.594-0.742,1.261-1.438,1.492-1.578 and 0.135-0.163μg·g~(-1),respectively.Conclusion:The method can be used to determine the content of six diterpenoid alkaloids in Fuzilizhong pills.The present study provides a scientific basis for the quality control of Fuzilizhong pills.
引文
[1]中华人民共和国药典2015年版.一部[S].2015:1013Ch P 2015.VolⅠ[S].2015:1013
[2]马光明.附子理中丸治疗特发性水肿[J].医药论坛杂志,2007,28(3):104MA GM.Treatment of idiopathic edema with Fuzi Lizhong pills[J].J Med Forum,2007,28(3):104
[3]张向力,张丽萍,王晓霞.附子理中丸中毒致心律失常一例[J].中国中医药信息杂志,1996,3(4):37ZHANG XL,ZHANG LP,WANG XX.One case of arrhythmia caused by Fuzi Lizhong pills[J].Chin J Inf Tradit Chin Med,1996,3(4):37
[4]王建凯,刘国良.附子理中丸致中毒反应2例报道[J].实用中医药杂志,2001,17(1):43WANG JK,LIU GL.Report of two cases about the toxic reaction of Fuzi Lizhong pills[J].J Pract Tradit Chin Med,2001,17(1):43
[5]张庆辉.附子理中丸中毒致心律失常1例[J].临床荟萃,2010,25(23):2082ZHANG QH.A case report about the arrhythmia caused by Fuzi Lizhong pills[J].Clin Focus,2010,25(23):2082
[6]聂黎行,张聿梅,鲁静,等.附子和附片质量标准研究[J].中国药学杂志,2010,45(15):1182NIE LX,ZHANG YM,LU J,et al.Study on quality standard of Fuzi and Fupian[J].Chin Pharm J,2010,45(15):1182
[7]刘小鸣,张振凌,梁君,等.《国家中成药标准汇编》中含乌头碱类中药制剂质量标准的分析[J].中国实验方剂学杂志,2015,21(8):230LIU XM,ZHANG ZL,LIANG J,et al.Analysis quality standards of Chinese drug preparations that contain aconitine in Compilation of National Standard for Traditional Chinese Medicines[J].Chin J Exp Tradit Med Form,2015,21(8):230
[8]王勇,邓晓春,夏博.三种中成药中乌头生物碱飞行时间质谱分析[J].深圳大学学报(理工版),2009,26(3):257WANG Y,DENG XC,XIA B.Analysis of Aconitum alkaloids in three kinds of proprietary Chinese medicines by MALDITOF mass spectrometry[J].J Shenzhen Univ Sci Eng,2009,26(3):257
[9]周超,张聿梅,何轶,等.HPLC-MS法同时测定益母草及其颗粒剂中盐酸水苏碱与盐酸益母草碱的含量[J].药物分析杂志,2016,36(5):830ZHOU C,ZHANG YM,HE Y,et al.Simultaneous content determination of stachydrine hydrochloride and leonurine hydrochloride in Herba Leonuri and Yimucao granules by HPLC-MS[J].Chin J Pharm Anal,2016,36(5):830
[10]朱瑞龙,景明,柴国林,等.HPLC测定附子理中丸中双酯型乌头碱含量[J].中成药,1996,18(10):13ZHU RL,JING M,CHAI GL,et al.Determination of diester aconitine in Fuzi Lizhong pills by HPLC[J].Chin Tradit Pat Med,1996,18(10):13
[11]钟碧华,陆帅.HPLC法测定附子理中丸中乌头碱的含量[J].海峡药学,2014,26(11):115ZHONG BH,LU S.Determination of aconitine in Fuzi Lizhong pills by HPLC[J].Strait Pharm J,2014,26(11):115
[12]邵峰,俞瑜,任刚,等.HPLC同时测定附子理中丸中3种单酯型生物碱含量[J].中国实验方剂学杂志,2012,18(16):58SHAO F,YU Y,REN G,et al.Simultaneous determination of benzoylmesaconitine,benzoylaconitine and benzoylhypaconitine in Fuzi Lizhong pills by HPLC[J].Chin J Exp Tradit Med Form,2012,18(16):58
[13]索志荣,徐敏,秦海燕,等.LC-MS测定附子理中丸中3种双酯型生物碱含量[J].药物分析杂志,2010,30(12):2279SUO ZR,XU M,QIN HY,et al.LC-MS determination of three diester diterpenoid alkaloids in Fuzi Lizhong pills[J].Chin J Pharm Anal,2010,30(12):2279
[14]刘岚,范智超,张志琪.HPLC同时测定成药中4种乌头类生物碱含量[J].药物分析杂志,2010,30(2):236LIU L,FAN ZC,ZHANG ZQ.HPLC simultaneous determination of four aconitine alkaloids in patent medicine[J].Chin J Pharm Anal,2010,30(2):236
[15]张盼盼,魏文跃,李岑,等.附子理中丸中乌头碱类毒性成分的富集及检测[J].华西药学杂志,2012,27(5):563ZHANG PP,WEI WY,LI C,et al.Enrichment and detection of aconitine-type toxic composition in Fuzi Lizhong pills[J].West China J Pharm Sci,2012,27(5):563
[2]马光明.附子理中丸治疗特发性水肿[J].医药论坛杂志,2007,28(3):104MA GM.Treatment of idiopathic edema with Fuzi Lizhong pills[J].J Med Forum,2007,28(3):104
[3]张向力,张丽萍,王晓霞.附子理中丸中毒致心律失常一例[J].中国中医药信息杂志,1996,3(4):37ZHANG XL,ZHANG LP,WANG XX.One case of arrhythmia caused by Fuzi Lizhong pills[J].Chin J Inf Tradit Chin Med,1996,3(4):37
[4]王建凯,刘国良.附子理中丸致中毒反应2例报道[J].实用中医药杂志,2001,17(1):43WANG JK,LIU GL.Report of two cases about the toxic reaction of Fuzi Lizhong pills[J].J Pract Tradit Chin Med,2001,17(1):43
[5]张庆辉.附子理中丸中毒致心律失常1例[J].临床荟萃,2010,25(23):2082ZHANG QH.A case report about the arrhythmia caused by Fuzi Lizhong pills[J].Clin Focus,2010,25(23):2082
[6]聂黎行,张聿梅,鲁静,等.附子和附片质量标准研究[J].中国药学杂志,2010,45(15):1182NIE LX,ZHANG YM,LU J,et al.Study on quality standard of Fuzi and Fupian[J].Chin Pharm J,2010,45(15):1182
[7]刘小鸣,张振凌,梁君,等.《国家中成药标准汇编》中含乌头碱类中药制剂质量标准的分析[J].中国实验方剂学杂志,2015,21(8):230LIU XM,ZHANG ZL,LIANG J,et al.Analysis quality standards of Chinese drug preparations that contain aconitine in Compilation of National Standard for Traditional Chinese Medicines[J].Chin J Exp Tradit Med Form,2015,21(8):230
[8]王勇,邓晓春,夏博.三种中成药中乌头生物碱飞行时间质谱分析[J].深圳大学学报(理工版),2009,26(3):257WANG Y,DENG XC,XIA B.Analysis of Aconitum alkaloids in three kinds of proprietary Chinese medicines by MALDITOF mass spectrometry[J].J Shenzhen Univ Sci Eng,2009,26(3):257
[9]周超,张聿梅,何轶,等.HPLC-MS法同时测定益母草及其颗粒剂中盐酸水苏碱与盐酸益母草碱的含量[J].药物分析杂志,2016,36(5):830ZHOU C,ZHANG YM,HE Y,et al.Simultaneous content determination of stachydrine hydrochloride and leonurine hydrochloride in Herba Leonuri and Yimucao granules by HPLC-MS[J].Chin J Pharm Anal,2016,36(5):830
[10]朱瑞龙,景明,柴国林,等.HPLC测定附子理中丸中双酯型乌头碱含量[J].中成药,1996,18(10):13ZHU RL,JING M,CHAI GL,et al.Determination of diester aconitine in Fuzi Lizhong pills by HPLC[J].Chin Tradit Pat Med,1996,18(10):13
[11]钟碧华,陆帅.HPLC法测定附子理中丸中乌头碱的含量[J].海峡药学,2014,26(11):115ZHONG BH,LU S.Determination of aconitine in Fuzi Lizhong pills by HPLC[J].Strait Pharm J,2014,26(11):115
[12]邵峰,俞瑜,任刚,等.HPLC同时测定附子理中丸中3种单酯型生物碱含量[J].中国实验方剂学杂志,2012,18(16):58SHAO F,YU Y,REN G,et al.Simultaneous determination of benzoylmesaconitine,benzoylaconitine and benzoylhypaconitine in Fuzi Lizhong pills by HPLC[J].Chin J Exp Tradit Med Form,2012,18(16):58
[13]索志荣,徐敏,秦海燕,等.LC-MS测定附子理中丸中3种双酯型生物碱含量[J].药物分析杂志,2010,30(12):2279SUO ZR,XU M,QIN HY,et al.LC-MS determination of three diester diterpenoid alkaloids in Fuzi Lizhong pills[J].Chin J Pharm Anal,2010,30(12):2279
[14]刘岚,范智超,张志琪.HPLC同时测定成药中4种乌头类生物碱含量[J].药物分析杂志,2010,30(2):236LIU L,FAN ZC,ZHANG ZQ.HPLC simultaneous determination of four aconitine alkaloids in patent medicine[J].Chin J Pharm Anal,2010,30(2):236
[15]张盼盼,魏文跃,李岑,等.附子理中丸中乌头碱类毒性成分的富集及检测[J].华西药学杂志,2012,27(5):563ZHANG PP,WEI WY,LI C,et al.Enrichment and detection of aconitine-type toxic composition in Fuzi Lizhong pills[J].West China J Pharm Sci,2012,27(5):563