LC-MS/MS同时测定复方救必应胶囊中3种同分异构体的含量
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摘要
目的建立液相色谱-串联质谱(LC-MS/MS)分析方法,以同时分离测定复方救必应胶囊中3种互为同分异构体的有效成分(α-香附酮、香附烯酮、圆柚酮),并对随机抽样的3批复方救必应胶囊进行质量分数比较。方法采用Simpsil C18色谱柱(250 mm×4.6 mm,5μm),以乙腈-0.1%甲酸水溶液(体积比95∶5)为流动相,25 min内进行等度洗脱。流速为0.3 mL/min,柱温为室温,进样量为20.0μL。选用电喷雾离子源(ESI),正离子模式下多反应监测扫描(MRM),优化相关质谱参数,对3种有效成分进行检测和定量分析。结果α-香附酮、香附烯酮在质量浓度0.45～200μg/L范围内线性关系良好(r≥0.996 6);圆柚酮在质量浓度3～1 500μg/L范围内线性关系良好(r=0.996 0)。α-香附酮、香附烯酮、圆柚酮的平均加样回收率皆在95%～105%,RSD<3.5%(n=6)。3批复方救必应胶囊中α-香附酮、香附烯酮、圆柚酮的平均质量分数分别为437.17、847.39、42.92μg/g,RSD<15%。结论本方法准确可靠、结果稳定、重复性良好、灵敏度高,可用于复方救必应胶囊中3种同分异构体有效成分的质量分数测定。
Objective To establish an LC-MS method for the simultaneous determination of the three effective components(α-cyperone,cyperotundone and nootkatone) in compound Jiubiying capsules,and to compare the mass fractions of three batches of compound Jiubiying capsules sampled randomly.Methods A Simpsil C18 column(250 mm×4.6 mm,5 μm) was used with a mobile phase containing acetonitrile and0.5% formic acid water(95∶5),the separation was carried out in isocratic mode within 25 minutes.The flow rate was 0.3 mL/min,the column temperature was room temperature,and the injection volume was20.0 μL.The electrospray ionization source(ESI) was used to perform multi-reaction monitoring scanning(MRM) in positive ion mode,and the relevant mass spectrometry parameters were optimized to detect and quantify the three active components.Results α-cyperone and cyperotundone had a good linear relationship(r≥0.996 6) in the concentration range of 0.45-200 μg/L,and the linear relationship of nootkatone was good(r=0.996 0) in the concentration range of 3-1 500 μg/L.The average recovery(n=6) of α-cyperone,cyperotundone and nootkatone was between 95% and 105%,RSD<3.5%.The average content ofα-cyperone,cyperotundone,and nootkatone in three batches of compound Jiubiying capsules was 437.17,847.39,42.92 μg/g,RSD<15%.Conclusion The method is accurate,reliable,stable,reproducible and sensitive,which can be used to determine the content of three isomers in compound Jiubiying capsules.
Objective To establish an LC-MS method for the simultaneous determination of the three effective components(α-cyperone,cyperotundone and nootkatone) in compound Jiubiying capsules,and to compare the mass fractions of three batches of compound Jiubiying capsules sampled randomly.Methods A Simpsil C18 column(250 mm×4.6 mm,5 μm) was used with a mobile phase containing acetonitrile and0.5% formic acid water(95∶5),the separation was carried out in isocratic mode within 25 minutes.The flow rate was 0.3 mL/min,the column temperature was room temperature,and the injection volume was20.0 μL.The electrospray ionization source(ESI) was used to perform multi-reaction monitoring scanning(MRM) in positive ion mode,and the relevant mass spectrometry parameters were optimized to detect and quantify the three active components.Results α-cyperone and cyperotundone had a good linear relationship(r≥0.996 6) in the concentration range of 0.45-200 μg/L,and the linear relationship of nootkatone was good(r=0.996 0) in the concentration range of 3-1 500 μg/L.The average recovery(n=6) of α-cyperone,cyperotundone and nootkatone was between 95% and 105%,RSD<3.5%.The average content ofα-cyperone,cyperotundone,and nootkatone in three batches of compound Jiubiying capsules was 437.17,847.39,42.92 μg/g,RSD<15%.Conclusion The method is accurate,reliable,stable,reproducible and sensitive,which can be used to determine the content of three isomers in compound Jiubiying capsules.
引文
[1]国家药典委员会.卫生部药品标准中药成方制剂:第10册(WS3-B-1986-95)[S].北京:化学工业出版社,1995:116.
[2]国家药典委员会.中华人民共和国药典:2015年版一部[M].北京:中国医药科技出版社,2015:附录10,34.
[3]欧润妹,邓远辉,李伟英,等.香附不同溶剂提取物解热镇痛效应的比较[J].山东中医杂志,2004,23(12):740-742.
[4]禹良艳,华永庆,刘培,等.香附四物汤对大鼠卵巢颗粒细胞增殖的影响及活性部位成分分析[J].南京中医药大学学报,2011,27(3):238-242.
[5]郭慧玲,董能峰,胡律江,等.基于成分敲出策略辨识四制香附抗痛经的主要效应成分[J].中国实验方剂学杂志,2017,23(10):7-11.
[6]范恺磊,蔡皓,刘晓,等.香附与艾叶对治疗原发性痛经作用机制的研究进展[J].中药新药与临床药理,2017,28(1):139-143.
[7]蒋莉娟,皮胜玲,方铁铮,等.复方救必应胶囊高效液相色谱指纹图谱研究[J].中南药学,2016,10(7):687-691.
[8]XUYan,ZHANG Hongwu,WAN Xiaochun,et al.Complete assignments of1H and13C NMR data for two new sesquiterpenes from Cyperus rotundus L.[J].Magn Reson Chem,2009,47(6):527-531.
[9]黄险峰.香附的化学成分及药理研究进展[J].中药材,2003,26(1):65-68.
[10]TSOYI K,JANG H J,LEE Y S,et al.(+)-Nootkatone and(+)-valencene from rhizomes of Cyperus rotundus increase survival rates in septic mice due to heme oxygenase-1induction[J].J Ethnoph armacol,2011,137(3):1311-1317.
[11]邓远辉,刘瑜彬,罗淑文,等.α-香附酮的分离及其解热镇痛作用研究[J].中药新药与临床药理,2012,23(6):620-623.
[12]卢君蓉,李文兵,傅超美,等.香附药材的质量控制方法的探讨[J].中国医院药学杂志,2015,35(5):389-392.
[13]贾宜军.HPLC内标法测定香附中α-香附酮的含量[J].中药材,2005,28(8):722-723.
[14]王世宇,李文兵,卢君蓉,等.HPLC法同时测定不同产地香附药材中香附烯酮、圆柚酮和α-香附酮[J].中成药,2015,37(3):588-591.
[15]孙维广,符素平,俞脉文,等.高效液相色谱法测定复方救必应胶囊中紫丁香苷含量[J].中国药业,2007,16(11):22-22.
[16]高佳,程晓昆,班璐,等.液质联用技术的应用与发展[J].当代化工研究,2016,10(5):86-87.
[17]周婷.液质联用在中药复方研究中的应用[J].山东化工,2016,45(11):63-64.
[2]国家药典委员会.中华人民共和国药典:2015年版一部[M].北京:中国医药科技出版社,2015:附录10,34.
[3]欧润妹,邓远辉,李伟英,等.香附不同溶剂提取物解热镇痛效应的比较[J].山东中医杂志,2004,23(12):740-742.
[4]禹良艳,华永庆,刘培,等.香附四物汤对大鼠卵巢颗粒细胞增殖的影响及活性部位成分分析[J].南京中医药大学学报,2011,27(3):238-242.
[5]郭慧玲,董能峰,胡律江,等.基于成分敲出策略辨识四制香附抗痛经的主要效应成分[J].中国实验方剂学杂志,2017,23(10):7-11.
[6]范恺磊,蔡皓,刘晓,等.香附与艾叶对治疗原发性痛经作用机制的研究进展[J].中药新药与临床药理,2017,28(1):139-143.
[7]蒋莉娟,皮胜玲,方铁铮,等.复方救必应胶囊高效液相色谱指纹图谱研究[J].中南药学,2016,10(7):687-691.
[8]XUYan,ZHANG Hongwu,WAN Xiaochun,et al.Complete assignments of1H and13C NMR data for two new sesquiterpenes from Cyperus rotundus L.[J].Magn Reson Chem,2009,47(6):527-531.
[9]黄险峰.香附的化学成分及药理研究进展[J].中药材,2003,26(1):65-68.
[10]TSOYI K,JANG H J,LEE Y S,et al.(+)-Nootkatone and(+)-valencene from rhizomes of Cyperus rotundus increase survival rates in septic mice due to heme oxygenase-1induction[J].J Ethnoph armacol,2011,137(3):1311-1317.
[11]邓远辉,刘瑜彬,罗淑文,等.α-香附酮的分离及其解热镇痛作用研究[J].中药新药与临床药理,2012,23(6):620-623.
[12]卢君蓉,李文兵,傅超美,等.香附药材的质量控制方法的探讨[J].中国医院药学杂志,2015,35(5):389-392.
[13]贾宜军.HPLC内标法测定香附中α-香附酮的含量[J].中药材,2005,28(8):722-723.
[14]王世宇,李文兵,卢君蓉,等.HPLC法同时测定不同产地香附药材中香附烯酮、圆柚酮和α-香附酮[J].中成药,2015,37(3):588-591.
[15]孙维广,符素平,俞脉文,等.高效液相色谱法测定复方救必应胶囊中紫丁香苷含量[J].中国药业,2007,16(11):22-22.
[16]高佳,程晓昆,班璐,等.液质联用技术的应用与发展[J].当代化工研究,2016,10(5):86-87.
[17]周婷.液质联用在中药复方研究中的应用[J].山东化工,2016,45(11):63-64.