高效液相色谱法同时测定青黛中色胺酮、靛蓝及靛玉红的含量
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摘要
为了建立高效液相色谱法同时测定青黛中色胺酮、靛蓝及靛玉红的含量,以色胺酮、靛蓝和靛玉红对照品为对照,对三批青黛药材中色胺酮、靛蓝及靛玉红的含量进行测定,采用色谱条件为:Shim-pack VP-ODS C18(4.6mm×150mm,5μm)色谱柱,流动相为乙腈-水(53∶47,V∶V),流速:1mL/min,检测波长290nm。实验结果表明色胺酮在0.02～0.16μg/mL时,线性关系良好(R2=0.999 5);靛蓝在8～64μg/mL时,线性关系良好(R~2=0.999 5);靛玉红在0.5～4μg/mL时,线性关系良好(R~2=0.999 1),方法的平均回收率为97.63%,相对标准偏差(RSD)=1.61%、回收率为100.4%,相对标准偏差(RSD)=2.21%、回收率为97.50%,相对标准偏差(RSD)=2.13%。三批青黛中色胺酮,靛蓝和靛玉红三种成分的质量浓度分别为0.065～0.10、17～31、2.4～3.9mg/g。实验操作简单,灵敏度高,重现性良好,结果准确可靠,可以同时测定青黛有效成分的含量。
In order to establish high pressure liquid chromatography(HPLC)method for simultaneous determination of tryptanamine,indigo and indirubin in indigo naturalis,the contents of the above three index components in three batches of indigo naturalis were determined by comparing with tryptophan,indigo and indirubin.The following chromatographic conditions were used:Shim-pack VP-ODS C18(4.6 mm×150 mm,5μm)column,mobile phase:acetonitrile-water(53∶47,V∶V),flow rate:1 mL/min,detection wavelength:290 nm.The experimental results showed that a linear relationship was very good between0.02～0.16μg/mL of tryptanthrin(R2=0.999 5),indigo was good at a linear range of 8～64μg/mL(R~2=0.999 5)and indirubin has a good linear relationship at 0.5～4μg/mL(R~2=0.999 1),The average recoveries were 97.63%(relative standard deviation=1.61%),100.4%(relative standard deviation=2.21%)and 97.50%(relative standard deviation=2.13%),respectively.The contents of tryptanthrin,indigo and indirubin were used in the three batches of indigo naturalis are 0.065～0.10,17～31 and 2.4～3.9 mg/g,respectively.The experiment is simple in operation,with high sensitivity,good reproducibility and accurate and reliable results,and it can determine the content of active ingredients in indigo naturalis.
In order to establish high pressure liquid chromatography(HPLC)method for simultaneous determination of tryptanamine,indigo and indirubin in indigo naturalis,the contents of the above three index components in three batches of indigo naturalis were determined by comparing with tryptophan,indigo and indirubin.The following chromatographic conditions were used:Shim-pack VP-ODS C18(4.6 mm×150 mm,5μm)column,mobile phase:acetonitrile-water(53∶47,V∶V),flow rate:1 mL/min,detection wavelength:290 nm.The experimental results showed that a linear relationship was very good between0.02～0.16μg/mL of tryptanthrin(R2=0.999 5),indigo was good at a linear range of 8～64μg/mL(R~2=0.999 5)and indirubin has a good linear relationship at 0.5～4μg/mL(R~2=0.999 1),The average recoveries were 97.63%(relative standard deviation=1.61%),100.4%(relative standard deviation=2.21%)and 97.50%(relative standard deviation=2.13%),respectively.The contents of tryptanthrin,indigo and indirubin were used in the three batches of indigo naturalis are 0.065～0.10,17～31 and 2.4～3.9 mg/g,respectively.The experiment is simple in operation,with high sensitivity,good reproducibility and accurate and reliable results,and it can determine the content of active ingredients in indigo naturalis.
引文
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[8]Chinese Pharmacopoeia Commission.Pharmacopoeia of the People’s Republic of China 2015Edition Volume 1[M].Beijing:China Medical Science Press,2015:199(in Chinese).(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,2015:199.)
[9]Yan W,Zhang N,Liu J L.Determination of Tryptanthrin,Indigo,Indirubin in Oral Ulcers by HPLC[J].Chin Tradit Pat Med,2009,31(5):743-745(in Chinese).(闫玮,张楠,刘建利.HPLC测定口腔溃疡散中色胺酮、靛蓝、靛玉红的含量[J].中成药,2009,31(5):743-745.)
[10]Xie Y L,He B Y,Li Y B,et al.Simultaneous Determination of Indigo and Indirubin in Medicine Material of Indigo Naturalis[J].Tradit Chin Drug Res Clin Pharmacol,2011,22(4):452-455(in Chinese).(谢友良,何百寅,李远彬,等.青黛药材中的靛蓝和靛玉红含量的同时测定[J].中药新药与临床药理,2011,22(4):452-455.)
[11]Zhu W,Yao L,Zhao H,et al.Identification and Content Determination for Indigo Naturalis in the Realgar-Indigo Naturalis Tablets[J].Asia-Pacific Tradit Med,2017,13(14):27-29(in Chinese).(朱伟,姚岚,赵赫,等.复方黄黛片中青黛鉴别与含量测定[J].亚太传统医药,2017,13(14):27-29.)
[12]Pei Y,Huang Z J,Huo Z P,et al.Simultaneous Determination of Indigo and Indirubin in Indigo Naturalis by HPLC with Multiple UV Wavelength Detection[J].Tianjin Pharm,2017,29(2):14-18(in Chinese).(裴玉,黄芝娟,霍志鹏,等.多波长HPLC法同时测定青黛药材中靛蓝和靛玉红的含量[J].天津药学,2017,29(2):14-18.)
[2]Liu Z L,Liu S.The Pharmacological Action and Clinical Application of the Indigo Naturalis[J].JAnhui Univ Chin Med,1997,(2):58-59(in Chinese).(刘宗林,刘圣.青黛药理作用与临床应用[J].安徽中医学院学报,1997,(2):58-59.)
[3]Hua H Q.The Ancient and Modern Application of Indigo Naturalis[J].China J Chin Mater Med,1992,(2):117-120(in Chinese).(华海清.青黛的古今应用[J].中国中药杂志,1992,(2):117-120.)
[4]Lai J L,Liu Y H,Liu C,et al.Research Progress of Mechanism and Clinical Application of Traditional Chinese Medicine Indirubin[J].J Tradit Chin Vet Med,2017,36(1):76-79(in Chinese).(赖金伦,刘玉辉,刘畅,等.中药靛玉红作用机理及其临床应用研究进展[J].中兽医医药杂志,2017,36(1):76-79.)
[5]Zou A D,Zhang Q,Zhao X C,et al.Determination of Indirubin in Indigo Naturalis by HPLC[J].Pract Pharm Clin Rem,2004,(3):13-14(in Chinese).(邹爱东,张群,赵晓春,等.HPLC法测定青黛中靛玉红的含量[J].实用药物与临床,2004,(3):13-14.)
[6]Miao S,Sun J Y,Xie Y H,et al.Research Progress of Tryptanthrin[J].Chin Pharmacol Bull,2008,(2):152-155(in Chinese).(缪珊,孙纪元,谢艳华,等.色胺酮的研究进展[J].中国药理学通报,2008,(2):152-155.)
[7]Bai Z L,Ren Y Z,Chen Y L,et al.Determination of Indigo and Indirubin in Indigo Naturalis by HPLC[J].Mod Chin Med,2010,12(8):27-29(in Chinese).(白宗利,任玉珍,陈彦琳,等.HPLC测定市售青黛中靛蓝和靛玉红的含量[J].中国现代中药,2010,12(8):27-29.)
[8]Chinese Pharmacopoeia Commission.Pharmacopoeia of the People’s Republic of China 2015Edition Volume 1[M].Beijing:China Medical Science Press,2015:199(in Chinese).(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,2015:199.)
[9]Yan W,Zhang N,Liu J L.Determination of Tryptanthrin,Indigo,Indirubin in Oral Ulcers by HPLC[J].Chin Tradit Pat Med,2009,31(5):743-745(in Chinese).(闫玮,张楠,刘建利.HPLC测定口腔溃疡散中色胺酮、靛蓝、靛玉红的含量[J].中成药,2009,31(5):743-745.)
[10]Xie Y L,He B Y,Li Y B,et al.Simultaneous Determination of Indigo and Indirubin in Medicine Material of Indigo Naturalis[J].Tradit Chin Drug Res Clin Pharmacol,2011,22(4):452-455(in Chinese).(谢友良,何百寅,李远彬,等.青黛药材中的靛蓝和靛玉红含量的同时测定[J].中药新药与临床药理,2011,22(4):452-455.)
[11]Zhu W,Yao L,Zhao H,et al.Identification and Content Determination for Indigo Naturalis in the Realgar-Indigo Naturalis Tablets[J].Asia-Pacific Tradit Med,2017,13(14):27-29(in Chinese).(朱伟,姚岚,赵赫,等.复方黄黛片中青黛鉴别与含量测定[J].亚太传统医药,2017,13(14):27-29.)
[12]Pei Y,Huang Z J,Huo Z P,et al.Simultaneous Determination of Indigo and Indirubin in Indigo Naturalis by HPLC with Multiple UV Wavelength Detection[J].Tianjin Pharm,2017,29(2):14-18(in Chinese).(裴玉,黄芝娟,霍志鹏,等.多波长HPLC法同时测定青黛药材中靛蓝和靛玉红的含量[J].天津药学,2017,29(2):14-18.)