一测多评法同时测定龙胆不同部位中3个环烯醚萜苷类成分的含量
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摘要
目的:建立一测多评法同时测定龙胆药材中马钱苷酸、獐牙菜苦苷及龙胆苦苷的含量,并比较龙胆药材上(根茎)、中(簇生根中部)、下(簇生根尾部)三部分中3个成分的含量差异。方法:采用HPLC法,使用Agilent Eclipse XDB-C_(18)(4.6 mm×250 mm,5μm)色谱柱,流动相A为0.1%磷酸水溶液,流动相B为乙腈,梯度洗脱(0～5 min,6%B;5～35 min,6%B→3.5%B;35～65 min,3.5%B→9%B;65～85 min,9%B→6%B),流速0.8 mL·min~(-1),柱温40℃,检测波长240 nm,进样量10μL。以龙胆苦苷为内参物,建立其与马钱苷酸、獐牙菜苦苷的相对校正因子,并计算2个成分分别在药材上、中、下三部分的含量,实现一测多评;同时采用外标法测定龙胆中3个环烯醚萜苷类成分分别在药材上、中、下三部分的含量,比较2种测定方法的差异,验证一测多评法的可行性,采用SPSS21.0软件分别对龙胆药材上、中、下三部分中3个成分的含量进行单因素方差分析。结果:建立的校正因子重复性良好,含量间无显著差异。龙胆药材不同部位中3个环烯醚萜苷类成分的含量差异显著,中、下部的马钱苷酸、獐牙菜苦苷及龙胆苦苷含量均高于上部(P<0.05,有显著性差异)。结论:建立的一测多评法可作为龙胆药材中3个环烯醚萜苷类化学成分的含量测定方法。通过分析龙胆药材上、中、下三部分环烯醚萜苷类成分的含量差异,初步判断龙胆药材质量优劣的合理性,为龙胆药材等级划分及制订龙胆药材的质量评价提供参考依据,在其生产及商品流通等方面也具有一定的指导意义。
Objective:To establish a QAMS method for content determination of three compositions(loganic acid,sweroside and gentiamarin)in Gentiana,and to compare the contents in upper(rhizome),middle(middle of tufted roots)and lower(tufted root tail)parts of the herb. Methods:The HPLC analysis was performed on an Agilent Eclipse XDB-C_(18) column(4.6 mm×250 mm,5 μm). The mobile phase A was 0.1% aqueous phosphate solution,the mobile phase B was acetonitrile,and gradient elution was used(0-5 min,6%B;5-35 min,6%B → 3.5% B;35-65 min,3.5%B → 9% B;65-85 min,9% B-6% B)at a flow rate of 0.8 mL·min~(-1).The detection wavelength was 254 nm,the column temperature was 30 ℃ and the injection amount was 10 μL. Using gentiamarin as the external standard,relative correction factors for loganic acid and sweroside were calculated. The contents of the two components in three parts of the herb were determined by QAMS method. At the same time,determination of the three iridoid glycosides in three parts of Gentiana was performed by the external standard method to evaluate the feasibility of the QAMS method. Single factor variance analysis was performed on contents of the three components in upper,middle and lower parts of Gentiana using SPSS 21.0 software. Results:The calibration factors were reproducible and there was no significant difference between the QAMS method and the external standard method. The contents of three iridoid glycosides in different parts of Gentiana were significantly different. The contents of loganin,swertiamarin and gentiopicroside in the middle and lower parts were higher than those in the upper parts(P<0.05,with significant difference). Conclusion:The established method can be used for the determination of three iridoid glycosides in Gentiana. Through the analysis of the difference in contents in the three parts of Gentiana,the rationality of quality of Gentiana is preliminarily judged,which can provide reference for the classification of Gentiana and the quality evaluation of Gentiana. It also has certain guiding significance in the production and commodity circulation of Gentianae scabra.
Objective:To establish a QAMS method for content determination of three compositions(loganic acid,sweroside and gentiamarin)in Gentiana,and to compare the contents in upper(rhizome),middle(middle of tufted roots)and lower(tufted root tail)parts of the herb. Methods:The HPLC analysis was performed on an Agilent Eclipse XDB-C_(18) column(4.6 mm×250 mm,5 μm). The mobile phase A was 0.1% aqueous phosphate solution,the mobile phase B was acetonitrile,and gradient elution was used(0-5 min,6%B;5-35 min,6%B → 3.5% B;35-65 min,3.5%B → 9% B;65-85 min,9% B-6% B)at a flow rate of 0.8 mL·min~(-1).The detection wavelength was 254 nm,the column temperature was 30 ℃ and the injection amount was 10 μL. Using gentiamarin as the external standard,relative correction factors for loganic acid and sweroside were calculated. The contents of the two components in three parts of the herb were determined by QAMS method. At the same time,determination of the three iridoid glycosides in three parts of Gentiana was performed by the external standard method to evaluate the feasibility of the QAMS method. Single factor variance analysis was performed on contents of the three components in upper,middle and lower parts of Gentiana using SPSS 21.0 software. Results:The calibration factors were reproducible and there was no significant difference between the QAMS method and the external standard method. The contents of three iridoid glycosides in different parts of Gentiana were significantly different. The contents of loganin,swertiamarin and gentiopicroside in the middle and lower parts were higher than those in the upper parts(P<0.05,with significant difference). Conclusion:The established method can be used for the determination of three iridoid glycosides in Gentiana. Through the analysis of the difference in contents in the three parts of Gentiana,the rationality of quality of Gentiana is preliminarily judged,which can provide reference for the classification of Gentiana and the quality evaluation of Gentiana. It also has certain guiding significance in the production and commodity circulation of Gentianae scabra.
引文
[1]中华人民共和国药典2015年版.一部[S].2015:96ChP 2015.VolⅠ[S].2015:96
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[3]匡海学,吴高松,刘华,等.龙胆苦苷和龙胆碱的生物转化和药理活性研究[J].天津中医药,2016,33(7):385KUANG HX,WU GS,LIU H,et al.Biotransformation and pharmacological activity of gentiopicroside and gentiopicrine[J].Tianjin Tra Chin Med,2016,33(7):385
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[5]郭建华,田成旺,刘晓,等.中药环烯醚萜类化合物研究进展[J].药物评价研究,2011,34(4):293-297GUO JH,TIAN CW,LIU X,et al.Research progress of iridoid terpenoids in traditional Chinese medicine[J].Drug Eval Res,2011,34(4):293
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[7]王晓燕,霍甜甜,王翀,等.一测多评法测定酒萸肉中5种环烯醚萜苷类成分的含量[J].中药材,2016,39(7):1586WANG XY,HUO TT,WANG C,et al.Determination of five iridoid glycosides in Cornus officinalis by one-step multi-evaluation method[J].Chin Herb Med,2016,39(7):1586
[8]昝珂,崔淦,过立农,等.一测多评法同时测定冬虫夏草中5个核苷类成分的含量[J].药物分析杂志,2018,38(4):630ZAN K,CUI G,GUO LN,et al.Simultaneous determination of five nucleosides in Cordyceps sinensis by a multi-evaluation method[J].Chin J Pharm Anal,2018,38(4):630
[9]黄华花,陈景海.UPLC法测定小叶榕不同部位样品中3个黄酮类成分的含量[J].药物分析杂志,2017,37(1):161HUANG HH,CHEN JH.UPLC determination of three flavonoids in different parts of Ficus microcarpa[J].Chin J Pharm Anal,2017,37(1):161
[10]冉柏光.中药饮片性状鉴别[J].实用中医药杂志,2013,29(6):488RAN BG.Character identification of Chinese herbal pieces[J].Chin J Prac Chin Med,2013,29(6):488
[11]孙紫薇,陈丽,辛宇,等.酒炙龙胆炮制工艺的研究[J].人参研究,2017,29(6):48SUN ZW,CHEN L,XIN Y,et al.Study on the processing technology of wine cellar gentian[J].Ginseng Res,2017,29(6):48
[12]张村,肖永庆,李丽,等.栀子不同饮片环烯醚萜苷类成分比较研究[J].中国中药杂志,2008,33(10):1138ZHANG C,XIAO YQ,LI L,et al.Comparative study on the content of iridoid glycosides in different decoction pieces of Chinese wolfberry[J].China J Chin Mater Med,2008,33(10):1138
[13]孙紫薇,张楠,翁丽丽,等.朝鲜龙胆的生药学研究[J].长春中医药大学学报,2017,33(5):720SUN ZW,ZHANG N,WENG LL,et al.Study on the pharmacology of Korean gentian[J].Changchun Univ J Tradit Chin Med,2017,33(5):720
[14]刘武占,范建伟,高艳红,等.HPLC同时测定栀子中8个环烯醚萜苷类成分的含量[J].中国中药杂志,2012,37(16):2418LIU WZ,FAN JW,GAO YH,et al.Simultaneous determination of eight iridoid glycosides in gardenia by HPLC[J].China J Chin Mater Med,2012,37(16):2418
[15]张楠,姜雨昕,任悦,等.龙胆及朝鲜龙胆中龙胆多糖与龙胆苦苷的含量测定[J].时珍国医国药,2016,27(10):2395ZHANG N,JIANG YX,REN Y,et al.Determination of gentian polysaccharide and gentiopicroside in gentian and Korean gentian[J].Lishizhen Med Mater Med Res,2016,27(10):2395
[16]罗文艳,段和祥,杨毅生,等.一测多评法测定蒌蒿药材中10种成分的含量[J].中药材,2016,39(4):822LUO WY,DUAN HX,YANG YS,et al.Determination of 10components in Artemisia scoparia by a multi-evaluation method[J].J Chin Med Mater,2016,39(4):822
[2]杨书彬,王承.龙胆化学成分和药理作用研究进展[J].中医药学报,2005,33(6):54YANY SB,WANG C.Advances in research on chemical constituents and pharmacological effects of gentian[J].Tradit J Chin Med,2005,33(6):54
[3]匡海学,吴高松,刘华,等.龙胆苦苷和龙胆碱的生物转化和药理活性研究[J].天津中医药,2016,33(7):385KUANG HX,WU GS,LIU H,et al.Biotransformation and pharmacological activity of gentiopicroside and gentiopicrine[J].Tianjin Tra Chin Med,2016,33(7):385
[4]董丽萍,倪梁红,赵志礼,等.龙胆属环烯醚萜类化学成分研究进展[J].中草药,2017,48(10):2117.DONG LP,NI LH,ZHAO ZL,et al.Advances in the study of iridoid terpenoids from gentiana[J].Chin Tradit Herb Drugs,2017,48(10):2117
[5]郭建华,田成旺,刘晓,等.中药环烯醚萜类化合物研究进展[J].药物评价研究,2011,34(4):293-297GUO JH,TIAN CW,LIU X,et al.Research progress of iridoid terpenoids in traditional Chinese medicine[J].Drug Eval Res,2011,34(4):293
[6]张敬莹.糙龙胆地上部分的化学成分研究[D].大连:大连理工大学,2008:10ZHANG JY.Studies on the Chemical Constituents of the Aerial Parts of Gentiana[D].Dalin:Dalian University of Technology,2008:10
[7]王晓燕,霍甜甜,王翀,等.一测多评法测定酒萸肉中5种环烯醚萜苷类成分的含量[J].中药材,2016,39(7):1586WANG XY,HUO TT,WANG C,et al.Determination of five iridoid glycosides in Cornus officinalis by one-step multi-evaluation method[J].Chin Herb Med,2016,39(7):1586
[8]昝珂,崔淦,过立农,等.一测多评法同时测定冬虫夏草中5个核苷类成分的含量[J].药物分析杂志,2018,38(4):630ZAN K,CUI G,GUO LN,et al.Simultaneous determination of five nucleosides in Cordyceps sinensis by a multi-evaluation method[J].Chin J Pharm Anal,2018,38(4):630
[9]黄华花,陈景海.UPLC法测定小叶榕不同部位样品中3个黄酮类成分的含量[J].药物分析杂志,2017,37(1):161HUANG HH,CHEN JH.UPLC determination of three flavonoids in different parts of Ficus microcarpa[J].Chin J Pharm Anal,2017,37(1):161
[10]冉柏光.中药饮片性状鉴别[J].实用中医药杂志,2013,29(6):488RAN BG.Character identification of Chinese herbal pieces[J].Chin J Prac Chin Med,2013,29(6):488
[11]孙紫薇,陈丽,辛宇,等.酒炙龙胆炮制工艺的研究[J].人参研究,2017,29(6):48SUN ZW,CHEN L,XIN Y,et al.Study on the processing technology of wine cellar gentian[J].Ginseng Res,2017,29(6):48
[12]张村,肖永庆,李丽,等.栀子不同饮片环烯醚萜苷类成分比较研究[J].中国中药杂志,2008,33(10):1138ZHANG C,XIAO YQ,LI L,et al.Comparative study on the content of iridoid glycosides in different decoction pieces of Chinese wolfberry[J].China J Chin Mater Med,2008,33(10):1138
[13]孙紫薇,张楠,翁丽丽,等.朝鲜龙胆的生药学研究[J].长春中医药大学学报,2017,33(5):720SUN ZW,ZHANG N,WENG LL,et al.Study on the pharmacology of Korean gentian[J].Changchun Univ J Tradit Chin Med,2017,33(5):720
[14]刘武占,范建伟,高艳红,等.HPLC同时测定栀子中8个环烯醚萜苷类成分的含量[J].中国中药杂志,2012,37(16):2418LIU WZ,FAN JW,GAO YH,et al.Simultaneous determination of eight iridoid glycosides in gardenia by HPLC[J].China J Chin Mater Med,2012,37(16):2418
[15]张楠,姜雨昕,任悦,等.龙胆及朝鲜龙胆中龙胆多糖与龙胆苦苷的含量测定[J].时珍国医国药,2016,27(10):2395ZHANG N,JIANG YX,REN Y,et al.Determination of gentian polysaccharide and gentiopicroside in gentian and Korean gentian[J].Lishizhen Med Mater Med Res,2016,27(10):2395
[16]罗文艳,段和祥,杨毅生,等.一测多评法测定蒌蒿药材中10种成分的含量[J].中药材,2016,39(4):822LUO WY,DUAN HX,YANG YS,et al.Determination of 10components in Artemisia scoparia by a multi-evaluation method[J].J Chin Med Mater,2016,39(4):822