HPLC-PDA法同时测定痛泻要方中五种有效成分
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摘要
目的:建立HPLC-PDA方法同时测定痛泻要方水煎液中白术内酯Ⅰ、Ⅱ、Ⅲ,芍药苷,芍药内酯苷五种有效成分的含量。方法:采用Thermo Fisher C18(4.6 mm×150 mm,5μm)色谱柱,白术内酯Ⅰ、Ⅱ、Ⅲ,芍药苷,芍药内酯苷以乙腈(A)-水(B)为流动相梯度洗脱,运行时间:32 min;柱温:30℃;流速:1.0 m L/min;检测波长:白术内酯Ⅰ及白术内酯Ⅲ为222 nm,白术内酯Ⅱ为278 nm;芍药苷、芍药内酯苷为232 nm;进样量:10μL。结果:对检测结果进行线性考察,以峰面积(Y)与浓度(X)进行线性回归,结果表明白术内酯Ⅰ、Ⅱ、Ⅲ在0.032~0.320μg之间线性关系良好,芍药苷、芍药内酯苷在0.6~6.0μg之间线性关系良好(r≥0.999 5),平均加样回收率分别为:100.16%(RSD=2.7%),100.01%(RSD=1.7%),99.95%(RSD=1.5%),99.57%(RSD=0.5%),99.34%(RSD=0.9%);供试品在24 h内稳定;痛泻要方中白术内酯Ⅰ、Ⅱ、Ⅲ,芍药苷,芍药内酯苷含量分别为(n=3):0.043 6、0.037 0、0.155 7、4.688 7、1.524 5mg/g。结论:建立HPLC-PDA方法可同时测定痛泻要方中白术内酯Ⅰ、Ⅱ、Ⅲ,芍药苷,芍药内酯苷的含量,该方法简单便捷,重复性好,可用于痛泻要方的质量控制评价。
Objective:To establish an HPLC-PDA method for the simultaneous determination of five effective constituents( atractylenolide I,atractylenolide II,atractylenolide III,paeoniflorin,and albiflorin) in Tongxie Yaofang decoction. Method:The column of Thermo Fisher C18(4.6 mm×150 mm,5 μm) was used,atractylenolide I,atractylenolide II,atractylenolide III,paeoniflorin and albiflorin were eluted with acetonitrile( A)-water( B) as the gradient mobile phase; running time was 32mins; column temperature was 30℃; and the flow rate was 1.0 m L/min; the detection wavelengths were set at 222 nm for atractylenolide I and atractylenolide III,278 nm for atractylenolide II,and 232 nm for paeoniflorin and albiflorin; and the sample size was 10μL. Results: Linear investigation of test results was performed with peak area( Y) and concentration( X),the result showed that the atractylenolide I,atractylenolide II,and atractylenolide III had good linearity in the range of 0.032 ~ 0.320 μg and paeoniflorin and albiflorin were at 0.6 ~ 6.0 μg( r≥0.999 5); The average recoveries were 100.16%( RSD = 2.7%),100.01%( RSD = 1.7%),99.95%( RSD = 1.5 %),99.57%( RSD = 0.5%),and 99.34%( RSD = 0.9%); the sample solution was stable within 24 h,and the contents were 0.0436,0.0370,0. 1557,4. 6887,1. 5245( mg/g). Conclusion:The established method for determination of atractylenolide I,atractylenolide II,atractylenolide III,Paeoniflorin,and Albiflorin was accurate and reliable,it can be used for quality control of Tongxie Yaofang.
Objective:To establish an HPLC-PDA method for the simultaneous determination of five effective constituents( atractylenolide I,atractylenolide II,atractylenolide III,paeoniflorin,and albiflorin) in Tongxie Yaofang decoction. Method:The column of Thermo Fisher C18(4.6 mm×150 mm,5 μm) was used,atractylenolide I,atractylenolide II,atractylenolide III,paeoniflorin and albiflorin were eluted with acetonitrile( A)-water( B) as the gradient mobile phase; running time was 32mins; column temperature was 30℃; and the flow rate was 1.0 m L/min; the detection wavelengths were set at 222 nm for atractylenolide I and atractylenolide III,278 nm for atractylenolide II,and 232 nm for paeoniflorin and albiflorin; and the sample size was 10μL. Results: Linear investigation of test results was performed with peak area( Y) and concentration( X),the result showed that the atractylenolide I,atractylenolide II,and atractylenolide III had good linearity in the range of 0.032 ~ 0.320 μg and paeoniflorin and albiflorin were at 0.6 ~ 6.0 μg( r≥0.999 5); The average recoveries were 100.16%( RSD = 2.7%),100.01%( RSD = 1.7%),99.95%( RSD = 1.5 %),99.57%( RSD = 0.5%),and 99.34%( RSD = 0.9%); the sample solution was stable within 24 h,and the contents were 0.0436,0.0370,0. 1557,4. 6887,1. 5245( mg/g). Conclusion:The established method for determination of atractylenolide I,atractylenolide II,atractylenolide III,Paeoniflorin,and Albiflorin was accurate and reliable,it can be used for quality control of Tongxie Yaofang.
引文
[1]章宸,刘斌,郑虎占,等.痛泻要方药理作用和临床应用研究概况[J].中国实验方剂学杂志,2010,16(7):223-226.
[2]滕超,徐惠娟,刘慧慧,等.痛泻要方及拆方对腹泻型肠易激综合征模型大鼠结肠组织水通道蛋白3表达的影响[J].中国中西医结合消化杂志,2011,19(5):290-294.
[3]旺建伟,殷越,隋方宇,等.痛泻要方对内脏高敏性大鼠结肠MC活化与5-HT相关性影响的研究[J].中医药学报,2013,41(1):82-85.
[4]朱向东,王燕,段永强,等.痛泻要方对大鼠结肠组织病理形态及抗炎机制研究[J].中医药学报,2012,40(6):8-11.
[5]马祥雪,王凤云,张北华,等.痛泻要方治疗腹泻型肠易激综合征的作用机制研究现状与思考[J].世界中医药,2015,10(7):977-981.
[6]陈建林,陈锦锋,邓健敏,等.加味痛泻要方对肝郁脾虚型肠易激综合征患者血清脑肠肽水平的影响[J].中医药信息,2016,33(4):82-83.
[7]刘良红,吴方评,金平,等.HPLC法同时测定菝葜和金刚藤糖浆中4种成分[J].中成药,2016,38(12):2598-2601.
[8]吴高松,王知斌,杨春娟,等.HPLC-ELSD法同时测定无梗五加果中3种3,4-裂环羽扇豆烷型三萜化合物的含量[J].中医药信息,2017,34(1):36-39.
[9]王学军,梁旭华,徐恒.HPLC法同时测定杜仲叶中4种成分的含量[J].中医药信息,2017,34(1):33-35.
[10]刘静,傅杰,丁舸.痛泻要方核心药组论证[J].中医杂志,2012,53(2):172-174.
[11]万德华,叶桂存.中药汤剂特点及质量影响研究[J].黄冈技术学院学报,2015,17(5):94-96.
[12]刘玉强,才谦.50批不同来源白术药材及饮片中白术内酯Ⅰ和白术内酯Ⅲ含量的HPLC法测定[J].药物分析杂志,2012,32(7):1249-1252.
[13]尹华,王知青,王玲,等.HPLC-DAD波长切换法同时测定白术中白术内酯Ⅰ、Ⅱ、Ⅲ和苍术酮的含量[J].中华中医药杂志,2013,28(1):233-236.
[14]余捷婧,吴金雄,梁亚凤,等.HPLC同时测定赤芍和白芍中没食子酸等6种成分的量[J].中草药,2015,46(11):1673-1677.
[15]张红,唐朝辉,李娜,等.桂枝茯苓胶囊主要成分含量变化对主要药效学的影响研究[J].世界科学技术—中医药现代化,2015,17(5):1066-1072.
[16]高峰.大承气汤方药配伍成分变化与药效相关性研究[D].广州:广州中医药大学,2015.
[17]吴梓春,何兆锦.不同产地白术药材白术内酯Ⅰ和白术内酯Ⅱ的含量测定[J].北方药学,2016,13(9):10-11.
[18]李鸿翔,彭腾,邓!,等.HPLC测定不同产地白术水煎液中的白术内酯Ⅲ[J].华西药学杂志,2013,28(2):197-198.
[2]滕超,徐惠娟,刘慧慧,等.痛泻要方及拆方对腹泻型肠易激综合征模型大鼠结肠组织水通道蛋白3表达的影响[J].中国中西医结合消化杂志,2011,19(5):290-294.
[3]旺建伟,殷越,隋方宇,等.痛泻要方对内脏高敏性大鼠结肠MC活化与5-HT相关性影响的研究[J].中医药学报,2013,41(1):82-85.
[4]朱向东,王燕,段永强,等.痛泻要方对大鼠结肠组织病理形态及抗炎机制研究[J].中医药学报,2012,40(6):8-11.
[5]马祥雪,王凤云,张北华,等.痛泻要方治疗腹泻型肠易激综合征的作用机制研究现状与思考[J].世界中医药,2015,10(7):977-981.
[6]陈建林,陈锦锋,邓健敏,等.加味痛泻要方对肝郁脾虚型肠易激综合征患者血清脑肠肽水平的影响[J].中医药信息,2016,33(4):82-83.
[7]刘良红,吴方评,金平,等.HPLC法同时测定菝葜和金刚藤糖浆中4种成分[J].中成药,2016,38(12):2598-2601.
[8]吴高松,王知斌,杨春娟,等.HPLC-ELSD法同时测定无梗五加果中3种3,4-裂环羽扇豆烷型三萜化合物的含量[J].中医药信息,2017,34(1):36-39.
[9]王学军,梁旭华,徐恒.HPLC法同时测定杜仲叶中4种成分的含量[J].中医药信息,2017,34(1):33-35.
[10]刘静,傅杰,丁舸.痛泻要方核心药组论证[J].中医杂志,2012,53(2):172-174.
[11]万德华,叶桂存.中药汤剂特点及质量影响研究[J].黄冈技术学院学报,2015,17(5):94-96.
[12]刘玉强,才谦.50批不同来源白术药材及饮片中白术内酯Ⅰ和白术内酯Ⅲ含量的HPLC法测定[J].药物分析杂志,2012,32(7):1249-1252.
[13]尹华,王知青,王玲,等.HPLC-DAD波长切换法同时测定白术中白术内酯Ⅰ、Ⅱ、Ⅲ和苍术酮的含量[J].中华中医药杂志,2013,28(1):233-236.
[14]余捷婧,吴金雄,梁亚凤,等.HPLC同时测定赤芍和白芍中没食子酸等6种成分的量[J].中草药,2015,46(11):1673-1677.
[15]张红,唐朝辉,李娜,等.桂枝茯苓胶囊主要成分含量变化对主要药效学的影响研究[J].世界科学技术—中医药现代化,2015,17(5):1066-1072.
[16]高峰.大承气汤方药配伍成分变化与药效相关性研究[D].广州:广州中医药大学,2015.
[17]吴梓春,何兆锦.不同产地白术药材白术内酯Ⅰ和白术内酯Ⅱ的含量测定[J].北方药学,2016,13(9):10-11.
[18]李鸿翔,彭腾,邓!,等.HPLC测定不同产地白术水煎液中的白术内酯Ⅲ[J].华西药学杂志,2013,28(2):197-198.