HPLC同时测定雪胆属植物中3种活性成分的含量
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摘要
建立了高效液相色谱法同时测定雪胆属植物中雪胆乙素、竹节参皂苷Ⅳa和雪胆甲素含量的方法,可用于11种雪胆属植物中3种活性成分的同时测定。采用InertSustain C18(250 mm×4.6 mm, 5μm)色谱柱;流动相为0.1%磷酸水溶液(A)-乙腈(B),梯度洗脱(0～20 min, 5%～20%B; 20～60 min, 20%～30%B; 60～80 min,30%～64%B);流速1.0 mL/min;柱温35℃;检测波长203 nm。经方法学考察,本方法具有准确快速、精密度高、重复性好的特点,雪胆乙素、竹节参皂苷Ⅳa和雪胆甲素分别在0.075～3.000μg、0.810～32.400μg和0.620～24.800μg范围内与峰面积呈现良好的线性关系(r≥0.999 5)。定量分析测得11种雪胆属植物中雪胆乙素含量在0.479～1.904 mg/g范围内,彭县雪胆中含量最高;竹节参皂苷Ⅳa含量在1.098～18.58 mg/g范围内,峨眉雪胆中含量最高;雪胆甲素含量在3.079～17.56 mg/g范围内,十一叶雪胆中含量最高。本方法可为雪胆属优质药用资源开发及综合利用提供参考。
A method for the simultaneous determination of CucurbitacinⅡb, Chikusetsu saponin Ⅳa and CucurbitacinⅡa in Hemsleya by HPLC was established. It can be used for the simultaneous determination of three active components in 11 species of Hemsleya. InertSustain C18(250 mm×4.6 mm, 5 μm) was used, with acetonitrile-0.1%phosphoric acid as the mobile phase in gradient elution mode. The column temperature was 35℃ with flow rate1.0 mL/min. The UV detection wavelength was 203 nm. The method has the characteristics of accurate and rapid,high precision and good repeatability. The CucurbitacinⅡb, Chikusetsu saponin Ⅳa and CucurbitacinⅡa showed a good linear relationship with the peak area in the range of 0.075～3.000 μg, 0.810～32.40 μg and 0.620～24.80 μg(r≥0.999 5). Quantitative analysis showed that the content of Cucurbitacin Ⅱb in 11 species of Hemsleya was in the range of 0.479～1.904 mg/g, and the content in H. pengxianensis was the highest. The content of Chikusetsu saponin IVa is in the range of 1.098～18.58 mg/g, the content of H. omeiensis is the highest. CucurbitacinⅡa content in the range of 3.079～17.56 mg/g, the content of H. endecaphylla is the highest. The method can provide a reference for the development and comprehensive utilization of high-quality medicinal resources of Hemsleya.
A method for the simultaneous determination of CucurbitacinⅡb, Chikusetsu saponin Ⅳa and CucurbitacinⅡa in Hemsleya by HPLC was established. It can be used for the simultaneous determination of three active components in 11 species of Hemsleya. InertSustain C18(250 mm×4.6 mm, 5 μm) was used, with acetonitrile-0.1%phosphoric acid as the mobile phase in gradient elution mode. The column temperature was 35℃ with flow rate1.0 mL/min. The UV detection wavelength was 203 nm. The method has the characteristics of accurate and rapid,high precision and good repeatability. The CucurbitacinⅡb, Chikusetsu saponin Ⅳa and CucurbitacinⅡa showed a good linear relationship with the peak area in the range of 0.075～3.000 μg, 0.810～32.40 μg and 0.620～24.80 μg(r≥0.999 5). Quantitative analysis showed that the content of Cucurbitacin Ⅱb in 11 species of Hemsleya was in the range of 0.479～1.904 mg/g, and the content in H. pengxianensis was the highest. The content of Chikusetsu saponin IVa is in the range of 1.098～18.58 mg/g, the content of H. omeiensis is the highest. CucurbitacinⅡa content in the range of 3.079～17.56 mg/g, the content of H. endecaphylla is the highest. The method can provide a reference for the development and comprehensive utilization of high-quality medicinal resources of Hemsleya.
引文
Boykin C.,Zhang G.,Chen Y.H.,Zhang R.W.,Fan X.E.,Yang W.M.,and Lu Q.,2011,CucurbitacinⅡa:a novel class of anti-cancer drug inducing non-reversible actin aggregation and inhibiting survivin independent of JAK2/STAT3 phosphorylation,Br.J.Cancer.,104(5):781-789
Chen J.C.,Zhang G.H.,Zhang Z.Q.,Qiu M.H.,Zheng Y.T.,Yang L.M.,and Yu K.B.,2008,Octanorcucurbitane and cucurbitane triterpenoids from the tubers of Hemsleya endecaphylla with HIV-1 inhibitory activity,J.Nat.Prod.,71(1):153-155
Chen X.J.,Wu Y.Y.,Kuang W.J.,and Zhou L.M.,2009,Experiment study on anti-inflammation and antitussive of Xuedansu tablet,Sichuan Shengli Kexue Zazhi(Sichuan Journal of Physiological Sciences),31(4):153-154(陈夏静,伍怡颖,匡文娟,缪世坤,周黎明,2009,雪胆素片抗炎镇咳作用的实验研究,四川生理科学杂志,31(4):153-154)
Chen Y.L.,Xian Q.,Xiao C.,Zhong Y.L.,Su X.M.,Xu L.,Luo Q.L.,Cheng P.,Wang T.,Liu J.,Zhang T.,Yang T.,Zou Q.,and Li H.,2017,Effects of cucurbitacinⅡa on apoptosis of human lung cancer cell lines NCI-H460 and A549 and its mechanism,Zhongguo Yaolixue Tongbao(Chinese Pharmacological Bulletin),33(7):922-927(陈育林,咸婧,肖翠,钟月玲,苏小妹,徐丽,罗巧丽,程朋,王涛,刘进,张涛,阳泰,邹强,李华,2017,雪胆素甲诱导非小细胞肺癌系NCI-H460和A549的凋亡效应及其作用机制初探,中国药理学通报,33(7):922-927)
Duan J.L.,Yin Y.,Cui J.,Yan J.J.,Zhu Y.R.,Guan Y.,Wei G.,Weng Y.,Wu X.X.,Guo C.,Wang Y.H.,Xi M.M.,and Wen A.D.,2015,Chikusetsu saponin IVa ameliorates cerebral ischemia reperfusion injury in diabetic mice via adiponectinmediated AMPK/GSK-3βpathway in vivo and in vitro,Mol.Neurobiol.,53(1):728-743
Editorial Committee of Flora of China,ed.,1979,Flora of China,Science Press,Beijing,China,pp.102-129(中国植物志编辑委员会,主编,1986,中国植物志,科学出版社,中国,北京,pp.102-129)
He J.,Xu L.H.,Qiao J.,Ouyang D.Y.,and He X.H.,2012,CucurbitacinⅡa induces apoptosis of LPS-stimulated RAW264.7cells through disrupting microfilament cytoskeleton,Mianyixue Zazhi(Immunological Journal),7:553-557(何健,徐丽慧,乔静,欧阳东云,何贤辉,2012,雪胆素甲通过破坏微丝结构诱导LPS活化的RAW264.7细胞凋亡,免疫学杂志,7:553-557)
He X.P.,Yang Q.F.,Wang X.Y.,and Liu L.Y.,2007,Determination of cucurbitacin in xuedan by HPLC,Chuanbei Yixueyuan Xuebao(Journal of North Sichuan Medical College),22(3):264-266(何效平,杨琼芳,王贤英,刘兰英,2007,高效液相色谱法测定雪胆中雪胆甲素的含量,川北医学院学报,22(3):264-266)
Liu A.H.,Shi M.,Bao F.K.,Zhang Y.B.,Duan X.Y.,Zhao G.P.,and Dai X.L.,2012,Hemsleyadine has protective effects on experimental liver injury in mice,Kunming Yike Daxue Xuebao(Journal of Kunming Medical University),33(9):8-10(柳爱华,石梅,宝福凯,张云波,段新亚,赵桂萍,代旭磊,2012,雪胆素对实验性肝损伤的保护作用,昆明医科大学学报,33(9):8-10)
Liu Y.L.,Li X.R.,Fan J.Y.,Xu Q.M.,and Yang S.L.,2010,Ex perimental research on effects of Xue Dan Wei Chang pill on gastric ulcer,Shizhen Guoyi Guoyao(Lishizhen Medicine and Materia Medica Research),21(2):301-302(刘艳丽,李笑然,范金胤,许琼明,杨世林,2010,雪胆胃肠丸对胃溃疡的作用实验研究,时珍国医国药,21(2):301-302)
Li Y.W.,Zhang T.J.,Cui J.,Jia N.,Wu Y.,Xi M.M.,and Wen A.D.,2015,Chikusetsu saponin IVa regulates glucose uptake and fatty acid oxidation:implications in antihyperglycemic and hypolipidemic effects,J.Pharm.Pharmacol.,67(7):997-1007
Nie R.L.,and Chen Z.L.,1986,The research history and present status an the chemical components of genus hemsleya(Cucurbitaceae),Zhiwu Fenlei Yu Ziyuan Xuebao(Plant Diversity),8(1):117-126(聂瑞麟,陈宗莲,1986,雪胆属植物化学研究历史和现状,植物分类与资源学报,8(1):117-126)
Ren S.,Xu L.H.,Zeng L.H.,Ouyang D.Y.,and He X.H.,2012,Inhibitory effect of cucurbitacinIIb on proliferation of human prostate cancer PC-3 cells through disruption of microfilamentsand upregulation of p21Cip1 expression,Zhongguo Yaolixue Yu Dulixue Zazhi(Cancer Chemotherapy and Pharmacology),26(6):835-841(任帅,徐丽慧,曾龙辉,欧阳东云,何贤辉,2012,雪胆素乙通过破坏微丝结构及上调p21Cip1抑制前列腺癌PC-3细胞的增殖,中国药理学与毒理学杂志,26(6):835-841)
Tian R.R.,Chen J.C.,Zhang G.H.,Qiu M.H.,Wang Y.H.,Du L.,Shen X.,Liu N.F.,and Zhang Y.T.,2008,Anti-HIV-1 activities of hemslecins A and B,Chinese Journal of Natural Medicines,6(3):214-218
Wang Y.,Liu K.P.,Song F.M.,Pan H.,Xu L.H.,Ouyang D.Y.,and He X.H.,2013,The effects of cucurbitacinⅡb on the in vitro activation and proliferation of mouse lymphocytes,Mianyixue Zazhi(Immunological Journal),29(3):190-194(王遥,刘坤鹏,宋方茗,潘浩,徐丽慧,欧阳东云,何贤辉,2013,雪胆素乙对小鼠淋巴细胞体外活化及增殖的影响,免疫学杂志,29(3):190-194)
Wan L.,and Qin R.A.,2003,Studies on effects of Xuedan on the experimental gastric ulcer,Zhongguo Zhongyao Zazhi(China Journal of Chinese Materia Medica),28(3):266-268(宛蕾,覃仁安,2003,雪胆提取物对实验性胃溃疡作用的研究,中国中药杂志,28(3):266-268)
Zhi B.Y.,Yan Y.,and Yi Z.L.,2008,Investigation of antimicrobial model of hemsleya pengxianensis Chang W.J.and its main active component by metabolomics technique,J.Ethnopharmacol.,116(1):89-95
Chen J.C.,Zhang G.H.,Zhang Z.Q.,Qiu M.H.,Zheng Y.T.,Yang L.M.,and Yu K.B.,2008,Octanorcucurbitane and cucurbitane triterpenoids from the tubers of Hemsleya endecaphylla with HIV-1 inhibitory activity,J.Nat.Prod.,71(1):153-155
Chen X.J.,Wu Y.Y.,Kuang W.J.,and Zhou L.M.,2009,Experiment study on anti-inflammation and antitussive of Xuedansu tablet,Sichuan Shengli Kexue Zazhi(Sichuan Journal of Physiological Sciences),31(4):153-154(陈夏静,伍怡颖,匡文娟,缪世坤,周黎明,2009,雪胆素片抗炎镇咳作用的实验研究,四川生理科学杂志,31(4):153-154)
Chen Y.L.,Xian Q.,Xiao C.,Zhong Y.L.,Su X.M.,Xu L.,Luo Q.L.,Cheng P.,Wang T.,Liu J.,Zhang T.,Yang T.,Zou Q.,and Li H.,2017,Effects of cucurbitacinⅡa on apoptosis of human lung cancer cell lines NCI-H460 and A549 and its mechanism,Zhongguo Yaolixue Tongbao(Chinese Pharmacological Bulletin),33(7):922-927(陈育林,咸婧,肖翠,钟月玲,苏小妹,徐丽,罗巧丽,程朋,王涛,刘进,张涛,阳泰,邹强,李华,2017,雪胆素甲诱导非小细胞肺癌系NCI-H460和A549的凋亡效应及其作用机制初探,中国药理学通报,33(7):922-927)
Duan J.L.,Yin Y.,Cui J.,Yan J.J.,Zhu Y.R.,Guan Y.,Wei G.,Weng Y.,Wu X.X.,Guo C.,Wang Y.H.,Xi M.M.,and Wen A.D.,2015,Chikusetsu saponin IVa ameliorates cerebral ischemia reperfusion injury in diabetic mice via adiponectinmediated AMPK/GSK-3βpathway in vivo and in vitro,Mol.Neurobiol.,53(1):728-743
Editorial Committee of Flora of China,ed.,1979,Flora of China,Science Press,Beijing,China,pp.102-129(中国植物志编辑委员会,主编,1986,中国植物志,科学出版社,中国,北京,pp.102-129)
He J.,Xu L.H.,Qiao J.,Ouyang D.Y.,and He X.H.,2012,CucurbitacinⅡa induces apoptosis of LPS-stimulated RAW264.7cells through disrupting microfilament cytoskeleton,Mianyixue Zazhi(Immunological Journal),7:553-557(何健,徐丽慧,乔静,欧阳东云,何贤辉,2012,雪胆素甲通过破坏微丝结构诱导LPS活化的RAW264.7细胞凋亡,免疫学杂志,7:553-557)
He X.P.,Yang Q.F.,Wang X.Y.,and Liu L.Y.,2007,Determination of cucurbitacin in xuedan by HPLC,Chuanbei Yixueyuan Xuebao(Journal of North Sichuan Medical College),22(3):264-266(何效平,杨琼芳,王贤英,刘兰英,2007,高效液相色谱法测定雪胆中雪胆甲素的含量,川北医学院学报,22(3):264-266)
Liu A.H.,Shi M.,Bao F.K.,Zhang Y.B.,Duan X.Y.,Zhao G.P.,and Dai X.L.,2012,Hemsleyadine has protective effects on experimental liver injury in mice,Kunming Yike Daxue Xuebao(Journal of Kunming Medical University),33(9):8-10(柳爱华,石梅,宝福凯,张云波,段新亚,赵桂萍,代旭磊,2012,雪胆素对实验性肝损伤的保护作用,昆明医科大学学报,33(9):8-10)
Liu Y.L.,Li X.R.,Fan J.Y.,Xu Q.M.,and Yang S.L.,2010,Ex perimental research on effects of Xue Dan Wei Chang pill on gastric ulcer,Shizhen Guoyi Guoyao(Lishizhen Medicine and Materia Medica Research),21(2):301-302(刘艳丽,李笑然,范金胤,许琼明,杨世林,2010,雪胆胃肠丸对胃溃疡的作用实验研究,时珍国医国药,21(2):301-302)
Li Y.W.,Zhang T.J.,Cui J.,Jia N.,Wu Y.,Xi M.M.,and Wen A.D.,2015,Chikusetsu saponin IVa regulates glucose uptake and fatty acid oxidation:implications in antihyperglycemic and hypolipidemic effects,J.Pharm.Pharmacol.,67(7):997-1007
Nie R.L.,and Chen Z.L.,1986,The research history and present status an the chemical components of genus hemsleya(Cucurbitaceae),Zhiwu Fenlei Yu Ziyuan Xuebao(Plant Diversity),8(1):117-126(聂瑞麟,陈宗莲,1986,雪胆属植物化学研究历史和现状,植物分类与资源学报,8(1):117-126)
Ren S.,Xu L.H.,Zeng L.H.,Ouyang D.Y.,and He X.H.,2012,Inhibitory effect of cucurbitacinIIb on proliferation of human prostate cancer PC-3 cells through disruption of microfilamentsand upregulation of p21Cip1 expression,Zhongguo Yaolixue Yu Dulixue Zazhi(Cancer Chemotherapy and Pharmacology),26(6):835-841(任帅,徐丽慧,曾龙辉,欧阳东云,何贤辉,2012,雪胆素乙通过破坏微丝结构及上调p21Cip1抑制前列腺癌PC-3细胞的增殖,中国药理学与毒理学杂志,26(6):835-841)
Tian R.R.,Chen J.C.,Zhang G.H.,Qiu M.H.,Wang Y.H.,Du L.,Shen X.,Liu N.F.,and Zhang Y.T.,2008,Anti-HIV-1 activities of hemslecins A and B,Chinese Journal of Natural Medicines,6(3):214-218
Wang Y.,Liu K.P.,Song F.M.,Pan H.,Xu L.H.,Ouyang D.Y.,and He X.H.,2013,The effects of cucurbitacinⅡb on the in vitro activation and proliferation of mouse lymphocytes,Mianyixue Zazhi(Immunological Journal),29(3):190-194(王遥,刘坤鹏,宋方茗,潘浩,徐丽慧,欧阳东云,何贤辉,2013,雪胆素乙对小鼠淋巴细胞体外活化及增殖的影响,免疫学杂志,29(3):190-194)
Wan L.,and Qin R.A.,2003,Studies on effects of Xuedan on the experimental gastric ulcer,Zhongguo Zhongyao Zazhi(China Journal of Chinese Materia Medica),28(3):266-268(宛蕾,覃仁安,2003,雪胆提取物对实验性胃溃疡作用的研究,中国中药杂志,28(3):266-268)
Zhi B.Y.,Yan Y.,and Yi Z.L.,2008,Investigation of antimicrobial model of hemsleya pengxianensis Chang W.J.and its main active component by metabolomics technique,J.Ethnopharmacol.,116(1):89-95
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