摘要
建立超高效液相色谱法(UPLC)同时测定纤毛婆婆纳中梓醇、梓苷、胡黄连苷Ⅱ和6-O-藜芦酰梓苷4种主要环烯醚萜苷类成分含量.采用Waters ACOUITY T3色谱柱(100 mm×2.1 mm,1.8μm);柱温为40℃;流动相为乙腈-水(0.1%甲酸+2.5 mmol甲酸铵);流速为0.4 mL/min;检测波长为260 nm.结果表明梓醇、梓苷、胡黄连苷Ⅱ和6-O-藜芦酰梓苷4种主要环烯醚萜苷类成分含量.结果表明梓醇、梓苷、胡黄连苷Ⅱ和6-O-藜芦酰梓苷分别在1.562~50μg/mL(R~2=0.9987)、1.562~50μg/mL(R~2=0.9989)、4.689~143μg/mL(R~2=0.9996)、1.562~50μg/mL(R~2=0.9998)范围内呈现良好的线性关系,平均回收率分别为100.21%(RSD=1.3%)、99.43%(RSD=1.8%)、100.6%(RSD=2.5%)、102.7%(RSD=0.65%).该方法简便、灵敏、经济、可靠,可用于纤毛婆婆纳的质量控制和评价.
A UPLC method has been developed in the current investigation for simultaneous determination of four main iridoid glycosides, which were verproside, catalposide, amphicoside and 6-O-veratroylcatalposide in Veronica ciliata Fisch. The separation was performed at 40 ℃ on Waters ACOUITY T3 column(100 mm×2.1 mm, 1.8 μm); the mobile phase was acetonitrile-water(0.1% formic acid+2.5 mmol ammoniumformate); the flow rate was 0.4 mL/min and the detection wavelength was 260 nm. The results showed that the calibration curves were the good linear relationships in the range of 1.562~50 μg/mL(R~2=0.9987) for verproside, 1.562~50 μg/mL(R~2=0.9989) for catalposide, 4.689~143 μg/mL(R~2=0.9996) for amphicoside, 1.562~50 μg/mL(R~2=0.9998) for 6-O-veratroylcatalposide, the average recovery rate were 100.21%(RSD=1.3%), 99.43%(RSD=1.8%), 100.6%(RSD=2.5%), 102.7%(RSD=0.65%),respectively. The method is simple, sensitive, economical and reliable, and can be used for quality control and evaluation of Veronica ciliata Fisch.
引文
[1] 西藏自治区革命委员会卫生局.西藏常用中草药[M].拉萨:西藏人民出版社,1971.
[2] 候宽昭.中国种子植物科属词典[M].北京:科学出版社,1982.
[3] Yin L,Wei L,Fu R,et al.Antioxidant and hepatoprotective activity of Veronica ciliata Fisch.extracts against carbon tetrachloride-induced liver injury in mice [J].Molecules,2014,19:7223.
[4] Sun Y,Lu Q,He L,et al.Active Fragment of Veronica ciliata Fisch.Attenuates t-BHP-Induced oxidative stress injury in HepG2 cells through antioxidant and antiapoptosis activities [J].Oxid Med Cell Longev,2017,2017:4727151.
[5] 高坤,李旭琴,刘安.西北高寒植物长果婆婆纳化学成分研究[J].西北植物学报,2003,23:633.
[6] 尹利.传统藏药纤毛婆婆纳生物有效成分与生物活性研究[D].成都:四川大学,2014.
[7] Li Y,Lu Q,Tan S,et al.Bioactivity-guided isolation of antioxidant and anti-hepatocarcinoma constituents from Veronica ciliata [J].Chem Cent J,2016,10:27.
[8] Tan S,Lu Q,Shu Y,et al.Iridoid glycosides fraction isolated from Veronica ciliata Fisch.protects against acetaminophen-induced liver injury in mice[J].Evid-Based Compl Alt,2017,2017:6106572.
[9] Lu Q,Sun Y,Shu Y,et al.HSCCC separation of the two iridoid glycosides and three phenolic compounds from Veronica ciliata and their in vitro antioxidant and anti-hepatocarcinoma activities[J].Molecules,2016,21:1234.
[10] Lee S U,Sung M H,Ryu H W,et al.Verproside inhibits TNF-α-induced MUC5AC expression through suppression of the TNF-α/NF-κB pathway in human airway epithelial cells[J].Cytokine,2016,77:168.
[11] 王奇志,管福琴,孙浩,等.梓苷的抗肿瘤活性研究[J].中成药,2012,34:2381.
[12] 顾伟,范昕建,吴疆,等.胡黄连苷Ⅱ对H2O2损伤L-02细胞的保护作用[J].世界华人消化杂志,2008,16:3274.
[13] Le M Q,Kim M S,Song Y S,et al.6-O-Veratroyl catalpol suppresses pro-inflammatory cytokines via regulation of extracellular signal-regulated kinase and nuclear factor-κB in human monocytic cells [J].Biochimie,2015,119:52.
[14] Quan J,Piao L,Xu H,et al.Protective effect of iridoid glucosides from Boschniakia rossica on acute liver injury induced by carbon tetrachloride in rats[J].Biosci Biotech Bioch,2009,73:849.
[15] Peng W,Qiu X Q,Shu Z H,et al.Hepatoprotective activity of total iridoid glycosides isolated from Paederia scandens (lour.) Merr.var.tomentosa[J].J Ethnopharmacol,2015,174:317.
[16] Liu Y F,Liang D,Luo H,et al.Hepatoprotective iridoid glycosides from the roots of Rehmannia glutinosa [J].J Nat Prod,2012,75:1625.
[17] Tundis R,Loizzo M R,Menichini F,et al.Biological and pharmacological activities of iridoids:recent developments[J].Mini-Rev Med Chem,2008,8:399.
[18] 余鸽,龙凤来,黄时伟.环烯醚萜药理作用研究进展[J].陕西林业科技,2009,2:69.
[19] Kwak J H,Kim H J,Lee K H,et al.Antioxidative iridoid glycosides and phenolic compounds from Veronica peregrina [J].Arch Pharm Res,2009,32:207.
[20] 罗思齐.中草药活性成分含量测定概述[J].中草药,1989,20:19.
[21] 罗国安,王义明,曹进,等.建立我国现代中药质量标准体系的研究[J].世界科学技术,2002,4:5.
[22] Swartz M E.UPLCTM:an introduction and review[J].J Liq Chromatogr R T,2005,28:1253.
[23] 高意,周光明,张彩虹,等.HPLC测定香椿中的有机酸和黄酮7种活性成分[J].四川大学学报:自然科学版,2017,54:1039.
[24] 李数数,王静,侯静,等.藏红花侧芽中L-瓜氨酸的提取与测定[J].四川大学学报:自然科学版,2018,55:1297.