多成分定性/定量分析结合模式识别分析3个主产区乌拉尔甘草水溶性特征组分
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摘要
采集了3个不同主产区32批种植或野生乌拉尔甘草药材,采用指纹图谱、LC-TOF/MS及HPLC多指标成分分析结合模式识别(PCA和OPLS-DA)对不同主产地甘草样品水溶性特征组分进行差异性分析。32批次药材指纹图谱相似度在0. 903～0. 999,相似度较高;采用模式识别可以将甘肃、新疆种植药材与内蒙古种植药材及野生药材进行区分,进一步以LCTOF/MS及对照品对照鉴定了指纹图谱中31个共有峰成分,并对其中4个黄酮苷及5个三萜皂苷特征组分含量进行了对比分析,结果表明3个产区种植甘草药材中5个三萜皂苷总含量差异不明显,但4个黄酮苷总含量呈现出内蒙古>甘肃≈新疆的趋势(P<0. 05),而野生药材与种植药材相比4个黄酮苷及5个三萜皂苷总含量均有显著提升(P<0. 01);其中,甘肃与新疆种植药材在甘草苷、异甘草苷、甘草皂苷A_3、22β-乙酰甘草酸及乌拉尔皂苷B上的含量均要低于内蒙古地区种植药材,但主要指标性成分甘草酸在3个产区药材中含量差异并不明显,而在内蒙古产区内的野生药材比种植药材在甘草苷、异甘草苷、甘草皂苷A_3、甘草皂苷G_2及甘草酸含量上均有显著升高。多指标成分定性/定量分析结合模式识别能有效评价不同区域种植及野生甘草的质量,有助于深入了解不同区域甘草的现状,可为甘草的资源开发与综合利用提供科学依据。
Thirty-two batches of cultivated and wild Glycyrrhiza uralensis were obtained from three geographical regions. Comparative study of water characteristic components of G. uralensis from three geographical origins was conducted by PCA,OPLS-DA chemical pattern recognition combined with LC-TOF/MS and muti-component analysis. The similarity of fingerprints of 32 batches of medicinal materials ranged from 0. 903 to 0. 999. Patterns recognition could be used to distinguish cultivated G. uralensis in Gansu and Xinjiang areas from cultivated and wild plants in Inner Mongolia. Then a total of thirty-one common constituents were identified by LC-TOF/MS analysis coupled with standard compounds information. The contents of four flavonoid glycosides and five saponins were determinated by HPLC and compared using One-way ANOVA. The results showed that there was no significant difference in the contents of 5 triterpenoid saponins among the three regions,but the contents of 4 flavonoid saponins showed the trend of Inner Mongolia >Gansu≈Xinjiang( P<0. 05). In the same Inner Mongolia region,the contents of 4 flavonoid glycosides and 5 triterpenoid saponins in wild plant was significantly higher than that in cultivated plants( P<0. 01). In addition,the contents of liquiritin,isoliquiritin,licorice-saponin A_3,22β-acetoxyl-glycyrrhizic acid and uralsaponin B in Gansu and Xinjiang were obviously lower than those in Inner Mongolia,but the contents of glycyrrhizic acid,the main component of G. uralensis,were not different in the three geographical regions. In Inner Mongolia,the contents of liquiritin,isoliquiritin,licorice-saponin A_3,licorice-saponin G_2 and glycyrrhizic acid in wild plants were significantly higher than those in cultivated plants. In conclusion,qualitative/quantitative analysis of multi-index components combined with pattern recognition could effectively evaluate the quality of cultivated and wild licorice in different regions. It was helpful for us to understand the reality of licorice in different regions,and provided scientific basis for the development and comprehensive utilization of licorice resources.
Thirty-two batches of cultivated and wild Glycyrrhiza uralensis were obtained from three geographical regions. Comparative study of water characteristic components of G. uralensis from three geographical origins was conducted by PCA,OPLS-DA chemical pattern recognition combined with LC-TOF/MS and muti-component analysis. The similarity of fingerprints of 32 batches of medicinal materials ranged from 0. 903 to 0. 999. Patterns recognition could be used to distinguish cultivated G. uralensis in Gansu and Xinjiang areas from cultivated and wild plants in Inner Mongolia. Then a total of thirty-one common constituents were identified by LC-TOF/MS analysis coupled with standard compounds information. The contents of four flavonoid glycosides and five saponins were determinated by HPLC and compared using One-way ANOVA. The results showed that there was no significant difference in the contents of 5 triterpenoid saponins among the three regions,but the contents of 4 flavonoid saponins showed the trend of Inner Mongolia >Gansu≈Xinjiang( P<0. 05). In the same Inner Mongolia region,the contents of 4 flavonoid glycosides and 5 triterpenoid saponins in wild plant was significantly higher than that in cultivated plants( P<0. 01). In addition,the contents of liquiritin,isoliquiritin,licorice-saponin A_3,22β-acetoxyl-glycyrrhizic acid and uralsaponin B in Gansu and Xinjiang were obviously lower than those in Inner Mongolia,but the contents of glycyrrhizic acid,the main component of G. uralensis,were not different in the three geographical regions. In Inner Mongolia,the contents of liquiritin,isoliquiritin,licorice-saponin A_3,licorice-saponin G_2 and glycyrrhizic acid in wild plants were significantly higher than those in cultivated plants. In conclusion,qualitative/quantitative analysis of multi-index components combined with pattern recognition could effectively evaluate the quality of cultivated and wild licorice in different regions. It was helpful for us to understand the reality of licorice in different regions,and provided scientific basis for the development and comprehensive utilization of licorice resources.
引文
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[2]中国药典.一部[S]. 2015:86.
[3]高晓娟,赵丹,赵建军,等.甘草的本草考证[J].中国实验方剂学杂志,2017,23(2):193.
[4]冯毓秀,林寿全.甘草的本草考证及研究概况[J].时珍国药研究,1993(2):43.
[5]黄明进,王文全,魏胜利.我国甘草药用植物资源调查及质量评价研究[J].中国中药杂志,2010,35(8):947.
[6] Li Y,Sun F,Jing Z,et al. Glycyrrhizic acid exerts anti-inflammatory effect to improve cerebral vasospasm secondary to subarachnoid hemorrhage in a rat model[J]. Neurol Res,2017,39(8):727.
[7] Rui F,Nan L,Jiang X,et al. HPLC-DAD-MS/MS identification and HPLC-ABTS+online antioxidant activity evaluation of bioactive compounds in liquorice(Glycyrrhiza uralensis Fisch.)extract[J]. Eur Food Res Technol,2015,240(5):1035.
[8] Cao L,Ding W,Jia R,et al. Anti-inflammatory and hepatoprotective effects of glycyrrhetinic acid on CCl4-induced damage in precision-cut liver slices from Jian carp(Cyprinus carpio var.jian)through inhibition of the NF-pathway[J]. Fish Shellfish Immunol,2017,64:234.
[9] Baltina L A,Zarubaev V V,Baltina L A,et al. Glycyrrhizic acid derivatives as influenza A/H1N1 virus inhibitors[J]. Bioorg Med Chem Lett,2015,25(8):1742.
[10] Han S,Sun L,He F,et al. Anti-allergic activity of glycyrrhizic acid on Ig E-mediated allergic reaction by regulation of allergy-related immune cells[J]. Sci Rep,2017,7(1):7222.
[11]闫永红,王文全,杨娜.栽培甘草中总黄酮的含量测定[J].中国中药杂志,2006,31(12):1021.
[12]冯薇,王文全,赵平然.栽培年限和采收期对甘草总皂苷、总黄酮含量的影响[J].中药材,2008,31(2):184.
[13] Zheng Y F,Qi L W,Cui X B,et al. Oleanane-type triterpene glucuronides from the roots of Glycyrrhiza uralensis Fischer.[J].Planta Med,2010,76(13):1457.
[14]闫永红.不同来源甘草的质量特征及评价研究[D].北京:北京中医药大学,2006.
[15]范铭,曹爱农,晋小军,等.陇中半干旱地区不同年限甘草生长与有效成分积累动态研究[J].西北农业学报,2016,25(10):1522.
[16]唐晓敏.水分和盐分处理对甘草药材质量的影响[D].北京:北京中医药大学,2008.
[17]张琪,王俊,彭励,等.中波紫外线辐射对甘草光合作用及有效成分积累的影响[J].农业科学研究,2008,29(1):11.
[18]樊国全,杜润清,马生军,等.Mn2+处理对甘草不同器官活性成分含量的比较研究[J].现代中药研究与实践,2017,31(3):9.
[19]林伟雄,乐智勇,车海燕,等.甘草饮片标准汤剂的研究[J].中国中药杂志,2017,42(5):830.
[20]杨明,伍振峰,郑琴,等.中药经典名方开发与制剂研究的关键问题[J].中草药,2010,41(10):1590.