不同加工竹节参中多元活性成分同时测定及灰色关联度分析
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摘要
建立超快速液相色谱-三重四极杆/线性离子阱质谱(UFLC-QTRAP-MS/MS)同时测定竹节参中皂苷、氨基酸及核苷类共33种成分含量的方法,考察不同加工方法对竹节参药材中目标成分的影响。采用XBridgeC_(18)(4.6 mm×100 mm, 3.5μm),以0.1%甲酸水-0.1%甲酸乙腈为流动相,梯度洗脱,流速0.8 mL·min~(-1),柱温30℃,多反应监测离子扫描模式(MRM)测定;用灰色关联度分析(GRA)对不同加工竹节参进行综合分析与评价。结果表明,33种成分在一定浓度范围内均呈现良好的线性关系,相关系数均大于0.999 0;精密度、重复性和稳定性良好;平均加样回收率为95.33%～101.8%,RSD均小于5%。GRA评价结果显示,微波干燥、完整根茎不去皮的竹节参样品综合质量较好。所建立的方法准确、可靠,可用于竹节参药材内在质量的综合评价,该研究可为竹节参适宜产地加工方法的优选及其加工方法的规范化和标准化提供基础资料。
A method, for determination of saponins, amino acids and nucleosides in Panacis Japonici Rhizoma of ultra fast liquid chromatography with triple quadrupole linear ion trap mass spectrometry(UFLC-QTRAP-MS/MS), was established to investigate the effect of different processing methods on the target components of Panacis Japonici Rhizoma. The chromatographic separation was performed on a XBridgeC_(18)(4.6 mm×100 mm, 3.5 μm) at 30 ℃ with a gradient elution of 0.1% formic acid solution-0.1% formic acid acetonitrile, and the flow rate was 0.8 mL·min~(-1), using multiple-reaction monitoring(MRM) mode. The grey relational analysis was adopted for the analysis of different processing samples. The results showed that the thirty-three constituents were in a good linear range and the correlation coefficient was greater than 0.999 0; the precision, repeatability and stability were good; the average recovery rates were between 95.33% and 101.8%, and the relative standard deviations were less than 5%. The result of grey relational analysis showed that the complete rhizomes without peeling, which were adopted for the microwave dried method, had the best quality. The established method was accurate and reliable, which could be used to appraise the quality of Panacis Japonici Rhizoma. Our study may lay the way for the processing method of Panacis Japonici Rhizoma in optimization,normalization and standardization.
A method, for determination of saponins, amino acids and nucleosides in Panacis Japonici Rhizoma of ultra fast liquid chromatography with triple quadrupole linear ion trap mass spectrometry(UFLC-QTRAP-MS/MS), was established to investigate the effect of different processing methods on the target components of Panacis Japonici Rhizoma. The chromatographic separation was performed on a XBridgeC_(18)(4.6 mm×100 mm, 3.5 μm) at 30 ℃ with a gradient elution of 0.1% formic acid solution-0.1% formic acid acetonitrile, and the flow rate was 0.8 mL·min~(-1), using multiple-reaction monitoring(MRM) mode. The grey relational analysis was adopted for the analysis of different processing samples. The results showed that the thirty-three constituents were in a good linear range and the correlation coefficient was greater than 0.999 0; the precision, repeatability and stability were good; the average recovery rates were between 95.33% and 101.8%, and the relative standard deviations were less than 5%. The result of grey relational analysis showed that the complete rhizomes without peeling, which were adopted for the microwave dried method, had the best quality. The established method was accurate and reliable, which could be used to appraise the quality of Panacis Japonici Rhizoma. Our study may lay the way for the processing method of Panacis Japonici Rhizoma in optimization,normalization and standardization.
引文
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[14] 杨守伟, 张志华, 谢贻芳. 腺苷在脊髓痛感受和调制中的作用[J]. 国外医学药理、病理科学与临床分册, 1994, 14(4): 204.
[15] 刘岱琳, 曲戈霞, 王乃利, 等. 瓜蒌的抗血小板聚集活性研究[J]. 中草药, 2004, 35(12): 1334.
[16] Shimizu A, Yano T, Saito Y, et al. Isolation of an inhibitor of platelet aggregation from a fungus, Ganoderma lucidum[J]. Chem Pharm Bull, 1985, 33(7): 3012.
[17] 吕爱娟, 吴皓. 中药中核苷类成分的研究进展[J]. 中国中医药信息杂志, 2006, 13(7): 94.
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[19] 任冰. 人参属药材竹节参和珠子参化学成分指纹图谱研究[D]. 武汉:武汉轻工大学, 2015.
[20] 胡远浪, 袁丁, 何毓敏, 等. 竹节参HPLC-ELSD指纹图谱和化学成分分析[J]. 中国中药杂志, 2010, 35(8): 1009.
[21] 郑菊艳, 邹坤, 陈强, 等. 竹节参质量标准研究[J]. 中药材, 2014, 37(12): 2192.
[22] 王小平,王芳,白吉庆, 等.HPLC同时测定藤珠胃康颗粒中4个皂苷类成分[J].中国现代应用药学,2017,34(4):553.
[23] 赵润怀, 段金廒, 高振江, 等. 中药材产地加工过程传统与现代干燥技术方法的分析评价[J]. 中国现代中药, 2013, 15(12): 1026.
[2] 武秋爽, 陈平, 张庆文. 竹节参化学成分、药理活性及分析方法研究进展[J]. 亚太传统医药, 2016, 12(6): 46.
[3] 李有贵. 竹节人参皂苷对乙醇性肝损伤的保护机理研究[D]. 杭州:浙江大学, 2011: 108.
[4] Dun Y Y, Liu M, Chen J, et al. Regulatory effects of saponins from Panax japonicus on colonic epithelial tight junctions in aging rats[J]. J Ginseng Res, 2017, 42(1):50.
[5] He H B, Xu J, Xu Y Q, et al. Cardioprotective effects of saponins from Panax japonicus on acute myocardial ischemia against oxidative stress-triggered damage and cardiac cell death in rats[J].J Ethnopharmacol, 2012, 140: 73.
[6] 张士善, 张丹参, 朱桐君, 等. 冬虫夏草氨基酸成分的药理分析[J]. 药学学报, 1991, 26(5): 326.
[7] Efron D T, Barbul A, Facs M D, et al. Arginine and immunonutrition: a reevaluation[J]. Nutrition, 2000, 16(1): 73.
[8] 周琼. 甘氨酸对心肌缺血性损伤的防治作用及机制研究[D]. 广州:暨南大学, 2002.
[9] Carini R, Bellomo G, Cesaris M D, et al. Glycine protects against hepatocyte killing by KCN or hypoxia by preventing intracellular Na+ overload in the rat[J]. Hepatology, 1997,26(1): 107.
[10] Schmidt A P, Lara D R, De F M J, et al. Guanosine and GMP prevent seizures induced by quinolinic acid in mice[J]. Brain Res, 2000, 864(1): 40.
[11] 艾中, 钱正明, 李文佳, 等. 冬虫夏草核苷类成分分析研究进展[J]. 菌物学报, 2016, 35(4): 388.
[12] Green P G, Basbaum A I, Helms C, et al. Purinergic regulation of bradykinin-induced plasma extravasation and adjuvant-induced arthritis in the rat[J]. Neurobiology, 1991, 88: 4162.
[13] 蒋激扬, 刘发. 腺苷的抗炎作用[J]. 中国药理学通报, 1998, 14(s1): 79.
[14] 杨守伟, 张志华, 谢贻芳. 腺苷在脊髓痛感受和调制中的作用[J]. 国外医学药理、病理科学与临床分册, 1994, 14(4): 204.
[15] 刘岱琳, 曲戈霞, 王乃利, 等. 瓜蒌的抗血小板聚集活性研究[J]. 中草药, 2004, 35(12): 1334.
[16] Shimizu A, Yano T, Saito Y, et al. Isolation of an inhibitor of platelet aggregation from a fungus, Ganoderma lucidum[J]. Chem Pharm Bull, 1985, 33(7): 3012.
[17] 吕爱娟, 吴皓. 中药中核苷类成分的研究进展[J]. 中国中医药信息杂志, 2006, 13(7): 94.
[18] Wu Q S, Wang C M, Lu J J, et al. Simultaneous determination of six saponins in Panax Japonici Rhizoma using quantitative analysis of multi-components with single-marker method[J]. Curr Pharm Anal, 2017(13): 289.
[19] 任冰. 人参属药材竹节参和珠子参化学成分指纹图谱研究[D]. 武汉:武汉轻工大学, 2015.
[20] 胡远浪, 袁丁, 何毓敏, 等. 竹节参HPLC-ELSD指纹图谱和化学成分分析[J]. 中国中药杂志, 2010, 35(8): 1009.
[21] 郑菊艳, 邹坤, 陈强, 等. 竹节参质量标准研究[J]. 中药材, 2014, 37(12): 2192.
[22] 王小平,王芳,白吉庆, 等.HPLC同时测定藤珠胃康颗粒中4个皂苷类成分[J].中国现代应用药学,2017,34(4):553.
[23] 赵润怀, 段金廒, 高振江, 等. 中药材产地加工过程传统与现代干燥技术方法的分析评价[J]. 中国现代中药, 2013, 15(12): 1026.