中国中药杂志

2020, v.45(16) 3890-3899

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基于UHPLC-QTRAP-MS/MS的石斛中氨基酸和核苷类成分分析与评价
Simultaneous determination and analysis of amino acids and nucleosides in Dendrobium by UHPLC-QTRAP-MS/MS

刘峻麟;俞年军;邢丽花;王瑞;徐君;彭代银;童小慧;欧金梅;何祥林;
LIU Jun-lin;YU Nian-jun;XING Li-hua;WANG Rui;XU Jun;PENG Dai-yin;TONG Xiao-hui;OU Jin-mei;HE Xiang-lin;Anhui University of Chinese Medicine;Institute of Traditional Chinese Medicine Resources Protection and Development, Anhui Academy of Chinese Medicine;Synergetic Innovation Center of Anhui Authentic Chinese Medicine Quality Improvement;Anhui Province Key Laboratory of Chinese Medicinal Formula;Huoshan Changchong Chinese Medicinal Material Development Co., Ltd.;

摘要(Abstract):

使用超高效液相色谱-三重四极杆离子阱质谱联用技术(UHPLC-QTRAP-MS/MS)快速、准确、高效直接测定8种不同石斛(共9批次)中21种氨基酸和10种核苷类成分的方法,结合多元统计分析进行综合质量评价,为石斛资源综合利用提供科学依据。使用Waters XBridge Amide色谱柱(2.1 mm×100 mm,3.5μm),以0.2%甲酸水-0.2%甲酸乙腈为流动相进行梯度洗脱,流速0.2 mL·min~(-1),柱温30℃;电喷雾离子源(ESI)正离子模式下,多反应监测(MRM)模式测定。采用主成分分析(PCA)和逼近理想点排序法(TOPSIS)对实验数据进行综合评价。21种氨基酸和10种核苷类被测成分在一定浓度范围内线性关系均良好,相关系数大于0.999,稳定性、精密度、重复性RSD均小于3.0%,加样回收率在93.31%~107.5%,RSD 1.1%~3.7%。PCA结果显示,以氨基酸为指标,评分最高为霍山石斛(S1);以核苷类成分为指标,评分最高为铁皮石斛(S6);TOPSIS法中C_i最大差异达68.7%,综合评价显示霍山石斛整体品质最优。该实验方法准确、快速、高效;熵权TOPSIS分析方法将多维问题转变为一维问题,提高了决策分析的准确性与科学性。为进一步探索评价石斛品种的内在质量及控制提供依据,同时为其资源的进一步开发与利用提供借鉴。
By using multivariate statistical analysis to evaluate essential quality, and provide scientific basis for their comprehensive utilization, we established an UHPLC-QTRAP-MS/MS method for the fast, precise, efficient determination of 21 kinds of amino acids and 10 kinds of nucleosides in different species of Dendrobium. The analysis was performed on a Waters XBridge Amide column(2.1 mm×100 mm,3.5 μm) with elution by mobile phase of 0.2% formic acid in water-0.2% formic acid in acetonitrile at a flow rate of 0.2 mL·min~(-1) with the column temperature at 30 ℃. The target compounds were analyzed by the positive ion multiple reaction monitoring(MRM) mode. The comprehensive evaluation of different species of Dendrobium was carried out by PCA and TOPSIS analysis. All 21 kinds of amino acids and 10 nucleosides showed good linearity among certain concentration range(r>0.999), the RSDs of the stability, precision, and repeatability tests were less than 3.0%. The recovery rate was in the range from 93.31% to 107.5%, and RSD was in the range of 1.1%-3.7%. The comprehensive evaluation index obtained with PCA showed that D. huoshanense was significantly higher than others regarding amino acids and D. officinale has higher nucleosides than other species. The biggest C_i difference of TOPSIS was 68.7%, and comprehensive evaluation showed that D. huoshanense produced the highest comprehensive quality. The method is precise, fast and efficient and can provide reliable basis for further researches and intrinsic quality control of Dendrobium.

关键词(KeyWords): 石斛;氨基酸;核苷;含量测定;UHPLC-QTRAP-MS/MS
Dendrobium;amino acids;nucleoside;content determination;UHPLC-QTRAP-MS/MS

Abstract:

Keywords:

基金项目(Foundation): 国家自然科学基金联合基金项目(U19A2009);; 安徽高校协同创新项目(GXXT-2019-043);; 安徽道地中药材品质提升协同创新中心项目(皖教科[2013]2号);; 安徽省教育厅项目(皖教秘科[2014]44号);; 安徽中医药大学自然重点项目(2019zrzd02);; 霍山石斛种苗繁育和栽培技术林业标准化示范区建设项目(Z175070050002)

作者(Author): 刘峻麟;俞年军;邢丽花;王瑞;徐君;彭代银;童小慧;欧金梅;何祥林;
LIU Jun-lin;YU Nian-jun;XING Li-hua;WANG Rui;XU Jun;PENG Dai-yin;TONG Xiao-hui;OU Jin-mei;HE Xiang-lin;Anhui University of Chinese Medicine;Institute of Traditional Chinese Medicine Resources Protection and Development, Anhui Academy of Chinese Medicine;Synergetic Innovation Center of Anhui Authentic Chinese Medicine Quality Improvement;Anhui Province Key Laboratory of Chinese Medicinal Formula;Huoshan Changchong Chinese Medicinal Material Development Co., Ltd.;

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