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[超高效液相色谱-四极杆飞行时间串联质谱法同时测定和分析石斛中的氨基酸和核苷]

[Simultaneous determination and analysis of amino acids and nucleosides in Dendrobium by UHPLC-QTRAP-MS/MS].

作者信息

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

机构信息

Anhui University of Chinese Medicine Hefei 230012, China Institute of Traditional Chinese Medicine Resources Protection and Development, Anhui Academy of Chinese Medicine Hefei 230012, China.

Anhui University of Chinese Medicine Hefei 230012, China Institute of Traditional Chinese Medicine Resources Protection and Development, Anhui Academy of Chinese Medicine Hefei 230012, China Synergetic Innovation Center of Anhui Authentic Chinese Medicine Quality Improvement Hefei 230012, China Anhui Province Key Laboratory of Chinese Medicinal Formula Hefei 230012, China.

出版信息

Zhongguo Zhong Yao Za Zhi. 2020 Aug;45(16):3890-3899. doi: 10.19540/j.cnki.cjcmm.20200513.201.

Abstract

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.

摘要

通过运用多元统计分析来评估内在品质,并为其综合利用提供科学依据,我们建立了一种超高效液相色谱-四极杆飞行时间串联质谱法(UHPLC-QTRAP-MS/MS),用于快速、准确、高效地测定不同种石斛中21种氨基酸和10种核苷。分析在Waters XBridge酰胺柱(2.1 mm×100 mm,3.5 μm)上进行,流动相为水-乙腈中的0.2%甲酸,以0.2 mL·min⁻¹的流速洗脱,柱温为30℃。目标化合物采用正离子多反应监测(MRM)模式进行分析。通过主成分分析(PCA)和逼近理想解排序法(TOPSIS)分析对不同种石斛进行综合评价。所有21种氨基酸和10种核苷在一定浓度范围内均呈现良好的线性关系(r>0.999),稳定性、精密度和重复性试验的相对标准偏差(RSD)均小于3.0%。回收率在93.31%至107.5%之间,RSD在1.1%至3.7%之间。PCA得到的综合评价指标表明,霍山石斛在氨基酸方面显著高于其他品种,而铁皮石斛在核苷方面高于其他品种。TOPSIS的最大Ci差异为68.7%,综合评价表明霍山石斛的综合品质最高。该方法准确、快速且高效,可为石斛的进一步研究和内在质量控制提供可靠依据。

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