目的 基于“蛛网”模式,采用UHPLC-Q-Exactive-Orbitrap-MS法建立甘松中6种倍半萜成分的含量测定方法。方法 采用“蛛网”模式,确定甘松中6种倍半萜的提取方法;采用Waters CORTECSTM C18色谱柱(4.6 mm×150 mm,2.7 μm);以乙腈为流动相A,体积分数0.05%甲酸溶液(含5 mmol·L-1甲酸铵)为流动相B,梯度洗脱,流速0.5 mL·min-1;柱温35 ℃;电喷雾正离子模式进行检测。结果 “蛛网”结果显示,最佳提取条件为甲醇提取,料液比1∶12.5,加热回流30 min。去氧甘松醇A、甘松新酮二醇、甘松香酮A、Axinysone B、甘松新酮、甘松香酮H分别在0.386 4~1.425 6、0.366 3~2.424 5、0.462 1~1.458 8、0.639 7~1.783 2、3.542 5~14.657 6和0.432 7~1.439 3 μg·mL-1内与峰面积积分值的线性关系良好,相关系数r均不小于0.999 3;6个成分的平均加样回收率为101.42%、99.48%、95.58%、97.10%、102.34%和97.75%;将结果通过聚类和偏最小二乘法(PLS-DA)进行分析评价,结果显示甘松的潜在差异标志物为甘松新酮、甘松香酮A和甘松香酮H。结论 本研究建立的方法能够实现甘松中6种倍半萜成分的快速、准确定量分析,可以为甘松的质量评价提供参考。
Abstract
OBJECTIVE To establish a UHPLC-Q-Exactive-Orbitrap-MS method to determine the contents of six sesquiterpenes in Nardostachys jatamansi DC. based on the ‘spider web' model. METHODS The extraction methods of six sesquiterpenes in Nardostachys jatamansi DC. were studied and determined by cobweb mode. Waters CORTECSTM C18 chromatographic column (4.6 mm×150 mm, 2.7 μm) was used. Acetonitrile was used as mobile phase A, and 0.05% formic acid solution containing 5 mmol·L-1 ammonium formate was used as mobile phase B. Gradient elution was conducted at a flow rate of 0.5 mL·min-1. The column temperature was maintained at 35 ℃. Detection was carried out in electrospray positive ion mode. RESULTS The results of spider web showed that the optimum extraction conditions were methanol extraction∶solid-liquid=1∶12.5, heating reflux for 30 min. Deoxyeugenol A, nardosinonediol, nardosinone A, axinysone B, nardosinone and nardosinone H had good linear relationship with the peak area integral value in the range of 0.386 4-1.425 6, 0.366 3-2.424 5, 0.462 1-1.458 8, 0.639 7-1.783 2, 3.542 5-14.657 6 and 0.432 7-1.439 3 μg·mL-1, respectively, and the correlation coefficients (r) were not less than 0.999 3. The average recoveries of the six components were 98.39%, 100.05%, 100.35%, 97.13%, 102.84% and 97.15%, respectively. The results were analyzed and evaluated by clustering and partial least squares (PLS-DA). The results showed that the potential differential markers of Nardostachys jatamansi DC were nardosinone, nardosinone A and nardosinone H. CONCLUSION The method established in this study can realize the rapid and accurate quantitative analysis of six sesquiterpenes in Nardostachys jatamansi DC, which can provide reference for the quality evaluation of Nardostachys jatamansi DC.
关键词
甘松 /
多成分含量测定 /
“蛛网”模式 /
多元统计分析 /
高分辨质谱
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Key words
Nardostachys jatamansi DC. /
multi-component content determination /
‘spider web' model /
multivariate statistical analysis /
high resolution mass spectrometry
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中图分类号：
R284
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脚注
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基金
国家重点研发计划(中医药现代化)课题资助(2023YFC3504102);“新高校20条”科研带头人工作室项目资助(202228096);泉城产业领军人才支持计划创新团队项目资助(MRJT2105)
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