一测多评法同时测定龙胆不同部位中3个环烯醚萜苷类成分的含量

目的：建立一测多评法同时测定龙胆药材中马钱苷酸、獐牙菜苦苷及龙胆苦苷的含量，并比较龙胆药材上（根茎）、中（簇生根中部）、下（簇生根尾部）三部分中3个成分的含量差异。方法：采用HPLC法，使用Agilent Eclipse XDB-C₁₈（4.6 mm&#215;250 mm，5 μm）色谱柱，流动相A为0.1%磷酸水溶液，流动相B为乙腈，梯度洗脱（0~5 min，6% B；5~35 min，6% B→3.5% B；35~65 min，3.5% B→9% B；65~85 min，9% B→6% B），流速0.8 mL&#183;min^-1，柱温40℃，检测波长240 nm，进样量10 μL。以龙胆苦苷为内参物，建立其与马钱苷酸、獐牙菜苦苷的相对校正因子，并计算2个成分分别在药材上、中、下三部分的含量，实现一测多评；同时采用外标法测定龙胆中3个环烯醚萜苷类成分分别在药材上、中、下三部分的含量，比较2种测定方法的差异，验证一测多评法的可行性，采用SPSS21.0软件分别对龙胆药材上、中、下三部分中3个成分的含量进行单因素方差分析。结果：建立的校正因子重复性良好，含量间无显著差异。龙胆药材不同部位中3个环烯醚萜苷类成分的含量差异显著，中、下部的马钱苷酸、獐牙菜苦苷及龙胆苦苷含量均高于上部（P<0.05，有显著性差异）。结论：建立的一测多评法可作为龙胆药材中3个环烯醚萜苷类化学成分的含量测定方法。通过分析龙胆药材上、中、下三部分环烯醚萜苷类成分的含量差异，初步判断龙胆药材质量优劣的合理性，为龙胆药材等级划分及制订龙胆药材的质量评价提供参考依据，在其生产及商品流通等方面也具有一定的指导意义。

Objective:To establish a QAMS method for content determination of three compositions(loganic acid,sweroside and gentiamarin)in Gentiana,and to compare the contents in upper(rhizome),middle(middle of tufted roots)and lower(tufted root tail)parts of the herb. Methods:The HPLC analysis was performed on an Agilent Eclipse XDB-C₁₈ column(4.6 mm&#215;250 mm,5 μm). The mobile phase A was 0.1% aqueous phosphate solution,the mobile phase B was acetonitrile,and gradient elution was used(0-5 min,6%B;5-35 min,6%B→3.5% B;35-65 min,3.5%B→9% B;65-85 min,9% B-6% B)at a flow rate of 0.8 mL&#183;min^-1. The detection wavelength was 254 nm,the column temperature was 30℃ and the injection amount was 10 μL. Using gentiamarin as the external standard,relative correction factors for loganic acid and sweroside were calculated. The contents of the two components in three parts of the herb were determined by QAMS method. At the same time,determination of the three iridoid glycosides in three parts of Gentiana was performed by the external standard method to evaluate the feasibility of the QAMS method. Single factor variance analysis was performed on contents of the three components in upper,middle and lower parts of Gentiana using SPSS 21.0 software. Results:The calibration factors were reproducible and there was no significant difference between the QAMS method and the external standard method. The contents of three iridoid glycosides in different parts of Gentiana were significantly different. The contents of loganin,swertiamarin and gentiopicroside in the middle and lower parts were higher than those in the upper parts(P<0.05,with significant difference). Conclusion:The established method can be used for the determination of three iridoid glycosides in Gentiana. Through the analysis of the difference in contents in the three parts of Gentiana,the rationality of quality of Gentiana is preliminarily judged,which can provide reference for the classification of Gentiana and the quality evaluation of Gentiana. It also has certain guiding significance in the production and commodity circulation of Gentianae scabra.

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