HPLC法同时测定蒙药秦艽花中龙胆苦苷、异荭草苷、牡荆素的含量及其指纹图谱的建立*

作者：牧其尔，白艳秋，乌吉斯古楞，齐珺伲，萨础拉

单位：内蒙古医科大学蒙医药学院，内蒙古 呼和浩特 010110

引用：引用：牧其尔，白艳秋，乌吉斯古楞，齐珺伲，萨础拉.HPLC法同时测定蒙药秦艽花中龙胆苦苷、异荭草苷、牡荆素的含量及其指纹图谱的建立[J].中医药导报,2025,31(12):83-89.

DOI：10.13862/j.cn43-1446/r.2025.12.013

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摘要：

目的：建立一种高效液相色谱（HPLC）法，同时测定蒙药秦艽花中龙胆苦苷、异荭草苷和牡荆素的含量，并结合指纹图谱技术对其质量进行综合评价。方法：色谱柱采用Agilent Eclipse XDB-C18柱（250.0 mm×4.6 mm，5.0 μm），流动相：0.1%磷酸水溶液-乙腈，梯度洗脱，流速：1.0 mL/min，柱温：30 ℃，进样量：10 μL。含量测定检测波长为270 nm（龙胆苦苷）、350 nm（异荭草苷、牡荆素），指纹图谱检测波长为240 nm。结果：蒙药秦艽花中龙胆苦苷、异荭草苷、牡荆素的质量浓度分别在0.018 4～0.294 0、0.003 1～0.049 9、0.003 1～0.049 2 mg/mL范围内与色谱峰面积呈良好的线性关系（r≥0.999 5），平均回收率分别为102.00%、106.72%，100.10%，RSD值分别为1.87%、1.10%、1.31%（n=3）。在所建立的蒙药秦艽花HPLC指纹图谱中，以S1号样品色谱图为参照图谱，通过多点矫正和全谱峰匹配，生成对照指纹图谱，共确定15个共有峰，经与对照品比对指认了其中7个成分。聚类分析、主成分分析、正交偏最小二乘法-判别分析（OPLS-DA）及VIP结果表明，陕西产地的秦艽花较其他产地样品存在显著差异，其中龙胆苦苷含量差异最大，异荭草苷与牡荆素含量也有所差异，但幅度不大。结论：本研究所建立的方法为蒙药秦艽花的质量控制、产地鉴别及药用价值评估提供了科学依据，并为其进一步的资源开发和利用奠定了基础。

关键词：秦艽花；含量测定；龙胆苦苷；异荭草苷；牡荆素；指纹图谱；高效液相色谱

Abstract：

Objective: To establish a high-performance liquid chromatographic (HPLC) method for the simultaneous determination of gentiopicroside, isoorientin and vitexin in the Mongolian medicinal Qinjiaohua (Gentiana Mmacrophylla Flower), and to integrate it with a fingerprinting technique for comprehensive quality assessment. Methods: Chromatographic separation was performed on an Agilent Eclipse XDB-C18 column (250.0 mm×4.6 mm, 5.0 μm) with a mobile phase consisting of 0.1% aqueous phosphoric acid solution-acetonitrile under gradient elution conditions. The flow rate was 1.0 mL/min, and the column temperature was maintained at 30 ℃, with the injection volume of 10 μL. The detection wavelength of the content test was 270 nm (gentiopicroside), and 350 nm (isoorientin and vitexin), and the detection wavelength of the fingerprint was 240 nm. Results: The mass concentrations of gentiopicroside, isoorientin, and vitexin in Mongolian medicine Qinjiaohua (Gentiana Mmacrophylla Flower) demonstrated excellent linearity with their respective chromatographic peak areas over the ranges of 0.018 4-0.294 0, 0.003 1-0.049 9, and 0.003 1-0.049 2 mg/mL respectively, with a good linear relationship with the chromatographic peak area (r≥0.999 5). The average recoveries were 102.00%, 106.72%, and 100.10%, and the relative standard deviations (RSD) were 1.87%, 1.10%, and 1.31%, respectively (n=3). The HPLC fingerprints of the Mongolian medicine Qinjiaohua (Gentiana Mmacrophylla Flower) were established. With the chromatogram of sample S1 as the reference chromatogram, the control fingerprints were generated through multi-point correction and full-spectrum peak matching, and a total of 15 common peaks were identified, of which 7 components were identified by comparison with the standards. The results of cluster analysis, principal component analysis, orthogonal partial least squares discriminant analysis (OPLS-DA), and variable importance in the projection (VIP) showed that the Qinjiaohua (Gentiana Mmacrophylla Flower) from Shaanxi origin was significantly different from the samples from other regions, with the most significant difference in the content of gentiopicroside, while the contents of isoorientin and isovitexin also showed differences, albeit to a lesser extent.  Conclusion: The method established in this study provided a scientific basis for the quality control, identification of producing area and evaluation of medicinal value of Mongolian medicine Qinjiaohua (Gentiana Mmacrophylla Flower) and laid a foundation for the further resource development and utilization.

Key words：Qinjiaohua (Gentiana Mmacrophylla Flower); content determination; gentiopicroside; isoorientin; vitexin; fingerprint; high-performance liquid chromatographic

发布时间：2026-01-30

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