目的:建立青海产龙胆科藏药材大花龙胆、蓝玉簪龙胆、线叶龙胆、喉毛花中8个活性成分的UPLC分析方法,比较其在4种藏药材中的差异。方法:超高效液相色谱法,采用InfinityLab Poroshell 120 SB-C8(150 mm×2.1 mm,2.7 μm)色谱柱,乙腈(B)-0.05%乙酸水溶液(C)梯度洗脱,流速0.2 mL·min-1,检测波长254 nm,柱温25 ℃。结果:马钱苷酸、獐芽菜苦苷、龙胆苦苷、獐芽菜苷、异荭草素、异金雀花素-2″-O-吡喃糖苷、异牡荆素及异金雀花素8个成分在线性范围内相关系数为0.999 8~1.000,平均回收率(n=6)98.4%~102.1%,RSD为0.77%~2.3%。8个活性成分在4种龙胆科植物中的总含量,喉毛花最高,为85.63 mg·g-1,其次是线叶龙胆龙胆66.01 mg·g-1和蓝玉簪龙胆42.11 mg·g-1,大花龙胆最低,为19.33 mg·g-1。4种龙胆科藏药都含有马钱苷酸、异牡荆素,但其含量均较低。蓝玉簪龙胆中龙胆苦苷和獐牙菜苷含量较高,均超过10 mg·g-1,线叶龙胆中龙胆苦苷含量最高,为(19.26±1.67) mg·g-1,均不含异金雀花素-2″-O-吡喃糖苷。大花龙胆中未检出獐芽菜苦苷、龙胆苦苷和獐芽菜苷,但异金雀花素-2″-O-吡喃糖苷和异金雀花素含量较高,达(7.99±0.09) mg·g-1和(6.77±0.03) mg·g-1;喉毛花中獐芽菜苦苷含量最高,为(76.79±7.42) mg·g-1;但未检出龙胆苦苷和异红草素。结论:所建立的方法简便、准确,可用于4种龙胆科藏药的质量评价并为其资源的深入研究和利用提供参考。
Objective: To establish an UPLC method for the analysis of eight active components Gentiana szechenyii Kanitz, Gentiana veitchiorum Hemsl, Gentiana lawrencei var. farreri and Comastoma pulmonarium, and to compare the differences of active components in the four Tibetan medicinal materials. Methods: A UPLC method was set up, an InfinifityLab Poroshell 120 SB-C8 (150 mm×2.1 mm,2.7 μm)chromatographic column was used, the mobile phase was acetonitrile (B)-0.05% acetic acid aqueous solution (C) with a flow rate of 0.2 mL·min-1, gradient elution, detection wavelength of 254 nm, and column temperature of 25 ℃. Results: The linear regression equation correlation coefficient of the eight components of loganic acid, swertiamarine, gentiopicroside, sweroside, isoorientin, isoscoparin-2″-O-glucopyranoside, isovitexin, isoscoparin was 0.999 8-1.000, the average recovery (n=6) was 98.4%-102.1%, and the RSD was 0.77%-2.3%. The total contents of 8 active components in 4 species of Gentianaceae plants were as below: the highest content was 85.63 mg·g-1 in Comastoma pulmonarium, followed by 66.01 mg·g-1 in Gentiana veitchiorum Hemsl, 42.11 mg·g-1 in P. Gentiana lawrencei var. farreri, and 19.33 mg·g-1 in Gentiana szechenyii Kanitz. All four gentianaceae Tibetan medicines contain loganic acid and isovitexin, but their contents were low. The contents of gentiopicroside and sweroside in Gentiana scabra were high, both exceeding 10 mg·g-1. The content of gentiopicroside in Gentiana rigescens was the highest of (19.26±1.67) mg·g-1, and they all did not contain isorkinin-2″-O-pyranoside. Swertiamarin, gentiopicroside. Sweroside were not detected in Gentiana macrophylla, but the contents of isoflorin-2″-O-pyranoside and isoflorin were higher, reaching (7.99±0.09) mg·g-1 and (6.77±0.03) mg·g-1, respectively. The highest content of swertiamarin was (76.79±7.42) mg·g-1, but gentiopicroside and isofraxidin were not detected. Conclusion: The established method is simple, accurate and can be used for the quality evaluation of the four gentianaceae Tibetan medicines. The content analysis results of eight active components in four gentianaceae Tibetan medicinal materials from Qinghai can provide reference for the in-depth study and utilization of their resources.
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