目的:建立以环糊精为流动相添加剂的高效液相色谱法测定山茱萸中熊果酸和齐墩果酸,为山茱萸的质量控制提供依据。方法:运用分子设计和效应面法优化山茱萸中熊果酸和齐墩果酸含量测定的方法,采用安捷伦Eclipse XDB C18(150 mm×4.6 mm,5 μm)色谱柱,以乙腈-3 mmol·L-1 γ-环糊精溶液(含0.05%磷酸)(60∶40)为流动相,流速1.0 mL·min-1,柱温25 ℃,检测波长210 nm。结果:熊果酸和齐墩果酸的分离度为3.81,熊果酸进样量在400~4 000 ng,齐墩果酸进样量在200~2 000 ng范围内线性关系良好(r=0.999 7和0.999 9),精密度RSD≤1.4%,重现性RSD≤1.8%,稳定性RSD≤1.7%,回收率分别为98.5%(RSD=1.7%)和99.1%(RSD=2.0%),3批山茱萸药材中熊果酸含量分别为2.66、2.35和2.91 mg·g-1,齐墩果酸含量分别为0.83、0.78和1.12 mg·g-1。结论:该方法简便,准确可靠,且经济环保,可作为山茱萸的质量控制方法。
Objective: To establish high-performance liquid chromatography method using cyclodextrin as a mobile phase additive for the determination of ursolic acid and oleanolic acid in Corni Fructus, providing a basis for quality control of Corni Fructus. Methods: Molecular design and response surface method were used to optimize the determination method of ursolic acid and oleanolic acid in Corni Fructus. Agilent Eclipse XDB C18 column (150 mm×4.6 mm, 5 μm) was adopted as the stationary phase, the mobile phase was 3 mmol·L-1 γ-cyclodextrin solution (contained 0.05% phosphoric acid)-acetonitrile (60∶40), the flow rate was 1.0 mL·min-1, the column temperature was 25 ℃ and the detecting wavelength was 210 nm. Results: The resolution of chromatographic peaks of ursolic acid and oleanolic acid was 3.81. Linearities were good in the range of 400-4 000 ng for ursolic acid and in the range of 200-2 000 ng for oleanolic acid. The RSDs for precision, reproducibility, and stability were less than 1.4%, 1.8%, and 1.7%, respectively. The recovery rates were 98.5% (RSD=1.7%) and 99.1% (RSD=2.0%). The contents of ursolic acid in three batches of Corni Fructus were 2.66 mg·g-1, 2.35 mg·g-1 and 2.91 mg·g-1, and those of oleanolic acid were 0.83 mg·g-1, 0.78 mg·g-1and 1.12 mg·g-1, respectively. Conclusion: This method is simple, accurate, reliable, and environmental friendly, and can be used as a quality control for Corni Fructus.
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