基于响应面法对藏药线叶龙胆不同部位中齐墩果酸和熊果酸的提取方法研究OA

Objective:This paper aims to establish a high-performance liquid chromatography(HPLC)method for determining the contents of oleanolic acid and ursolic acid in the Tibetan medicine Gentiana lawrencei var.farreri,optimize the extraction process,and provide scientific evidence for the quality evaluation and rational development and utilization of the medicine.Methods:The extraction process of oleanolic acid and ursolic acid from G.lawrencei var.farreri was optimized by Box-Behnken design response surface methodology,and the factors such as drug-liquid ratio,extraction temperature,and ultrasonic time were optimized.An Agilent Zorbax XDB-C18 chromatographic column(250 mm×4.6 mm,5 μm)was used with the mobile phase of methanol and 0.05 mol·L-1 ammonium acetate solution(88∶12)at a flow rate of 0.6 mL·min-1,a column temperature of 25℃,a detection wavelength of 215 nm,and an injection volume of 10 μL.Results:When methanol was used as the extraction solvent,the extraction efficiency was the highest at a solid-liquid ratio of 1∶40,an extraction temperature of 29℃,and an ultrasonic time of 24 min.Both components achieved baseline separation under the above chromatographic conditions.The linear equations for oleanolic acid and ursolic acid were Y=516 027X-25.425(r=0.999 9)and Y=489 438X-48.954(r=0.999 8),respectively.The average recoveries were 103.30%(RSD=0.81%)for oleanolic acid and 99.52%(RSD=2.43%)for ursolic acid.The results of comparison of oleanolic acid and ursolic acid contents in different parts of G.lawrencei var.farreri showed that the flower exhibits the highest mass fraction(23.302 mg·g-1),followed by stems and leaves(18.419 mg·g-1)and the whole herb(18.457 mg·g-1),and that the contents of the latter two were similar.Conclusions:The extraction process of G.lawrencei var.farreri optimized via response surface methodology is stable and reliable,and it is suitable for the extraction of oleanolic acid and ursolic acid from G.lawrencei var.farreri.At the same time,the contents of oleanolic acid and ursolic acid in G.lawrencei var.farreri were determined by the HPLC method.The method is accurate,rapid,and stable,and it can be used for the quality control of G.lawrencei var.farreri.