HPLC法测定紫花地丁中槲皮素、芹菜素、木犀草素含量

目的:建立高效液相色谱法测定紫花地丁中槲皮素、芹菜素和木犀草素的含量测定方法。方法:采用HPLC法对紫花地丁中的槲皮素、芹菜素、木犀草素进行含量测定。色谱条件为:C18柱(5μm,4.6 mm×250 mm),柱温30℃,流动相为乙腈-甲醇-0.2%磷酸水溶液梯度洗脱,流速0.8 mL.min-1,检测波长350 nm。结果:槲皮素、芹菜素和木犀草素色谱分离良好,浓度与峰面积呈良好线性关系,线性范围分别为槲皮素7.5~90.0 mg.L-1,r=0.999 9(n=6);芹菜素7.5~180.0 mg.L-1,r=0.999 8(n=6),木犀草素2.5~55.0 mg.L-1,r=0.999 9(n=6)。平均加样回收率分别为95.1%(n=9,RSD=1.0%)、95.7%(n=9,RSD=1.2%)、97.8%(n=9,RSD=0.7%)。结论:本方法测定了5批紫花地丁的槲皮素、芹菜素和木犀草素的含量,该方法分离度好、快速、简便、重现性好。

Determination of Quercetin, Apigenin and Luteolin in Viola yedoensis Makino by RP-HPLC

Objective： To establish an HPLC method for determination of Quercetin, Apigenin and Luteolin in Viola yedoensis Makino. Methods： HPLC analysis was performed on C18 column （4. 6 mm× 250 mm, 5 μm） with the methanol potassium dihydrogen phosphate as mobile phase in gradient mode, the column temperature was set at 30℃, the flow rate was 0. 8 mL ·min^-1 and the detecting wavelength was at 350 nm. Results ： The linear response ranges from 7.5 to 90.0 mg·L^-1 of quercetin （ r = 0. 999 9, n = 6 ） and from 7.5 to 180. 0 mg·L^-1 of apigenin （r =0. 999 8,n =6） and from 2. 5 to 55.0·L^-1 of luteolin（r =0. 999 9,n =6） ; the recoveries of quercetin, apigenin and luteolin were 95.1% （ n = 9, RSD = 1.0% ） , 95.7% （ n = 9, RSD = 1.2% ） , 97.8% （ n = 9, RSD = 0. 7% ）, respectively. Conclusion： All 5 samples were determined by HPLC. The method is a practical method in determination of Quereetin, Apigenin and Luteolin in Viola yedoensis Makino.