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目的:建立HPLC法同时测定萱草花蕾中芦丁、槲皮素、山柰酚3个黄酮类化合物的含量,并比较不同产地萱草中这3个化合物的含量。方法:采用Agilent SB-C18(4.6 mm×250 mm,5μm)色谱柱,以甲醇(A)-pH 3磷酸水溶液(B)为流动相,梯度洗脱(0～10 min,20%A;10～10.5 min,20%A→45%A;10.5～27.5 min,45%A;27.5～28 min,45%A→60%A;28～45min,60%A→20%A),流速0.7 mL.min-1,检测波长360 nm,柱温33℃,进样量20μL。结果:芦丁、槲皮素及山柰酚浓度分别在32.6～326μg.mL-1(r=0.9999)、1.06～10.6μg.mL-1(r=0.9997)和0.32～3.2μg.mL-1(r=0.9997)范围内线性关系良好;平均加样回收率(n=6)分别为99.53%(RSD=1.8%),96.96%(RSD=1.7%),98.75%(RSD=1.9%)。结论:该方法简便,操作简单,结果准确,重复性好,为评价萱草药材的质量提供了可靠的分析方法,可作为萱草中黄酮类化合物的含量测定方法。
OBJECTIVE: To establish a HPLC method for the simultaneous determination of three flavonoid compounds, rutin, quercetin and kaempferol in flower buds of Hemerocallis tuberosus and to compare the contents of these three compounds in Hemerocallis of different origins. Methods: The mobile phase was eluted with gradient elution (0-10 min, 20% A, 10%) with Agilent SB-C18 column (4.6 mm × 250 mm, ~ 10.5 min, 20% A → 45% A; 10.5-27.5 min, 45% A; 27.5-28 min, 45% A → 60% A; 28-45 min, 60% A → 20% .min-1, detection wavelength 360 nm, column temperature 33 ℃, injection volume 20μL. Results: The concentrations of rutin, quercetin and kaempferol were 32.6 ~ 326μg.mL-1 (r = 0.9999), 1.06-10.6μg.mL-1 (r = 0.9997) and 0.32 ~ 3.2μg.mL-1 (r = 0.9997). The average recoveries (n = 6) were 99.53% (RSD = 1.8%), 96.96% (RSD = 1.7%) and 98.75% (RSD = 1.9%) respectively. Conclusion: The method is simple, simple, accurate and reproducible. It provides a reliable analytical method for evaluating the quality of Hemerocallis tuberosus and can be used as a method for determination of flavonoids in.