[关键词]
[摘要]
目的 建立同时测定珍珠菜提取物中芦丁、异鼠李素-3-O-芸香糖苷、江户樱花苷、槲皮素、山柰酚的HPLC方法。方法 采用Chromasil C18色谱柱(250 mm×4.6 mm,5.0 μm),柱温35 ℃,以乙腈-0.1%磷酸水溶液为流动相,梯度洗脱,双波长检测(λ1=283 nm,λ2=370 nm),体积流量1.0 mL/min。结果 5个成分均能达到基线分离,线性回归方程分别为芦丁Y=14 958 X+179.22(r=0.999 3),异鼠李素-3-O-芸香糖苷Y=12 126 X+3.14(r=0.999 4),江户樱花苷Y=23 821 X+76.81(r=0.999 4),槲皮素Y=35 761 X-20.30(r=0.999 5),山柰酚Y=39 078 X+1.81(r=0.999 1),芦丁、异鼠李素-3-O-芸香糖苷、江户樱花苷、槲皮素、山柰酚进样量分别在228.60~1 143.00、99.60~498.00、232.20~1 161.00、22.08~110.40、15.12~75.60 ng与峰面积线性关系良好,平均回收率分别为97.8%、98.9%、102.4%、98.4%、92.2%。结论 本检测方法简便、准确,为珍珠菜提取物中黄酮类成分的质量控制提供了依据。
[Key word]
[Abstract]
Objective To develop an HPLC method for determination of five flavonoids (rutin, isorhamnetin-3-O-rutinoside, prunin, quercetin, and kaempferol) in extract of Lysimachia clethroides. Methods HPLC was performed on a Chromasil C18 analytical column (250 mm × 4.6 mm, 5.0 μm) at 35 ℃ with acetonitrile-0.1% phosphoric acid solution as the mobile phase by gradient elution. The detection wavelengths were 283 and 370 nm and the flow rate was 1.0 mL/min. Results Five flavonoids were separated perfectly. The equation linear regressions were rutin Y=14 958 X+179.22 (r=0.999 3), isorhamnetin-3-O-rutinoside Y=12 126 X+3.14 (r=0.999 4), prunin Y=23 821 X+76.81 (r=0.999 4), quercetin Y=35 761 X-20.30 (r=0.999 5), and kaempferol Y=39 078 X+1.81 (r=0.999 1). Linearities of rutin, isorhamnetin-3-O-rutinoside, prunin, quercetin, and kaempferol were good in ranges of 228.60―1143.00, 99.60―498.00, 232.20―1161.00, 22.08―110.40, and 15.10―75.60 ng, respectively. The average recoveries were 97.8%, 98.9%, 102.4%, 98.4%, and 92.2%, respectively. Conclusion The validated method is simple, accurate, and able to provide the basis for the quality control of flavonols in active fraction of L. clethroides.
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[基金项目]
国家自然科学基金资助项目(30873362);“重大新药创制”科技重大专项资助项目(2009ZX09102-135)