HPLC法测定山野豌豆与狭山野豌豆中4个黄酮成分

Determination of four flavonoids in Vicia amoena Fisch.and Vicia amoena Fisch.var.angusta Freyn.by HPLC

目的:建立HPLC法测定山野豌豆与狭山野豌豆中山柰酚-3-O-α-L-鼠李糖苷、槲皮素、苜蓿素、山柰酚的含量,为山野豌豆与狭山野豌豆的质量控制提供依据。方法:采用Diamonsil C_(18)色谱柱(200 mm×4.6 mm,5μm),以乙腈-0.1%磷酸水溶液梯度洗脱,流速1.0mL·min^(-1),检测波长350 nm,柱温25℃,进样量10μL。结果:山柰酚-3-O-α-L-鼠李糖苷、槲皮素、苜蓿素、山柰酚的检测限分别为0.20、0.21、0.21、0.20μg·mL^(-1),线性范围分别为6.85~1 369.60μg·mL^(-1)(r=0.999 9)、6.34~1 267.20μg·mL^(-1)(r=0.999 8)、3.33~665.60μg·mL^(-1)(r=0.999 9)、3.45~689.60μg·mL^(-1)(r=0.999 9),平均回收率(n=6)为98.1%~100.1%之间。山野豌豆样品中山柰酚-3-O-α-L-鼠李糖苷、槲皮素、苜蓿素、山柰酚含量范围分别为0.013~0.232、0.007~0.165、0.006~0.122、0.008~0.153 mg·g^(-1);狭山野豌豆样品中山柰酚-3-O-α-L-鼠李糖苷、槲皮素、苜蓿素、山柰酚含量范围分别为0.010~0.231、0.022~0.196、0.012~0.200、0.010~0.208 mg·g^(-1)。结论:为检测山野豌豆与狭山野豌豆中山柰酚-3-O-α-L-鼠李糖苷、槲皮素、苜蓿素、山柰酚含量提供了可信方法。

Objective：To establish an HPLC method for determination of kaempferol-3-O-α-L-rhamnoside,quercetin,tricin and kaempferol in Vicia amoena Fisch.and Vicia amoena Fisch.var.angusta Freyn.,and to provide basis for quality control of the herbs.Methods：Analysis was carried out on a Diamonsil C_（18） column（250 mm×4.6 mm,5 μm）using acetonitrile-0.1% phosphoric acid as mobile phase at the flow rate of 1.0mL·min（-1） for gradient elution and the detection wavelength was 350 nm.The column temperature was set at 25 ℃ and the injection volume was 10 μL.Results：The lowest detection limits of kaempferol-3-O-α-L-rhamnoside,quercetin,tricin and kaempferol were 0.20,0.21,0.21 and 0.20 μg·mL（-1）,respectively.The linear ranges of the analytes were 6.85（-1） 369.60（r=0.999 9）,6.34（-1） 267.20（r=0.999 8）,3.33-665.60（r=0.999 9） and 3.45-689.60（r=0.999 9） mg·mL（-1）,respectively.The average recoveries of the components were within 98.1% and 100.1%.Ranges of the content of kaempferol-3-O-α-L-rhamnoside,quercetin,tricin and kaempferol in Vicia amoena Fisch.were 0.013-0.232,0.007-0.165,0.006-0.122 and 0.008-0.153 mg·g（-1）,respectively.Ranges of those in Vicia amoena Fisch.var.angusta Freyn.were 0.010-0.231,0.022-0.196,0.012-0.200 and 0.010-0.208 mg·g（-1）,respectively.Conclusion：Reliable method was offered for determination of kaempferol-3-O-α-L-rhamnoside,quercetin,tricin and kaempferol in the two herbs.