HPLC法测定山野豌豆与狭山野豌豆中4个黄酮成分

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 C₁₈ 色谱柱（200mm×4.6 mm，5 μm），以乙腈-0.1% 磷酸水溶液梯度洗脱，流速1.0 mL·min^-1，检测波长350 nm，柱温25 ℃，进样量10 μL。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、0.20 μg·mL^-1，线性范围分别为6.85~1 369.60 μg·mL^-1（r=0.999 9）、6.34~1 267.20 μg·mL^-1（r=0.999 8）、3.33~665.60 μg·mL^-1（r=0.999 9）、3.45~689.60 μg·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 C₁₈ column(250 mm×4.6 mm, 5 μm)using acetonitrile-0.1% phosphoric acid as mobile phase at the flow rate of 1.0 mL·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.

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