HPLC法同时测定民族药芒萁中5个成分的含量

目的:建立同时测定民族药芒萁中芦丁、槲皮苷、山柰酚-3-O-α-L-鼠李糖苷、槲皮素和山柰酚的RP-HPLC测定方法。方法:采用Pntulips^TM BP-C₁₈色谱柱(4.6 mm×250 mm,5 μm),以乙腈(A)-0.5%磷酸(B)为流动相,梯度洗脱(0~10 min,15%A;10~11 min,15%A→25%A;11~20 min,25%A→55%A;20~30 min,55%A→15%A),流速1 mL·min^-1,检测波长360 nm,柱温30 ℃。结果:在选定的实验条件下,芦丁、槲皮苷、山柰酚-3-O-α-L-鼠李糖苷、槲皮素和山柰酚得到良好的分离,质量浓度分别在77.2~694.8 μg(r=0.999 8,n=6)、43.6~392.4 μg(r=0.999 9,n=6)、288.0~2 592.0 μg(r=0.999 9,n=6)、38.4~354.6 μg(r=0.999 9,n=6)和43.8~394.2 μg(r=0.999 2,n=6)范围内呈良好线性关系,平均回收率分别为97.6%(RSD=1.94%)、98.6%(RSD=1.11%)、96.9%(RSD=1.22%)、97.1%(RSD=1.97%)和95.3%(RSD=2.01%)。4个不同产地芒萁的含量测定结果表明,山柰酚-3-O-α-L-鼠李糖苷为芒萁的主要成分,其次为槲皮苷。不同产地的芒萁中5种成分的含量差异变化不大,芦丁、槲皮苷、山柰酚-3-O-α-L-鼠李糖苷、槲皮素和山柰酚的含量分别为0.109~0.176、0.584~0.686、2.416~2.627、0.088~0.131和0.018~0.031 mg·g^-1。结论:该方法简单易行,高效快速,可用于民族药芒萁的品质评价及质量控制。

Objective:To establish an HPLC-DAD method for simultaneous determination of the content of five main active ingredients-rutin,quercitrin,kaempferol-3-O-α-L-rhamnoside,quercetin and kaempferol in the medicine plant of Dicranopteris pedata.Methods:The HPLC separation was performed on a Pntulips^TM BP-C₁₈ column(4.6 mm×250 mm,5 μm),using acetonitrile(A)-0.5% phosphoric acid solution(B)as the mobile phase in a gradient mode(0-10 min,15%A;10-11 min,15%A→25%A;11-20 min,25%A→55%A;20-30 min,55%A→15%A)at a flow rate of 1 mL·min^-1.The detection wavelength was 360 nm,and the column temperature was 30 ℃.Results:A good separation of five compounds was achieved under the selected experimental conditions,showing a good linearity relationship separately within the range of 77.2-694.8 μg(r=0.999 8,n=6)for rutin,43.6-392.4 μg(r=0.999 9,n=6)for quercitrin,288.0-2 592.0 μg(r=0.999 9,n=6)for kaempferol-3-O-α-L-rhamnosides,38.4-354.6 μg(r=0.999 9,n=6)for quercetin and 43.8-394.2 μg(r=0.999 2,n=6)for kaempferol.The respective average recoveries were 97.6%(RSD=1.94%),98.6%(RSD=1.11%),96.9%(RSD=1.22%),97.1%(RSD=1.97%),and 95.3%(RSD=2.01%).Determination results of Dicranopteris pedata from four different producing areas showed that kaempferol-3-O-α-L-rhamnoside was the main component of Dicranopteris pedata,followed by quercetin.The content of five kinds of compositions was similar to Dicranopteris pedata from different producing area.The contents of rutin,quercitrin,kaempferol-3-O-a-L-rhamnoside,quercetin and kaempferol ranged in 0.109-0.176,0.584-0.686,2.416-2.627,0.088-0.131 and 0.018-0.031 mg·g^-1,respectively.Conclusion:The established method is easy and efficient,and can be applied to determine the contents of five main active ingredients of the national drug Dicranopteris pedata.