HPLC法同时测定新疆鼠尾草中5个酚酸类成分的含量

目的:建立HPLC同时测定新疆鼠尾草中原儿茶醛、丹参素钠、原儿茶酸、咖啡酸、迷迭香酸的含量。方法:采用Phenomennex Luna C₁₈(250 mm×4.6 mm,5 μm)色谱柱,以甲醇(A)-0.5%甲酸水溶液(B)为流动相,梯度洗脱(0~10 min,20%A→30%A;10~20 min,30%A→35%A;20~30 min,35%A→36%A;30~35 min,36%A→47%A),流速1 mL·min^-1,检测波长为285 nm,柱温40 ℃,进样量20 μL。结果:新疆鼠尾草5个成分在各自的线性范围内(原儿茶醛0.080 96~4.048 0 μg·mL^-1,丹参素钠0.803 2~40.16 μg·mL^-1,原儿茶酸0.172 8~8.640 μg·mL^-1,咖啡酸0.166 4~8.320 μg·mL^-1,迷迭香酸0.800 8~40.40 μg·mL^-1)均呈良好的线性关系(r=0.999 8),平均加样回收率、精密度、重现性和稳定性均符合有关规定。10批样品中5种酚酸类成分的含量分别为原儿茶醛0.024~0.140 mg·g^-1,丹参素钠0.676~1.940 mg·g^-1,原儿茶酸0.017~0.276 mg·g^-1,咖啡酸0.032~0.264 mg·g^-1,迷迭香酸0.199~6.319 mg·g^-1。结论:该方法经方法学验证,可用于新疆鼠尾草药材的质量评价。

Objective:To establish an HPLC method for the simultaneous determination of 5 salvianolic acid constituents,including protocatechuic aldehyde,Danshensu sodium,protocatechuic acid,caffeic acid,and rosmarinic acid,in the roots and rhizomes of Salvia deserta Schang.Methods:Phenomennex Luna C₁₈ column(4.6 mm×250 mm,5 μm)was adopted.The mobile phase was composed of methanol(A)and 0.5% formic acid aqueous(B)with a gradient elution(0-10 min,20%A→30%A;10-20 min,30%A→35%A;20- 30 min,35%A→36%A;30-35 min,36%A→47%A)at the flow rate of 1.0 mL·min^-1.The detection wavelength was 285 nm,the column temperature was 40 ℃,and the injection volume was 20 μL.Results:The standard curve revealed a good linear relationship(r=0.999 8)over the range of 0.080 96-4.048 0 μg·mL^-1 for protocatechuic aldehyde,0.803 2-40.16 μg·mL^-1 for Danshensu sodium,0.172 8-8.640 μg·mL^-1 for protocatechuic acid,0.166 4-8.320 μg·mL^-1 for caffeic acid,0.800 8-40.40 μg·mL^-1 for rosmarinic acid,respectively.The contents of 5 salvianolic acid constituents were 0.024-0.140 mg·g^-1 for protocatechuic aldehyde,0.676-1.940 mg·g^-1 for Danshensu sodium,0.017-0.276 mg·g^-1 for protocatechuic acid,0.032-0.264 mg·g^-1 for caffeic acid,0.199-6.319 mg·g^-1 for rosmarinic acid,respectively.Conclusion:The established determination method,which is rapid,sensitive,reproducible and stable,can be used for qualitative evaluation of Salvia deserta Schang.