HPLC法同时测定新疆鼠尾草中5个酚酸类成分的含量

Simultaneous determination of 5 salvianolic acid constituents in the roots and rhizomes of Salvia deserta Schang by HPLC

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,5μm)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 m L·min^(-1),检测波长为285 nm,柱温40℃,进样量20μL。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 0μg·m L^(-1),丹参素钠0.803 2～40.16μg·m L^(-1),原儿茶酸0.172 8～8.640μg·m L^(-1),咖啡酸0.166 4～8.320μg·m L^(-1),迷迭香酸0.800 8～40.40μg·m L^(-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（-1）0 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 m L·min（-1）.The detection wavelengthwas 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·m L（-1） for protocatechuic aldehyde,0.803 2-40.16 μg·m L（-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·m L（-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.