HPLC测定不同产地马尾松等松叶中莽草酸的含量

目的: 建立松叶中莽草酸的HPLC含量测定方法,为松叶药材质量标准的建立提供参考.同时对不同产地马尾松等松叶中莽草酸的含量进行比较研究. 方法: 采用HPLC法,使用Ultimate XB-C₁₈(4.6 mm×250 mm,5 μm)色谱柱,以甲醇-0.5%磷酸(2:98)为流动相,流速1.0 mL·min^-1,柱温25 ℃,检测波长213 nm.通过优化参数(溶剂体积、浸泡时间、提取次数)确定莽草酸含量测定前处理的最佳条件,并测定不同产地马尾松及同一产地不同种属松叶中莽草酸的含量. 结果: 莽草酸的含量测定前处理方法为加水60 mL超声提取50 min;松叶莽草酸在0.094~1.875 μg范围内线性关系良好(r=0.999 8);平均回收率为98.8%(RSD=0.56%);方法精密度及重复性良好,RSD均小于2%.不同种属松叶中莽草酸的含量为2.62%~0.07%,其中红松、马尾松较高,落叶松最低;在所测的不同产地马尾松中莽草酸的含量为4.28%~2.00%,其中广西马尾松中莽草酸含量最高,河南产马尾松含量最低. 结论: 本方法操作简便,定量准确,重现性好,可用于松叶药材中莽草酸的含量测定;不同种属松叶中,马尾松含量较高;不同产地马尾松中莽草酸含量有一定差异,其中广西、四川、浙江、江苏产马尾松中莽草酸含量较高.

Objective: To establish an HPLC method for determining shikimic acid contained in pine needles,so as to provide preliminary experimental data for quality control standards,and to compare the contents of Pinus massoniana Lamb.from different areas and different species of pine needles from Jilin province. Methods: HPLC and Ultimate XB-C₁₈ column(4.6 mm×250 mm,5 μm)were adopted,with methanol-0.5% phosphoric acid(2:98)as the mobile phase at the flow rate of 1.0 mL·min^-1.The column temperature was set at 25 ℃,and the UV detection wavelength was 213 nm.The optimum condition for determining shikimic acid in pine needles was decided by optimizing the parameters of preparation(decoloration,extraction time,solvent volume,immersion time and extraction frequency)and the contents of Pinus massoniana Lamb.from different areas and different species were determined. Results: The pretreatment of shikimic acid was ultrasonic extraction for 50 min with addition of 60 mL water.Shikimic acid showed a good linear relationship within the range of 0.094-1.875 μg(r=0.999 8),with the average recovery of 98.8%(RSD=0.56%).Good precision and reproducibility was achieved with the RSDs smaller than 2.0%.The content range was 2.62%-0.07% for different species of pine needles,and species with the relatively high contents were Pinus koraiensis Sieb.et Zucc.,and Pinus massoniana Lamb.,those with the lowest contents were Larix gmelinii(Rupr.)Rupr..The content range was 4.28%-2.00% for Pinus massoniana Lamb.from different areas,and the highest and the lowest contents appeared in those from Guangxi and Henan. Conclusion: The method is simple,accurate,reproducible and can be used to determine the content of shikimic acid in pine needles. The content of shikimic acid in Pinus massoniana Lamb.is higher than that in other species.The content of shikimic acid is different among Pinus massoniana Lamb.from different areas and is higher in those from Guangxi,Sichuan,Zhejiang,and Jiangsu.

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