枞酸的HPLC内标法定量分析研究

Determination of Abietic Acid by HPLC with Internal Standard Method

建立了高效液相色谱-内标法测定枞酸含量的方法。以苯甲酸为内标物，ZORBAXSB-C18为分析柱；进样量：0．5μL；流动相：甲醇：水=90：10（v／v），流速0．5mL/min；检测器：紫外检测波长240nm；柱温室温。在枞酸浓度0．0101mg／L-0．1388mg／L范围内，枞酸样品与内标物苯甲酸的色谱峰面积比呈良好的线性关系：y=54．50264x＋0．12674；r=0．99919；重现性试验RSD为0．4594％（n=5）；精密度RSD为0．3222％（n=5）。平均回收率为99．85％，RSD为0．2799％（n=5）。稳定性实验，放置1、2、3d的同一测试样品，其稳定性RSD值为0．1899％（n=9）。验证了该方法准确、快速、灵敏度高、重现性好。

The determination method of abietic acid was established utilizing high performance liquid chromatography （HPLC） by taking benzoic acid as internal standard. The HPLC was conducted on ZORBAX SB-C18 （5μ） with methanoL/water = 90：10（v/v） as mobile phase. The flow rate of mobile phase was 0.5mL/min. The detective wavelength was set to 240 nm and room temperature as column temperature. The chromatography peak area ratio of standard to the inter- nal standard was very well correlated with the concentration of abietie acid in the range of 0.0101 mg/L - 0. 1388 mg/ L（y =54. 50264 x ＋0. 12674;r =0.99919 ）. The stability experiment was also investigated. The RSD of the reproduc- ibility, precision and stability test was 0. 4594% （ n = 5 ） ,0.3222% （ n = 5 ）, and 0.1899 % （ n = 9 ）, respectively. The average recovery rate for abietic acid was 99.85% and RSD was 0.2799% （ n = 5 ）. The method is simple, accurate and sensitive ,which can be used for the determination of the content of abietic acid.