高效液相色谱法测定刺糖多胞菌发酵液多杀菌素含量

Determination of Spinosad from Saccharopolyspora Spinosa Fermentation Solution by HPLC

为了建立刺糖多胞菌（Saccharopolyspora spinosa）发酵液中多杀菌素不同组分的分离与定量分析的高效液相色谱检测方法.以AQ12S05-1546WT（150×4.6 mmL.D.S-5μm,12 nm）为分析柱,以甲醇-乙腈-2%醋酸铵溶液（45∶45∶10,v/v/v）为流动相,流速1.5 mL/min,检测波长250 nm.结果表明多杀菌素在25-1000 mg/L范围内线性关系良好,本方法标准偏差为1.486 9,变异系数为0.79%,相关系数为0.9992,平均回收率为99.58%.多杀菌素化合物A,B,C,D,E,F的保留时间分别为7.01,3.59,3.21,8.33,5.72,4.81 min.本方法能准确测定刺糖多胞菌发酵液中多杀菌素含量,有效分离多杀菌素不同组分.

To establish a method for the separation and quantitative determination of different kinds of spinosads from felmentation solution of Saccharopolyspora spinosa by HPLC. The AQ12S05-1546WT （150 × 4. 6 mmL. D. S -5 um,12 nm） column was used. The mobile phase consisted of methanol, acetonitrile and 2% ammonium acetate solutions （45：45： 10, v/v/v）, and the flow rate was 1.5 mL/min. Detection wavelength was 250 nm. The results showed that the good linear relationship was obtained in the concentration range of 25 - 1 000 mg/ L. Standard deviation of the method was 1. 486 9. Coefficient of variation was 0. 79%. Linear correlation was 0. 999 2. Average recovery was 99.58%. The retention time for spinosad A, B, C, D, E, F were 7.01,3.59, 3.21,8.33,5. 72,4. 81 min, respectively. The method is simple, sensitive, and reproducible for determination of spinosads from fermentation of Saccharopolyspora spinosa.