高效液相色谱法测定农用抗生素武夷菌素的方法优化

Optimization of the determination methodology of wuyiencin by high performance liquid chromatography

目的建立并优化了高效液相色谱法测定发酵液中武夷菌素的方法。方法选用ZORBAXSB-Aq色谱柱，优化了流动相组成、磷酸二氢钾浓度、流动相pH，考察了方法的线性范围、检测限、稳定性、精密度和回收率。结果实验表明，武夷菌素在205nm有最大吸收，确定为液相检测波长，最优的流动相比例为磷酸二氢钾溶液（0．02mol／L）：甲醇=95：5，pH7．0，线性范围为0．2-1．0mg／mL，线性相关系数为0．9999，平均回收率为99．10％，检出限为0．01mg／mL，日内和日间稳定性良好，RSD值分别为1．44％和0．51％。结论该方法准确，灵敏，能够有效分离并监测发酵液中武夷菌素和杂质。

Objective To establish and optimize a method for the determination of wuyiencin titer in the fermentation broth by high performance liquid chromatography. Methods The phosphate concentration, pH and composition ratio of mobile phase were optimized using ZORBAX SB-Aq column. The range of linearity, detection limit, stability, precision and recovery were investigated. Results The results showed that the optimal detection wavelength was 205nm, and the best composition of mobile phase was methanol and 0.02mol/L phosphate solution （5：95, V/V, pH 7.0）. The method yielded good linear correlation for 0.2-1.0mg/mL of wuyiencin （R2=0.9999）. The average recovery was 99.10%, and the detection limit was 0.01mg/mL. The stability of wuyiencin within 72h was detected and showed good results, and its RSD value is 1.44% and 0.51%, respectively. Conclusion The method is accurate and sensitive, and is suitable for the determination of wuyiencin in the fermentation.