菌渣中截短侧耳素含量的高效液相色谱法测定

Determination of Pleuromutilin in fermentation fungi residues by HPLC

目的:建立截短侧耳素的提纯及含量测定方法。方法:采用甲醇从发酵菌渣中提纯截短侧耳素后,用高效液相色谱法(HPLC)测定其含量。色谱分析条件:采用Agilent-ODSC18色谱柱(5μm,4.6mm×250mm),流动相为甲醇-体积分数为0.5%甲酸溶液(体积比为60:40),流速1mL/min,检测波长为205nm,进样量20μL,柱温30℃。结果:截短侧耳素在341～2730mg/L范围内,线性关系良好(r=0.9999);平均加样回收率为99.2%,相对标准偏差为0.54%(n=6)。结论:此方法简便、灵敏、稳定、可靠,可用于截短侧耳素的含量测定。

Aim：To establish a method for purification and determination of pleuromutilin in fermentation fungi residues. Methods：The fermentation fungi residues were extracted with methanol and the extraction was analyzed by HPLC. The Agilent-ODS Cj8（5 μm,4.6 mm ×250 mm） column was used with a UV detector at 205 nm;The mobile phase was methanol-0, 5% formic acid solution （60 ： 40） at a flow rate of 1.0 mL/min. Results：The calibrated linear curve of pleuromutilin concentration was within 341 - 2 730 mg/L（ r = 0. 999 9） , the mean recovery was 99.2% with RSD 0. 54% （ n = 6 ）. Conclusion ： This method is simple, sensitive, stable, reliable and suitable to be applied in determination of pleuromutilin.