高效液相色谱法测定发酵液中截短侧耳素的含量

建立了测定发酵液中截短侧耳素含量的高效液相色谱法。发酵样品用甲醇萃取后进样,色谱柱ShimpackVPODSC18(5μm,4.6mm×200mm),流动相为甲醇∶0.1mol/LKH2PO4(50∶50,体积比),流速为1.00mL/min,紫外检测波长205nm和柱温25℃。结果表明,截短侧耳素在106.25～2500μg/mL浓度范围内线性关系良好,回归方程为y=1790.3x+145405,r=0.9996,平均回收率为96.5%,最低检测限为3.28ng。该法具有操作简单、快速、回收率好和准确性好等优点,可有效地检测发酵液中截短侧耳素的含量。

Determination of Pleuromutilin in Fermentation Broth by HPLC

A RP-HPLC method for the determination of pleuromutilin fermentation broth was developed.The fermentation broth was extracted by methanol and the extraction was analysed by HPLC.The samle was separated at temperature 25℃ on a Shim-pack VP-ODSC18(5 μm,4.6 mm×200 mm)eluted at a flow rate of 1.00 mL/min with mobile phase of methanol-0.1mol/L KH_2PO_4(50∶50,V/V),pleuromutilin was detected and quantified by the absorbance at 205 nm. The linearity was obtained over the range of 106.25～2500μg/mL for pleuromutilin with the coefficient being 0.9996 and the regression equation being y=1790.3x+145405. The average recovery was 96.5% and the detective limit was 3.28ng. This method is simple,accurate, thighly specific and reliable for the determination of pleuromutilin.