高效液相色谱法同时测定发酵液中胡椒醛、胡椒酸以及黄樟油素

Determination of piperonal,piperonylic acid and safrole in fermentation broth by high performance liquid chromatography

建立了高效液相色谱法同时测定发酵液中胡椒醛、胡椒酸以及黄樟油素的分析方法。采用梯度洗脱程序,流动相为A:乙腈,流动相B:0.1%甲酸水溶液,Amethyst C_(18)-H反相色谱柱(4.6 mm×250 mm,5μm),柱温30℃,检测波长270 nm,胡椒醛、胡椒酸以及底物黄樟油素能够良好分离,且平均加标回收率在98.0%~101.0%,精密度偏差在0.5%~2.0%范围内。用此方法测得液化沙雷氏菌CCTCC No:M2016170发酵液中含有182mg/L胡椒醛和9.8 mg/L胡椒酸。

A high performance liquid chromatography method was developed for the determination of piperonal, piperonylic acid and safrole in fermentation broth. The separation was performed on Amethyst C18-H C18 column （250 mm × 4.6 mm,5μm） using gradient elution,the mobile phase A was acetonitrile and B was 0.1% formic acid solution. The column temperature was chosen for 30℃ and the detection wave length was 270 nm. It was found that piperonal,piperonylic acid and substrate safrole could be well separated in the conditions,which showed that average recoveries were within the range of 98.0% - 101.0% , and the relative standard deviations ranging from 0.5% to 2.0% in samples. This method was also applied for the determination of fermentation broth of Serratia liquefaciens M2016170, which contained 182 mg/L piperonal and 9.8 mg/L piperonylic acid.