采用高效液相色谱法(HPLC)同时检测丁二酸发酵液中的有机酸和水溶性维生素。以RP—HPLC为分离模式,选用HederaODS-2色谱柱(5Ixm,4.6×250mm),20mm01/L的磷酸盐缓冲溶液(pH2.9)和乙腈为流动相,梯度洗脱,检测波长为210nm,建立了同时测定分析8种水溶性维生素和3种有机酸的方法,15min内所有组分都洗脱完全。8种水溶性维生素和3种有机酸在线性范围内峰面积与浓度呈良好的线性关系,相关系数为0。9989~0。9997;加标回收率在92.5%~100.3%之间;RSD(n=5):0.9%~1.9%。本方法采用同一种流动相既可以测定发酵液中的有机酸又可以测定水溶性维生素的含量:
An innovative system of RP-HPLC method was proposed and optimized for detecting the organic acids and hydrosoluble vitamins in succinic acid fermentation broth. Under the optimal conditions, a Hedera ODS-2column (5lxm, 4.6 mm x 250 ram) was used, the detection wavelength was 210 nm. Flow rate was 1.0 mL/min; Mobile phase A:20 mmolfL sodium dihydrogen phosphate(pH2.9) , Mobile phase B:ACN; the gradient protocol was applied. All compounds were completely eluted in 15 rain. All the correlation coefficients (r) of regression equations were above 0. 9989, and the recoveries ranged from 92.5% to 100.3%. The relative standard deviations were in the range from 0.9% to 1.9%. By this method, organic acids and vitamins can be easily, rapidly and effectively determined.