高效液相色谱法检测发酵液中二羟基丙酮和甘油的含量

目的建立二羟基丙酮(DHA)发酵过程中产物和残余甘油底物的高效液相色谱检测方法。方法流动相为乙腈-水(90:10),色谱柱为Lichrospher 5-NH2(4.6 mm×250 mm),用紫外检测器在271 nm处检测DHA,用示差折光检测器检测甘油。结果DHA的线性范围为2.00~12.00 mg/mL,最低检出限(LOD)和最低定量限(LOQ)分别为0.30和1.20 mg/mL。甘油的线性范围为1.00~10.00 mg/mL,LOD和LOQ分别为0.22和0.50 mg/mL。结论此文建立的DHA和甘油的检测方法精密度好,准确性高,不受发酵液中其他杂质的影响,可以应用于DHA发酵过程中底物和产物的检测。

Quantitative determination of dihydroxyacetone and glycerol in the fermentation broth by HPLC

Purpose To develop high performance liquid chromatographic(HPLC) methods for the quantitative determination of dihydroxyacetone(DHA) and glycerol in the fermentation broth.Methods The analytes were separated on a Lichrospher 5-NH2 column(4.6 mm×250 mm) with the mobile phase of acetonitrile-water(90∶10) at a flow rate of 1 mL/min.DHA was detected by UV detector at 271 nm and glycerol was detected by refractometer.Validation parameters such as linearity,precision,accuracy,specificity,limit of detection(LOD) and limit of quantitation(LOQ) were determined.Results The linearity range for the determination of DHA was 2.00～12.00 mg/mL with a correlation coefficient(r) of 0.999 4.The LOD and LOQ were 0.30 and 1.20 mg/mL respectively.The linearity range for the determination of glycerol was 1.00～10.00 mg/mL with a correlation coefficient of 0.999 8.The LOD and LOQ were 0.22 and 0.50 mg/mL respectively.Conclusion The established methods in this paper were simple and rapid and there is no interference from other constitutes in the fermentation broth on using the methods.The methods were applicable for DHA and glycerol determination in the fermentation process.