高效液相色谱法同时测定香灰菌发酵液中L-苯丙氨酸和2-苯基乙醇

为了检测香灰菌发酵液中L-苯丙氨酸的利用情况和2-苯基乙醇的生成量,建立了一种利用HPLC外标法同时检测香灰菌发酵液中L-苯丙氨酸和2-苯基乙醇的方法。方法色谱条件为:色谱柱:C₁₈(5 μm,250 mm×4.6 mm);柱温30 ℃;进样量10 μL;流速0.7 mL/min;DAD检测器,检测波长258 nm;流动相A:0.6%乙酸水溶液;流动相B:甲醇。结果表明:两种物质的浓度分别在1~5和1.02~5.1 g/L范围内与峰面积有良好的线性关系,相关系数r均大于0.999,L-苯丙氨酸检出限和定量限分别为1.023和3.409 mg/L,2-苯基乙醇检出限和定量限分别为0.567和1.890 mg/L,L-苯丙氨酸加标回收率为99.16%~101.58%,RSD(n=6)为0.67%,2-苯基乙醇的加标回收率为98.53%~101.06%,RSD(n=6)为1.22%。该方法操作简便快速、准确度和灵敏度高,可满足对香灰菌发酵液中L-苯丙氨酸和2-苯基乙醇风味物质的同时检测。

Simultaneous Detection of L-phenylalanine and 2-phenylethanol in Hypoxylon sp.Fermented Liquid by HPLC

In order to simultaneously detect the usage of L-phenylalanine and the production of 2-phenyl-ethanol in Hypoxylon sp. fermented liquid, an HPLC external standard method was developed. The method chromatographic conditions were as follows:Chromatographic column:C₁₈ (5 μm, 250 mm×4.6 mm), Column temperature 30℃, the injection volume 10 μL, flow rate 0.7 mL/min, DAD detector, detection wavelength 258 nm, mobile phase A 0.6% acetic acid solution, mobile phase B methanol.The results indicated that good linearities between concentration and peak area were exist for L-phenylalanine and 2-phenylethanol with correlation coefficients greater than 0.999 in the range of 1~5 and 1.02~5.1 g/L, respectively.The detection limit and the quantitative limit of L-phenylalanine were 1.023 and 3.409 mg/L, respectively. The detection limit and the quantitative limit of 2-phenylethanol were 0.567 and 1.890 mg/L, respectively. The spiked recoveries of L-phenylalanine were in the range of 99.16%~101.58% with relative standard deviations (n=6) of 0.67%. The spiked recoveries of 2-phenylethanol were in the range of 98.53%~101.06% with relative standard deviations (n=6) of 1.22%.The method was simple width high accuracy and sensitivity. It was suitable for simultaneously determining the L-phenylalanine and 2-phenylethanol in the Hypoxylon sp. fermented liquid.