反相高效液相色谱法测定发酵液中的风味强化肽

Determination of the Content of Enhancing Flavor Peptide in Fermentation by Reverse Phase High Performance Liquid Chromatography

采用反相高效液相色谱法对发酵液中的风味强化肽进行定量分析。色谱条件 :色谱柱为日本shimadzu公司的ShimadzuVP ODSC18(5 μm ,1 5 0mm× 4 6mmi d )不锈钢柱 ,流动相为乙腈 水 (含体积分数为 0 1 %的三氟乙酸 )溶液 ,采用线性梯度洗脱方式 ,流动相B(含 0 1 %三氟乙酸的乙腈溶液 )体积分数在 3 0min内由 5 %线性升至 5 0 % ,流速为 1 0mL/min ,检测波长为 2 3 0nm ,室温测定。结果 :RP HPLC法测定浓度在 0 1 2 5～ 2 0 0 0mg/mL时线性关系良好 ,r =0 9998。回收率为 86 2 %～ 1 0 3 4% ,最低检测限量为 1 2 5ng ,该方法精密度高 ,相对标准偏差 (RSD)为1 4%。此方法准确、灵敏、重现性好 。

A reverse phase high performance liquid chromatographic method was developed for the quantitative analysis of enhancing flavor peptide in fermentation. A shimadzu VP-ODS C 18(5?μm,150?mm×4.6?mm i.d.)column was used as the analytical column and the analysis was carried out with linear gradient elution of acetonitrile-water solution with 0.1% trifluoroacetate at a flow rate of 1.0?mL/min. The analysis was carried out at room temperature and the sample was detected at 230?nm. The mobile phase B was increased from 5% to 50% in thirty?minutes. It was found that the standard curves for enhancing flavor peptide were linear, r=0.9998, the recovery was 86.2%～103.4%,and the detection limit was 125ng. The precision of this method is good and the RSD is 1.4%. The results showed that this method is accurate, sensitive and reproducible, and suitable for quantitative analysis of enhancing flavor peptide in complex mixtures.