反相高效液相色谱法测定猪脑提取物中神经生长因子的含量

Determination of Nerve Growth Factor in Pig Brain Extracts by Reverse Phase High Performance Liquid Chromatography

目的 建立猪脑提取物中神经生长因子含量测定的方法。方法 采用高效液相色谱法,色谱柱为Dia monsil C18(150 mm×4.6 mm,5 μm),流动相为乙腈:(0.5％三氟乙酸水溶液:甲醇=52:48)=30:70,10 min后 梯度升至50:50。柱温30℃,流速1.0 mL／min,波长280 nin,进样量20μL。标准曲线法计算含量。结果 在 该色谱条件下,测得神经生长因子的线性良好(r=0.999 4);平均回收率100.9％-102.4％;日内精密度RSD 1.36％-1.77％,日间精密度RSD 1.85％-2.40％;最低检测浓度0.25 mg／L;3批样品中神经生长因子含量分 别为0.326％、0.321％0.332％(mg／100 g)。结论 该方法灵敏、准确、可靠,适用于本品中神经生长因子含量的 测定。

Purpose To establish a method for the determination of the content of nerve growth factor (NGF) in pig brain extracts.Methods A simple high performance liquid chromatography method was established with C18 column. The mobile phase was acetonitrile:(0.5% trifluoroaceticacid:methanol=52:48)=30:70 and was then programmed to 50:50 in 10 minute. The column temperature was 30℃. The flow rate was 1.0 mL/min. The detection wavelength was 280 nm and the injection volume was 20 μL. NGF was calculated by calibration curve method. Results The linearity was good (r=0.999 4). The average recoveries were 100.9%-102.4% with within day RSD of 1.36%-1.77% and between-day RSD of 1.85%-2. 40%. The limit of detection was 0.25mg/L. The contents of nerve growth factor in three batches of samples were 0.326% ,0.321% ,0.332% (mg/100 g). Conclusions This method was proved to be accurate,sensitive and reliable. It is suitable to be adopted in the analysis of nerve growth factor.