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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
【摘要】 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。
【Abstract】 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.
【Key words】 high performance liquid chromatography; assay; pig brain extracts; nerve growth factor;
- 【文献出处】 复旦学报(医学版) ,Journal of Shanghai Medica(University) , 编辑部邮箱 ,2005年01期
- 【分类号】TQ464
- 【被引频次】2
- 【下载频次】177