液相色谱法测定大鼠血清中IgG的研究

目的建立一种用于测定大鼠血清中IgG的液相色谱法。方法根据被分析物质的性质,以HI-Trap Protein G柱为分离柱,以磷酸盐缓冲液和甘氨酸盐酸缓冲液为流动相,采用梯度洗脱方式,选择0.8mL/min流速,检测波长为280nm。结果浓度在0.5～1.5mg/mL浓度范围内IgG与峰面积呈良好的线性关系,以信噪比为10计算最低检测浓度为0.31mg/mL,平均回收率为90.5%。结论试验结果表明该方法可行,快速简便,准确可靠。

Determination of IgG in Rat Serum by Liquid Chromatography

Objective To establish a method for separation and determination of rat serum IgG using the high performance liquid chromatography （HPLC）. Methods A HI-Trap Protein G column was used as the analytical column, and gradient elution was proformed using the mixture of phosphate buffer and Tris-Glycine buffer as the mobile phase according to the characteristics of the samples. The target compound was detected at the adjustable wavelength of 280nm. Results The detection limit was in the range of 0.5～1.5 mg/mL. Taking the signal-to-noise ratio （S/N） of 10 as the detection limit, the lowest examination density was 0.31 mg/mL, and the average rate of recovery in rat serum was 90.5%. Conclusion The results show that HPLC method is feasible, rapid, simple and accurate for separation and determination of serum IgG.