高效液相色谱法测定人血白蛋白制品中乙酰色氨酸的含量

Determination of Acetyltryptophan Content in Human Albumin by High Performance Liquid Chromatography

目的应用高效液相色谱法测定人血白蛋白制品中乙酰色氨酸的含量。方法参照《中国药典》三部(2005版)附录ⅢD,采用凝胶色谱柱,以紫外吸收检测器在280nm波长处检测,按照《中国药典》三部(2005版)附录ⅢB中的公式计算人血白蛋白制品中乙酰色氨酸的含量。分析线性关系,计算加标回收率,并验证该方法的重复性。结果乙酰色氨酸含量在0.16～0.64mmol/L的范围内与峰面积有良好的线性关系,相关系数为0.99992;3个浓度梯度(0.16、0.32和0.48mmol/L)乙酰色氨酸的加标回收率分别为100.4%、103.9%和102.8%;连续5次检测人血白蛋白制品中乙酰色氨酸的含量,变异系数为0.46%。结论应用高效液相色谱法测定人血白蛋白制品中乙酰色氨酸的含量,结果准确可靠。

Objective To determine the acetyltryptophan content in human albumin by high performance liquid chromatography（HPLC）.Methods The test samples were determined by the method described in Appendix Ⅲ D of Chinese Pharmacopoeia（Volume Ⅲ,2005）using gel chromatographic column and ultraviolet absorption detector at a wavelength of 280 nm,based on which the acetyltryptophan content in human albumin was calculated by the formula in Appendix Ⅲ B of Chinese Pharmacopoeia（Volume Ⅲ,2005）.The linear relationship between acetyltryptophan content and peak area was analyzed,the standardized recovery rate was calculated,and the reproducibility of the method was verified.Results Acetyltryptophan at a content of 0.16 ~ 0.64 mmol /L showed good linear relationship to peak area,with a correlation coefficient of 0.999 92.The standardized recovery rates of acetyltryptophan at 3 concentration densities（0.16,0.32 and 0.48 mmol /L）were 100.4%,103.9% and 102.8% respectively.The coefficient of variation of acetyltryptophan contents in human albumin in five tests was 0.46%.Conclusion The determination result of acetyltryptophan content in human albumin by HPLC was accurate and reliable.