摘要
目的:建立利用毛细管电泳法测定人血白蛋白和人免疫球蛋白类血液制品纯度的方法,并进行方法学验证。方法:采用次氨基三亚甲基膦酸(NTMP)试剂盒,未涂层石英毛细管,有效长度为20 cm,总长度为30 cm,内径为50μm;检测波长200 nm;0.5psi压力进样8 s;分离电压+10.0 k V,升压时间1 min,分离20 min;毛细管温度25℃;样品温度25℃;按面积归一法计算纯度结果。结果:系统适应性结果显示人血浆在此条件下分离良好,在0.01~10 mg·ml^-1浓度范围内,人血白蛋白、人免疫球蛋白的峰面积与其相对应的浓度有良好的线性关系,r分别为0.999 4,0.999 2;人血白蛋白、人免疫球蛋白迁移时间的RSD分别为1.42%,0.18%(n=10),纯度RSD分别为0.07%,0.26%(n=10),检测限分别为2.0,2.4μg·ml^-1,定量限分别为6.7,7.9μg·ml^-1;在添加量为0.1%~25%范围内,回收率分别为101.8%~105.9%(RSD=1.7%,n=8)、97.6%~101.6%(RSD=1.25%,n=8)。结论:该方法快速、准确度高,可用于人血白蛋白和人免疫球蛋白类血液制品纯度检测,作为中国药典收载方法的补充。
Objective: To establish and verify a method for detecting the purities of human albumin and human immunoglobulin by capillary electrophoresis.Methods: A CEofix kit was used with a uncoated quartz capillary tube with the total length of 30 cm,the effective length of 20 cm,and the inner diameter of 50 mm.The detection wavelength was 200 nm,0.5 psi pressure injecting time was8 s,the separation voltage was + 10.0 k V,the boost time was 1 min,the separation time was 20 min,the capillary temperature was25℃,and the sample temperature was 25℃.The purity results were calculated by area normalization method.Results: The results of system suitability showed that human plasma was well separated under the conditions.Within the concentration range of 0.01-10 mg·ml^-1,the peak area of human serum albumin and human immunoglobulins had a good linear relationship with the corresponding concentration,and the correlation coefficient was 0.999 4 and 0.999 2,respectively.The relative standard deviation of migration time of human serum albumin and human immunoglobulin was 1.42% and 0.18%(n=10),and the relative standard deviation of purity results was 0.07% and 0.26%,respectively(n=10).The detection limit was 2.0 μg·ml^-1 and 2.4 μg·ml^-1,and the quantification limit was 6.7 μg · ml^-1 and 7.9 μg · ml^-1,respectively.Within the addition range of 0.1%-25%,the recovery rate was101.8%-105.9%(RSD=1.7%,n=8) and 97.6%-101.6%(RSD=1.25%,n=8),respectively.Conclusion: The method is fast and accurate,and can be used for the purity determination of albumin and human immunoglobulin as a supplement to the Chinese Pharmacopoeia.
作者
李炎
林涛
邹剑
马晶
王叔桥
Li Yan;Lin Tao;Zou Jian;Ma Jing;Wang Shuqiao(Sichuan Institute for Food and Drug Control,Chengdu 611731,China)
出处
《中国药师》
CAS
2019年第8期1552-1555,共4页
China Pharmacist
关键词
人血白蛋白
人免疫球蛋白
纯度
毛细管电泳
Human albumin
Human immunoglobulin
Purity
Capillary electrophoresis
