抗人T淋巴细胞猪免疫球蛋白中戊二醛残留量的检测

Determination of Residual Glutaraldehyde Content in Anti-human T Lymphocyte Porcine Immunoglobulin

目的建立高效液相色谱测定抗人T淋巴细胞猪免疫球蛋白中戊二醛残留量的方法。方法取供试品,与2,4-二硝基苯肼柱前衍生化,过滤,进样。色谱条件:色谱柱采用十八烷基硅烷键合硅胶填充柱;以50%乙腈(乙腈∶水=1∶1)溶液为流动相,流速为0.9 ml/min,检测波长为360 nm,柱温30℃。用该方法检测2批供试品中戊二醛的含量,并确定该方法的检测限、定限量、准确度和精密度。结果该方法检测的2批供试品中戊二醛的含量分别为9.8和7.5μg/ml;该方法的检测限为0.61μg/ml,定量限为2.04μg/ml,在2～20μg/ml范围内,其线性相关系数为0.998 2,检测5、10、15μg/ml戊二醛的回收率分别为102.3%、101.6%和99.2%,相对标准偏差均小于4.0%。结论高效液相色谱法可以简便、准确地检测抗人T淋巴细胞猪免疫球蛋白中戊二醛的残留量,为其生产工艺参数的确定提供了实验依据。

Objective To develop a high performance liquid chromatography（HPLC） method for determination of residual glutaraldehyde content in anti-human T lymphocyte porcine immunoglobulin.Methods Test samples were subjected to pre-column derivation with 2,4-dinitrophenylhydrazine（DNPH）,then filtered and injected into column.The condition for chromatography was as follows： chromatographic column was filled with octadecylsilane bonded silica gel;50% acetonitrile solution（acetonitrile ∶ water = 1 ∶ 1） was served as mobile phase;the flow rate was 0.9 ml / min;the detection wavelength was 360 nm;the temperature of column was 30℃.The glutaraldehyde contents in two batches of test samples were determined by the developed method,based on which the detection limit,quantitative detection limit,accuracy and precision of the method were determined.Results The glutaraldehyde contents in two batches of test samples determined by the developed method were 9.8 and 7.5 μg / ml respectively.The detection limit,quantitative detection limit and linear range of the method were 0.61,2.04 and 2 ～ 20 μg / ml（r = 0.998 2） respectively.The recovery rates of 5,10 and 15 μg / ml of glutaraldehyde determined by the method were 102.3%,101.6% and 99.2% respectively,of which the relative standard deviations（RSDs） were less than 4.0%.Conclusion HPLC method was suitable for simple and accurate determination of residual glutaraldehyde content in anti-human T lymphocyte porcine immunoglobulin,which provided an experimental basis for determination of production procedure parameters.