人凝血酶原复合物中磷酸三丁酯残留量检测方法验证

Validation of detection method for tributyl phosphate residue in human prothrombin complex

目的建立气相色谱法测定人凝血酶原复合物中磷酸三丁酯残留量的方法,并进行方法验证。方法采用酸改性聚二乙醇(20M)毛细管柱,正己烷为溶剂,各项色谱参数为:气化室温度220℃、柱温箱温度155℃、检测器温度220℃、柱流量为2.0 mL/min、载气为氮气、检测器FID、采集时间10 min,进行准确性、重复性、线性、专属性、中间精密度、检测限、定量限、范围、耐用性的方法验证。结果验证结果显示该方法专属性良好;标准曲线的直线回归相关系数为0.99990;在50%、100%、150%(100%水平浓度为30μg/mL)三个水平浓度的的回收率分别是98.4%、97.5%、95.7%,平均回收率为97.2%,相对偏差为2.15%;在水平浓度为100%重复性为2.08%,中间精密度的相对偏差为1.63%;检测限为0.255μg/mL,定量限为0.511μg/mL,在改换同一厂家同一型号不同批号的毛细色谱柱之后系统适用性实验满足要求,方法耐用性良好。结论该方法能满足实验室人凝血酶原复合物磷酸三丁酯残留量的测定。

Objective To establish gas chromatography for the determination of tributyl phosphate(TBP)residues in human prothrombin complex and then verify it.Methods Acid modified polyethylene glycol(PEG)(20 M)capillary column was used with n-hexane as solvent.The chromatographic parameters were as follows:gasification chamber temperature at 220℃,column temperature at 155℃,detector temperature at 220℃,column flow rate at 2.0 mL/min,carrier gas as N2,detector as FID,and collection time for 10 min.The accuracy,repeatability,linearity,specificity,intermediate precision,detection limit,quantitative limit,range and durability were verified.Results The verification results showed that the method had good specificity.The linear regression correlation coefficient of standard curve was 0.99990.The recovery rate were 98.4%,97.5%and 95.7%when the concentration at 50%,100%(30μg/mL)and 150%,respectively,with an average recovery of 97.2%and a relative deviation of 2.15%.When the concentration was 100%,the repeatability was 2.08%,and the relative deviation of intermediate precision was 1.63%.The detection limit was 0.255μg/mL,and the quantitative limit was 0.511μg/mL.After changing capillary chromatographic columns with different batch numbers but the same types and manufacturer,the applicability test of the system met the requirements,and the method had good durability.Conclusion This method can be used for the determination of TBP residues of human prothrombin complex in laboratory.