总有机碳检测法在血液制品清洁验证中的应用

Application of total organic carbon method to cleaning validation of blood products

目的利用总有机碳（total organic carbon,TOC）分析仪建立血液制品清洁验证分析方法。方法制备碳浓度为5、10、15 ppm的牛血清白蛋白（bovine serum albumin,BSA）溶液,检测TOC值,并计算回收率,验证方法的准确性;制备6份碳浓度为10 ppm的BSA溶液,检测TOC值,计算相对标准偏差（relative standard deviation,RSD）,验证方法的重复性;制备碳浓度分别为0、1、2.5、5、10、15和20 ppm的BSA溶液,检测TOC值,验证方法的线性;测定10次TOC检测用水的TOC值,计算SD,确定方法的检测限（limit of detection,LOD）和定量限（limit of quantitation,LOQ）;制备碳浓度分别为0、1、2.5、5、10、15和20 ppm的BSA溶液,先以从低到高浓度,再以从高到低浓度顺序分析,检测TOC值,分别计算回归曲线决定系数,确定残留效应;分别在不锈钢和玻璃材质上擦拭取样,检测TOC值,计算回收率及RSD。对清洁后的血液制品生产用设备和器具擦拭取样,检测TOC值,确认清洁效果。结果 3个碳浓度的BSA溶液TOC值回收率均在90%~110%之间,RSD值均小于5%;6份BSA溶液的TOC值的SD为0.202 ppm,RSD为2.13%;碳浓度在0-20 ppm范围内的BSA溶液的加样TOC值与测得TOC值呈良好的线性关系,R2=0.999 5;检测用水的10次TOC检测值的LOD和LOQ值均明显低于10 ppm;两条回归曲线的决定系数的RSD为0.03%;不锈钢和玻璃材质擦拭取样的TOC检测值最低回收率分别为74%和77%,不同工作人员检测结果 RSD均≤20%。血液制品TOC检测擦拭法清洁验证结果均低于2μg/cm2。结论 TOC检测法具有良好的准确性、重复性、灵敏度、线性,血液制品清洁残留量均低于残留限度,达到预计的清洁效果。该方法快捷简便,适用于血液制品的擦拭法清洁验证。

Objective To develop and validate a reliable method for cleaning validation of blood products with total organic carbon（TOC） analyzer. Methods The bovine serum albumin（BSA） solutions at carbon concentrations of 5, 10 and 15 ppm were prepared, of which the TOC levels were determined, and recovery rates were calculated to validate the accuracy of the method. Six BSA solutions each at a carbon concentration of 10 ppm were prepared and determined for TOC level, and the relative standard deviation（RSD）of determination results was calculated to validate the reproducibility of the method. The BSA solutions at carbon concentrations of 0, 1, 2. 5, 5, 10, 15 and 20 ppm were prepared and determined for TOC level to validate the linearity of the method. The TOC level of water for determination of TOC was determined for 10 times, of which the standard deviation（SD） was calculated to define the limit of detection（LOD） and limit of quantitation（LOQ）of the method. The BSA solutions at carbon concentrations of 0, 1, 2. 5, 5, 10, 15 and 20 ppm were prepared, and determined for TOC levels in the order of low to high concentrations at first then high to low concentrations, of which the determination coefficients of regression curves were calculated respectively to evaluate the residual effect. The samples of BSA at a carbon concentration of 25 ppm were collected by swabbing stainless steel and glass, of which the TOC levels were determined to calculate the recovery rates and RSDs. Samples were collected from the equipments and apparatuses for production of blood products, after being cleaned, by swabbing, and determined for TOC level to validate the cleaning effect. Results The recovery rates of TOC levels of BSA solutions at three carbon concentrations were 90% ~ 110%, of which the RSDs were less than 5%. The SD and RSD of TOC levels of six BSA solutions were 0. 202 ppm and 2. 13% respectively. The TOC levels of samples of BSA solutions at a carbon concentration range of 0 ~ 20 ppm showed good linearity to the measured TOC levels（R2 = 0. 999 5）. Both the LOD and LOQ of 10 determination results of TOC levels in water for determination were less than 10 ppm. The RSD of determination coefficients of two regression curves was 0. 03%. The minimum recovery rates of TOC levels in samples collected from the surface of stainless steel and glass were 74% and 77% respectively, of which the RSDs of determination results by different staff were not more than 20%. The validation results of cleaning effect showed that the residual blood product levels were less than 2 μg / cm2. Conclusion TOC method was of high accuracy, reproducibility, sensitivity and linearity. The residual level of blood products was less than the limit of residue, indicating an expected cleaning effect. The method was rapid, simple, and suitable for the cleaning validation of blood products by surface swabbing procedure.