采用细胞毒性试验对23个药用包装材料进行生物安全性评价

目的：用体外细胞毒性试验对23个药用包装材料进行生物安全性评价，探讨样品浸提液制备方法对检测结果的影响。方法：设定样品质量/浸提液体积比值为0.2 g/mL，药用包装材料分别用MEM培养基(37℃、24 h)、氯化钠注射液和去离子水(121℃、1 h)制备浸提液。小鼠成纤维细胞L929培养于96孔板中，每孔加入样品浸提液100μL培养48 h后，用酶标仪测定吸光度值，根据细胞的相对增殖度(RGR)进行细胞毒性反应分级，≤1级为阴性，2级为可疑阳性，≥3级为阳性。结果：23个受试样品的MEM培养基浸提液均为0～1级；氯化钠注射液浸提液检出9个受试样品为2级以上，其中3个为3级以上；用去离子水浸提液复核9个受试样品可疑阳性结果，仅3个为2级以上，其中1个为3级。结论：体外试验在排除渗透压、pH值等的干扰，用最大溶出度浸提液作为溶剂配制培养基进行试验，能够明显增加药用包装材料毒性反应阳性结果的检出率。

Biosafety evaluation of 23 pharmaceutical packaging materials by cytotoxicity test

OBJECTIVE： In vitro cytotoxicity test was used to evaluate biological safety of 23 pharmaceutical packaging materials. METHODS： The sample mass/extract volume ratio was set to be 0.2 g/mL. Extracts of pharmaceutical packaging materials were prepared with MEM medium (37 ℃，24 h)，sodium chloride injection and deionized water (121 ℃，1 h). Mouse fibroblasts L929 were cultured in a 96-well plate. 100 μL of sample extracts was added to each well and cultured for 48 hours. The absorbance was measured with a microplate reader. According to the relative proliferation of the cells (RGR) for grading，grade ≤ 1 was negative，grade 2 was suspicious positive，and grade ≥ 3 was positive. RESULTS：The 23 samples of MEM medium extracts ranged between 0-1 grade. Nine samples tested in sodium chloride injection extracts were suspicious positive above grade 2，of which 3 were positive above grade 3. The suspected positive results of 9 test samples were rechecked with deionized water extract，only 3 were suspected positive above grade 2，of which 1 was positive at grade 3. CONCLUSION：In vitro test，excluding the interference of osmotic pressure and pH value， was conducted by culture medium prepared with the maximum dissolution extract as the solvent. The approach significantly increased the detection of positive results of pharmaceutical packaging materials.