摘要
目的:比较不同来源甲硫氨酸维B_1注射剂体外细胞毒性的差别并寻找差别形成的主要原因。方法:将不同来源甲硫氨酸维B_1注射剂不同浓度的稀释液与小鼠成纤维细胞L-929接触培养,通过倒置相差显微镜观察细胞形态,采用四甲基偶氮唑盐比色法(MTT法)量化细胞毒性,计算相对增殖率,并进行细胞毒性评价。建立甲硫氨酸维B_1注射剂中枸橼酸含量的高效液相色谱检测方法,考察枸橼酸含量与制剂细胞毒性的相关性。结果:7个生产厂家的甲硫氨酸维B_1注射液细胞毒性无明显差别。12个生产厂家的注射用甲硫氨酸维B_1中有11个厂家的样品不同浓度组对L-929的细胞毒性级别为1级或2级。但J厂家的注射用甲硫氨酸维B_1样品(20 mg·mL^(-1)和10 mg·mL^(-1))对L-929的细胞毒性级别为4级,表现为重度毒性。枸橼酸质量浓度3,1.5,0.75 mg·mL^(-1)的相对增殖率分别为7%、45%、74%,而对应的含等量枸橼酸的制剂浓度(以甲硫氨酸计)20,10,5 mg·mL^(-1)的相对增殖率分别为6%、28%、53%,细胞毒性试验结果吻合。结论:不同厂家生产的甲硫氨酸维B_1注射剂由于处方工艺不同等,其细胞毒性存在差别。枸橼酸作为注射剂的常用辅料,在高于0.188 mg·mL^(-1)的浓度下存在细胞毒性,枸橼酸含量与细胞毒性呈正相关性,J厂家样品的重度细胞毒性可能主要源于过高浓度的辅料枸橼酸。建议生产厂家降低枸橼酸添加量,以减少制剂的细胞毒性,提高药品的安全性。
OBJECTIVE To compare the in vitro cytotoxic responses of methionine and vitamin B1 injections with different sources,and to find the causes for the differences.METHODS The mice fibroblast line L-929 was cultured with various methionine and vitamin B1 injections at different doses.The cellular morphology was observed under an inverted phase contrast microscope and proliferation of the cells was examined by using methyl thiazolyl tetrazolium(MTT)assay.Relative growth rate(RGR)was calculated.Moreover,cytotoxicity was evaluated.HPLC method for the determination of citric acid in methionine and vitamin B1 injection was established,and the correlation between the content of citric acid and cytotoxicity of the preparation was investigated.RESULTS There was no significant difference in cytotoxicity between methionine and vitamin B1 injections from 7 manufacturers,and L-929 cytotoxicity of the samples from 11 out of 12 manufacturers in the different concentration groups was grade 1 or 2.However,the samples of methionine and vitamin B1(20 mg·mL-1 and 10 mg·mL-1)from J manufacturers showed cytotoxic response of grade 4,indicating severe toxicity.Relative growth rates of the citric acid concentrations of 3 mg·mL-1,1.5 mg·mL-1 and 0.75 mg·mL-1 were 7%,45%and 74%,respectively.Relative growth rates of the corresponding preparation concentrations(calculated with reference to methionine)of 20 mg·mL-1,10 mg·mL-1 and 5 mg·mL-1 containing equivalent citric acid were 6%,28%,53%,respectively,which was consistent with the results of cytotoxicity tests.CONCLUSION Methionine and vitamin B1 injections produced by different manufacturers have different in cytotoxicity due to different prescriptions and processes.As a common excipient for injections,citric acid at a concentration higher than 0.188 mg·mL-1may present cytotoxicity.The content of citric acid exhibits positive correlation with cytotoxicity grades.The severe cytotoxicity of samples from J manufacturers may be mainly due to the too high concentration of citric acid.It is
作者
许雷鸣
王莉丽
李翔
孙备
顾倩
XU Lei-ming;WANG Li-li;LI Xiang;SUN Bei;GU Qian(Anhui Institute for Food and Drug Control,Anhui Hefei 230051,China;Wannan Medical College,Anhui Wuhu 241000,China)
出处
《中国医院药学杂志》
CAS
北大核心
2019年第10期1046-1051,共6页
Chinese Journal of Hospital Pharmacy
