乳胶制品中亚硝胺的分析及生物安全性评价

【作者】 封棣；

【导师】 王介东； 杨全力；

【作者基本信息】 中国协和医科大学 ， 细胞生物学， 2009， 博士

【摘要】 亚硝胺是一类具有-N-N=O结构的强致癌性有机化合物。研究人员已在广泛应用于人们日常生活和工作中的乳胶制品(奶嘴、气球、避孕套等)中检测出了致癌物亚硝胺的存在。由于在使用过程中与人体的皮肤、体液甚至组织器官密切接触,因此对乳胶制品中致癌物亚硝胺的检测分析及生物安全性评价对于维护人们的健康具有重要意义。首先,本论文对乳胶制品中的亚硝胺的迁移进行了分析研究:首次用EN12868方法(婴儿奶嘴中亚硝胺迁移量的检测方法)对中国市售的37种乳胶避孕套、10种乳胶手套、5种乳胶气球和4种硅胶奶嘴中亚硝胺的迁移量进行了检测分析。结果显示,除硅胶奶嘴中未检出亚硝胺迁移外,乳胶避孕套、手套和气球中亚硝胺迁移超标(93/11/EEC)情况严重。首次建立了乳胶手套中亚硝胺迁移量的检测方法。该方法模拟了人们在使用乳胶手套半个工作日(4小时)后亚硝胺从乳胶手套迁移至人造汗液中的情况。乳胶手套经人造汗液浸提4小时后,用二氯甲烷进行液液萃取,气相色谱-热能分析仪检测。该方法简单可靠,合理可行。中国市售的27种乳胶手套中NDMA、NDEA和NDBA被检出,总的亚硝胺迁移量为18.89～244.51μg/kg,全部超标。建立了乳胶避孕套中亚硝胺迁移量的检测方法。该方法模拟了人们在使用乳胶避孕套进行一次性交活动(10分钟)后亚硝胺从乳胶避孕套迁移至人造汗液中的情况。该方法简单可靠,合理可行。中国市场上的27种乳胶避孕套中NDMA、NDEA和NDBA被检出,总的亚硝胺迁移量为15.62～792.89μg/kg,全部超标。其次,本论文对乳胶制品及亚硝胺迁移进行了部分安全性评价:对天然乳胶制品进行了体外细胞毒性评价。通过采用浸提液法及MTT比色法对L929小鼠成纤维细胞的增殖和抑制影响进行测试,首次对国内乳胶手套进行细胞毒性评价并对其细胞毒性和亚硝胺迁移量进行了相关性分析。结果表明部分所检样品(包括乳胶避孕套)有较高的细胞毒性,并且乳胶制品的细胞毒性与亚硝胺的迁移量具有显著相关性。首次利用遗传毒理学试验(Ames试验和MN试验)评价乳胶制品中亚硝胺迁移量的安全性。根据模拟日常使用乳胶手套E05和乳胶避孕套C02后亚硝胺的迁移量设计试验浓度,对乳胶制品中迁移出的亚硝胺的诱变性进行定量鉴定。结果表明乳胶手套和避孕套中亚硝胺迁移在Ames试验和MN试验中具有较高的遗传毒性。首次对亚硝胺(NDMA、NDEA和NDBA)的小鼠阴道急毒试验进行了探索性的研究,就小鼠阴道给药方式、阴道急毒的临床表现及对组织器官的病理变化的影响作了初步研究,为小鼠亚硝胺阴道给药的毒性及致癌性试验提供依据。更多还原

【Abstract】 Nitrosamines are a family of carcinogenic compounds.Nitrosamines in latex products such as nipples,balloons and condoms have been detected by researchers.In view of their close contact with skin,body fluids and tissues,it is very important to research the release of nitrosamines from natural latex products and to evaluate their potential toxicity.Firstly,nitrosamines migration from latex products were studied in the dissertation:Nitrosamines migration from 37 latex condoms,10 latex gloves,5 latex balloons and 4 sillicon nipples collected from China market were detected by EN12868(Methods for determining the release of N-Nitrosamines from elastomer or rubber teats and soothers) for the first time.The result showed nitrosamines migration from condoms, gloves and balloons were beyond seriously the 93/11/EEC limits.It was established a method to detect the migration of nitrosamines from nature latex gloves for the first time.After the gloves were soaked in artificial sweat for 4h,the nitrosamines were extracted from artificial sweat by dichloromethane and detected by GC-TEA.The method is effective,quick and accurate.NDMA,NDEA and NEBA were found in almost 27 gloves from China market.The total migration of nitrosamines ranged from 18.89 to 244.51μg/kg,which were all above the 93/11/EEC limits.It was established a method to detect the migration of nitrosamines from nature latex condoms.After the condoms were soaked in artificial sweat for 10min,the nitrosamines were extracted from artificial sweat by dichloromethane and detected by GC-TEA.The method is effective,quick and accurate.NDMA,NDEA and NEBA were found in almost 27 condoms from China market.The total migration of nitrosamines ranged from 15.62 to 792.89μg/kg,which were all above the 93/11/EEC limits.Secondly,the biological safety evaluation of latex products and nitrosamines migration from them were studied in the dissertation:The cytotoxic potential of the gloves and condoms collected from China market was evaluated by a MTT assay with a L929 cell line.Extracts from partial gloves and condoms were found to be cytotoxic and there was a positive significant correlation between cytotoxicity and the the release level of nitrosamines.The mutagenicity of migration of nitrosamines from latex gloves and condoms were evaluated by a Salmonella typhimurium mutation assay with the strains TA98, TA97,TA100 and TA102 and by a micronucleus test(MN test) using ICR mice for the first time.The migration level of nitrosamines from glove E05 and condom C02 were Selected to design the doses in two tests to evaluate quantitatively.The high mutagenicity toxicity was found in Ames assay and MN test.An exposure of NDMA,NDEA and NDBA in ICR mice by vaginal administration to observe acute toxicity was undertaken,respectively.The vaginal administration mode, the clinical manifestations and pathomorphological change of tissues and organs was researched preliminarily.更多还原