药品包装材料中全氟辛烷磺酸类物质检测与安全评价技术
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摘要
药品包装的质量安全是药品质量安全的重要组成部分。全氟辛烷磺酸类物质(PFOS)因其疏水、疏油的特性,在药品包装领域得到了广泛应用。近期的研究结果表明,PFOS具有毒性、持久性和生物蓄积性,已被许多国家政府和国际组织列为限用物质。因此,开展药品包装材料中PFOS的检测技术和毒理学行为研究,并开展PFOS的生态风险和健康风险评价研究具有重要的意义。
本文首次应用先进的快速溶剂萃取技术(ASE),以甲醇为溶剂萃取药品包装材料样品中的全氟辛烷磺酸类物质,利用响应面法得到了萃取的优化条件。萃取液经N,O-双(三甲基硅烷基)三氟乙酰胺(BSTFA)衍生化后,用气相色谱/质谱联用(GC/MS)仪进行测定。在优化的实验条件下,测得药品包装材料中PFOS检测方法的线性范围为50.0 ng/mL～0.5×103 ng/mL,线性相关系数为0.9913,最小检出限13.9 ng/mL,样品的加标回收率为98.4%。对药品包装材料的测试结果为,聚四氟乙烯材质的药品包装材料中检测出PFOS的含量范围为31.6ng/g～33.7 ng/g之间。
本文同时应用超高效液相色谱-质谱/质谱联用技术(UPLC-MS/MS)对药品包装材料中PFOS的检测进行了研究。药品包装材料以甲醇为溶剂,采用快速溶剂萃取技术萃取其中的PFOS,萃取液经过滤、浓缩处理后,进行液相色谱-质谱/质谱分析。在优化的实验条件下,测得方法的线性范围:0.5μg/L～10μg/L,线性相关系数0.9996;方法的回收率在88%～105%之间,最低检出限1.0 ng/g。测得聚四氟乙烯包装材料中PFOS含量为34.1 ng/g,表明得到了与GC/MS相近的结果。
本文首次系统开展了PFOS急性毒性、皮肤刺激/腐蚀性、生殖毒性和神经毒性等毒理学行为研究,结果表明昆明种小鼠PFOS的经口急性毒性LD50值为501 mg/kg,按照联合国《全球化学品分类和标签协调制度》(GHS)的对于急性毒性物质的分类标准,分类定级为类型Ⅳ。急性吸入毒性的实验结果说明PFOS不属于急性吸入毒性物质。斑马鱼PFOS的水生生物急性毒性LC50值为37.31 mg/L,在GHS体系中,PFOS为危害水环境的毒性物质,类型为急性Ⅲ型。皮肤刺激性/腐蚀性实验中,PFOS皮肤刺激评分为0.6分,根据刺激强度分类为轻微刺激性物质。PFOS生殖毒性实验中,骨髓微核实验结果显示PFOS有诱导小鼠骨髓细胞微核数升高的作用,提示PFOS能够诱导染色体的损伤和畸变,具有一定的遗传毒性。PFOS的精子畸形实验结果显示PFOS染毒对小鼠精子数量没有明显影响,但对精子活动力有影响并且致使小鼠精子畸形率增加,提示PFOS对雄性小鼠生殖细胞具有潜在的损伤作用。PFOS的神经毒性实验提示PFOS对脑组织有直接毒性损害或者其他继发性损害,且存在一定的剂量—效应关系。
本文首次应用商值法对PFOS在中国部分水域、亚洲沿海和美国部分城市生态环境风险进行了评价,结果表明,亚洲沿海和我国部分地区含PFOS的鱼类和水生生物生态风险和藻类生长抑制风险属于低风险水平;美国部分城市水环境PFOS对绿藻和水华鱼腥藻等藻类的生长抑制风险属于低风险水平;对于肋骨条藻具有中等风险,对斑马鱼、虹鳟鱼、红鲈鱼、水蚤等的生态风险属低风险水平;对于大鳍鳞鳃太阳鱼、黑头软口鲦糠虾和牡蛎具有中等风险。
应用人体暴露模型对PFOS在中国部分水域、亚洲沿海和美国部分城市的健康风险进行了评价。以人群终生超额风险度(R)作为评价指标时,亚洲沿海和我国部分地区水域的人群终生超额风险度R均小于<10-6,表示风险度不明显。美国部分城市水体人群终生超额风险度R介于10-6～10-4之间,存在一定的风险。
The safety of drug packaging materials is an important part of the drug safety. Perfluorooctane sulfonate (PFOS) is widely used in production of drug packaging materials, due to its non-hydrophilic and non-lipophilic properties. Recent research demonstrated that PFOS is toxic, bioaccumulative and persistent in the environment, and was listed as restricted substance by many countries and international organization. It is of great significance to determine the PFOS in packaging materials and to assess the ecological and health risk of PFOS.
Accelerated solvent extraction technique was used to effectively extract PFOS with methanol from drug packaging materials, of which the parameters were optimized by response surface methodology. A silylation step was then carried out for the extract by adding N,O-bis(trimethylsilyl)trifluoroacetamide (BSTFA). The silylated PFOS were identified and quantified by gas chromatograph/mass (GC/MS). The test results showed that the line range is from 50.0 ng/mL to 0.5×103 ng/mL; the correlation coefficient is 0.9913; the low limit of detection (LOD) is 13.9 ng/mL; and the average recovery is 98.4%. The samples test results showed that the concentrations of PFOS in a kind of polytertfluoroethylene (PTFE) drug packaging materials is from 31.6 ng/g to 33.7 ng/g.
The ultra proficiency liquid chromatograph-mass/mass (UPLC-MS/MS) method was also developed to determine the trace amount of PFOS in the drug packaging materials. The methanol was used to extract PFOS from the samples. After filtration and concentration, the extract was quantified by LC-MS/MS. The test results showed that the line range is from 0.5μg/mL to 1.0×102μg/mL; the correlation coefficient is 0.9996; the low limit of detection (LOD) is 1.0 ng/g, and the average recovery is between 88% - 105%. The samples test results showed that the concentration of PFOS in a kind of polytertfluoroethylene (PTFE) drug packaging materials is 34.1 ng/g, indicating that the similar determination result was obtained compared to GC/MS method.
The acute toxicity, skin irritation / corrosion, reproductive toxicity and neurotoxicity of Perfluorooctanesulfonate (PFOS) were studied in this thesis. The resulted show that acute toxicity LD50 value of mice is 501 mg/kg. PFOS is classified as 4 type graded according to the document of“Globally Harmonized System of Classification and Labelling of Chemicals”(GHS). Results of acute inhalation toxicity test showed that PFOS does not belong to acute inhalation of toxic substances. LC50 value of aquatic acute toxicity (zebrafish) is 37.31 mg/L. In the GHS system, PFOS is aquatic toxicity substance type 3. Skin irritation/corrosion test results showed that PFOS is not irritation substance but minor irritant substance. Bone marrow micronucleus test results showed that PFOS induced micronuclei in mouse bone marrow cells elevated, which means PFOS has a certain degree of genetic toxicity. And the sperm abnormality test results also prompted that PFOS has potential damage effect on male reproductive cells. Finally the neurotoxicity test results indicated that PFOS has a direct toxic damage or other damages on the brain tissue with certain dose - response relationship.
Based on the published data of PFOS levels from China and Asia coast areas as well as from the aquatic environment in the USA, ecological risk assessment (ERA) of PFOS was performed using hazard quotient method. The results showed that, for both China and Asia coast, the ecological risk index Q(f,c) is lower than 0.001, and the growth inhibition risk index Q(a) is lower than 0.003, of which the risk level is relatively low. The growth inhibition risk for selenastrum capricornutum and anabaena flosaquae is low. While for skeletonema costatum, the growth inhibition risk is moderate. For zebra fish, rainbow traut, sheepshead minnow and water flea, the ecological risk is low in America. And for bluegill sunfish, pimephales promelas, mysidopsis bahia and oyster, the ecological risk is moderate.
Mathematical model was established and applied for human health risk assessment (HRA) of PFOS for China water area, Asia coastal area and USA water area. And the human lifetime excess risk index (R) is lower than 10-6 for both China water area and Asia coastal area, and is of low risk level. The human lifetime excess risk index (R) of some USA cities is between 10-6 and 10-4, and reaches the low risk level.
本文首次应用先进的快速溶剂萃取技术(ASE),以甲醇为溶剂萃取药品包装材料样品中的全氟辛烷磺酸类物质,利用响应面法得到了萃取的优化条件。萃取液经N,O-双(三甲基硅烷基)三氟乙酰胺(BSTFA)衍生化后,用气相色谱/质谱联用(GC/MS)仪进行测定。在优化的实验条件下,测得药品包装材料中PFOS检测方法的线性范围为50.0 ng/mL～0.5×103 ng/mL,线性相关系数为0.9913,最小检出限13.9 ng/mL,样品的加标回收率为98.4%。对药品包装材料的测试结果为,聚四氟乙烯材质的药品包装材料中检测出PFOS的含量范围为31.6ng/g～33.7 ng/g之间。
本文同时应用超高效液相色谱-质谱/质谱联用技术(UPLC-MS/MS)对药品包装材料中PFOS的检测进行了研究。药品包装材料以甲醇为溶剂,采用快速溶剂萃取技术萃取其中的PFOS,萃取液经过滤、浓缩处理后,进行液相色谱-质谱/质谱分析。在优化的实验条件下,测得方法的线性范围:0.5μg/L～10μg/L,线性相关系数0.9996;方法的回收率在88%～105%之间,最低检出限1.0 ng/g。测得聚四氟乙烯包装材料中PFOS含量为34.1 ng/g,表明得到了与GC/MS相近的结果。
本文首次系统开展了PFOS急性毒性、皮肤刺激/腐蚀性、生殖毒性和神经毒性等毒理学行为研究,结果表明昆明种小鼠PFOS的经口急性毒性LD50值为501 mg/kg,按照联合国《全球化学品分类和标签协调制度》(GHS)的对于急性毒性物质的分类标准,分类定级为类型Ⅳ。急性吸入毒性的实验结果说明PFOS不属于急性吸入毒性物质。斑马鱼PFOS的水生生物急性毒性LC50值为37.31 mg/L,在GHS体系中,PFOS为危害水环境的毒性物质,类型为急性Ⅲ型。皮肤刺激性/腐蚀性实验中,PFOS皮肤刺激评分为0.6分,根据刺激强度分类为轻微刺激性物质。PFOS生殖毒性实验中,骨髓微核实验结果显示PFOS有诱导小鼠骨髓细胞微核数升高的作用,提示PFOS能够诱导染色体的损伤和畸变,具有一定的遗传毒性。PFOS的精子畸形实验结果显示PFOS染毒对小鼠精子数量没有明显影响,但对精子活动力有影响并且致使小鼠精子畸形率增加,提示PFOS对雄性小鼠生殖细胞具有潜在的损伤作用。PFOS的神经毒性实验提示PFOS对脑组织有直接毒性损害或者其他继发性损害,且存在一定的剂量—效应关系。
本文首次应用商值法对PFOS在中国部分水域、亚洲沿海和美国部分城市生态环境风险进行了评价,结果表明,亚洲沿海和我国部分地区含PFOS的鱼类和水生生物生态风险和藻类生长抑制风险属于低风险水平;美国部分城市水环境PFOS对绿藻和水华鱼腥藻等藻类的生长抑制风险属于低风险水平;对于肋骨条藻具有中等风险,对斑马鱼、虹鳟鱼、红鲈鱼、水蚤等的生态风险属低风险水平;对于大鳍鳞鳃太阳鱼、黑头软口鲦糠虾和牡蛎具有中等风险。
应用人体暴露模型对PFOS在中国部分水域、亚洲沿海和美国部分城市的健康风险进行了评价。以人群终生超额风险度(R)作为评价指标时,亚洲沿海和我国部分地区水域的人群终生超额风险度R均小于<10-6,表示风险度不明显。美国部分城市水体人群终生超额风险度R介于10-6～10-4之间,存在一定的风险。
The safety of drug packaging materials is an important part of the drug safety. Perfluorooctane sulfonate (PFOS) is widely used in production of drug packaging materials, due to its non-hydrophilic and non-lipophilic properties. Recent research demonstrated that PFOS is toxic, bioaccumulative and persistent in the environment, and was listed as restricted substance by many countries and international organization. It is of great significance to determine the PFOS in packaging materials and to assess the ecological and health risk of PFOS.
Accelerated solvent extraction technique was used to effectively extract PFOS with methanol from drug packaging materials, of which the parameters were optimized by response surface methodology. A silylation step was then carried out for the extract by adding N,O-bis(trimethylsilyl)trifluoroacetamide (BSTFA). The silylated PFOS were identified and quantified by gas chromatograph/mass (GC/MS). The test results showed that the line range is from 50.0 ng/mL to 0.5×103 ng/mL; the correlation coefficient is 0.9913; the low limit of detection (LOD) is 13.9 ng/mL; and the average recovery is 98.4%. The samples test results showed that the concentrations of PFOS in a kind of polytertfluoroethylene (PTFE) drug packaging materials is from 31.6 ng/g to 33.7 ng/g.
The ultra proficiency liquid chromatograph-mass/mass (UPLC-MS/MS) method was also developed to determine the trace amount of PFOS in the drug packaging materials. The methanol was used to extract PFOS from the samples. After filtration and concentration, the extract was quantified by LC-MS/MS. The test results showed that the line range is from 0.5μg/mL to 1.0×102μg/mL; the correlation coefficient is 0.9996; the low limit of detection (LOD) is 1.0 ng/g, and the average recovery is between 88% - 105%. The samples test results showed that the concentration of PFOS in a kind of polytertfluoroethylene (PTFE) drug packaging materials is 34.1 ng/g, indicating that the similar determination result was obtained compared to GC/MS method.
The acute toxicity, skin irritation / corrosion, reproductive toxicity and neurotoxicity of Perfluorooctanesulfonate (PFOS) were studied in this thesis. The resulted show that acute toxicity LD50 value of mice is 501 mg/kg. PFOS is classified as 4 type graded according to the document of“Globally Harmonized System of Classification and Labelling of Chemicals”(GHS). Results of acute inhalation toxicity test showed that PFOS does not belong to acute inhalation of toxic substances. LC50 value of aquatic acute toxicity (zebrafish) is 37.31 mg/L. In the GHS system, PFOS is aquatic toxicity substance type 3. Skin irritation/corrosion test results showed that PFOS is not irritation substance but minor irritant substance. Bone marrow micronucleus test results showed that PFOS induced micronuclei in mouse bone marrow cells elevated, which means PFOS has a certain degree of genetic toxicity. And the sperm abnormality test results also prompted that PFOS has potential damage effect on male reproductive cells. Finally the neurotoxicity test results indicated that PFOS has a direct toxic damage or other damages on the brain tissue with certain dose - response relationship.
Based on the published data of PFOS levels from China and Asia coast areas as well as from the aquatic environment in the USA, ecological risk assessment (ERA) of PFOS was performed using hazard quotient method. The results showed that, for both China and Asia coast, the ecological risk index Q(f,c) is lower than 0.001, and the growth inhibition risk index Q(a) is lower than 0.003, of which the risk level is relatively low. The growth inhibition risk for selenastrum capricornutum and anabaena flosaquae is low. While for skeletonema costatum, the growth inhibition risk is moderate. For zebra fish, rainbow traut, sheepshead minnow and water flea, the ecological risk is low in America. And for bluegill sunfish, pimephales promelas, mysidopsis bahia and oyster, the ecological risk is moderate.
Mathematical model was established and applied for human health risk assessment (HRA) of PFOS for China water area, Asia coastal area and USA water area. And the human lifetime excess risk index (R) is lower than 10-6 for both China water area and Asia coastal area, and is of low risk level. The human lifetime excess risk index (R) of some USA cities is between 10-6 and 10-4, and reaches the low risk level.
引文
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