适于蛋白类食品重金属酶联免疫检测的样品前处理研究
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摘要
酶联免疫技术是一种快速的检测重金属的方法,它可以检测水中的重金属离子,若要实现农产品中重金属的检测,需将样品中结合态的重金属转化为离子态的重金属。传统处理农产品及水产品中重金属的前处理技术虽然能准确提取样品中的重金属,但相对费时费力。针对酶联免疫检测重金属的要求,开发研究出一种能满足酶联免疫技术检测重金属要求的快速、高效和简便的农产品前处理技术具有十分现实的意义。
本文以近江牡蛎、翡翠贻贝和基围虾等代表水产类蛋白质食品为研究对象,用稀盐酸、稀硫酸、稀硝酸对三种样品进行浸提,探索水产品中重金属铜、铅、镉的最佳浸提条件。结果表明,稀盐酸浸提条件为浓度2M-4M,反应时间为10min以上,浸提温度范围为20-90℃；稀硫酸浸提条件为浓度2M-4M,反应时间为10min以上,浸提温度范围为10-90℃；稀硝酸浸提条件为浓度2M-4M,反应时间为15min以上,浸提温度范围为10-90℃。三种酸浸提水产品中重金属铜、铅、镉的效果为稀硝酸>稀盐酸>稀硫酸,尤其是稀硝酸浸提近江牡蛎中的铜、铅、镉,可以代替国标法。
本文还以猪肉、牛肉、鸡肉和鸭肉等代表禽畜类蛋白质食品为研究对象,用稀盐酸、稀硫酸、稀硝酸对四种样品进行浸提,了解禽畜类重金属铜、铅、镉残留的最佳浸提条件。结果表明,稀盐酸浸提条件为浓度1M-4M,反应时间为10min以上,浸提温度范围为10-90℃；稀硫酸浸提条件为浓度2M-4M,反应时间为10min以上,浸提温度范围为10-90℃;稀硝酸浸提条件为浓度2M-4M,反应时间为15min以上,浸提温度范围为10-90℃。稀酸浸提法三种酸的效果依次为稀硝酸>稀盐酸>稀硫酸,尤其是稀硝酸浸提鸭肉中重金属的方法,可以代替传统的重金属提取技术。
最后,本文用最适的的稀酸浸条件对水产品及禽畜类中的重金属进行了浸提,分别用ICP和酶联免疫试剂盒检测样品中的重金属含量,统计结果表明,两种检测方法的结果具有良好的线性相关关系,本文所建立的稀酸浸提蛋白类农产品的前处理方法能完全满足酶联免疫试剂盒检测重金属的要求。
Enzyme-Linked Immunosorbent Assay (ELISA) was a kind of rapid and high effective method detecting heavy metals ions in water. Heavy metals exist in agricultural products as combinated state. In order to detect the content of heavy metal in foods, it was necessary to transformed combinated metals into free ions. Heavy metals content in agricultural products or aquatic products could be determined accurately by traditional detective methods, but the methods applied in field were impeded by time-consuming in pretreatment foods sample and the large equipment. It would be of a profound significance to develop a new method applied for fast and efficient sample pretreatment according to the request of ELISA detection.
In the present, the perfect pretreatment conditions for aquatic products protein foods, such as oyster (Ostrea rivularis), mussel (Perna viridis) and Metapenaeus ensi, investigated with soak and extraction of three dilute acid (hydrochloric acid, sulfuric acid, nitric acid). The results show that the pretreatment conditions become perfect with 2M-4M concentration, 10min reaction time and 20-90℃temperature for HCl and 2M-4M concentration, 10min reaction time and 10-90℃temperature for H2SO4, and 2M-4M concentration,15min reaction time,10-90℃temperature for HNO3. The order of effection which dilute acid extract heavy metals (copper, lead, cadmium) of aquatic protein samples was HNO3> HC> H2SO4. The effection of methods extracted heavy metals (copper, lead, cadmium) of oyster with dilute nitric acid was the best in all the methods, which could replace the traditional pretreatment methods.
The perfect pretreatment conditions for livestork and poultry protein foods, such as pork meat, beef meat, chichen meat and duck meat, investigated with soak and extraction of three dilute acid (hydrochloric acid, sulfuric acid, nitric acid). The results show that the pretreatment conditions become perfect with 1M-4M concen-tration, 10min reaction time and 10-90℃temperature for HCl and 1M-4M concentration, 10min reaction time and 10-90℃temperature for H2SO4, and 2M-4M concentration,15min reaction time,20-90℃temperature for HNO3. The order of effection which dilute acid extract heavy metals (copper, lead, cadmium) of aquatic protein samples was HNO3>HCl>H2SO4. The effection of methods extracted heavy metals (copper, lead, cadmium) of duck meat with dilute nitric acid was the best in all the methods, which could replace the traditional pretreatment method.
Finally, heavy metal concentration of extractions of the aquatic products protein and livestock and poultry, obtained by the soak and extractive perfect conditions, was determined with ICP and ELISA,respectively. Statistical results show that there was a linear relationship between the two groups of concentration detected with the two methods. Therefore, the pretreatment methods of dilute acid extract could meet the requirements of detecting heavy metals (copper, lead, cadmium) of protein foods with ELISA.
本文以近江牡蛎、翡翠贻贝和基围虾等代表水产类蛋白质食品为研究对象,用稀盐酸、稀硫酸、稀硝酸对三种样品进行浸提,探索水产品中重金属铜、铅、镉的最佳浸提条件。结果表明,稀盐酸浸提条件为浓度2M-4M,反应时间为10min以上,浸提温度范围为20-90℃；稀硫酸浸提条件为浓度2M-4M,反应时间为10min以上,浸提温度范围为10-90℃；稀硝酸浸提条件为浓度2M-4M,反应时间为15min以上,浸提温度范围为10-90℃。三种酸浸提水产品中重金属铜、铅、镉的效果为稀硝酸>稀盐酸>稀硫酸,尤其是稀硝酸浸提近江牡蛎中的铜、铅、镉,可以代替国标法。
本文还以猪肉、牛肉、鸡肉和鸭肉等代表禽畜类蛋白质食品为研究对象,用稀盐酸、稀硫酸、稀硝酸对四种样品进行浸提,了解禽畜类重金属铜、铅、镉残留的最佳浸提条件。结果表明,稀盐酸浸提条件为浓度1M-4M,反应时间为10min以上,浸提温度范围为10-90℃；稀硫酸浸提条件为浓度2M-4M,反应时间为10min以上,浸提温度范围为10-90℃;稀硝酸浸提条件为浓度2M-4M,反应时间为15min以上,浸提温度范围为10-90℃。稀酸浸提法三种酸的效果依次为稀硝酸>稀盐酸>稀硫酸,尤其是稀硝酸浸提鸭肉中重金属的方法,可以代替传统的重金属提取技术。
最后,本文用最适的的稀酸浸条件对水产品及禽畜类中的重金属进行了浸提,分别用ICP和酶联免疫试剂盒检测样品中的重金属含量,统计结果表明,两种检测方法的结果具有良好的线性相关关系,本文所建立的稀酸浸提蛋白类农产品的前处理方法能完全满足酶联免疫试剂盒检测重金属的要求。
Enzyme-Linked Immunosorbent Assay (ELISA) was a kind of rapid and high effective method detecting heavy metals ions in water. Heavy metals exist in agricultural products as combinated state. In order to detect the content of heavy metal in foods, it was necessary to transformed combinated metals into free ions. Heavy metals content in agricultural products or aquatic products could be determined accurately by traditional detective methods, but the methods applied in field were impeded by time-consuming in pretreatment foods sample and the large equipment. It would be of a profound significance to develop a new method applied for fast and efficient sample pretreatment according to the request of ELISA detection.
In the present, the perfect pretreatment conditions for aquatic products protein foods, such as oyster (Ostrea rivularis), mussel (Perna viridis) and Metapenaeus ensi, investigated with soak and extraction of three dilute acid (hydrochloric acid, sulfuric acid, nitric acid). The results show that the pretreatment conditions become perfect with 2M-4M concentration, 10min reaction time and 20-90℃temperature for HCl and 2M-4M concentration, 10min reaction time and 10-90℃temperature for H2SO4, and 2M-4M concentration,15min reaction time,10-90℃temperature for HNO3. The order of effection which dilute acid extract heavy metals (copper, lead, cadmium) of aquatic protein samples was HNO3> HC> H2SO4. The effection of methods extracted heavy metals (copper, lead, cadmium) of oyster with dilute nitric acid was the best in all the methods, which could replace the traditional pretreatment methods.
The perfect pretreatment conditions for livestork and poultry protein foods, such as pork meat, beef meat, chichen meat and duck meat, investigated with soak and extraction of three dilute acid (hydrochloric acid, sulfuric acid, nitric acid). The results show that the pretreatment conditions become perfect with 1M-4M concen-tration, 10min reaction time and 10-90℃temperature for HCl and 1M-4M concentration, 10min reaction time and 10-90℃temperature for H2SO4, and 2M-4M concentration,15min reaction time,20-90℃temperature for HNO3. The order of effection which dilute acid extract heavy metals (copper, lead, cadmium) of aquatic protein samples was HNO3>HCl>H2SO4. The effection of methods extracted heavy metals (copper, lead, cadmium) of duck meat with dilute nitric acid was the best in all the methods, which could replace the traditional pretreatment method.
Finally, heavy metal concentration of extractions of the aquatic products protein and livestock and poultry, obtained by the soak and extractive perfect conditions, was determined with ICP and ELISA,respectively. Statistical results show that there was a linear relationship between the two groups of concentration detected with the two methods. Therefore, the pretreatment methods of dilute acid extract could meet the requirements of detecting heavy metals (copper, lead, cadmium) of protein foods with ELISA.
引文
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