分子印迹技术及其与毛细管电泳联用在复杂样品中双酚A检测的应用研究
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摘要
双酚A(BPA)是一种具有生物毒性和内分泌干扰作用的环境雌激素,广泛的痕量存在于环境、生物和食品中。所以需要建立一种快速方便、灵敏度高、选择性好、价廉的分析方法来分离检测复杂样品中的双酚A。
分子印迹技术是基于分子识别理论而发展起来的,其核心为分子印迹聚合物(MIPs)的制备。MIPs也称人工抗体,能够特异性的结合目标分子,且其机械稳定性和化学稳定性好,在环境、生物、临床等领域都有广阔的应用前景。毛细管电泳(CE)由于其快速、高效、进样量少等特点,目前已成为分析化学领域重要的分离工具。
本文把分子印迹技术和固相萃取技术结合起来,得到了一种高选择性、快速有效的新型样品前处理技术——分子印迹固相萃取(MISPE),并首次把MISPE与CE技术联用,对复杂样品中的双酚A进行了检测。主要内容如下:
一、对分子印迹技术作了较为全面和系统的综述,重点对分子印迹聚合物的制备方法和分子印迹技术在分析化学当中的应用作了详细的评述。
二、用沉淀聚合方法制备了双酚A的分子印迹聚合物(MIPs),并研究了致孔剂溶解度参数,致孔剂用量的影响,功能单体、模板用量对吸附容量和选择性的影响。最优化的条件是4-VP为功能单体,60ml乙腈为致孔剂,模板双酚A用量6mmol。该优化条件下制备的MIPs为规则的球形微球,最大吸附容量Bmax=1.37×10~(-5)mol/g,印迹因子I=6.53。
三、CE的优化分离条件为pH=11.5,电压为18kv,此时双酚A和其结构类似物在15min内得到了基线分离。对MISPE柱的淋洗和洗脱条件进行了优化,在最佳条件下,考察了MISPE回收率和准确度。实验表明,不同上样体积对MISPE影响回收率不大,RSD<5.8。在不同样品基质下,回收率略有变化,为71.20%~86.23%。通过和C18柱比较,MISPE对双酚A有更高的选择性和回收率。最后,采用MISPE与CE联用对自来水、化工废水、长江水、长江土壤、虾、人尿样品中的双酚A进行了检测,加标回收率为93.69%~105.4%,在水样、固体样和尿样中的检测限分别为3.0~6.9μg/l,1.8~2.1μg/g和84μg/l,结果满意。
Bisphenol A (BPA), a well known environmental estrogen, has biotoxic and endocrine-disrupting effects, wildly exits in environmental, biological and food samples at a trace amount. So it is necessary to develop a fast, sensitive, high selective and cheap method for determination BPA in complex sample matrices.
Molecular imprinted technology is based on the molecular recognition theory,and its main procedure is to synthesize molecular imprinted polymers(MIPs). Because MIPs have chemical stability and selective combination with the target analyte, they have been widely used in environmental, biological and clinical samples. Capillary electrophoresis (CE) is a noble analytical technique in analytical chemistry which has some advantages, such as highly efficient and rapid analysis.
In this thesis, a new high selective, fast and effective sample pretreatment technique-molecular imprinted solid-phase extraction (MISPE) was achieved by combination of molecular imprinted technology and solid-phase extraction (SPE). It was the first time to determine BPA in complex sample matrices by MISPE coupled with CE. The main contents in this thesis are described as follows:
1. In this paper, the molecular imprinted technology was systematically summed up. Synthetic methods of molecularly imprinted polymers and application of the molecular imprinted technology in analytical chemistry were particularly outlined.
2. Bisphenol A imprinted polymers were synthesized by precipitation polymerization, and effects of the preparing condition, such as the ratio and volume of the porogenic solvent, the sorts of the functional monomers and the amount of template on the maximum absorption capacity and imprinted effect of the polymers were investigated. At last, 4-VP and 60ml acetonitrile were used as functional monomer and porogenic solvent, respectively, the amount of the bisphenol A was 6mmol. Under this optimal condition, regulary microspheres were obtained, the values of imprinted factor and Bmax were 6.53 and 1.37×10~(-5)mol/g, respectively.
3. The optimal separation condition of CE were pH=11.5, voltage=18kv. Under the optimal condition, BPA and its analogues were separated in 15 min. The washing and eluting protocols of MISPE were optimized, then the recoveries and the relative standard deviations (RSD) were investigated. It was showed the recoveries were basically same when samples of different volumes were loaded, RSD<5.8, and the recoveries changed from 71.20% to 86.23% for different sample matrices. Compared with C18 SPE, the MISPE has higher selectivity and recovery for BPA. At last, BPA was determined in tap water, chemical plant waste water, Yangtse River water, Yangtse River soil, shrimp and human urine samples by MISPE coupled with CE, and good recovery and limit of detection (LOD) were obtained, spiked recoveries was from 93.69% to 105.4%, LOD in aqueous sample, solid sample and human urine sample were 3.0~6.9μg/l,1.8~2.1μg/g and 84μg/l, respectively.
分子印迹技术是基于分子识别理论而发展起来的,其核心为分子印迹聚合物(MIPs)的制备。MIPs也称人工抗体,能够特异性的结合目标分子,且其机械稳定性和化学稳定性好,在环境、生物、临床等领域都有广阔的应用前景。毛细管电泳(CE)由于其快速、高效、进样量少等特点,目前已成为分析化学领域重要的分离工具。
本文把分子印迹技术和固相萃取技术结合起来,得到了一种高选择性、快速有效的新型样品前处理技术——分子印迹固相萃取(MISPE),并首次把MISPE与CE技术联用,对复杂样品中的双酚A进行了检测。主要内容如下:
一、对分子印迹技术作了较为全面和系统的综述,重点对分子印迹聚合物的制备方法和分子印迹技术在分析化学当中的应用作了详细的评述。
二、用沉淀聚合方法制备了双酚A的分子印迹聚合物(MIPs),并研究了致孔剂溶解度参数,致孔剂用量的影响,功能单体、模板用量对吸附容量和选择性的影响。最优化的条件是4-VP为功能单体,60ml乙腈为致孔剂,模板双酚A用量6mmol。该优化条件下制备的MIPs为规则的球形微球,最大吸附容量Bmax=1.37×10~(-5)mol/g,印迹因子I=6.53。
三、CE的优化分离条件为pH=11.5,电压为18kv,此时双酚A和其结构类似物在15min内得到了基线分离。对MISPE柱的淋洗和洗脱条件进行了优化,在最佳条件下,考察了MISPE回收率和准确度。实验表明,不同上样体积对MISPE影响回收率不大,RSD<5.8。在不同样品基质下,回收率略有变化,为71.20%~86.23%。通过和C18柱比较,MISPE对双酚A有更高的选择性和回收率。最后,采用MISPE与CE联用对自来水、化工废水、长江水、长江土壤、虾、人尿样品中的双酚A进行了检测,加标回收率为93.69%~105.4%,在水样、固体样和尿样中的检测限分别为3.0~6.9μg/l,1.8~2.1μg/g和84μg/l,结果满意。
Bisphenol A (BPA), a well known environmental estrogen, has biotoxic and endocrine-disrupting effects, wildly exits in environmental, biological and food samples at a trace amount. So it is necessary to develop a fast, sensitive, high selective and cheap method for determination BPA in complex sample matrices.
Molecular imprinted technology is based on the molecular recognition theory,and its main procedure is to synthesize molecular imprinted polymers(MIPs). Because MIPs have chemical stability and selective combination with the target analyte, they have been widely used in environmental, biological and clinical samples. Capillary electrophoresis (CE) is a noble analytical technique in analytical chemistry which has some advantages, such as highly efficient and rapid analysis.
In this thesis, a new high selective, fast and effective sample pretreatment technique-molecular imprinted solid-phase extraction (MISPE) was achieved by combination of molecular imprinted technology and solid-phase extraction (SPE). It was the first time to determine BPA in complex sample matrices by MISPE coupled with CE. The main contents in this thesis are described as follows:
1. In this paper, the molecular imprinted technology was systematically summed up. Synthetic methods of molecularly imprinted polymers and application of the molecular imprinted technology in analytical chemistry were particularly outlined.
2. Bisphenol A imprinted polymers were synthesized by precipitation polymerization, and effects of the preparing condition, such as the ratio and volume of the porogenic solvent, the sorts of the functional monomers and the amount of template on the maximum absorption capacity and imprinted effect of the polymers were investigated. At last, 4-VP and 60ml acetonitrile were used as functional monomer and porogenic solvent, respectively, the amount of the bisphenol A was 6mmol. Under this optimal condition, regulary microspheres were obtained, the values of imprinted factor and Bmax were 6.53 and 1.37×10~(-5)mol/g, respectively.
3. The optimal separation condition of CE were pH=11.5, voltage=18kv. Under the optimal condition, BPA and its analogues were separated in 15 min. The washing and eluting protocols of MISPE were optimized, then the recoveries and the relative standard deviations (RSD) were investigated. It was showed the recoveries were basically same when samples of different volumes were loaded, RSD<5.8, and the recoveries changed from 71.20% to 86.23% for different sample matrices. Compared with C18 SPE, the MISPE has higher selectivity and recovery for BPA. At last, BPA was determined in tap water, chemical plant waste water, Yangtse River water, Yangtse River soil, shrimp and human urine samples by MISPE coupled with CE, and good recovery and limit of detection (LOD) were obtained, spiked recoveries was from 93.69% to 105.4%, LOD in aqueous sample, solid sample and human urine sample were 3.0~6.9μg/l,1.8~2.1μg/g and 84μg/l, respectively.
引文
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