原柱富集-高效液相色谱法测定水体中痕量污染物
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摘要
高效液相色谱法因具有分离效率高、分析速度快等特点而被广泛应用于药物分析及环境分析等领域。然而面对复杂基质样品和痕量组分样品,常常需要在分析前对样品进行预处理以达到净化和富集的目的。固相萃取、液液萃取等经典的离线预处理方法不但会消耗大量溶剂,操作繁琐、费时,还会造成组分的损失。于是,在线样品预处理技术和高效液相色谱联用的分析方法应运而生,其中固相萃取-高效液相色谱在线联用方法结合了二者的优势,在生物样品及环境污染物分析中得到广泛的应用。本文建立了一种简便的在线固相萃取-液相色谱方法,在分析柱上完成对待测组分的富集,因而称其为原柱富集-高效液相色谱法。对新方法进行了性能评价并将其用于水体中几种痕量污染物的分析研究。本论文的主要研究内容如下:
论文第2章在总结在线固相萃取-液相色谱分析方法特点的基础上,通过改变常规液相色谱操作模式,提出并建立了原柱富集-液相色谱分析方法。通过考察发现,在改变富集时间及样品浓度两个条件下,新方法测得的峰面积和变化因素之间线性关系良好,R2分别为0.9996和0.9840,说明新方法具备对痕量组分进行定量富集的能力。
论文第3章采用原柱富集-液相色谱方法对水体中邻苯二甲酸酯类物质进行了分析,通过对各影响因素的考察,得到邻苯二甲酸二甲酯(DMP)的最佳分析条件为:水样添加5%体积分数的甲醇,以0.6 mL/min的流速富集30 min,用60%甲醇水溶液洗脱,在240 nm波长下检测。在最佳分析条件下,DMP的检测限为20 ng/L,通过外标工作曲线法测得校内一处湖水中DMP浓度为0.3μg/L。另外,在单组分分析基础上,进行了新方法多组分分析的尝试。
论文第4章和第5章采用原柱富集-液相色谱方法对自来水中硝基苯、苯、氯苯、溴苯四种芳香化合物和对硝基酚、对甲基酚、邻硝基酚、对氯酚、2,4-二氯酚五种酚类物质进行了分析,得到最佳分析条件分别为:水样中添加5%体积分数的甲醇,以0.6 mL/min的流速富集20 min,用75%甲醇水溶液进行洗脱,在260 nm波长下检测;水样中添加5%体积分数的甲醇,以0.6 mL/min的流速富集20 min,用65%甲醇水溶液(添加0.5%醋酸)洗脱,在280 nm波长下检测。在最佳分析条件下,四种芳香化合物的检测限分别为0.024,3.346,1.268和1.189μg/L,五中酚类物质的检测限分别为20.8,34.17,18.96,54.74和74.63 ng/L。建立了两类污染物的工作曲线,并对加标自来水中四种芳香化合物和五种酚类物质进行了检测。另外,第5章还考察了新方法对样品中可能含有的表面活性剂和盐的抗干扰能力,最后对富集时间、富集流速及样品浓度三个因素进行详细考察为以后的富集实验提出了理论上的指导。
本文建立的原柱富集-液相色谱方法操作简便、富集效果好,分离度高,能够满足水体中痕量污染物检测的要求。
High performance liquid chromatography (HPLC) method is widely used in pharmaceutical and environmental analysis for its high resolution and fast analyzing. But when it comes to those samples with complicate base or contain trace component, a pretreatment step will usually be imperative for purifying and enrichment. Traditional offline pretreatment method such as SPE or LLE consumes lots of solvent and takes long time. In addition either method will lead to partial loss of the compounds. So on-line pretreatment technique coupled with HPLC methods are created. On-ling SPE-HPLC method combines the advantages of both SPE and HPLC and gains extensive application in the analysis of organism sample and environmental pollutant.
One convenient on-line SPE-HPLC method was established in this paper that enriching compounds by the analytical column. Thus this method was called on-column enrichment-HPLC. The new method was used for the analysis of several trace pollutant in water after its performance was assessed. The main content of this paper is summarized as follows:
In chapter 2, On-column Enrichment-HPLC was established based on the summarizing of characters of On-ling SPE-HPLC methods and changing operation mode of HPLC. When enriching time and sample concentration was changed, good linear correlation was found between peak area and influence factors through investigation. And the correlation coefficient was 0.9996 and 0.9840 which indicate the method was able to enrich trace compounds quantificationally. In chapter 3, PAEs in water were analyzed by On-column Enrichment-HPLC
method. The optimum analyzing condition for DMP was obtained after the investigation of each influence factor: water sample added with 5% methanol, enriched for 30 min under the flow rate of 0.6 mL/min, eluted with 60% methanol and detected under 240 nm. The LOD of DMP is 20ng/L under optimum analyzing condition. The concentration of DMP in lake water on the campus was detected as 0.3μg/L. On the other side, multi-component analysis has been attempt on the base of single component analysis.
In chapter 4 and 5, four aromatic compounds and five phenols in tap water were analyzed by On-column Enrichment-HPLC method. The optimum analyzing condition for four aromatic compounds was: water sample added with 5% methanol, enriched for 20 min under the flow rate of 0.6 mL/min, eluted with 75% methanol and detected under 260 nm. And the optimum analyzing condition for five phenols was: water sample added with 5% methanol, enriched for 20 min under the flow rate of 0.6 mL/min, eluted with 65% methanol (added with 0.5% HAc) and detected under 280nm. LODs for nitrobenzene, benzene, chlorlbenzene and bromobenzene were 0.024, 3.346, 1.268 and 1.189μg/L. And LODs for p-nitrophenol, p-hydroxytoluene, o-nitrophenol, p-chlorophenol and 2,4-dichlorophenol were 20.8, 34.17, 18.96, 54.74 and 74.63 ng/L. Calibration curves were established and spiked tap water was detected. By the way, the anti-jamming capability towards surfactant and salt possibly contained in the sample of the new method was tested in chapter 5. At last, the investigation of three influence factors (enriching time, enriching flow rate and sample concentration) gave some directions for the enriching experiment.
In short, the On-column Enrichment-HPLC method is convenient to operate, possesses good enriching effect and high resolution and is capable for the determination of trace pollutant in water.
论文第2章在总结在线固相萃取-液相色谱分析方法特点的基础上,通过改变常规液相色谱操作模式,提出并建立了原柱富集-液相色谱分析方法。通过考察发现,在改变富集时间及样品浓度两个条件下,新方法测得的峰面积和变化因素之间线性关系良好,R2分别为0.9996和0.9840,说明新方法具备对痕量组分进行定量富集的能力。
论文第3章采用原柱富集-液相色谱方法对水体中邻苯二甲酸酯类物质进行了分析,通过对各影响因素的考察,得到邻苯二甲酸二甲酯(DMP)的最佳分析条件为:水样添加5%体积分数的甲醇,以0.6 mL/min的流速富集30 min,用60%甲醇水溶液洗脱,在240 nm波长下检测。在最佳分析条件下,DMP的检测限为20 ng/L,通过外标工作曲线法测得校内一处湖水中DMP浓度为0.3μg/L。另外,在单组分分析基础上,进行了新方法多组分分析的尝试。
论文第4章和第5章采用原柱富集-液相色谱方法对自来水中硝基苯、苯、氯苯、溴苯四种芳香化合物和对硝基酚、对甲基酚、邻硝基酚、对氯酚、2,4-二氯酚五种酚类物质进行了分析,得到最佳分析条件分别为:水样中添加5%体积分数的甲醇,以0.6 mL/min的流速富集20 min,用75%甲醇水溶液进行洗脱,在260 nm波长下检测;水样中添加5%体积分数的甲醇,以0.6 mL/min的流速富集20 min,用65%甲醇水溶液(添加0.5%醋酸)洗脱,在280 nm波长下检测。在最佳分析条件下,四种芳香化合物的检测限分别为0.024,3.346,1.268和1.189μg/L,五中酚类物质的检测限分别为20.8,34.17,18.96,54.74和74.63 ng/L。建立了两类污染物的工作曲线,并对加标自来水中四种芳香化合物和五种酚类物质进行了检测。另外,第5章还考察了新方法对样品中可能含有的表面活性剂和盐的抗干扰能力,最后对富集时间、富集流速及样品浓度三个因素进行详细考察为以后的富集实验提出了理论上的指导。
本文建立的原柱富集-液相色谱方法操作简便、富集效果好,分离度高,能够满足水体中痕量污染物检测的要求。
High performance liquid chromatography (HPLC) method is widely used in pharmaceutical and environmental analysis for its high resolution and fast analyzing. But when it comes to those samples with complicate base or contain trace component, a pretreatment step will usually be imperative for purifying and enrichment. Traditional offline pretreatment method such as SPE or LLE consumes lots of solvent and takes long time. In addition either method will lead to partial loss of the compounds. So on-line pretreatment technique coupled with HPLC methods are created. On-ling SPE-HPLC method combines the advantages of both SPE and HPLC and gains extensive application in the analysis of organism sample and environmental pollutant.
One convenient on-line SPE-HPLC method was established in this paper that enriching compounds by the analytical column. Thus this method was called on-column enrichment-HPLC. The new method was used for the analysis of several trace pollutant in water after its performance was assessed. The main content of this paper is summarized as follows:
In chapter 2, On-column Enrichment-HPLC was established based on the summarizing of characters of On-ling SPE-HPLC methods and changing operation mode of HPLC. When enriching time and sample concentration was changed, good linear correlation was found between peak area and influence factors through investigation. And the correlation coefficient was 0.9996 and 0.9840 which indicate the method was able to enrich trace compounds quantificationally. In chapter 3, PAEs in water were analyzed by On-column Enrichment-HPLC
method. The optimum analyzing condition for DMP was obtained after the investigation of each influence factor: water sample added with 5% methanol, enriched for 30 min under the flow rate of 0.6 mL/min, eluted with 60% methanol and detected under 240 nm. The LOD of DMP is 20ng/L under optimum analyzing condition. The concentration of DMP in lake water on the campus was detected as 0.3μg/L. On the other side, multi-component analysis has been attempt on the base of single component analysis.
In chapter 4 and 5, four aromatic compounds and five phenols in tap water were analyzed by On-column Enrichment-HPLC method. The optimum analyzing condition for four aromatic compounds was: water sample added with 5% methanol, enriched for 20 min under the flow rate of 0.6 mL/min, eluted with 75% methanol and detected under 260 nm. And the optimum analyzing condition for five phenols was: water sample added with 5% methanol, enriched for 20 min under the flow rate of 0.6 mL/min, eluted with 65% methanol (added with 0.5% HAc) and detected under 280nm. LODs for nitrobenzene, benzene, chlorlbenzene and bromobenzene were 0.024, 3.346, 1.268 and 1.189μg/L. And LODs for p-nitrophenol, p-hydroxytoluene, o-nitrophenol, p-chlorophenol and 2,4-dichlorophenol were 20.8, 34.17, 18.96, 54.74 and 74.63 ng/L. Calibration curves were established and spiked tap water was detected. By the way, the anti-jamming capability towards surfactant and salt possibly contained in the sample of the new method was tested in chapter 5. At last, the investigation of three influence factors (enriching time, enriching flow rate and sample concentration) gave some directions for the enriching experiment.
In short, the On-column Enrichment-HPLC method is convenient to operate, possesses good enriching effect and high resolution and is capable for the determination of trace pollutant in water.
引文
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