离子液体气浮溶剂浮选分离/富集环境中四环素类抗生素残留的研究
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摘要
四环素类(tetracyclines,TCs)抗生素是一类重要的广谱抗菌性药物,包括四环素(tetracycline,TC)、金霉素(chlotetracycline,CTC)、土霉素(oxytetracycline,OTC)、甲烯土霉素(metacycline,MTC)等,被广泛用于临床治疗和养殖业中,是动物源性食品和自然环境中抗生素残留的主要来源,严重威胁人类健康和生态平衡。对于环境样品的分析,样品的前处理是重要环节,液.液萃取、固相萃取是常用的环境样品前处理方法,但是这两种方法或使用挥发性有机溶剂造成二次污染;或处理成本高;或操作繁琐、富集倍数有限,使其应用受到限制。因而,建立高效、无毒、无污染的绿色分离/富集方法尤为重要。
本研究建立了一种新的绿色分离/富集体系——离子液体气浮溶剂浮选(Ionic liHquid gas solvent sublation,ILGSS)(包括离子液体双水相气浮溶剂浮选Ionic liquid aquatic two phases gas solvent sublation,ILATPGSS)),利用该体系分离/富集环境样品中四环素类抗生素残留并与分子荧光光谱法、紫外-可见吸收光谱法和高效液相色谱法联用,为四环素类抗生素残留分析开辟了新途径。该体系首次将离子液体应用于气浮溶剂浮选,首次提出双水相气浮溶剂浮选,为浮选技术的研究拓展了新方向,具有广阔的发展前景。主要研究内容和结论如下:
(1)研究了TCs-La(Ⅲ)体系离子液体[Bmim]PF_6气浮溶剂浮选环境水样中TC、OTC、CTC三种四环素类抗生素的形为,以La(Ⅲ)为捕集剂,[Bmim]PF_6-EA(V/V=1:1)为浮选溶剂,分离/富集了环境水样中TCs,用紫外-可见分光光度法对其总含量测定,加标回收率为96.4%～101.8%,RSD＜4.31%,并与溶剂萃取法进行了比较研究,红外光谱分析离子液体只起到溶剂作用,没有和TCs-La(Ⅲ)配合物发生反应。
(2)研究了TC-Ce(Ⅲ)体系离子液体[Hmim]PF_6气浮溶剂浮选TC抗生素的形为,以[Hmim]PF_6-EA(V/V=1:0.9)浮选溶剂,以Ce(Ⅲ)为捕集剂,对鸡肉、鸡肝脏及排泄物中TC进行分离/富集,并用分光光度法测定,加标回收率为84.6%～94.0%,RSD为1.49%～3.46%。
(3)研究了TCs-Ca(Ⅱ)体系离子液体[Bmim]PF_6气浮溶剂浮选荧光光谱法分离/富集环境废水中痕量四环素类抗生素的形为,以[Bmim]PF_6-EA(V/V=1:1)为浮选剂,Ca(Ⅱ)为捕集剂,分离/富集了养殖场废水中TCs,用荧光光谱法对其总含量测定,并考察了离子液体重复使用情况,结果表明离子液体可以回收再利用,红外光谱分析表明,TCs-Ca(Ⅱ)只是溶解于离子液体中并没有和离子液体发生反应。
(4)研究了TCs-Fe(Ⅲ)体系离子液体[Omim]PF_6气浮溶剂浮选荧光光谱法分离/富集环境样品中痕量TCs抗生素的形为,以Fe(Ⅲ)为捕集剂,离子液体[Omim]PF_6-EA(V/V=5:4)为浮选溶剂,对鱼塘表面水体和沉积物中TCs分离/富集,并用荧光光谱法对总含量测定,回收率为94.2%～100.4%,RSD＜3.2%。
(5)研究了TCs-Al(Ⅲ)体系[Bmim]PF_6气浮溶剂浮选分离/富集TC、OTC、CTC三种四环素类抗生素残留的形为,以[Bmim]PF_6-EA(V/V=1:1)为浮选剂,以Al(Ⅲ)为捕集剂,分离/富集牛奶样品中三种四环素残留,经HPLC分析,测得平均回收率在88.5%～104.0%之间,相对标准偏差在1.36%～3.76%之间,红外光谱分析结果表明离子液体只起到溶剂作用。
(6)研究了离子液体双水相气浮溶剂浮选体系,与离子液体双水相萃取相比,具有无毒、无污染、选择性高,富集倍数大,分相快,无乳化现象等优点。实验以[Bmim]BF_4溶液(V/V=1:1)为浮选溶剂,以NaH_2PO_4为分相盐,分离/富集了肉制品中TC、OTC、CTC三种四环素类抗生素残留,经HPLC测定,加标回收率为94.4%～106.4%,RSD为1.23%～4.53%。
研究结果表明,离子液体气浮溶剂浮选(包括离子液体双水相气浮溶剂浮选)集离子液体萃取和气浮溶剂浮选优点于一体,具有无毒、无污染、选择性好、富集倍数高等优点,适合于环境样品中痕量四环素类抗生素残留的分离/富集/分析。
Tetracyclines(TCs) antibiotics,especial oxytetracycline(OTC),tetracycline(TC) and chlortetracycline(CTC) have been used extensively due to their broad range of activity against bacteria and their economic advantages.Liquid-liquid extraction and Solid-phase extraction(SPE) have been most commonly used to pre-concentrate for determining TCs.However,the methods are still undesirable for most analyses due to consumption of time and a large amount of volatile,toxic,pollution organic solvents. Additionally,these method can cause secondary pollution and high cost.
The aim of the project was to develop a fast and reliable method for the separation/enrichment of TCs antibiotic in environment samples.The method is based on ionic liquid solvent sublation combined with spectrophotometric spectrometry or fluorescence spectrometry or high performance liquid chromatography.It is the first time that ionic liquids and aqueous two phases applied to gas solvent sublation,the method takes a new direction for the flotation process with vast development foreground.The main research contents and conclusion are as follows:
(1) Studied on separation/enrichment of TCs antibiotic based on the system of TCs-La(Ⅲ) by ionic liquid gas solvent sublation,lanthanum nitrate as the trapping agent,the mixture of[Bmim]PF_6 and EA(volume ratio 1/1) as solvent, separated/enriched the TCs residue in the water sample and determinated by spectrophotometric spectrometry,the recoveries ranged from 96.4%to 101.8%,RSD was less than 4.31%.Comparative studied on solvent extraction and gas solvent sublation.
(2) Studied on separation/enrichment of TC antibiotic based on the system of TC-Ce(Ⅲ) by ionic liquid gas solvent sublation,cerium nitrate as the trapping agent, the mixture of[Hmim]PF_6 and EA(volume ratio 1/0.9) as solvent,separated/enriched the TC residue in the chicken,liver and the excrement of the poultry which have been determinated by spectrophotometric spectrometry,the recoveries ranged from 84.6%to 94.0%,RSD was 1.49%to 3.46%.
(3) Studied on separation/enrichment of TCs antibiotic based on the system of TCs-Ca(Ⅱ) by ionic liquid gas solvent sublation,calcium chloride as the trapping agent,the mixture of[Bmim]PF_6 and EA(volume ratio 1/1) as solvent,separated/ enriched the TCs residue in the sample and detected total content by fluorescence spectrometry and moreover researched on the reuse of ionic liquid which can be used repeatedly.
(4) Studied on separation/enrichment of TCs antibiotic based on the system of TCs-Fe(Ⅲ) by ionic liquid gas solvent sublation,ferric chloride as the trapping agent, the mixture of[Omim]PF6 and EA(volume ratio 5/4) as solvent,separated/enriched the TCs residue in the surface water and the sediment of fish pond,the analyte have been detected by fluorescence spectrometry,the recoveries ranged from 94.2%to 100.4%, RSD was less than 3.2%.
(5) Studied on separation/enrichment of TCs antibiotic based on the system of TCs-Al(Ⅲ) by ionic liquid gas solvent sublation,Al(Ⅲ) ion as the trapping agent,the mixture of[Bmim]PF_6 and EA(volume ratio 1/1) as solvent,separated/enriched the TCs residue in the milk samples detected by high performance liquid chromatography (HPLC).The recoveries of three tetracyclines ranged from 88.5%to 104.0%,the relative standard deviations were between 1.36%and 3.76%.
(6) Studied on ionic liquid aqueous two phases gas solvent sublation, [Bmim]BF_4 solution(VN=1) was used as solvent in this experiment with sodium phosphateand as the salt of phase separation,directed trapping and separated/enriched the residue of TCs in pork,detected by HPLC,The recoveries of three tetracyclines ranged from 94.4%to 106.4%,the relative standard deviations were between 1.23%and 4.53%.
The results show that ionic liquid gas solvent sublation(ionic liquid aqueous two phases gas solvent sublation) may be thought of a combined method of flotation and solvent extraction(aqueous two phases extraction) with the advantages of both techniques,the method has many merits,such as,non toxic,non pollution,high enrichment factor,high selectivity,negligible emulsion phenomenon,etc.The method is suitable for determining the Tetracyclines residue in environment sample.
本研究建立了一种新的绿色分离/富集体系——离子液体气浮溶剂浮选(Ionic liHquid gas solvent sublation,ILGSS)(包括离子液体双水相气浮溶剂浮选Ionic liquid aquatic two phases gas solvent sublation,ILATPGSS)),利用该体系分离/富集环境样品中四环素类抗生素残留并与分子荧光光谱法、紫外-可见吸收光谱法和高效液相色谱法联用,为四环素类抗生素残留分析开辟了新途径。该体系首次将离子液体应用于气浮溶剂浮选,首次提出双水相气浮溶剂浮选,为浮选技术的研究拓展了新方向,具有广阔的发展前景。主要研究内容和结论如下:
(1)研究了TCs-La(Ⅲ)体系离子液体[Bmim]PF_6气浮溶剂浮选环境水样中TC、OTC、CTC三种四环素类抗生素的形为,以La(Ⅲ)为捕集剂,[Bmim]PF_6-EA(V/V=1:1)为浮选溶剂,分离/富集了环境水样中TCs,用紫外-可见分光光度法对其总含量测定,加标回收率为96.4%～101.8%,RSD＜4.31%,并与溶剂萃取法进行了比较研究,红外光谱分析离子液体只起到溶剂作用,没有和TCs-La(Ⅲ)配合物发生反应。
(2)研究了TC-Ce(Ⅲ)体系离子液体[Hmim]PF_6气浮溶剂浮选TC抗生素的形为,以[Hmim]PF_6-EA(V/V=1:0.9)浮选溶剂,以Ce(Ⅲ)为捕集剂,对鸡肉、鸡肝脏及排泄物中TC进行分离/富集,并用分光光度法测定,加标回收率为84.6%～94.0%,RSD为1.49%～3.46%。
(3)研究了TCs-Ca(Ⅱ)体系离子液体[Bmim]PF_6气浮溶剂浮选荧光光谱法分离/富集环境废水中痕量四环素类抗生素的形为,以[Bmim]PF_6-EA(V/V=1:1)为浮选剂,Ca(Ⅱ)为捕集剂,分离/富集了养殖场废水中TCs,用荧光光谱法对其总含量测定,并考察了离子液体重复使用情况,结果表明离子液体可以回收再利用,红外光谱分析表明,TCs-Ca(Ⅱ)只是溶解于离子液体中并没有和离子液体发生反应。
(4)研究了TCs-Fe(Ⅲ)体系离子液体[Omim]PF_6气浮溶剂浮选荧光光谱法分离/富集环境样品中痕量TCs抗生素的形为,以Fe(Ⅲ)为捕集剂,离子液体[Omim]PF_6-EA(V/V=5:4)为浮选溶剂,对鱼塘表面水体和沉积物中TCs分离/富集,并用荧光光谱法对总含量测定,回收率为94.2%～100.4%,RSD＜3.2%。
(5)研究了TCs-Al(Ⅲ)体系[Bmim]PF_6气浮溶剂浮选分离/富集TC、OTC、CTC三种四环素类抗生素残留的形为,以[Bmim]PF_6-EA(V/V=1:1)为浮选剂,以Al(Ⅲ)为捕集剂,分离/富集牛奶样品中三种四环素残留,经HPLC分析,测得平均回收率在88.5%～104.0%之间,相对标准偏差在1.36%～3.76%之间,红外光谱分析结果表明离子液体只起到溶剂作用。
(6)研究了离子液体双水相气浮溶剂浮选体系,与离子液体双水相萃取相比,具有无毒、无污染、选择性高,富集倍数大,分相快,无乳化现象等优点。实验以[Bmim]BF_4溶液(V/V=1:1)为浮选溶剂,以NaH_2PO_4为分相盐,分离/富集了肉制品中TC、OTC、CTC三种四环素类抗生素残留,经HPLC测定,加标回收率为94.4%～106.4%,RSD为1.23%～4.53%。
研究结果表明,离子液体气浮溶剂浮选(包括离子液体双水相气浮溶剂浮选)集离子液体萃取和气浮溶剂浮选优点于一体,具有无毒、无污染、选择性好、富集倍数高等优点,适合于环境样品中痕量四环素类抗生素残留的分离/富集/分析。
Tetracyclines(TCs) antibiotics,especial oxytetracycline(OTC),tetracycline(TC) and chlortetracycline(CTC) have been used extensively due to their broad range of activity against bacteria and their economic advantages.Liquid-liquid extraction and Solid-phase extraction(SPE) have been most commonly used to pre-concentrate for determining TCs.However,the methods are still undesirable for most analyses due to consumption of time and a large amount of volatile,toxic,pollution organic solvents. Additionally,these method can cause secondary pollution and high cost.
The aim of the project was to develop a fast and reliable method for the separation/enrichment of TCs antibiotic in environment samples.The method is based on ionic liquid solvent sublation combined with spectrophotometric spectrometry or fluorescence spectrometry or high performance liquid chromatography.It is the first time that ionic liquids and aqueous two phases applied to gas solvent sublation,the method takes a new direction for the flotation process with vast development foreground.The main research contents and conclusion are as follows:
(1) Studied on separation/enrichment of TCs antibiotic based on the system of TCs-La(Ⅲ) by ionic liquid gas solvent sublation,lanthanum nitrate as the trapping agent,the mixture of[Bmim]PF_6 and EA(volume ratio 1/1) as solvent, separated/enriched the TCs residue in the water sample and determinated by spectrophotometric spectrometry,the recoveries ranged from 96.4%to 101.8%,RSD was less than 4.31%.Comparative studied on solvent extraction and gas solvent sublation.
(2) Studied on separation/enrichment of TC antibiotic based on the system of TC-Ce(Ⅲ) by ionic liquid gas solvent sublation,cerium nitrate as the trapping agent, the mixture of[Hmim]PF_6 and EA(volume ratio 1/0.9) as solvent,separated/enriched the TC residue in the chicken,liver and the excrement of the poultry which have been determinated by spectrophotometric spectrometry,the recoveries ranged from 84.6%to 94.0%,RSD was 1.49%to 3.46%.
(3) Studied on separation/enrichment of TCs antibiotic based on the system of TCs-Ca(Ⅱ) by ionic liquid gas solvent sublation,calcium chloride as the trapping agent,the mixture of[Bmim]PF_6 and EA(volume ratio 1/1) as solvent,separated/ enriched the TCs residue in the sample and detected total content by fluorescence spectrometry and moreover researched on the reuse of ionic liquid which can be used repeatedly.
(4) Studied on separation/enrichment of TCs antibiotic based on the system of TCs-Fe(Ⅲ) by ionic liquid gas solvent sublation,ferric chloride as the trapping agent, the mixture of[Omim]PF6 and EA(volume ratio 5/4) as solvent,separated/enriched the TCs residue in the surface water and the sediment of fish pond,the analyte have been detected by fluorescence spectrometry,the recoveries ranged from 94.2%to 100.4%, RSD was less than 3.2%.
(5) Studied on separation/enrichment of TCs antibiotic based on the system of TCs-Al(Ⅲ) by ionic liquid gas solvent sublation,Al(Ⅲ) ion as the trapping agent,the mixture of[Bmim]PF_6 and EA(volume ratio 1/1) as solvent,separated/enriched the TCs residue in the milk samples detected by high performance liquid chromatography (HPLC).The recoveries of three tetracyclines ranged from 88.5%to 104.0%,the relative standard deviations were between 1.36%and 3.76%.
(6) Studied on ionic liquid aqueous two phases gas solvent sublation, [Bmim]BF_4 solution(VN=1) was used as solvent in this experiment with sodium phosphateand as the salt of phase separation,directed trapping and separated/enriched the residue of TCs in pork,detected by HPLC,The recoveries of three tetracyclines ranged from 94.4%to 106.4%,the relative standard deviations were between 1.23%and 4.53%.
The results show that ionic liquid gas solvent sublation(ionic liquid aqueous two phases gas solvent sublation) may be thought of a combined method of flotation and solvent extraction(aqueous two phases extraction) with the advantages of both techniques,the method has many merits,such as,non toxic,non pollution,high enrichment factor,high selectivity,negligible emulsion phenomenon,etc.The method is suitable for determining the Tetracyclines residue in environment sample.
引文
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