载体辅助液相微萃取-HPLC联用分析复杂流体中的药物研究
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摘要
液相微萃取(LPME)是近年来发展起来的一种环境友好的新型的前处理技术。该技术集采样、萃取和富集于一体,具有操作简单、富集效果好、抗干扰能力强等特点。目前,LPME主要与气相色谱(GC)、气相色谱-质谱(GC-MS)、高效液相色谱(HPLC)、液相色谱-质谱(LC-MS)、毛细管电泳(CE)联用,广泛的应用于分析化学、药学、生物化学、临床医学和环境化学等领域,对于复杂流体中药物,特别是亲水性药物的研究报道还较少,本文就液相微萃取-高效液相色谱联用技术在复杂流体中违禁药物的分离分析方面进行了较为系统的研究。主要研究内容如下：
1.对样品前处理技术的发展进行了介绍,重点介绍了液液液微萃取(LLLPME)和分散液相微萃取(DLPME)。
2.载体辅助液相微萃取-HPLC联用检测人体尿样中的β-阻断剂。考察了有机相、载体种类和浓度、料液相pH值、搅拌速度、接受相浓度、萃取时间等因素对富集因子的影响,得到了萃取溶剂为0.005mol/L的TOAB-甲苯溶液,接受相为1.4μL0.5 mol/L HCl,料液相pH值为12,搅拌速度为700 rpm,萃取时间为30 min的最佳实验条件。在此实验条件下,索它洛尔、卡替洛尔、比索洛尔、普洛萘尔的富集比分别为235.7、152、253.7、182.5,线性范围分别为索它洛尔0.05-10.0 mg／L、卡替洛尔0.05-10.0 mg／L、比索洛尔0.05-8.0 mg／L、普洛萘尔0.05-8.0 mg／L,检测限除普萘洛尔为0.005 mg／L外,其余的为0.01 mg／L(S／N=3)。对浓度分别为1 mg／L和5 mg／L的加标尿样进行回收率分析,相应的方法回收率分别为安非他明：索它洛尔104.7%和91.7%、卡替洛尔104.2%和109.4%、比索洛尔100.8%和1 00.3%、普洛萘尔99.3%和97.9%说明该方法用于尿样中索它洛尔、卡替洛尔、比索洛尔、普洛萘尔的测定,能获得准确的结果。
3.载体辅助液相微萃取-HPLC法测定水样中的磺胺类药物。通过对实验条件的优化,我们选择用0.001 mol/L四辛基溴化铵的甲苯溶液作为有机相,0.5 mol/L的盐酸作为的接受相,料液相的pH值为12,搅拌速度为700 rpm,萃取时间为40 min的实验条件。在最优的实验条件下得到较高的富集因子,磺胺嘧啶、磺胺甲噁唑、磺胺二甲嘧啶方法的线性范围为：均为0.01-8 mg／L；检出限分别为：0.005 mg／L、0.001 mg／L和0.005 mg／L。相对标准偏差均小于7.2%,对浓度分别为5μg／mL和1μg／mL的加标水样进行回收率分析,相应的方法回收率分别为磺胺嘧啶100.1%和110.7%、磺胺甲噁唑100.2%和109.5%、磺胺二甲嘧啶108.6%和107.8&(信噪比为3)。
4.纳米金载体辅助液相微萃取-HPLC联用检测人体尿样中的四种局部麻醉剂。对各实验条件进行了优化,获得了优化的实验条件为：以0.79 mg／L的AuNPs-TOAB-甲苯溶液作为有机相,0.1 mol/L的盐酸作为接受相,搅拌速度为600 rpm,料液相的pH为11,萃取时间为25 min。在最佳萃取条件下,获得了较高的富集因子。方法的线性范围均为0.01-8 mg／L,相对标准偏差均小于6.0%,对浓度分别为1 mg／L和5 mg／L的加标尿样进行回收率分析,相应的方法回收率分别为：利多卡因100.1%和100.3%、布比卡因99.1%和101.1%、普鲁卡因107.4%和100.3%、丁卡因108.6%和100.9%。检测限普鲁卡因的为0.0001 mg/L,其余均为0.005 mg／L。
5.载体辅助分散液相微萃取-HPLC联用检测环境水中的β-阻断剂。考察了有机相、载体种类和浓度、料液相pH值、接受相浓度、分散萃取时间、三相微萃取的搅拌速度、三相微萃取的萃取时间等因素对富集因子的影响,得到了萃取溶剂为0.01 mol/L的TOAB-甲苯溶液,接受相为1.4μL 0.1 mol/L HCl,料液相pH值为13.7,分散萃取的搅拌速度为1700 rpm,分散萃取时间为30 S,三相微萃取搅拌速度为800 rpm,萃取时间为15 min的最佳实验条件。在此实验条件下,索它洛尔、卡替洛尔、美托洛尔、比索洛尔、普洛萘尔的线性范围分别为：索它洛尔0.01-10.0 mg／L、卡替洛尔0.01-10.0 mg／L、美托洛尔0.005-8.0 mg／L、比索洛尔0.01-8.0 mg／L、普洛萘尔0.01-8.0 mg／L,检测限除比索洛尔为0.001 mg／L外,其余的为0.005 mg／L(S／N=3)。对浓度分别为1 mg／L和4 mg／L的加标尿样进行回收率分析,相应的方法回收率分别为安非他明：索它洛尔94.5%和102.9%、卡替洛尔102.4%和101.2%、美托洛尔96.2%和98.7%、比索洛尔99.7%和103.6%、普洛萘尔107.3%和105.8%说明该方法用于尿样中索它洛尔、卡替洛尔、美托洛尔、比索洛尔、普洛萘尔的测定,能获得准确的结果。
Liquid phase microextraction (LPME) is a new and environment friendly pretreatment method. LPME integrates the sampling, extraction, and enrichment in one process and owns the merits of simple, excellent selectivity, and high enrichment. It has been applied widely in analytical chemistry, pharmacy, biochemistry, clinical medicine and environmental chemistry coupled with gas chromatography (GC), gas chromatography-mass spectrometry (GC-MS), high performance liquid chromatography (HPLC), high performance liquid chromatography-mass spectrometry (LC-MS) and capillary electrophoresis(CE). However, the studies focus mainly the hydrophobic compounds because the high enrichment with hydrophobic compounds extraction is obtained easily. In the present paper, some drugs, especial hydrophilic drugs, in complex fluids were separated and enriched by using carrier-mediated liquid phase microextraction (CM-LPME) and dispersed liquid-phase microextraction (DLPME).
The details are summarized as follows:
1. The review of several sample pretreatment methods, especially liquid phase microextraction (LPME) and dispersed liquid-phase microextraction (DLPME).
2. A CM-LPME-HPLC method was developed for the simultaneous determination of four kinds ofβ-blockers, sotalol, bisoprolol, carteolol, and propranolol in human urine. The effects of the organic phase, the pH in the donor solution, the composition and the concentration of the acceptor phase, the stirring rate, and the extraction time on the enrichment factors of analytes were investigated. Under the optimal conditions, high enrichment factors were reached. The linear ranges were from 0.1 to 10.0 mg·L-1 for sotalol and carteolol, from 0.1 to 8.0 mg·L-1 for bisoprolol and propranolol. The limits of detection (S/N=3) were 0.01 mg·L-1 for sotalol, carteolol and bisoprolol,0.005 mg·L-1 for propranolol. The relative standard deviations were lower than 5.9%. The method with little solvent consumption may provide high analyte pre-concentration and excellent sample clean-up, and it is a sensitive and suitable method for simultaneous determining of above four drugs in human urine.
3. A CM-LPME-HPLC method was developed for the determination of sulfadiazine, sulfamethoxazole, and sulfadimidine in environmental water. The effects of the organic phase, the pH in the donor solution, the composition and the concentration of the acceptor phase, the stirring rate, and the extraction time on the enrichment factors of analytes were investigated. Under the optimal conditions, high enrichment factors were reached. The linear ranges were from 0.1 to 8.0 mg-L-1 for the three drugs. The limits of detection (S/N=3) were 0.005 mg-L-1 for sulfadiazine and sulfadimidine,0.001 mg-L-1 for sulfamethoxazole. The relative standard deviations were lower than 7.2%. Analyse We determined the recovery of the sample by adding the standard samples, the recovery of water sample is 100.2～110.7%. he method with little solvent consumption may provide high analyte pre-concentration and excellent sample clean-up, and it is a sensitive and suitable method for simultaneous determining of above four drugs in environmental water.
4. Gold nanoparticles carrier mediated (AuNPs-CM)-LPME-HPLC was developed for the simultaneous determination of procaine, lidocaine, bupivacaine, and tetracainel in human urine. Relevant parameters were optimized. The organic phase was 0.79 mg/L AuNPs-TOAB-toluene, the acceptor was 0.1 mol/L HCl, the stirring speed is 600 rpm, pH in the donor phase was 11, the extraction time was 25 min. Under the optimal conditions, high enrichment factors were reached. The linear ranges were from 0.01 to 8.0 mg·L-1 for the four drugs. The limits of detection (S/N=3) were 0.005 mg·L-1 for lidocaine, bupivacaine, and tetracainel,0.0001 mg·L-1 for procaine. The relative standard deviations were lower than 6.0%. We determined the recovery of the sample by adding the standard samples, the recovery of water sample is 99.1～110.7%. The results indicate that AuNPs-CM-LPME-HPLC can be successfully applied to the separation and enrichment of procaine, lidocaine, bupivacaine, and tetracainel.
5. DLPME-HPLC was developed for the simultaneous determination of five kinds ofβ-blockers, sotalol, bisoprolol, metoprolol, carteolol, and propranolol in environmental water. The kind of the organic solvent, the concentration of carrier in the organic phase, pH in the donor phase, the composition of the acceptor solution, the DLPME time, the stirring rate and the back-extraction time were optimized. The organic phase was 0.01 mol/L TOAB-toluene, the acceptor was 0.1 mol/L HCl, the stirring speed in DLPME is 1700 rpm, pH in the donor phase was 13.7, the stirring speed in LLLME is 800 rpm, the DLPME time was 30 s, the extraction time was 15 min. Under the optimal conditions, high enrichment factors were reached. The linear ranges were from 0.01 to 10.0 mg·L-1 for sotalol and carteolol,0.005-8.0 mg/L for metoprolol,0.01-8.0 mg/L for bisoprolol and propranolol. The limits of detection (S/N=3) were 0.001 mg-L-1 for bisoprolol, and 0.005 mg·L-1 for the other four drugs. We determined the recovery of the sample by adding the standard samples, the recovery of water sample is 94.5～107.3%. The method was proved to be a fast and efficient extraction method, which can be applied to simultaneous determine sotalol, bisoprolol, metoprolol, carteolol, and propranolol in tap water and four environmental waters.
1.对样品前处理技术的发展进行了介绍,重点介绍了液液液微萃取(LLLPME)和分散液相微萃取(DLPME)。
2.载体辅助液相微萃取-HPLC联用检测人体尿样中的β-阻断剂。考察了有机相、载体种类和浓度、料液相pH值、搅拌速度、接受相浓度、萃取时间等因素对富集因子的影响,得到了萃取溶剂为0.005mol/L的TOAB-甲苯溶液,接受相为1.4μL0.5 mol/L HCl,料液相pH值为12,搅拌速度为700 rpm,萃取时间为30 min的最佳实验条件。在此实验条件下,索它洛尔、卡替洛尔、比索洛尔、普洛萘尔的富集比分别为235.7、152、253.7、182.5,线性范围分别为索它洛尔0.05-10.0 mg／L、卡替洛尔0.05-10.0 mg／L、比索洛尔0.05-8.0 mg／L、普洛萘尔0.05-8.0 mg／L,检测限除普萘洛尔为0.005 mg／L外,其余的为0.01 mg／L(S／N=3)。对浓度分别为1 mg／L和5 mg／L的加标尿样进行回收率分析,相应的方法回收率分别为安非他明：索它洛尔104.7%和91.7%、卡替洛尔104.2%和109.4%、比索洛尔100.8%和1 00.3%、普洛萘尔99.3%和97.9%说明该方法用于尿样中索它洛尔、卡替洛尔、比索洛尔、普洛萘尔的测定,能获得准确的结果。
3.载体辅助液相微萃取-HPLC法测定水样中的磺胺类药物。通过对实验条件的优化,我们选择用0.001 mol/L四辛基溴化铵的甲苯溶液作为有机相,0.5 mol/L的盐酸作为的接受相,料液相的pH值为12,搅拌速度为700 rpm,萃取时间为40 min的实验条件。在最优的实验条件下得到较高的富集因子,磺胺嘧啶、磺胺甲噁唑、磺胺二甲嘧啶方法的线性范围为：均为0.01-8 mg／L；检出限分别为：0.005 mg／L、0.001 mg／L和0.005 mg／L。相对标准偏差均小于7.2%,对浓度分别为5μg／mL和1μg／mL的加标水样进行回收率分析,相应的方法回收率分别为磺胺嘧啶100.1%和110.7%、磺胺甲噁唑100.2%和109.5%、磺胺二甲嘧啶108.6%和107.8&(信噪比为3)。
4.纳米金载体辅助液相微萃取-HPLC联用检测人体尿样中的四种局部麻醉剂。对各实验条件进行了优化,获得了优化的实验条件为：以0.79 mg／L的AuNPs-TOAB-甲苯溶液作为有机相,0.1 mol/L的盐酸作为接受相,搅拌速度为600 rpm,料液相的pH为11,萃取时间为25 min。在最佳萃取条件下,获得了较高的富集因子。方法的线性范围均为0.01-8 mg／L,相对标准偏差均小于6.0%,对浓度分别为1 mg／L和5 mg／L的加标尿样进行回收率分析,相应的方法回收率分别为：利多卡因100.1%和100.3%、布比卡因99.1%和101.1%、普鲁卡因107.4%和100.3%、丁卡因108.6%和100.9%。检测限普鲁卡因的为0.0001 mg/L,其余均为0.005 mg／L。
5.载体辅助分散液相微萃取-HPLC联用检测环境水中的β-阻断剂。考察了有机相、载体种类和浓度、料液相pH值、接受相浓度、分散萃取时间、三相微萃取的搅拌速度、三相微萃取的萃取时间等因素对富集因子的影响,得到了萃取溶剂为0.01 mol/L的TOAB-甲苯溶液,接受相为1.4μL 0.1 mol/L HCl,料液相pH值为13.7,分散萃取的搅拌速度为1700 rpm,分散萃取时间为30 S,三相微萃取搅拌速度为800 rpm,萃取时间为15 min的最佳实验条件。在此实验条件下,索它洛尔、卡替洛尔、美托洛尔、比索洛尔、普洛萘尔的线性范围分别为：索它洛尔0.01-10.0 mg／L、卡替洛尔0.01-10.0 mg／L、美托洛尔0.005-8.0 mg／L、比索洛尔0.01-8.0 mg／L、普洛萘尔0.01-8.0 mg／L,检测限除比索洛尔为0.001 mg／L外,其余的为0.005 mg／L(S／N=3)。对浓度分别为1 mg／L和4 mg／L的加标尿样进行回收率分析,相应的方法回收率分别为安非他明：索它洛尔94.5%和102.9%、卡替洛尔102.4%和101.2%、美托洛尔96.2%和98.7%、比索洛尔99.7%和103.6%、普洛萘尔107.3%和105.8%说明该方法用于尿样中索它洛尔、卡替洛尔、美托洛尔、比索洛尔、普洛萘尔的测定,能获得准确的结果。
Liquid phase microextraction (LPME) is a new and environment friendly pretreatment method. LPME integrates the sampling, extraction, and enrichment in one process and owns the merits of simple, excellent selectivity, and high enrichment. It has been applied widely in analytical chemistry, pharmacy, biochemistry, clinical medicine and environmental chemistry coupled with gas chromatography (GC), gas chromatography-mass spectrometry (GC-MS), high performance liquid chromatography (HPLC), high performance liquid chromatography-mass spectrometry (LC-MS) and capillary electrophoresis(CE). However, the studies focus mainly the hydrophobic compounds because the high enrichment with hydrophobic compounds extraction is obtained easily. In the present paper, some drugs, especial hydrophilic drugs, in complex fluids were separated and enriched by using carrier-mediated liquid phase microextraction (CM-LPME) and dispersed liquid-phase microextraction (DLPME).
The details are summarized as follows:
1. The review of several sample pretreatment methods, especially liquid phase microextraction (LPME) and dispersed liquid-phase microextraction (DLPME).
2. A CM-LPME-HPLC method was developed for the simultaneous determination of four kinds ofβ-blockers, sotalol, bisoprolol, carteolol, and propranolol in human urine. The effects of the organic phase, the pH in the donor solution, the composition and the concentration of the acceptor phase, the stirring rate, and the extraction time on the enrichment factors of analytes were investigated. Under the optimal conditions, high enrichment factors were reached. The linear ranges were from 0.1 to 10.0 mg·L-1 for sotalol and carteolol, from 0.1 to 8.0 mg·L-1 for bisoprolol and propranolol. The limits of detection (S/N=3) were 0.01 mg·L-1 for sotalol, carteolol and bisoprolol,0.005 mg·L-1 for propranolol. The relative standard deviations were lower than 5.9%. The method with little solvent consumption may provide high analyte pre-concentration and excellent sample clean-up, and it is a sensitive and suitable method for simultaneous determining of above four drugs in human urine.
3. A CM-LPME-HPLC method was developed for the determination of sulfadiazine, sulfamethoxazole, and sulfadimidine in environmental water. The effects of the organic phase, the pH in the donor solution, the composition and the concentration of the acceptor phase, the stirring rate, and the extraction time on the enrichment factors of analytes were investigated. Under the optimal conditions, high enrichment factors were reached. The linear ranges were from 0.1 to 8.0 mg-L-1 for the three drugs. The limits of detection (S/N=3) were 0.005 mg-L-1 for sulfadiazine and sulfadimidine,0.001 mg-L-1 for sulfamethoxazole. The relative standard deviations were lower than 7.2%. Analyse We determined the recovery of the sample by adding the standard samples, the recovery of water sample is 100.2～110.7%. he method with little solvent consumption may provide high analyte pre-concentration and excellent sample clean-up, and it is a sensitive and suitable method for simultaneous determining of above four drugs in environmental water.
4. Gold nanoparticles carrier mediated (AuNPs-CM)-LPME-HPLC was developed for the simultaneous determination of procaine, lidocaine, bupivacaine, and tetracainel in human urine. Relevant parameters were optimized. The organic phase was 0.79 mg/L AuNPs-TOAB-toluene, the acceptor was 0.1 mol/L HCl, the stirring speed is 600 rpm, pH in the donor phase was 11, the extraction time was 25 min. Under the optimal conditions, high enrichment factors were reached. The linear ranges were from 0.01 to 8.0 mg·L-1 for the four drugs. The limits of detection (S/N=3) were 0.005 mg·L-1 for lidocaine, bupivacaine, and tetracainel,0.0001 mg·L-1 for procaine. The relative standard deviations were lower than 6.0%. We determined the recovery of the sample by adding the standard samples, the recovery of water sample is 99.1～110.7%. The results indicate that AuNPs-CM-LPME-HPLC can be successfully applied to the separation and enrichment of procaine, lidocaine, bupivacaine, and tetracainel.
5. DLPME-HPLC was developed for the simultaneous determination of five kinds ofβ-blockers, sotalol, bisoprolol, metoprolol, carteolol, and propranolol in environmental water. The kind of the organic solvent, the concentration of carrier in the organic phase, pH in the donor phase, the composition of the acceptor solution, the DLPME time, the stirring rate and the back-extraction time were optimized. The organic phase was 0.01 mol/L TOAB-toluene, the acceptor was 0.1 mol/L HCl, the stirring speed in DLPME is 1700 rpm, pH in the donor phase was 13.7, the stirring speed in LLLME is 800 rpm, the DLPME time was 30 s, the extraction time was 15 min. Under the optimal conditions, high enrichment factors were reached. The linear ranges were from 0.01 to 10.0 mg·L-1 for sotalol and carteolol,0.005-8.0 mg/L for metoprolol,0.01-8.0 mg/L for bisoprolol and propranolol. The limits of detection (S/N=3) were 0.001 mg-L-1 for bisoprolol, and 0.005 mg·L-1 for the other four drugs. We determined the recovery of the sample by adding the standard samples, the recovery of water sample is 94.5～107.3%. The method was proved to be a fast and efficient extraction method, which can be applied to simultaneous determine sotalol, bisoprolol, metoprolol, carteolol, and propranolol in tap water and four environmental waters.
引文
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