解吸附电晕束离子源（DCBI）的研制与应用
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摘要
质谱仪是现代科学的基柱工具,质谱分析方法已经成为自然科学的重要实验手段。离子源是质谱仪器的核心部件,根据其发展可以大致分为真空下、大气压下、常压式(敞开式)离子化技术三个阶段。常压式质谱技术也曾经被一些学者称为直接分析质谱法、大气压解吸附质谱法、敞开式质谱法等。常压式离子源具有典型的特征,如样品原型分析,直接离子化,通用质谱仪接口,软电离方式等。不过,现有常压式离子源也存在定量分析精密度较差、样品表面定位不确定、空间分辨率较低等问题。
本论文中一种新的常压式离子化技术——解吸附电晕束离子源(DCBI)被研制,该源具有可视电晕束；进而DCBI定量分析的精密度得以研究改善,该方法亦可供类似离子化技术借鉴；联用其他技术扩展了DCBI源的适用对象和范围；实际应用中快速分析了降压类保健食品及中成药中7种芳氧丙醇胺类p-受体阻滞剂；高通量筛查了减肥类保健食品添加剂和茶包中的违禁化合物,并对这些违禁添加物进行了半定量检测分析。具体研究包括以下6部分内容。
1.开发了解吸附电晕束离子化(DCBI)技术,并以之构建了一新型常压式质谱用离子源,成功应用于多种物质的直接分析。该源具有独特的可视电晕束,为常压式质谱法开创了新的应用领域。正负离子化模式下的电离机理均得到研究与解释。利用程序升温分析复杂样品,降低了结果谱图中的化学噪声,使离子峰专属性加强。较之DESI, DCBI源结构紧凑,且不需辅助溶剂；较之DART, DCBI源工作于电晕放电区域,电流小,且源温度可程序扫描。
2.利用室温离子液体1-丁基-3甲基咪唑四氟硼酸盐([BMIM]BF4)作为基质,辅助DCBI质谱检测方法,显著改善了被分析物的定量精密度。通过红外热成像与质谱仪的同步监测,观察到了挥发性基质在DCBI离子源中的双峰现象,显示了DCBI源的解吸附、电离两步机理假设的合理性。研究发现样品进给过程中的共挥发效应是常压式敞开质谱法中定量结果出现波动的主要原因。当使用不挥发的室温离子液体作为基质时,其形成一个微缓释体系(MSRS),降低了解吸附波动,提高了定量精密度。其他相关的类APCI常压式离子化技术亦可参考该方法,以提高定量准确率与重现性。该研究也为液相微体积的过程研究与化学表征提供了可能。
3.利用聚二甲基硅氧烷(PDMS)膜获得浓缩系数1000倍以上的富集效率,还可以使不溶解于基质的化合物被富集,并应用DCBI对其进行分析。PDMS结合DCBI直接分析样品扩展了它们的应用范围,提高了分析的灵敏度。该联用技术对水样中农残进行定量分析,结果表明5种被分析农药的标准曲线都有很好的线性,检测限(1μg/L)。此研究表明联用PDMS基底与DCBI源,分析微量的液相样品快速、结果准确。这个方法可应用于分析各种不同溶液中的有机物,气态物,使得低浓度微量液体样品的直接质谱法分析成为可能。
4.联用近红外激光(976nm)与电晕束电离手段,开发了一种新型激光解吸附辅助电晕束离子化(LD-CBI)技术,考察了该技术方法特性,并成功将其应用于液体、粉末等无定形样品的直接分析。结果显示,因一般物质对近红外光的范频吸收特性,LD-CBI不但对样品前处理的要求更少,而且灵敏度更高。方法学验证中,19种农药样品的检测限在0.02-1ng之间。选择离子监测模式下,马拉硫磷的检测限可以低至10pg约合30fMol的水平,而且在样品浓度0.2μg/mL到100μg/mL的近三个数量级之间线性关系良好R2=0.9907。
5.利用DCBI源结合离子阱质谱仪,开发了一种快速、低耗的方法,快速筛选分析7种,芳氧丙醇胺类β-受体阻滞剂。在降血压类保健食品及中成药的复杂基质条件下,无需复杂的样品前处理或者传统的色谱分离,7种β-受体阻滞剂都成功地获得了快速定性分析。经HPLC-ESI/MS方法验证,DCBI-MS定性分析的结果准确可信,专属性高,分析速度快,有望成为通用工具并发挥重要作用。
6.开发了一种新的DCBI-MS分析方法,应用于减肥类保健食品添加剂和茶包中违禁化合物的快速筛查以及半定量检测。通过半自动进样装置,实现了减肥类保健食品中违禁添加物的高通量筛选。芬氟拉明、单去甲基西布曲明、双去甲基西布曲明、西布曲明、酚酞这5种常见非法添加成分在该方法中被成功检出。结果表明离子抑制等基质效应不影响DCBI-MS方法中对5种目标化合物的检出。然而,常压式质谱方法中存在时间歧视现象,故定量分析应以离子流谱图中峰面积为准。DCBI-MS不仅实现了高通量筛选分析,而且完成了半定量分析,使产品的现场普查得以实现。
Mass spectrometry (MS) is a powerful tool and has become one of the most important analytical methods in modern science. Ion source is the kernel of the mass spectrometer. There are3generations of ionization techniques, which are vacuum ionization, atmospheric ionization and ambient (open-air) ionization respectively. These ionization techniques for MS ever be called ambient desorption ionization, direct analysis MS, atmospheric desorption ionization, open air desorption ionization (OADI) and so forth. Ambient MS techniques have some unique characterizations. They exam objects of unusual shape and perform direct ionization without sample preparation. They generate ions softly and are compatible to most type of mass spectrometers. Although many ambient MS techniques have been reported, there are general shortcomings. Most of ambient ionization source suffers from insufficient quantification, poor precision, low space resolution, and uncertain location of sample surface.
In this paper, we report a new ambient ionization technique which termed Desorption Corona Beam Ionization (DCBI). The unique property of DCBI is a visible corona beam. The mechanism and improvement studies were carried out in order to get good quantification precision. Study on hyphenated technique or hybrid of other materials has broadened the sample range of DCBI. The concrete application of DCBI were demonstrated, such as rapid determination of (3-Receptor Blockers that illegally added in traditional medicines and dietary supplements, and high-throughput analysis of illicit additives in weight loss products. The following components are included in this study.
1. A novel Desorption Corona Beam Ionization (DCBI) source for direct analysis of samples from surface in mass spectrometry was developed, which is in accordance of ambient ionization techniques. The unique characteristic of DCBI is a visible corona beam that can extended from the source probe. A variety of samples has been analyzed by DCBI-MS approach. The ionization mechanism of DCBI was studied and along with the mechanism of dissociation in tandem mass spectrometry analysis. Program heating in desorption process was achieved to roughly separate the individual components in a complex mixture. Hence the mass spectrum is simple and specific. Compared with DESI, DCBI is a solvent free method though solvent can be added optionally. DCBI works in the corona discharge region whereas DART works in both corona and glow discharge regions. It implies that DCBI can be adopted to the commercial mass spectrometer easily due to commercial instrument supplies the power within corona current generally.
2. The room temperature ionic liquids (RTILs),1-butyl-3-methylimidazolium tetrafluoroborate ([BMIM]BF4) was employed as matrix to assist DCBI analysis. The quantification precision of the DCBI method was improved greatly. The thermal desorption process was recorded with thermal imaging and mass spectrometry simultaneously. When in a volatile liquid matrix, the analyte shows double peaks in ion chromatogram. It proved the hypothesis of two separated steps in desorption/ionization process. The unstable co-evaporation of analyte and matrix caused the deviation of quantitative result clearly. When non-volatile RTILs matrix applied, a micro slow release system (MSRS) is formed to smooth the evaporation fluctuation. Sharing the same thermal desorption mechanism, other APCI-related ambient MS methods may also be benefited from the RTILs matrix. The RTILs-DCBI-MS method also demonstrates the great potential for probing solution-phase droplet systems.
3. The poly(dimethylsiloxane)(PDMS) substrate enables capture of volatile and nonvolatile compounds from the liquid phase of a sample, offering large concentration factors of1000or more. Therefore, the soluble or insoluble analyte in matrix can both be detected with DCBI method since PDMS extraction results in matrixless/solventless sample introduction. Combined with PDMS substrate, the application range of DCBI analysis was broadened. The sensitive of method was enhanced significantly. In3orders of concentration, the calculated correlation coefficients for5pesticides were all satisfied. The limits of detection were all achieved on concentration of1μg/L. This PDMS/DCBI combination approach allows more organic compounds to be detected in complex mixtures with less time consuming. It provides the possibility of directly analyzing the trace level solution samples as well.
4. A novel hyphenated technique was proposed by coupling the near-infrared laser (976nm) with corona beam (LD-CBI) for direct analysis of formless samples. The samples can be liquid, powder and so forth. The LD-CBI method was characterized with broad region, non-specific absorption by common compounds. Thereby the heating is efficiency even without special substrate. There is less sample pretreatment in LD-CBI analysis. The sensitivity of this method is better than original DCBI approach. A variety of samples was detected and LD showed not only the better desorption efficiency but also necessity for some semi-volatile and non-volatile sample. The LOD of19pesticides were between0.02and1ng. For example, under selected ion mornitoring (SIM) mode, the detection limit of malathion was about10pg (30fMol) and the dynamic range is between0.2and100μg/mL along with a good correlation coefficient R2=0.9907.
5. A new rapid and economic method was developed by mounting DCBI source to an ion trap mass spectrometer. Seven p-receptor blockers were checked out in the complex matrix of antihypertensive traditional medicines and dietary supplements. The rapid screening was achieved without any tedious sample pretreatment and traditional chromatographic separation. The qualitative and semi-quantitative results were confirmed with the traditional HPLC-MS. Both in quality control and illicit additive screening of the β-receptor blockers, the DCBI-MS technique can be a powerful tool due to its accuracy, convenience and specificity.
6. The illicit additives in weight loss food were screened with the relative novel ambient MS technique, DCBI. Fenfluramine, N-di-desmethyl sibutramine, N-mono-desmethyl sibutramine, sibutramine, and phenolphthalein, the five abused chemicals were detected successfully with the developed DCBI-MS method. The matrix effect such as ion suppression did not influence the identification of target compounds. However, the time discrimination is significant in ambient MS analysis. Therefore, the quantification based on peak area in ion chromatogram is more accurate than ion intensity in the MS spectrum. Not only fast single-sample and high-throughput analysis were demonstrated but also the semi-quantitation was accomplished. The results implied that the on-site screening of illicit additives in weight-loss product can be achieved with DCBI-MS approach.
本论文中一种新的常压式离子化技术——解吸附电晕束离子源(DCBI)被研制,该源具有可视电晕束；进而DCBI定量分析的精密度得以研究改善,该方法亦可供类似离子化技术借鉴；联用其他技术扩展了DCBI源的适用对象和范围；实际应用中快速分析了降压类保健食品及中成药中7种芳氧丙醇胺类p-受体阻滞剂；高通量筛查了减肥类保健食品添加剂和茶包中的违禁化合物,并对这些违禁添加物进行了半定量检测分析。具体研究包括以下6部分内容。
1.开发了解吸附电晕束离子化(DCBI)技术,并以之构建了一新型常压式质谱用离子源,成功应用于多种物质的直接分析。该源具有独特的可视电晕束,为常压式质谱法开创了新的应用领域。正负离子化模式下的电离机理均得到研究与解释。利用程序升温分析复杂样品,降低了结果谱图中的化学噪声,使离子峰专属性加强。较之DESI, DCBI源结构紧凑,且不需辅助溶剂；较之DART, DCBI源工作于电晕放电区域,电流小,且源温度可程序扫描。
2.利用室温离子液体1-丁基-3甲基咪唑四氟硼酸盐([BMIM]BF4)作为基质,辅助DCBI质谱检测方法,显著改善了被分析物的定量精密度。通过红外热成像与质谱仪的同步监测,观察到了挥发性基质在DCBI离子源中的双峰现象,显示了DCBI源的解吸附、电离两步机理假设的合理性。研究发现样品进给过程中的共挥发效应是常压式敞开质谱法中定量结果出现波动的主要原因。当使用不挥发的室温离子液体作为基质时,其形成一个微缓释体系(MSRS),降低了解吸附波动,提高了定量精密度。其他相关的类APCI常压式离子化技术亦可参考该方法,以提高定量准确率与重现性。该研究也为液相微体积的过程研究与化学表征提供了可能。
3.利用聚二甲基硅氧烷(PDMS)膜获得浓缩系数1000倍以上的富集效率,还可以使不溶解于基质的化合物被富集,并应用DCBI对其进行分析。PDMS结合DCBI直接分析样品扩展了它们的应用范围,提高了分析的灵敏度。该联用技术对水样中农残进行定量分析,结果表明5种被分析农药的标准曲线都有很好的线性,检测限(1μg/L)。此研究表明联用PDMS基底与DCBI源,分析微量的液相样品快速、结果准确。这个方法可应用于分析各种不同溶液中的有机物,气态物,使得低浓度微量液体样品的直接质谱法分析成为可能。
4.联用近红外激光(976nm)与电晕束电离手段,开发了一种新型激光解吸附辅助电晕束离子化(LD-CBI)技术,考察了该技术方法特性,并成功将其应用于液体、粉末等无定形样品的直接分析。结果显示,因一般物质对近红外光的范频吸收特性,LD-CBI不但对样品前处理的要求更少,而且灵敏度更高。方法学验证中,19种农药样品的检测限在0.02-1ng之间。选择离子监测模式下,马拉硫磷的检测限可以低至10pg约合30fMol的水平,而且在样品浓度0.2μg/mL到100μg/mL的近三个数量级之间线性关系良好R2=0.9907。
5.利用DCBI源结合离子阱质谱仪,开发了一种快速、低耗的方法,快速筛选分析7种,芳氧丙醇胺类β-受体阻滞剂。在降血压类保健食品及中成药的复杂基质条件下,无需复杂的样品前处理或者传统的色谱分离,7种β-受体阻滞剂都成功地获得了快速定性分析。经HPLC-ESI/MS方法验证,DCBI-MS定性分析的结果准确可信,专属性高,分析速度快,有望成为通用工具并发挥重要作用。
6.开发了一种新的DCBI-MS分析方法,应用于减肥类保健食品添加剂和茶包中违禁化合物的快速筛查以及半定量检测。通过半自动进样装置,实现了减肥类保健食品中违禁添加物的高通量筛选。芬氟拉明、单去甲基西布曲明、双去甲基西布曲明、西布曲明、酚酞这5种常见非法添加成分在该方法中被成功检出。结果表明离子抑制等基质效应不影响DCBI-MS方法中对5种目标化合物的检出。然而,常压式质谱方法中存在时间歧视现象,故定量分析应以离子流谱图中峰面积为准。DCBI-MS不仅实现了高通量筛选分析,而且完成了半定量分析,使产品的现场普查得以实现。
Mass spectrometry (MS) is a powerful tool and has become one of the most important analytical methods in modern science. Ion source is the kernel of the mass spectrometer. There are3generations of ionization techniques, which are vacuum ionization, atmospheric ionization and ambient (open-air) ionization respectively. These ionization techniques for MS ever be called ambient desorption ionization, direct analysis MS, atmospheric desorption ionization, open air desorption ionization (OADI) and so forth. Ambient MS techniques have some unique characterizations. They exam objects of unusual shape and perform direct ionization without sample preparation. They generate ions softly and are compatible to most type of mass spectrometers. Although many ambient MS techniques have been reported, there are general shortcomings. Most of ambient ionization source suffers from insufficient quantification, poor precision, low space resolution, and uncertain location of sample surface.
In this paper, we report a new ambient ionization technique which termed Desorption Corona Beam Ionization (DCBI). The unique property of DCBI is a visible corona beam. The mechanism and improvement studies were carried out in order to get good quantification precision. Study on hyphenated technique or hybrid of other materials has broadened the sample range of DCBI. The concrete application of DCBI were demonstrated, such as rapid determination of (3-Receptor Blockers that illegally added in traditional medicines and dietary supplements, and high-throughput analysis of illicit additives in weight loss products. The following components are included in this study.
1. A novel Desorption Corona Beam Ionization (DCBI) source for direct analysis of samples from surface in mass spectrometry was developed, which is in accordance of ambient ionization techniques. The unique characteristic of DCBI is a visible corona beam that can extended from the source probe. A variety of samples has been analyzed by DCBI-MS approach. The ionization mechanism of DCBI was studied and along with the mechanism of dissociation in tandem mass spectrometry analysis. Program heating in desorption process was achieved to roughly separate the individual components in a complex mixture. Hence the mass spectrum is simple and specific. Compared with DESI, DCBI is a solvent free method though solvent can be added optionally. DCBI works in the corona discharge region whereas DART works in both corona and glow discharge regions. It implies that DCBI can be adopted to the commercial mass spectrometer easily due to commercial instrument supplies the power within corona current generally.
2. The room temperature ionic liquids (RTILs),1-butyl-3-methylimidazolium tetrafluoroborate ([BMIM]BF4) was employed as matrix to assist DCBI analysis. The quantification precision of the DCBI method was improved greatly. The thermal desorption process was recorded with thermal imaging and mass spectrometry simultaneously. When in a volatile liquid matrix, the analyte shows double peaks in ion chromatogram. It proved the hypothesis of two separated steps in desorption/ionization process. The unstable co-evaporation of analyte and matrix caused the deviation of quantitative result clearly. When non-volatile RTILs matrix applied, a micro slow release system (MSRS) is formed to smooth the evaporation fluctuation. Sharing the same thermal desorption mechanism, other APCI-related ambient MS methods may also be benefited from the RTILs matrix. The RTILs-DCBI-MS method also demonstrates the great potential for probing solution-phase droplet systems.
3. The poly(dimethylsiloxane)(PDMS) substrate enables capture of volatile and nonvolatile compounds from the liquid phase of a sample, offering large concentration factors of1000or more. Therefore, the soluble or insoluble analyte in matrix can both be detected with DCBI method since PDMS extraction results in matrixless/solventless sample introduction. Combined with PDMS substrate, the application range of DCBI analysis was broadened. The sensitive of method was enhanced significantly. In3orders of concentration, the calculated correlation coefficients for5pesticides were all satisfied. The limits of detection were all achieved on concentration of1μg/L. This PDMS/DCBI combination approach allows more organic compounds to be detected in complex mixtures with less time consuming. It provides the possibility of directly analyzing the trace level solution samples as well.
4. A novel hyphenated technique was proposed by coupling the near-infrared laser (976nm) with corona beam (LD-CBI) for direct analysis of formless samples. The samples can be liquid, powder and so forth. The LD-CBI method was characterized with broad region, non-specific absorption by common compounds. Thereby the heating is efficiency even without special substrate. There is less sample pretreatment in LD-CBI analysis. The sensitivity of this method is better than original DCBI approach. A variety of samples was detected and LD showed not only the better desorption efficiency but also necessity for some semi-volatile and non-volatile sample. The LOD of19pesticides were between0.02and1ng. For example, under selected ion mornitoring (SIM) mode, the detection limit of malathion was about10pg (30fMol) and the dynamic range is between0.2and100μg/mL along with a good correlation coefficient R2=0.9907.
5. A new rapid and economic method was developed by mounting DCBI source to an ion trap mass spectrometer. Seven p-receptor blockers were checked out in the complex matrix of antihypertensive traditional medicines and dietary supplements. The rapid screening was achieved without any tedious sample pretreatment and traditional chromatographic separation. The qualitative and semi-quantitative results were confirmed with the traditional HPLC-MS. Both in quality control and illicit additive screening of the β-receptor blockers, the DCBI-MS technique can be a powerful tool due to its accuracy, convenience and specificity.
6. The illicit additives in weight loss food were screened with the relative novel ambient MS technique, DCBI. Fenfluramine, N-di-desmethyl sibutramine, N-mono-desmethyl sibutramine, sibutramine, and phenolphthalein, the five abused chemicals were detected successfully with the developed DCBI-MS method. The matrix effect such as ion suppression did not influence the identification of target compounds. However, the time discrimination is significant in ambient MS analysis. Therefore, the quantification based on peak area in ion chromatogram is more accurate than ion intensity in the MS spectrum. Not only fast single-sample and high-throughput analysis were demonstrated but also the semi-quantitation was accomplished. The results implied that the on-site screening of illicit additives in weight-loss product can be achieved with DCBI-MS approach.
引文
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