质谱法用微波等离子体常压解吸电离源的研制
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摘要
以常压解吸电离源为代表的直接离子化技术是近年来质谱领域一次革命性发展。目前已经发展形成了20种以上的常压直接离子化技术,并广泛应用在食品、药品、环境、活体分析、代谢组学、蛋白质组学以及生物组织质谱成像等多个领域,具有非常重要的影响力及实用价值。基于此,本文开发了一种具有自主知识产权的基于微波等离子体的常压离子化技术,并对其进行了应用研究,具体为：
(1)在微波等离子体炬的基础上,构建了微波等离子体常压解吸电离源。并成功与质谱仪(Consair TOF MS)进行结合联用,开发了固体、液体、气体不同样品的直接检测方法。该方法无需样品预处理,可以直接对样品进行分析。
(2)研究了微波等离子体常压解吸电离源两种重要初级离子H3O+、NH4+及它们与H2O的团簇离子的产生条件及形成机理。认为质谱仪干燥氮气的流速及温度对这些初级离子及相关产物离子[M+H]+及[M+NH4]+的形成有很大的影响。而微波等离子体的解吸过程受等离子体中粒子的动量及等离子体温度的影响很大。
(3)采用本电离源比较研究了两种进样方式(即大气压挥发进样及传统的内管进样)下烷烃的电离及裂解规律。发现内管进样更容易使直链烷烃发生裂解,丢失亚甲基—CH2—后,一些碎片离子又会重新结合,致使碳链延长。得到减少或增加n个—CH2—的新烷烃(n不固定)。而大气压挥发进样更容易保证分子的完整性,对于碳数大于等于4的直链饱和烷烃,能够形成[M+13]+的产物离子,据此,提出了一种快速辨别物理性质相似的直链饱和烷烃的新方法。
(4)通过大气压挥发进样及内管进样两种进样方式研究了几种芳香族化合物在微波等离子体中发生的化学反应,发现芳香族化合物在这两种进样方式下都能够发生复杂多样的化学反应,如还原、氧化、质子化等,大气成分如氧气、氮气等也会参与其中。而在发生还原反应时,能够得到还原程度不同的加氢产物,该过程在微波等离子的参与下进行,无需外加催化剂及化学试剂。
(5)通过微波等离子体常压解吸电离质谱法对药物中活性成分进行检测研究,开发了一种对片状、膏状及胶囊类药物单一或多种主要成分的快速定性分析的方法。分析方法简单,无需样品预处理,无需有毒有害化学试剂。分析速度快,在手动进样的情况下可以达到360次/小时。一级谱图中所获得的碎裂离子信息有助于对活性成分的定性分析。
(6)对现有的微波等离子体常压解吸电离源进行了小型化改进,新的小型电离源操作更为灵活方便,功耗进一步降低。通过应用试验证明该电离源的解吸电离性能能够得到充分的保障。
Mass spectrometry has recently undergone a contemporary revolution with the introduction of a new group of ambient desorption/ionization (D1) sources known collectively as direct ionization techniques. To date, more than20ionization techniques have been developed and they are widely used in the field of food, medicine, environment, in vivo analysis, metabolomics, proteomics and bio-tissue mass spectrometry imaging, etc. The techniques show very important influences and the practical values. In this study, an ambient ionization technique with the independent intellectual property rights was developed based on microwave plasma, and the mass spectrometric applications of this technique were done, the main contents are as follows:
(1) The microwave plasma atmospheric pressure desorption/ionization (MWPAPDI) source was developed based on the microwave plasma torch (MPT), then by coupled it with a mass spectrometer (Consair TOF MS), the direct analysis methods for the various samples of solid phase, liquid phase and gaseous phase were developed. These methods do not need sample pretreatment and the samples can be analyzed directly.
(2) From the MWPAPDI, the conditions and the mechanisms for the generation of the two important primary ions H3O+and NH4+with their water clusters were studied. The results shows that the flowing rate and the temperature of the drying nitrogen from the mass spectrometer have important effects on these primary ions and the corresponding product ions [M+H]+and [M+NH4]+. While the momentum of the particles in the plasma and the plasma temperature are the important factors to influence the microwave plasma desorption process.
(3) The rules of dissociation and ionization for the alkanes were studied by MWPAPDI through the two sample introduction ways:the atmospheric pressure-volatizing sampling and the traditional central tube sampling. The results show that the straight-chain alkanes are more easily to dissociate by losing—CH2—then the generated fragment ions will recombine to prolong the carbon chain lengths. The end products are the new alkanes by losing or adding n (—CH2—)(nis not fixed) from the original alkanes through the central tube sampling. For the straight-chain alkanes through the atmospheric pressure-volatizing sampling, their molecules are more easily to be kept intact, and when the number of carbon atoms is equal to or more than4, the straight-chain alkanes can form the product ions of [M+13]+. Based on this rule, a novel direct analysis method for quick-identification of saturated straight-chain alkanes with similar physical properties was proposed.
(4) The reactions of the aromatic compounds in the microwave plasma were investigated through the atmospheric pressure-volatizing sampling and the central tube sampling. It was found that the complex and various reactions, such as reduction, oxidation and protonation, etc., could occur in either sampling ways. The atmosphereic components, such as oxygen and nitrogen, also participate in the reactions. For the reduction reactions, the hydrogenated products with different deduction degree can be obtained. These processes are facilitated by the microwave plasma and require no additional catalysts.
(5) Based on the analysis for medicines by MWPAPDI-MS, a rapid qualitative detection method for one or more active pharmaceutical ingredients (APIs) in medicines with different forms (tablets, ointments, and capsules) was developed. This method is simple without the need for chemical reagents and sample pretreatments. It also has high throughput that the analysis speed can reach360samples per hour with manual sampling. In this method, the fragment ion signals exist in the full-scan spectra, these information can assist to identify the APIs.
(6) The miniaturization was made for the current MWPAPDI source. The new miniature source is more easily to operate and the power consumption reduces. While the abilities for the desorption/ionization of this miniature source are still adequately kept and shown by the observation of the application experiments.
(1)在微波等离子体炬的基础上,构建了微波等离子体常压解吸电离源。并成功与质谱仪(Consair TOF MS)进行结合联用,开发了固体、液体、气体不同样品的直接检测方法。该方法无需样品预处理,可以直接对样品进行分析。
(2)研究了微波等离子体常压解吸电离源两种重要初级离子H3O+、NH4+及它们与H2O的团簇离子的产生条件及形成机理。认为质谱仪干燥氮气的流速及温度对这些初级离子及相关产物离子[M+H]+及[M+NH4]+的形成有很大的影响。而微波等离子体的解吸过程受等离子体中粒子的动量及等离子体温度的影响很大。
(3)采用本电离源比较研究了两种进样方式(即大气压挥发进样及传统的内管进样)下烷烃的电离及裂解规律。发现内管进样更容易使直链烷烃发生裂解,丢失亚甲基—CH2—后,一些碎片离子又会重新结合,致使碳链延长。得到减少或增加n个—CH2—的新烷烃(n不固定)。而大气压挥发进样更容易保证分子的完整性,对于碳数大于等于4的直链饱和烷烃,能够形成[M+13]+的产物离子,据此,提出了一种快速辨别物理性质相似的直链饱和烷烃的新方法。
(4)通过大气压挥发进样及内管进样两种进样方式研究了几种芳香族化合物在微波等离子体中发生的化学反应,发现芳香族化合物在这两种进样方式下都能够发生复杂多样的化学反应,如还原、氧化、质子化等,大气成分如氧气、氮气等也会参与其中。而在发生还原反应时,能够得到还原程度不同的加氢产物,该过程在微波等离子的参与下进行,无需外加催化剂及化学试剂。
(5)通过微波等离子体常压解吸电离质谱法对药物中活性成分进行检测研究,开发了一种对片状、膏状及胶囊类药物单一或多种主要成分的快速定性分析的方法。分析方法简单,无需样品预处理,无需有毒有害化学试剂。分析速度快,在手动进样的情况下可以达到360次/小时。一级谱图中所获得的碎裂离子信息有助于对活性成分的定性分析。
(6)对现有的微波等离子体常压解吸电离源进行了小型化改进,新的小型电离源操作更为灵活方便,功耗进一步降低。通过应用试验证明该电离源的解吸电离性能能够得到充分的保障。
Mass spectrometry has recently undergone a contemporary revolution with the introduction of a new group of ambient desorption/ionization (D1) sources known collectively as direct ionization techniques. To date, more than20ionization techniques have been developed and they are widely used in the field of food, medicine, environment, in vivo analysis, metabolomics, proteomics and bio-tissue mass spectrometry imaging, etc. The techniques show very important influences and the practical values. In this study, an ambient ionization technique with the independent intellectual property rights was developed based on microwave plasma, and the mass spectrometric applications of this technique were done, the main contents are as follows:
(1) The microwave plasma atmospheric pressure desorption/ionization (MWPAPDI) source was developed based on the microwave plasma torch (MPT), then by coupled it with a mass spectrometer (Consair TOF MS), the direct analysis methods for the various samples of solid phase, liquid phase and gaseous phase were developed. These methods do not need sample pretreatment and the samples can be analyzed directly.
(2) From the MWPAPDI, the conditions and the mechanisms for the generation of the two important primary ions H3O+and NH4+with their water clusters were studied. The results shows that the flowing rate and the temperature of the drying nitrogen from the mass spectrometer have important effects on these primary ions and the corresponding product ions [M+H]+and [M+NH4]+. While the momentum of the particles in the plasma and the plasma temperature are the important factors to influence the microwave plasma desorption process.
(3) The rules of dissociation and ionization for the alkanes were studied by MWPAPDI through the two sample introduction ways:the atmospheric pressure-volatizing sampling and the traditional central tube sampling. The results show that the straight-chain alkanes are more easily to dissociate by losing—CH2—then the generated fragment ions will recombine to prolong the carbon chain lengths. The end products are the new alkanes by losing or adding n (—CH2—)(nis not fixed) from the original alkanes through the central tube sampling. For the straight-chain alkanes through the atmospheric pressure-volatizing sampling, their molecules are more easily to be kept intact, and when the number of carbon atoms is equal to or more than4, the straight-chain alkanes can form the product ions of [M+13]+. Based on this rule, a novel direct analysis method for quick-identification of saturated straight-chain alkanes with similar physical properties was proposed.
(4) The reactions of the aromatic compounds in the microwave plasma were investigated through the atmospheric pressure-volatizing sampling and the central tube sampling. It was found that the complex and various reactions, such as reduction, oxidation and protonation, etc., could occur in either sampling ways. The atmosphereic components, such as oxygen and nitrogen, also participate in the reactions. For the reduction reactions, the hydrogenated products with different deduction degree can be obtained. These processes are facilitated by the microwave plasma and require no additional catalysts.
(5) Based on the analysis for medicines by MWPAPDI-MS, a rapid qualitative detection method for one or more active pharmaceutical ingredients (APIs) in medicines with different forms (tablets, ointments, and capsules) was developed. This method is simple without the need for chemical reagents and sample pretreatments. It also has high throughput that the analysis speed can reach360samples per hour with manual sampling. In this method, the fragment ion signals exist in the full-scan spectra, these information can assist to identify the APIs.
(6) The miniaturization was made for the current MWPAPDI source. The new miniature source is more easily to operate and the power consumption reduces. While the abilities for the desorption/ionization of this miniature source are still adequately kept and shown by the observation of the application experiments.
引文
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