Dissertation > Mathematical sciences and chemical > Chemistry > Analytical Chemistry > Instrument analysis ( physics and physical chemistry ) > Mass Spectrometry

Characterization of Mercury-Binding Metallothioneins (MTs) and Measurement of Lead Isotopic Ratios with Inductively Coupled Plasma Mass Spectrometry (ICP-MS)

Author HuangZhiYong
Tutor WangXiaoRu
School Xiamen University
Course Analytical Chemistry
Keywords Metallothionein (MTs) Cinnabar Combined with analytical techniques Isotope ratios Isotope dilution
CLC O657.63
Type PhD thesis
Year 2003
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Cinnabar, as a natural mineral drugs of traditional Chinese medicine (TCM) has been widely used to prove that it has a significant effect after two thousand years of clinical practice, but the toxicity of its use of cinnabar security debate has always never interruption. Based on this, the paper by fed with cinnabar rat organs after mercury accumulation behavior and characteristics of the in vivo induction of metallothionein to the toxicity of the drugs and the use of security research. This thesis consists of two parts, the first part from the first to the fourth chapter, several characterization fed with the cinnabar rat metallothionein (MTs); second part including from the fifth to the eighth chapter, inductively coupled plasma mass spectrometry (ICP-MS) method to study the determination of lead isotope ratios and its application. MTs are a class of small molecule cysteine-rich protein that has anti detoxifying effect of heavy metals in organisms, and to take advantage of this feature can be used as the evaluation of irrigation fed the cinnabar rats mercury toxicity biomarkers. Chapter by more than 100 references MTs basic knowledge, including: the origin of MTs, structural characteristics, biological functions, the traditional methods of measuring and modern combined with technology characterization methods. In the second chapter, respectively, by inductively coupled plasma mass spectrometry and mercury metal saturation were determined cinnabar in rats fed with various organs such as the liver, kidney, heart, brain and testis heavy metals (including mercury, copper and zinc) and MTs total and organ in the correlation between total heavy metal content and MTs for discussion. The results show that: fed with rat organs after cinnabar role in the accumulation of mercury have a certain degree, especially the level of mercury accumulation in the liver, kidney and testicular particularly significant. When fed with high-dose cinnabar or for a long time fed with the cinnabar mercury toxicity will be very significant, even fatal. The study found that there is a high degree of correlation between the rat liver and kidney mercury content and MTs total, therefore, MTs as biomarkers of cinnabar toxicity and safety studies. Third chapter describes fed with cinnabar and HgCl 2 rats induced metal generated using the combined method based gel chromatography (SEC) and ICP-MS hyphenated techniques, combined MTs The component line separation analysis. The results show that: fed with Cinnabar rats generated in the various organs Hg combined MTs components than those fed with HgCl 2 group less; With the extension of the time were fed and fed with dosage increase , the Hg-MTs components cinnabar rats kidney Abstract: Charaeterization ofmereury a bindingmetallothio indulge ins (MTs) andmeasurementofleadisotoPie ratios with induetively couP! which printed Lang said Plasmam me wonderful the (Ic BU MS) induced amount also will be significantly increased At the same time, the kidney, the amount of Cu-MTS induction significantly increased. MTs change plays an important role in detoxification in the kidney of heavy metal poisoning. Hg MTs component induced in the liver seems to be fed with the size of the dose (2.5 5.0 g / Kghw) and fed with a duration of 4 weeks (winter) has nothing to do, but the induction of cinnabar group were compared with the control group increased significantly. This chapter also compare Cinnabar group fed with the, H solution l: group and a control group of other organs of the rat Hg MTs component induced and variation. Chapter IV, the establishment of a reversed-phase liquid chromatography (rp · HPLc) together with the inductance curse plasma mass spectrometry (IcP MS) and electrospray ionization mass spectrometry (ESI-MS) MS rats of mercury combine different existing form of metallothionein characterization. Samples by dextran gel column (SePhadex G-75) after separation by dialysis (M lt; l 000 Da) desalination, rat kidney, liver MTs extract small inside diameter of reversed-phase column separated, and the gradient elution. Eluate by ICP-MS detection analysis combined with different metals MTs of subtypes, times subtype structure information. RpHPLC Esl Ms associated with the availability of the MTs multi-form ultraviolet (UV) and ESI-MS total ion current (TIC) chromatogram. Of Hg a MTs MTs morphology related Hg before of Esl-MS analysis reported rarely, and the lack of a standard reference material of Hg MTs and its subtypes, subtypes and sub-subtypes molecular structure only through Tlc chromatogram The mass of each fragment peaks corresponding mass number and reference literature related demetallization the MTs (aPO a MTs) inferred. The experimental results show that rats fed with Hgcl: kidney induced metallothionein generated mercury combined, and the presence of several multi-metal binding MTs morphology. The the liver Hg MTs induced less Esl a Ms detection sensitivity is relatively low and the number of unknown factors, and therefore unable to the liver Hg MTs morphological structure characterization. Isotope ratio measurements usually by thermal ionization mass spectrometry (TIMS) and spark source ionization mass spectrometry (sSMs) or secondary ionization source mass spectrometry (SIMS), but based on quadrupole separate inductors curse aggregate plasma mass spectrometry (ICP-QMS) its advantages of simple, accurate, ideal measuring tool element isotope ratios isotope dilution (ID) method, which can be used to accurately measure the concentration of the element. In the fifth chapter, through access to a large number of references, introduced the same affect ICP a MS determination of isotope ratio accuracy and precision of the main factors, such as generated by the plasma systems and quadrupole mass analyzer and detector isobaric ions, polyatomic molecular ion, quality discrimination, the dead time and the instrument operating parameters and calibration eliminate Abstract: mercury binding metallothionein inductive coupling plasma mass spectrometry characterization of isotopic ratios and lead server bit The corresponding interference factors taken method. Based on the JCP an MS accurate measurement of isotope ratios, the use of isotope dilution method to determine the value of the element concentrations in fine. To research the advantages of isotope dilution measurement method, measured IcP a Ms plants, human hair standard reference material lead isotope ratios in which the lead content is given, and then using isotope dilution method barium l experimental measurements with standard substances lead reference The value is very consistent. Chapter 6 describes the isotope dilution ICP-MS method for determination of the consumption of trace amounts of lead in CaC03. Sample after anion exchange method to remove the CaCO3 matrix, and then Tl as internal standard calibration quality discrimination, Sichuan isotope dilution and the JCP an MS determination of lead content in the samples. Blank three times the standard deviation of the measured values ??(ie 3X.) As the minimum detection limit of the measuring method, as 2.944ng/mL. The recovery was

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