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Determination of monophosphate nucleotides, sulfur-containing amino acids, arsenic species and various oxidation states of iron, vanadium and chromium by capillary electrophoresis inductively coupled plasma mass spectrometry

by Yeh, Ching-fen

Abstract (Summary)
Capillary electrophoresis (CE) is in comparison with other chromatographic techniques, CE has several advantages such as high resolving power, small sample volume requirement, minimal buffer consumption and high sample throughtput. As a detection technique, inductively coupled plasma mass spectrometry (ICPMS) provides the advantages of low detection limit, multielement detection, and element- and isotope-specific detection capabilities. Therefore, the use of CE as a high resolution separation technique with ICP-MS as a sensitive element specific detector is of growing interest for analytical research. Four studies in our research are described below, respectively. A preliminary study of a modified microconcentric nebulizer (CEI-100, CETAC) as the sample introduction device of capillary electrophoresis inductively coupled plasma mass spectrometry (CE-ICP-MS) for the determination of monophosphate nucleotides is described. The monophosphate nucleotides studied include adenosine 5¡¦-monophosphate (AMP), guanosine 5¡¦-monophosphate (GMP), uridine 5¡¦-monophosphate (UMP) and inosine 5¡¦-monophosphate (IMP). The species studied were well separated using a 70 cm length ¡Ñ 75 £gm id fused silica capillary while the applied voltage was set at -22 kV and a 20 mmol/L ammonium citrate/citric acid buffer (pH 4.0) containing 0.1% m/v cationic polymer (hexadimethrine bromide, Polybrene) was used as the electrophoretic buffer. The electroosmotic flow was reversed by flushing the fused silica capillary with 0.2% m/v Polybrene to accelerate separation. The detection limit of various species studied was in the range of 0.036~0.054 £gg P/mL, which corresponded to the absolute detection limit of 1.1~1.6 pg P based on the injection volume of 30 nl. We determined the concentrations of nucleotides in two IG-enriched monosodium glutamates purchased from the local market. The recovery was in the range of 100~112% for various species, and the concentrations of IMP and GMP in these samples were in the range of 0.15¡V0.18% m/m. Capillary electrophoresis dynamic reaction cellTM inductively coupled plasma mass spectrometry (CE-DRC-ICP-MS) for the determination of sulfur-containing amino acids is described. The sulfur-containing amino acids studied include L-cysteine, L-cystine, DL-homocystine and L-methionine. The species studied were well separated using a 70 cm length ¡Ñ 75 £gm i.d. fused silica capillary while the applied voltage was set at +22 kV and a 10 mmol/L disodium tetraborate buffer (pH 9.8) containing 0.1 mmol/L EDTA and 0.5 mmol/L Triton X-100 was used as the electrophoretic buffer. The sulfur-selective electropherogram was determined at m/z 48 as 32S16O+ by using its reaction with O2 in the reaction cell. The method avoided the effect of polyatomic isobaric interferences at m/z 32 caused by 16O16O+ and 14N18O+ on 32S+ by detecting 32S+ as the oxide ion 32S16O+ at m/z 48, which is less interfered. The detection limit of various species studied was in the range of 0.047~0.058 £gg S/mL, which corresponded to the absolute detection limit of 1.3~1.6 pg S based on the injection volume of 27 nl. We determined the concentrations of selected sulfur-containing amino acids in urine and nutritive complement samples. The recovery was in the range of 92~128% for various species. Capillary electrophoresis-dynamic reaction cell inductively coupled plasma mass spectrometry (CE-DRC-ICP-MS) for the speciation of iron (III/II), vanadium (V/IV) and chromium (VI/III) is described. Two different CE migration modes were employed for separating the six metal ions using pre-capillary complexation. One is counter-electroosmotic mode in which iron (III/II) and vanadium (V/IV) ions were well separated using a 60 cm ¡Ñ 75 £gm i.d. fused silica capillary. The voltage was set at +22 kV and a 15 mmol/L tris(hydroxymethyl)aminomethane (Tris) buffer (pH 8.75) containing 0.5 mmol/L ethylenediaminetetraacetic acid (EDTA) and 0.5 mmol/L ortho-phenanthroline (phen) was used as the electrophoretic buffer. The other is co-electroosmotic mode in which chromium (VI/III) ions were well separated while the applied voltage was set at ?22 kV and a 10 mmol/L ammonium citrate buffer (pH 7.7) containing 0.5 mmol/L diethylenetriaminepentaacetic acid (DTPA) and 0.01% polybrene was used as the electrophoretic buffer. The mass spectra were measured at m/z 51, 52 and 56 for V, Cr and Fe, respectively. The interfering polyatomic ions of 35Cl16O+, 40Ar12C+ and 40Ar16O+ on 51V+, 52Cr+ and 56Fe+ determination were reduced in intensity significantly by using NH3 as the reaction cell gas in the DRC. The detection limits were in the range of 0.1~0.5, 0.4~1.3 and 1.2~1.7 £gg/L for V, Cr and Fe, respectively. Applications of the method for the speciation of V, Cr and Fe in wastewater were demonstrated. The recoveries were in the range of 92~120% for various species. A capillary electrophoresis-inductively coupled plasma-mass spectrometric (CE-ICPMS) method for the speciation of six arsenic compounds, namely arsenite [As(III)], arsenate [As(V)], monomethyl arsonic acid, dimethylarsinic acid, arsenobetaine and arsenocholine is described. The separation has been achieved on a 70 cm length ¡Ñ 75 £gm ID fused-silica capillary. The electrophoretic buffer used was 15 mmol/L Tris (pH 9.0) containing 15 mmol/L sodium dodecyl sulfate (SDS), while the applied voltage was set at +22 kV. The arsenic species in biological tissues were extracted into 80% v/v methanol-water mixture, put in a closed centrifuge tube and kept in a water bath, using microwaves at 80¢J for 3 min. The extraction efficiencies of individual arsenic species added to the sample at 0.5 mg As/g level were between 96% and 107%, except for As(III), for which it was 89% and 77% for oyster and fish samples, respectively. The detection limits of the species studied were in the range 0.3~0.5 £gg As/L. The procedure has been applied for the speciation analysis of two reference materials, namely dogfish muscle tissue (NRCC DORM-2) and oyster tissue (NIST SRM 1566a), and two real-world samples.
Bibliographical Information:

Advisor:Hsuan-Jung Huang; Shu-Hui Chen; Tai-Sung Hsi; Chang-Huan Tsung; Shiuh-Jen Jiang; Jen-Taie Shiea

School:National Sun Yat-Sen University

School Location:China - Taiwan

Source Type:Master's Thesis

Keywords:monophosphate nucleotides vanadium capillary electrophoresis arsenic species chromium iron sulfur containing amino acids inductively coupled plasma mass spectrometry

ISBN:

Date of Publication:07/15/2005

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