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Production of WE43 magnesium alloy by powder metallurgy and the effect of glucose on wear resistance in biocorrosive wear. Biointerphases 2023; 18:2878718. [PMID: 37096903 DOI: 10.1116/6.0002270] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 10/10/2022] [Accepted: 02/22/2023] [Indexed: 03/19/2023] Open
Abstract
In this study, WE43 magnesium alloy was produced by the powder metallurgy method. Microstructural analyses of the produced samples were carried out using the scanning electron microscopy method. X-ray fluorescence, energy dispersive x-ray (EDS) analysis, and hardness tests were also implemented to investigate the physical and chemical properties of the alloys. The volumetric hardness was measured to be approximately 53 HV. The microstructural analysis and EDS results indicated the presence of Mg24Y5 and Mg41Nd5 phases in the alloys. Reciprocating-type experiments were carried out in dry and corrosive environments to evaluate the wear resistance. Hanks’s solution containing 2% g/l glucose was used as the corrosive environment. Gluconic acid resulting from the oxidation of glucose in the Hanks’s solution formed a new thin layer on the alloy surface, which was observed in the worn surface images. The formation of the thin film on the alloy surface resulted in an increase in wear resistance by 37%. The results unraveled the potential of the WE43 alloys as implant materials in areas in contact with glucose.
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Rageh AH, Pyell U. “Pseudostationary Ion-Exchanger” Sweeping as an Online Enrichment Technique in the Determination of Nucleosides in Urine via Micellar Electrokinetic Chromatography. Chromatographia 2018. [DOI: 10.1007/s10337-018-3570-9] [Citation(s) in RCA: 4] [Impact Index Per Article: 0.6] [Reference Citation Analysis] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 10/28/2022]
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3
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Buzatto AZ, de Oliveira Silva M, Poppi RJ, Simionato AVC. Assessment of nucleosides as putative tumor biomarkers in prostate cancer screening by CE–UV. Anal Bioanal Chem 2017; 409:3289-3297. [DOI: 10.1007/s00216-017-0297-7] [Citation(s) in RCA: 10] [Impact Index Per Article: 1.3] [Reference Citation Analysis] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 12/20/2016] [Revised: 02/14/2017] [Accepted: 03/03/2017] [Indexed: 10/19/2022]
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4
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Jiang M, Prokhorova AF, Rozhmanova NB, Shpigun OA. Electrophoretic separation of some nucleosides for the diagnosis of mastopathy and fibroadenoma. JOURNAL OF ANALYTICAL CHEMISTRY 2017. [DOI: 10.1134/s1061934816120091] [Citation(s) in RCA: 2] [Impact Index Per Article: 0.3] [Reference Citation Analysis] [Track Full Text] [Subscribe] [Scholar Register] [Indexed: 11/23/2022]
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5
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Detection of 1,N2-propano-2′-deoxyguanosine in human urine by stable isotope dilution UHPLC–MS/MS analysis. J Chromatogr B Analyt Technol Biomed Life Sci 2016; 1023-1024:68-71. [DOI: 10.1016/j.jchromb.2016.04.029] [Citation(s) in RCA: 3] [Impact Index Per Article: 0.3] [Reference Citation Analysis] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 02/22/2016] [Revised: 04/15/2016] [Accepted: 04/16/2016] [Indexed: 11/18/2022]
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6
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Buzatto AZ, Guedes SF, de Oliveira Silva M, Gallafrio JM, Simionato AVC. Higher detectability method for the analysis of nucleosides, putative tumor biomarkers, in blood serum samples by CE-UV with reversed EOF. Electrophoresis 2015; 36:2968-75. [DOI: 10.1002/elps.201500161] [Citation(s) in RCA: 5] [Impact Index Per Article: 0.5] [Reference Citation Analysis] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 03/26/2015] [Revised: 06/03/2015] [Accepted: 07/23/2015] [Indexed: 11/08/2022]
Affiliation(s)
- Adriana Zardini Buzatto
- Laboratory of Biomolecules Analysis Tiselius (LABi Tiselius), Department of Analytical Chemistry, Chemistry Institute; Campinas State University (UNICAMP); Campinas SP Brazil
| | - Sumaya Ferreira Guedes
- Laboratory of Biomolecules Analysis Tiselius (LABi Tiselius), Department of Analytical Chemistry, Chemistry Institute; Campinas State University (UNICAMP); Campinas SP Brazil
| | - Mariana de Oliveira Silva
- Laboratory of Biomolecules Analysis Tiselius (LABi Tiselius), Department of Analytical Chemistry, Chemistry Institute; Campinas State University (UNICAMP); Campinas SP Brazil
| | - Jéssica Mirela Gallafrio
- Laboratory of Biomolecules Analysis Tiselius (LABi Tiselius), Department of Analytical Chemistry, Chemistry Institute; Campinas State University (UNICAMP); Campinas SP Brazil
| | - Ana Valéria Colnaghi Simionato
- Laboratory of Biomolecules Analysis Tiselius (LABi Tiselius), Department of Analytical Chemistry, Chemistry Institute; Campinas State University (UNICAMP); Campinas SP Brazil
- National Institute of Science and Technology in Bioanalytics (INCTBio), Chemistry Institute; Campinas State University (UNICAMP); Campinas SP Brazil
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7
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Determination of urinary nucleosides via borate complexation capillary electrophoresis combined with dynamic pH junction-sweeping-large volume sample stacking as three sequential steps for their on-line enrichment. Anal Bioanal Chem 2014; 406:5877-95. [DOI: 10.1007/s00216-014-8022-2] [Citation(s) in RCA: 18] [Impact Index Per Article: 1.6] [Reference Citation Analysis] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 04/25/2014] [Revised: 06/20/2014] [Accepted: 07/07/2014] [Indexed: 01/14/2023]
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8
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Simionato AVC, Carrilho E, Maggi Tavares MF. CE-MS and related techniques as a valuable tool in tumor biomarkers research. Electrophoresis 2010; 31:1214-1226. [DOI: 10.1002/elps.200900671] [Citation(s) in RCA: 19] [Impact Index Per Article: 1.3] [Reference Citation Analysis] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 12/12/2022]
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9
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Tuytten R, Lemière F, Esmans EL, Herrebout WA, van der Veken BJ, Maes BUW, Witters E, Newton RP, Dudley E. Role of Nitrogen Lewis Basicity in Boronate Affinity Chromatography of Nucleosides. Anal Chem 2007; 79:6662-9. [PMID: 17672481 DOI: 10.1021/ac0709089] [Citation(s) in RCA: 44] [Impact Index Per Article: 2.4] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 11/29/2022]
Abstract
Urinary modified nucleosides have a potential role as cancer biomarkers, and most of the methods used in their study have utilized low-pressure phenylboronate affinity chromatography materials for the purification of the cis-diol-containing nucleosides. In this study, a boronate HPLC column was surprisingly shown not to trap the nucleosides as would be expected from experience with the classic Affigel 601 resin but showed only partial selectivity toward cis-diol groups while other groups exhibited better retention. In aprotic conditions, trapping of nucleosides was possible; however, the selectivity toward cis-diol-containing compounds was lost with the Lewis basicity of available nitrogens being the main determinant of retention. The experimental findings are compared to and confirmed by DFT calculations.
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Affiliation(s)
- Robin Tuytten
- Department of Chemistry, Nucleoside Research and Mass Spectrometry Unit, and Center for Proteomics and Mass Spectrometry, University of Antwerp, Antwerp, Belgium
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10
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Bond A, Dudley E, Lemière F, Tuytten R, El-Sharkawi S, Brenton AG, Esmans EL, Newton RP. Analysis of urinary nucleosides. V. Identification of urinary pyrimidine nucleosides by liquid chromatography/electrospray mass spectrometry. RAPID COMMUNICATIONS IN MASS SPECTROMETRY : RCM 2006; 20:137-50. [PMID: 16331740 DOI: 10.1002/rcm.2266] [Citation(s) in RCA: 15] [Impact Index Per Article: 0.8] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Subscribe] [Scholar Register] [Indexed: 05/05/2023]
Abstract
Modified urinary nucleosides are potentially invaluable in cancer diagnosis, as they reflect altered RNA turnovers. High-performance liquid chromatography (HPLC) was combined with full-scan mass spectrometry, tandem mass spectrometry, MS(n) analysis and accurate mass measurements in order to identify pyrimidine nucleosides purified from urine. Potential nucleosides were assessed by their evident UV absorbance in the HPLC chromatogram and then further examined by the various mass spectrometric techniques. In this manner numerous pyrimidine nucleosides were identified in the urine samples from cancer patients including pseudouridine, cytidine, two methylcytidines and an acetylcytidine. Furthermore, a number of novel modified pyrimidine nucleosides were tentatively identified via critical interpretation of the combined mass spectrometric data.
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Affiliation(s)
- Alison Bond
- Biomolecular Analysis Mass Spectrometry (BAMS) Facility, Grove Building, University of Wales Swansea, Singleton Park, Swansea SA2 8PP, UK
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11
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Yang J, Xu G, Zheng Y, Kong H, Wang C, Zhao X, Pang T. Strategy for metabonomics research based on high-performance liquid chromatography and liquid chromatography coupled with tandem mass spectrometry. J Chromatogr A 2005; 1084:214-21. [PMID: 16114257 DOI: 10.1016/j.chroma.2004.10.100] [Citation(s) in RCA: 57] [Impact Index Per Article: 2.9] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 10/26/2022]
Abstract
Metabonomics, the study of metabolites and their roles in various disease states, is a novel methodology arising from the post-genomics era. This methodology has been applied in many fields. Current metabonomic practice has relied on mass spectrometry (MS), gas chromatography-mass spectrometry (GC-MS), and nuclear magnetic resonance (NMR) to analyze metabolites. In this study, a strategy was developed for applying high-performance liquid chromatography (HPLC) and LC-MS-MS to metabonomics research. One of the key problems to be solved in this strategy is to match the peaks between the chromatograms. A peak alignment algorithm has been developed to match the chromatograms before the pattern recognition. As an application example, the strategy described above was applied to metabonomics research on liver diseases, and the false-positive result of live cancer diagnosis from the hepatocirrhosis and hepatitis diseases was effectively reduced to 7.40%. Based on the pattern recognition, several potential biomarkers were found and further identified by the following LC-MS-MS experiments. The structures of eight potential biomarkers were given for distinguishing the liver cancer from the hepatocirrhosis and hepatitis diseases.
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Affiliation(s)
- Jun Yang
- National Chromatographic R&A Center, Dalian Institute of Chemical Physics, Chinese Academy of Sciences, 116011 Dalian, China
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12
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Zheng YF, Yang J, Zhao XJ, Feng B, Kong HW, Chen YJ, Lv S, Zheng MH, Xu GW. Urinary nucleosides as biological markers for patients with colorectal cancer. World J Gastroenterol 2005; 11:3871-6. [PMID: 15991285 PMCID: PMC4504888 DOI: 10.3748/wjg.v11.i25.3871] [Citation(s) in RCA: 54] [Impact Index Per Article: 2.7] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Download PDF] [Journal Information] [Submit a Manuscript] [Subscribe] [Scholar Register] [Indexed: 02/06/2023] Open
Abstract
AIM: Fourteen urinary nucleosides, primary degradation products of tRNA, were evaluated to know the potential as biological markers for patients with colorectal cancer.
METHODS: The concentrations of 14 kinds of urinary nucleosides from 52 patients with colorectal cancer, 10 patients with intestinal villous adenoma and 60 healthy adults were determined by column switching high performance liquid chromatography method.
RESULTS: The mean levels of 12 kinds of urinary nucleosides (except uridine and guanosine) in the patients with colorectal cancer were significantly higher than those in patients with intestinal villous adenoma or the healthy adults. Using the levels of 14 kinds of urinary nucleosides as the data vectors for principal component analysis, 71% (37/52) patients with colorectal cancer were correctly classified from healthy adults, in which the identification rate was much higher than that of CEA method (29%). Only 10% (1/10) of patients with intestinal villous adenoma were indistinguishable from patients with colorectal cancer. The levels of m1G, Pseu and m1A were positively related with tumor size and Duke’s stages of colorectal cancer. When monitoring the changes in urinary nucleoside concentrations of patients with colorectal cancer associated with surgery, it was found that the overall correlations with clinical assessment were 84% (27/32) and 91% (10/11) in response group and progressive group, respectively.
CONCLUSION: These findings indicate that urinary nucleosides determined by column switching high performance liquid chromatography method may be useful as biological markers for colorectal cancer.
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Affiliation(s)
- Yu-Fang Zheng
- National Chromatographic R and A Center, Dalian Institute of Chemical Physics, Chinese Academy of Sciences, Dalian 116023, Liaoning Province, China
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13
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Gong YX, Li SP, Wang YT, Li P, Yang FQ. Simultaneous determination of anthraquinones in Rhubarb by pressurized liquid extraction and capillary zone electrophoresis. Electrophoresis 2005; 26:1778-82. [PMID: 15800969 DOI: 10.1002/elps.200400001] [Citation(s) in RCA: 35] [Impact Index Per Article: 1.8] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 11/12/2022]
Abstract
Rhubarb, a well-known Chinese herbal medicine, is also used in Europe and other places of the world. Anthraquinones derivatives are thought to be the major active components. A pressurized liquid extraction (PLE) and capillary zone electrophoresis (CZE) separation were developed for simultaneous determination of five anthraquinones including aloe-emodin, emodin, chrysophanol, physcion, and rhein in Rhubarb. The effects of the experimental variables on PLE and CZE have been optimized. The optimum conditions of PLE were: solvent, methanol; temperature, 140 degrees C; particle size, 0.13-0.2 mm; static extraction time, 5 min; pressure, 1500 psi; and one extraction. The best separation of the five anthraquinones could be obtained using 50 mM borate buffer (pH 8.2) containing 25% isopropyl alcohol and 25% acetontrile as modifier, while the separation voltage was 25 kV and the temperature was at 20 degrees C. The method developed is accurate, simple, and reproducible, and could be used for quality control of Rhubarb and its medical preparations.
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Affiliation(s)
- Yuan X Gong
- Institute of Chinese Medical Sciences, University of Macau, Taipa, Macau SAR, China
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14
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Liebich HM, Müller-Hagedorn S, Klaus F, Meziane K, Kim KR, Frickenschmidt A, Kammerer B. Chromatographic, capillary electrophoretic and matrix-assisted laser desorption ionization time-of-flight mass spectrometry analysis of urinary modified nucleosides as tumor markers. J Chromatogr A 2005; 1071:271-5. [PMID: 15865203 DOI: 10.1016/j.chroma.2004.12.055] [Citation(s) in RCA: 67] [Impact Index Per Article: 3.4] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 11/27/2022]
Abstract
Modified nucleosides are formed posttranscriptionally in RNA. During RNA turnover free modified nucleosides are formed which circulate in the blood stream and are excreted in the urine. Their levels are increased in a number of malignant diseases, and they can be used in clinical chemistry as tumor markers. The analysis includes the isolation of the nucleosides from urine with phenylboronate gel and their separation and quantitation by HPLC on C18 columns or by capillary electrophoresis on uncoated columns applying a sodium dodecyl sulfate-borate-phosphate buffer. Identification of the nucleosides is performed with matrix-assisted laser desorption ionization time-of-flight mass spectrometry including post source decay spectra. In two clinical studies the diagnostic value of urinary modified nucleosides is investigated, in a study on children with leukemia and other malignant diseases and a study on women with breast cancer. Candidate markers are pseudouridine, 1-methylguanosine, N2-methylguanosine, 3-methyluridine and 1-methyl-inosine.
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Affiliation(s)
- H M Liebich
- a Medizinische Klinik, Universität Tübingen, Zentrallaboratorium, D-72076 Tübingen, Germany.
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15
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Zheng YF, Kong HW, Xiong JH, Lv S, Xu GW. Clinical significance and prognostic value of urinary nucleosides in breast cancer patients. Clin Biochem 2005; 38:24-30. [PMID: 15607313 DOI: 10.1016/j.clinbiochem.2004.09.021] [Citation(s) in RCA: 72] [Impact Index Per Article: 3.6] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 11/17/2003] [Revised: 02/26/2004] [Accepted: 09/16/2004] [Indexed: 12/30/2022]
Abstract
OBJECTIVE Thirteen urinary nucleosides, primarily degradation products of tRNA, were evaluated as potential tumor markers for breast cancer patients. DESIGN AND METHODS The micellar electrokinetic chromatography (MEKC) method has been used to analyze the urinary nucleosides in 41 healthy controls, 20 patients with benign breast tumors, and 26 breast cancer patients. RESULTS Urinary nucleoside concentrations of breast cancer patients were found to increase significantly compared to those of patients with benign breast tumors and healthy controls. By using 13 nucleoside concentrations as data vectors for principal component analysis (PCA), 73% (19/26) of breast cancer patients were correctly identified from healthy controls, while only 20% (4/20) of patients with benign breast tumors were indistinguishable from breast cancer patients. The mean level of all forms of urinary nucleosides in patients with metastatic breast cancer was higher than that in patients with primary breast cancer. The levels of modified nucleosides tended to decrease and return to normal after surgery. CONCLUSION The results indicate that urinary nucleosides may be useful as tumor markers for breast cancer.
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Affiliation(s)
- Yu-Fang Zheng
- National Chromatographic R&A Center, Dalian Institute of Chemical Physics, Chinese Academy of Sciences, Dalian 116011, PR China
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16
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Dudley E, Tuytten R, Bond A, Lemière F, Brenton AG, Esmans EL, Newton RP. Study of the mass spectrometric fragmentation of pseudouridine: comparison of fragmentation data obtained by matrix-assisted laser desorption/ionisation post-source decay, electrospray ion trap multistage mass spectrometry, and by a method utilising electrospray quadrupole time-of-flight tandem mass spectrometry and in-source fragmentation. RAPID COMMUNICATIONS IN MASS SPECTROMETRY : RCM 2005; 19:3075-85. [PMID: 16206154 DOI: 10.1002/rcm.2151] [Citation(s) in RCA: 24] [Impact Index Per Article: 1.2] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Subscribe] [Scholar Register] [Indexed: 05/04/2023]
Abstract
Many nucleosides and their modified forms have been studied by mass spectrometry elaborating the detailed fragmentation pathways under MS2 and MS(n) conditions. Although the C-nucleoside pseudouridine has been fragmented and studied briefly, usually amongst many other nucleosides, it has not been investigated to the same extent as other nucleosides. In this report a number of different mass spectrometric techniques are applied to obtain a fuller picture of pseudouridine fragmentation. At the same time this study is used to compare different tandem mass spectrometric techniques, including a novel methodology utilising a quadrupole time-of-flight (Q-ToF) instrument for MS(n) analysis comparable with that available with an ion trap mass spectrometer.
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Affiliation(s)
- Edward Dudley
- Biomolecular Analysis Mass Spectrometry (BAMS) Facility, Grove Building, University of Wales Swansea, Singleton Park, Swansea SA2 8PP, UK.
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17
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Dudley E, Lemière F, Van Dongen W, Tuytten R, El-Sharkawi S, Brenton AG, Esmans EL, Newton RP. Analysis of urinary nucleosides. IV. Identification of urinary purine nucleosides by liquid chromatography/electrospray mass spectrometry. RAPID COMMUNICATIONS IN MASS SPECTROMETRY : RCM 2004; 18:2730-2738. [PMID: 15499664 DOI: 10.1002/rcm.1685] [Citation(s) in RCA: 24] [Impact Index Per Article: 1.1] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Subscribe] [Scholar Register] [Indexed: 05/24/2023]
Abstract
Modified urinary nucleosides are potentially invaluable in cancer diagnosis. High-performance liquid chromatography (HPLC) was combined with full scan mass spectrometry (MS), tandem mass spectrometry and MSn analysis in order to identify purine nucleosides purified from urine. UV peaks evident in the chromatogram were examined by the various mass spectrometric techniques and adenosine, 1-methyladenosine, xanthosine, N1-methylguanosine, N2-methylguanosine, N2,N2-dimethylguanosine, N2,N2,N7-trimethylguanosine, inosine, and 1-methylinosine were each identified in the urine samples from cancer patients. The benefits of the use of LC/MS compared with HPLC alone are discussed.
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Affiliation(s)
- Edward Dudley
- Biomolecular Analysis Mass Spectrometry (BAMS) Facility, Grove Building, University of Wales Swansea, Singleton Park, Swansea SA2 8PP, UK
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18
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La S, Cho J, Kim JH, Kim KR. Capillary electrophoretic profiling and pattern recognition analysis of urinary nucleosides from thyroid cancer patients. Anal Chim Acta 2003. [DOI: 10.1016/s0003-2670(03)00473-2] [Citation(s) in RCA: 60] [Impact Index Per Article: 2.7] [Reference Citation Analysis] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 01/09/2023]
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McKeown AP, Shaw PN, Barrett DA. Electrophoretic behaviour of oligonucleotides and mono-, di- and triphosphate nucleotides by capillary zone electrophoresis. Electrophoresis 2001; 22:1119-26. [PMID: 11358136 DOI: 10.1002/1522-2683()22:6<1119::aid-elps1119>3.0.co;2-x] [Citation(s) in RCA: 14] [Impact Index Per Article: 0.6] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 11/06/2022]
Abstract
A systematic investigation has been made into the mechanisms of the capillary zone electrophoresis (CZE) separation of 12 common nucleotides (mono-, di- and triphosphorylated) and polydeoxythymidylic acid oligonucleotides (pd(T)5-18) using electrophoretic mobility values calculated from migration time data. Relationships between electrophoretic mobility and the physicochemical characteristics of the analytes (charge, dissociation constants, charge-to-mass ratio) and the background electrolyte conditions (buffer strength, percentage organic modifier and buffer pH) were characterised. Nucleotide migration was dominated by the negatively charged phosphate groups. Additionally, there were important contributions to migration behaviour from the ionised amide groups of the nucleobases guanine and uracil at higher buffer pH values or with the presence of methanol in the electrolyte. Calculated electrophoretic mobility values for the nucleotides showed a substantially improved (5-fold) inter-run repeatability compared with migration time data. These studies show the value of representing nucleotide migration data as electrophoretic mobility in CZE for obtaining a more thorough analysis of separation mechanisms and to compensate for variation in migration time data caused by small changes in electrosmotic flow. Oligonucleotides pd(T)5-11 could be adequately resolved from their nearest neighbour, but the limit of single-base separation was pd(T)10 from pd(T)11 under the conditions used. It was calculated that a difference in charge-to-mass ratio of 2.64 x 10(-5) was required for resolution under the CZE conditions used.
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Affiliation(s)
- A P McKeown
- AstraZeneca R & D Charnwood, Pharmaceutical and Analytical R & D, Loughborough, Leicestershire, UK
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Strein TG, Morris D, Palmer J, Landers JP. Discontinuous electrophoretic stacking system for cholate-based electrokinetic chromatographic separation of 8-hydroxy-2'-deoxyguanosine from unmodified deoxynucleosides. JOURNAL OF CHROMATOGRAPHY. B, BIOMEDICAL SCIENCES AND APPLICATIONS 2001; 763:71-8. [PMID: 11710585 DOI: 10.1016/s0378-4347(01)00367-x] [Citation(s) in RCA: 6] [Impact Index Per Article: 0.3] [Reference Citation Analysis] [Abstract] [MESH Headings] [Grants] [Track Full Text] [Subscribe] [Scholar Register] [Indexed: 11/20/2022]
Abstract
The stacking and baseline-resolved separation of the oxidative damage marker, 8-hydroxy-2'-deoxyguanosine (8-OHdG), from unmodified deoxynucleosides in under 4 min is reported. Separations of 8-OHdG from 2'-deoxyadenosine, 2'-deoxycytosine, 2'-deoxyguanosine, and thymidine are accomplished using micellar electrokinetic capillary chromatography with sodium cholate. Importantly, the use of sulfate, intentionally added to the sample matrix, results in effective stacking of 8-OHdG and other analytes. This work extends electrokinetic stacking injection of neutral analytes to include deoxynucleosides. The procedure works well with either electrokinetic or hydrodynamic injection. The separation buffer and sample matrix composition were optimized to effect stacking conditions with an uncoated 50 microm fused-silica capillary. The lower limit of detection for the analytes is in the nanomolar range, and is more than an order of magnitude lower than without stacking. With 30 s (5.7 cm) electrokinetic injections, stacking and baseline separation of 8-hydroxy-2'-deoxyguanosine from the unmodified nucleosides is accomplished, even in the presence of a 400-fold excess of unmodified deoxynucleosides.
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Affiliation(s)
- T G Strein
- Department of Chemistry, Bucknell University, Lewisburg, PA 17837, USA
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Kim KR, La S, Kim A, Kim JH, Liebich HM. Capillary electrophoretic profiling and pattern recognition analysis of urinary nucleosides from uterine myoma and cervical cancer patients. JOURNAL OF CHROMATOGRAPHY. B, BIOMEDICAL SCIENCES AND APPLICATIONS 2001; 754:97-106. [PMID: 11318432 DOI: 10.1016/s0378-4347(00)00585-5] [Citation(s) in RCA: 41] [Impact Index Per Article: 1.7] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Subscribe] [Scholar Register] [Indexed: 12/15/2022]
Abstract
Capillary electrophoretic (CE) profiling analysis combined with pattern recognition methods is described for the correlation between urinary nucleoside profiles and uterine cervical cancer. Nucleosides were extracted from urine specimens by solid-phase extraction in affinity mode using phenylboronic acid gel. CE separation was carried out with an uncoated fused-silica capillary (570 mm x 50 microm I.D.) maintained at 20 degrees C, using 25 mM borate-42.5 mM phosphate buffer (pH 6.7) containing 200 mM sodium dodecyl sulfate as the run buffer under the applied voltage of 20 kV. A total of 15 nucleosides were positively identified in urine samples (2 ml) from eight uterine myoma (benign tumor group), 10 uterine cervical cancer (malignant tumor group) patients and 10 healthy females (normal group) studied. The star symbol plots drawn based on each mean concentration of nucleosides normalized to that in normal group enabled one to discriminate malignant and benign groups from normal group. In addition, canonical discriminant analysis performed on the nucleoside data of 28 individual urine specimens correctly classified into three separate clusters according to groups in the canonical plot.
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Affiliation(s)
- K R Kim
- College of Pharmacy, Sungkyunkwan University, Suwon, South Korea.
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22
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Li SP, Li P, Dong TT, Tsim KW. Determination of nucleosides in natural Cordyceps sinensis and cultured Cordyceps mycelia by capillary electrophoresis. Electrophoresis 2001; 22:144-50. [PMID: 11197164 DOI: 10.1002/1522-2683(200101)22:1<144::aid-elps144>3.0.co;2-t] [Citation(s) in RCA: 78] [Impact Index Per Article: 3.3] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 11/07/2022]
Abstract
Cordyceps sinensis is a well-known traditional Chinese medicine, and some of the active components are nucleosides. The analysis of nucleosides in Cordyceps material has been performed by reversed-phase high-performance liquid chromatography (HPLC) with gradient elution or by spectrometry. Here, we have explored the possibility of using capillary electrophoresis to determine the content of three major nucleosides (adenosine, guanosine and uridine) in Cordyceps. Capillary electrophoresis needs no gradients, and it provides a better separation due to its higher efficiency. In order to optimize the resolution, the separation of adenosine, guanosine and uridine was determined in Cordyceps with respect to the variation of buffer concentration, pH, temperature, and voltage. By using the calibrated electrophoresis system, the separation was achieved for the three nucleosides in less than 10 min with a background electrolyte consisting of 0.2 M boric acid-sodium hydroxide buffer, pH 8.5. The nucleoside contents of various types of natural Cordyceps and cultured Cordyceps mycelia were determined and compared. There was a great variation of nucleoside content in different sources of Cordyceps; the cultured Cordyceps mycelia, however, contains a much higher concentration than the natural Cordyceps.
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Affiliation(s)
- S P Li
- China Pharmaceutical University, Ji Xiang An, Nanjing, China
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23
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Dudley E, Lemiere F, Van Dongen W, Langridge JI, El-Sharkawi S, Games DE, Esmans EL, Newton RP. Analysis of urinary nucleosides. II. Comparison of mass spectrometric methods for the analysis of urinary nucleosides. RAPID COMMUNICATIONS IN MASS SPECTROMETRY : RCM 2001; 15:1701-1707. [PMID: 11555869 DOI: 10.1002/rcm.428] [Citation(s) in RCA: 19] [Impact Index Per Article: 0.8] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Subscribe] [Scholar Register] [Indexed: 05/23/2023]
Abstract
Qualitative and quantitative analyses of urinary nucleosides have diagnostic potential as tumour markers. We have developed separation techniques linked to mass spectrometric detection in order to overcome the problems associated with past identification and quantitation methods. The three methods of analysis utilised were: gas chromatography/mass spectrometry (GC/MS), high-performance liquid chromatography/ion-trap mass spectrometry (HPLC/ITMS) and capillary liquid chromatography/triple quadrupole mass spectrometry (CapLC/TQMS). Here we compare the relative effectiveness of each of the techniques for subsequent application in the systematic study of urinary nucleoside profiles in cancer patients. All three methods proved to be valuable techniques for such urinary nucleoside analyses, and a combination rather than one single choice is concluded as the ideal.
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Affiliation(s)
- E Dudley
- Biochemistry Group, School of Biological Sciences, Wallace Building, University of Wales Swansea, Singleton Park, Swansea SA2 8PP, UK
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24
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Xu G, Schmid HR, Lu X, Liebich HM, Lu P. Excretion pattern investigation of urinary normal and modified nucleosides of breast cancer patients by RP-HPLC and factor analysis method. Biomed Chromatogr 2000; 14:459-63. [PMID: 11113924 DOI: 10.1002/1099-0801(200011)14:7<459::aid-bmc7>3.0.co;2-k] [Citation(s) in RCA: 44] [Impact Index Per Article: 1.8] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 11/08/2022]
Abstract
Modified nucleosides, formed post-transcriptionally in RNA by a number of modification enzymes, are excreted in abnormal levels in the urine of patients with malignant tumors. To test their usefulness as tumor markers, and to compare them with the conventional tumor markers, a reversed-phase high-performance liquid chromatographic (RP-HPLC) method and a factor analysis method have been used to study the excretion pattern of nucleosides of breast cancer patients. A clear cut differentiation of the breast cancer group and the healthy individuals in two clusters without overlapping was obtained.
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Affiliation(s)
- G Xu
- National Chromatographic R. & A. Center, Dalian Institute of Chemical Physics, the Chinese Academy of Sciences, 116011 Dalian, People's Republic of China.
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25
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Liebich HM, Lehmann R, Xu G, Wahl HG, Häring HU. Application of capillary electrophoresis in clinical chemistry: the clinical value of urinary modified nucleosides. JOURNAL OF CHROMATOGRAPHY. B, BIOMEDICAL SCIENCES AND APPLICATIONS 2000; 745:189-96. [PMID: 10997714 DOI: 10.1016/s0378-4347(00)00263-2] [Citation(s) in RCA: 46] [Impact Index Per Article: 1.8] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Subscribe] [Scholar Register] [Indexed: 11/25/2022]
Abstract
Urinary modified nucleosides were determined by capillary electrophoresis using a 300 mM SDS-25 mM sodium tetraborate-50 mM sodium dihydrogenphosphate buffer. The nucleosides were extracted from urine by phenylboronate affinity gel chromatography. In cancer patients the levels of the modified nucleosides are generally elevated. By an artificial neural network method breast cancer patients were differentiated from normal individuals, which indicates that the modified nucleosides could be of clinical value as tumor markers.
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Affiliation(s)
- H M Liebich
- Medizinische Universitaetsklinik, Abteilung IV, Zentrallabor, Tübingen, Germany
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26
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27
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Weiss DJ, Lunte CE. Detection of a urinary biomaker for oxidative DNA damage 8-hydroxydeoxyguanosine by capillary electrophoresis with electrochemical detection. Electrophoresis 2000; 21:2080-5. [PMID: 10879970 PMCID: PMC2519811 DOI: 10.1002/1522-2683(20000601)21:10<2080::aid-elps2080>3.0.co;2-6] [Citation(s) in RCA: 53] [Impact Index Per Article: 2.1] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Grants] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 11/11/2022]
Abstract
8-Hydroxydeoxyguanosine (8-OHdG) is present in urine as a result of oxidative DNA damage associated with age-related diseases such as cancer. In this report a method is presented for the detection of 8-OHdG in human morning urine utilizing capillary electrophoresis with electrochemical detection (CEEC). The limit of detection for a aqueous standard of 8-OHdG is 50 nM (signal to noise ratio S/N = 3). A single solid-phase extraction (SPE) step with a C18 column is used for sample cleanup and 20-fold preconcentration of the urine before analysis by CEEC. Optimized conditions for analysis of extracted urine are E(app) = 0.5 V vs. Ag/AgCl with 20 mM sodium borate/20% MeOH v/v, pH 9, as the background electrolyte, and a separation voltage of 22 kV. The concentration of 8-OHdG varied from 6 to 86 nM with an average value of 42 +/- 26.9 nM for four healthy female and four healthy male subjects between the ages of 23 and 43.
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Affiliation(s)
- D J Weiss
- Department of Chemistry, University of Kansas, Lawrence 66045, USA
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28
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Dudley E, El-Sharkawi S, Games DE, Newton RP. Analysis of urinary nucleosides. I. Optimisation of high performance liquid chromatography/electrospray mass spectrometry. RAPID COMMUNICATIONS IN MASS SPECTROMETRY : RCM 2000; 14:1200-1207. [PMID: 10918368 DOI: 10.1002/1097-0231(20000730)14:14<1200::aid-rcm10>3.0.co;2-i] [Citation(s) in RCA: 29] [Impact Index Per Article: 1.2] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Subscribe] [Scholar Register] [Indexed: 05/23/2023]
Abstract
In order to optimise the analysis of urinary nucleosides by high performance liquid chromatography/mass spectrometry (HPLC/MS), the HPLC separation of these compounds was performed at different 'flow rates' and 0.2mL/min was found to give both a better separation and ionisation. The ionisation conditions were optimised to give the best intensity of the molecules quasi-molecular ions. The ion distribution profile and ionisation in both positive and negative mode were examined and the detection of the protonated molecule in positive mode chosen for further analysis. The limits of detection of the method developed are reported and representative LC/MS and LC/MS/MS spectra shown. Typical urinary nucleoside chromatograms are presented.
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Affiliation(s)
- E Dudley
- Biochemistry Group, School of Biological Sciences, University of Wales, Singleton Park, Swansea SA2 8PP, UK
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29
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Thormann W, Wey AB, Lurie IS, Gerber H, Byland C, Malik N, Hochmeister M, Gehrig C. Capillary electrophoresis in clinical and forensic analysis: recent advances and breakthrough to routine applications. Electrophoresis 1999; 20:3203-36. [PMID: 10596826 DOI: 10.1002/(sici)1522-2683(19991001)20:15/16<3203::aid-elps3203>3.0.co;2-e] [Citation(s) in RCA: 54] [Impact Index Per Article: 2.1] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 12/15/2022]
Abstract
This paper is a comprehensive review article on capillary electrophoresis (CE) in clinical and forensic analysis. It is based upon the literature of 1997 and 1998, presents CE examples in major fields of application, and provides an overview of the key achievements encountered, including those associated with the analysis of drugs, serum proteins, hemoglobin variants, and nucleic acids. For CE in clinical and forensic analysis, the past two years witnessed a breakthrough to routine applications. As most coauthors of this review are associated with diagnostic or forensic laboratories now using CE on a routine basis, this review also contains data from routine applications in drug, protein, and DNA analysis. With the first-hand experience of providing analytical service under stringent quality control conditions, aspects of quality assurance, assay specifications for clinical and forensic CE and the pros and cons of this maturing, cost-and pollution-controlled age technology are also discussed.
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Affiliation(s)
- W Thormann
- Department of Clinical Pharmacology, University of Bern, Switzerland.
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30
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Xu G, Di Stefano C, Liebich HM, Zhang Y, Lu P. Reversed-phase high-performance liquid chromatographic investigation of urinary normal and modified nucleosides of cancer patients. JOURNAL OF CHROMATOGRAPHY. B, BIOMEDICAL SCIENCES AND APPLICATIONS 1999; 732:307-13. [PMID: 10517352 DOI: 10.1016/s0378-4347(99)00296-0] [Citation(s) in RCA: 30] [Impact Index Per Article: 1.2] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Subscribe] [Scholar Register] [Indexed: 12/15/2022]
Abstract
Post-transcriptional modifications in RNA give rise to free modified ribonucleosides circulating in the blood stream and excreted in urine. Due to their abnormal levels in conjunction with several tumor diseases, they have been suggested as possible tumor markers. The developed RP-HPLC method has been applied to analyze the urinary nucleosides in 34 urinary samples from 15 kinds of cancer patients. The statistical analyses showed the urinary nucleoside excretion, especially modified nucleoside levels, in cancer patients were significantly higher than those in normal healthy volunteers. Factor analysis was used to classify the patients with cancer and normal healthy humans. It was found that using 15 urinary nucleoside levels or only five modified nucleoside levels as data vectors the factor analysis plot displayed two almost separate clusters representing each group.
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Affiliation(s)
- G Xu
- National Chromatographic R&A Center, Dalian Institute of Chemical Physics, The Chinese Academy of Sciences.
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31
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Zhao R, Xu G, Yue B, Liebich HM, Zhang Y. Artificial neural network classification based on capillary electrophoresis of urinary nucleosides for the clinical diagnosis of tumors. J Chromatogr A 1998; 828:489-96. [PMID: 9916327 DOI: 10.1016/s0021-9673(98)00589-5] [Citation(s) in RCA: 43] [Impact Index Per Article: 1.6] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 12/15/2022]
Abstract
Nucleosides in human urine have been studied frequently as a possible biomedical marker for cancers, acquired immune deficiency syndrome (AIDS) and the whole-body turnover of RNAs. A capillary electrophoretic method that can quantitatively analyze urinary normal and modified nucleosides in less than 40 min with a good resolution and sufficient sensitivity has been developed. Twelve kinds of normal and modified nucleosides were determined in urine samples from 25 healthy persons and 25 cancer patients of 14 kinds of cancers. Artificial neural networks have been used as a powerful pattern recognition tool to distinguish cancer patients from healthy persons. The recognition rate for the training set reached to 100% and above 85% of the members in the predicting set were correctly classified. In addition, the neural network technique was compared with methods of the principal component analysis and the canonical discriminant analysis. The results demonstrate that the predictive ability of the artificial neural network is stronger than the others in this study.
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Affiliation(s)
- R Zhao
- National Chromatographic R. & A. Centre, Dalian Institute of Chemical Physics, Chinese Academy of Sciences, China
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32
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Liebich HM, Xu G, Di Stefano C, Lehmann R. Capillary electrophoresis of urinary normal and modified nucleosides of cancer patients. J Chromatogr A 1998; 793:341-7. [PMID: 9474787 DOI: 10.1016/s0021-9673(97)00915-1] [Citation(s) in RCA: 42] [Impact Index Per Article: 1.6] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 02/06/2023]
Abstract
This paper gives a capillary electrophoretic method for the separation of 15 urinary normal and modified nucleosides from cancer patients in less than 40 min. A 500 mm x 50 microns uncoated capillary column (437.5 mm to window) was used. The effects of the voltage and the sodium dodecyl sulfate (SDS) concentration in the buffer on the separation were studied. With reproducibilities of migration times better than 1.2% (R.S.D.) and determined concentrations better than 5-25%, depending on the concentrations of nucleosides in the urine, the analytical characteristics of the method were good. Using this developed method, the concentrations of 13 normal and modified nucleosides, extracted on a phenyl boronic acid affinity chromatography column, in 25 urines from patients of 14 kinds of cancer were determined. The levels (nmol/mumol creatinine) of modified nucleosides in urines from cancer patients were increased as compared with those in normal urines.
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Affiliation(s)
- H M Liebich
- Medizinische Universitätsklinik, Department of Clinical Chemistry, Tübingen, Germany
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