Separation of dideoxyribonucleosides in trace amounts by automated liquid chromatography and capillary electrophoresis

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dc.contributor Wichita State University. Department of Chemistry en_US
dc.contributor.author Singhal, Ram P. en_US
dc.contributor.author Hughbanks, D. en_US
dc.contributor.author Xian, Jun en_US
dc.date.accessioned 2012-02-06T17:15:17Z
dc.date.available 2012-02-06T17:15:17Z
dc.date.issued 1992-09-18 en_US
dc.identifier 1430040 en_US
dc.identifier 0427043 en_US
dc.identifier.citation Journal of chromatography. 1992 Sep 18; 609(1-2): 147-61. en_US
dc.identifier.issn 0021-9673 en_US
dc.identifier.uri http://hdl.handle.net/10057/4221
dc.description Full text of this article is not available in SOAR. en_US
dc.description.abstract No satisfactory high-performance liquid chromatographic (HPLC) method is currently available for the separation of the major dideoxyribonucleosides (ddNs) and their derivatives. A method involving HPLC has been developed for the separation of five major ddNs [ddA, ddC, ddI, azT and 2',3'-dideoxy-2',3'-didehydrothymidine (d4T)]. Elution of the common and modified components of DNA was also examined under the selected separation conditions of HPLC. The elution characteristics of these compounds were studied using serum plasma samples spiked with ddN derivatives. In addition, capillary electrophoresis (CE) was investigated for the separation of ddNs and their derivatives. Picomolar amounts of the five major ddNs and the metabolic product of azT [5'-O-glucuronide-3'-azido-3'-deoxythymidine (Glo-azT)] were satisfactorily resolved in 10 min by using a modification of CE. The spectral properties of the ddNs were characterized under different pH conditions and compared with those of their parent deoxyribonucleosides (dNs) because these compounds are commonly measured in HPLC by their spectral properties. The spectra of ddC and ddT derivatives resemble very closely those of dC and dT, but those of ddA and ddI differ to some extent from their parent dNs. The HPLC method was extensively examined for satisfactory resolutions of these compounds. For example, an isocratic elution method, although simple, failed to resolve these compounds and ion-pair chromatography did not offer any advantage. Gradient elution involving buffered solutions and increasing amounts of an organic modifier yielded satisfactory results. Methanol appeared to be the organic modifier of choice. A reversed-phase matrix with smaller than octadecyl alkyl chains did not produce the necessary interactions. Uniform spherical beads of smaller diameter produced superior resolutions. The separation of these compounds on three commercially available columns is discussed. The separation of human plasma samples spiked with dideoxynucleoside derivatives by HPLC was accomplished in ca. 16 min. The presence of the dNs did not interfere in their separations. en_US
dc.format.extent 147-61 en_US
dc.language.iso eng en_US
dc.publisher Elsevier en_US
dc.relation.ispartofseries Journal of chromatography en_US
dc.relation.ispartofseries J. Chromatogr. en_US
dc.source NLM en_US
dc.subject.mesh Capillary Action en_US
dc.subject.mesh Chromatography, High Pressure Liquid/methods en_US
dc.subject.mesh DNA/isolation & purification en_US
dc.subject.mesh Dideoxynucleosides/isolation & purification en_US
dc.subject.mesh Electrophoresis/methods en_US
dc.subject.mesh Hydrogen-Ion Concentration en_US
dc.subject.mesh Ions en_US
dc.subject.mesh Micelles en_US
dc.subject.mesh Spectrophotometry en_US
dc.subject.mesh Temperature en_US
dc.title Separation of dideoxyribonucleosides in trace amounts by automated liquid chromatography and capillary electrophoresis en_US
dc.type Article en_US
dc.coverage.spacial Netherlands en_US
dc.description.version peer reviewed en_US
dc.rights.holder Copyright © 1992, Elsevier en_US

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