Analysis of oxidative DNA damage, Part II: Synthesis of the internal standard 8-[18O]hydroxy-2'-deoxyguanosine

2.50
Hdl Handle:
http://hdl.handle.net/10029/255623
Title:
Analysis of oxidative DNA damage, Part II: Synthesis of the internal standard 8-[18O]hydroxy-2'-deoxyguanosine
Authors:
Hermanns RCA; Zomer G; Stavenuiter JFC; Westra G; Visser T; van de Werken G
Other Titles:
[Analyse van oxidatieve DNA schade, deel II: Synthese van de interne standaard 8-[18O]hydroxy-2'-deoxyguanosine.]
Abstract:
In the project 'Oxidative DNA Damage' the first aim is to develop a mass spectrometric method for the quantification of 8-hydroxy-2'- deoxyguanosine (oh8dG). The required precision of the method requires the application of a labeled analogue as an internal standard. This report describes the syntheses of 8-[18O]hydroxy-2'-deoxyguanosine (18Oh8dG), which is to be used as internal standard in the quantitative GC-MS analysis of 8-hydroxy-2'deoxyguanosine. 8-[18O]Hydroxy-2'- deoxyguanosine was synthesised starting from 2'-deoxyguanosine (dG) in three steps. In the first step dG was brominated at the C-8 position, resulting in 8-bromo-2'-deoxyguanosine (8-Br-dG). 8-Br-dG was converted into 8-[18O]benzyloxy-2'-deoxyguanosine (8-[18O]OBz-dG) by treatment of 8-Br-dG with the sodium salt of [18O]benzyl alcohol. Finally 8-[18O] OBz-dG was hydrogenated resulting in 18Oh8dG. The sodium salt of [18O]benzyl alcohol was synthesised in three steps starting from ethyl benzimidate hydrochloride. In the first step ethyl benzimidate hydrochloride was converted in ethyl [18O]carboxy benzoate by treatment with H218O (95%). In the second step ethyl benzoate was reduced by lithium aluminium hydride to [18O]benzyl alcohol. In the last step [18O]benzyl alcohol was converted into the sodium salt of [18O] benzyl alcohol by treatment with sodium hydride. Starting with dG (5 g, 17 mmol), the overall yield of the total procedure was approximately 18 mg (0.11 mmol, 4%). The chemical purity of [18O]oh8dG was estimated to be 89%, based on HPLC analysis. The isotopic purity of 18Oh8dG was determined to be 93.4% based on GC-MS analysis. The mentioned chemical and isotopic purity comply with the planned application.<br>
Publisher:
Rijksinstituut voor Volksgezondheid en Milieu RIVM
Issue Date:
31-Aug-1993
URI:
http://hdl.handle.net/10029/255623
Additional Links:
http://www.rivm.nl/bibliotheek/rapporten/511004002.html
Type:
Onderzoeksrapport
Language:
en
Sponsors:
RIVM; NKI
Appears in Collections:
RIVM official reports

Full metadata record

DC FieldValue Language
dc.contributor.authorHermanns RCA-
dc.contributor.authorZomer G-
dc.contributor.authorStavenuiter JFC-
dc.contributor.authorWestra G-
dc.contributor.authorVisser T-
dc.contributor.authorvan de Werken G-
dc.date.accessioned2012-12-12T12:13:05Z-
dc.date.available2012-12-12T12:13:05Z-
dc.date.issued1993-08-31-
dc.identifier511004002-
dc.identifier.urihttp://hdl.handle.net/10029/255623-
dc.description.abstractIn the project 'Oxidative DNA Damage' the first aim is to develop a mass spectrometric method for the quantification of 8-hydroxy-2'- deoxyguanosine (oh8dG). The required precision of the method requires the application of a labeled analogue as an internal standard. This report describes the syntheses of 8-[18O]hydroxy-2'-deoxyguanosine (18Oh8dG), which is to be used as internal standard in the quantitative GC-MS analysis of 8-hydroxy-2'deoxyguanosine. 8-[18O]Hydroxy-2'- deoxyguanosine was synthesised starting from 2'-deoxyguanosine (dG) in three steps. In the first step dG was brominated at the C-8 position, resulting in 8-bromo-2'-deoxyguanosine (8-Br-dG). 8-Br-dG was converted into 8-[18O]benzyloxy-2'-deoxyguanosine (8-[18O]OBz-dG) by treatment of 8-Br-dG with the sodium salt of [18O]benzyl alcohol. Finally 8-[18O] OBz-dG was hydrogenated resulting in 18Oh8dG. The sodium salt of [18O]benzyl alcohol was synthesised in three steps starting from ethyl benzimidate hydrochloride. In the first step ethyl benzimidate hydrochloride was converted in ethyl [18O]carboxy benzoate by treatment with H218O (95%). In the second step ethyl benzoate was reduced by lithium aluminium hydride to [18O]benzyl alcohol. In the last step [18O]benzyl alcohol was converted into the sodium salt of [18O] benzyl alcohol by treatment with sodium hydride. Starting with dG (5 g, 17 mmol), the overall yield of the total procedure was approximately 18 mg (0.11 mmol, 4%). The chemical purity of [18O]oh8dG was estimated to be 89%, based on HPLC analysis. The isotopic purity of 18Oh8dG was determined to be 93.4% based on GC-MS analysis. The mentioned chemical and isotopic purity comply with the planned application.<br>en
dc.description.sponsorshipRIVM-
dc.description.sponsorshipNKI-
dc.format.extent26 p-
dc.language.isoen-
dc.publisherRijksinstituut voor Volksgezondheid en Milieu RIVM-
dc.relation.ispartofRIVM Rapport 511004002-
dc.relation.urlhttp://www.rivm.nl/bibliotheek/rapporten/511004002.html-
dc.subject07nl
dc.subjectdna schadenl
dc.subjectoxydatienl
dc.subjectvrije radicalennl
dc.subjectanalysenl
dc.subjectreferentiematerialennl
dc.subjectdna damageen
dc.subjectoxidationen
dc.subjectfree radicalsen
dc.subjectreactive oxygen speciesen
dc.subjectreference materialsen
dc.titleAnalysis of oxidative DNA damage, Part II: Synthesis of the internal standard 8-[18O]hydroxy-2'-deoxyguanosineen
dc.title.alternative[Analyse van oxidatieve DNA schade, deel II: Synthese van de interne standaard 8-[18O]hydroxy-2'-deoxyguanosine.]nl
dc.typeOnderzoeksrapport-
dc.date.updated2012-12-12T12:13:05Z-
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