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| Content Provider | World Health Organization (WHO)-Global Index Medicus |
|---|---|
| Author | Rodríguez-Gonzalo, Encarnación Herrero-Herrero, Leticia García-Gómez, Diego |
| Description | Author Affiliation: Rodríguez-Gonzalo E ( Departamento de Química Analítica, Nutrición y Bromatología, Facultad de Química, Universidad de Salamanca, 37008 Salamanca, Spain. Electronic address: erg@usal.es.); Herrero-Herrero L ( Departamento de Química Analítica, Nutrición y Bromatología, Facultad de Química, Universidad de Salamanca, 37008 Salamanca, Spain.); García-Gómez D ( Departamento de Química Analítica, Nutrición y Bromatología, Facultad de Química, Universidad de Salamanca, 37008 Salamanca, Spain.) |
| Abstract | In the present work we report the development and validation of a fast liquid chromatography-mass spectrometry method for the simultaneous determination of endogenous nucleosides derived from DNA and RNA in urine. The target compounds were 2'-deoxyguanosine and 8-hydroxy-2'-deoxyguanosine, derived from DNA, and the analogue 8-hydroxyguanosine, derived from RNA, together with adenosine, 1-methyladenosine, 7-methylguanosine and inosine. The method is based on the use of a chromatographic column packed with superficially porous particles for high-efficiency separation; further detection by MS/MS was accomplished with a triple quadrupole-mass spectrometer for analyte identification and accurate quantification. As a preliminary purification step, we developed a new procedure based on solid-phase extraction (SPE) with a mixed-sorbent prepared from three polymeric materials that facilitated the isolation of modified nucleosides, such as 2-deoxynucleosides, that are not retained by phenylboronic acid-based SPE. The proposed approach (SPE prior to LC-MS/MS) was validated in human urine in terms of linearity, the limit of detection, the limit of quantification, accuracy, recovery, repeatability, reproducibility and matrix-effects. For the SPE step, intra-day and inter-cartridge reproducibility were evaluated in natural and spiked urine samples, being ± 16.9% or below, with recoveries in the 74-125% range. No significant matrix effects were found in further MS/MS detection. The application of the present method to urine from healthy smoker and non-smoker volunteers is also reported in order to test its usefulness as a tool for clinical and toxicological trials. |
| File Format | HTM / HTML |
| ISSN | 15700232 |
| Journal | Journal of Chromatography B |
| Volume Number | 1019 |
| e-ISSN | 1873376X |
| Language | English |
| Publisher | Elsevier |
| Publisher Date | 2016-04-15 |
| Publisher Place | Netherlands |
| Access Restriction | One Nation One Subscription (ONOS) |
| Content Type | Text |
| Resource Type | Article |
| Subject | Cell Biology Medicine Analytical Chemistry Clinical Biochemistry Biochemistry |
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