Molecularly imprinted monolith in-tube solid-phase microextraction coupled with HPLC/UV detection for determination of 8-hydroxy-2'-deoxyguanosine in urine

• Published in: Analytical and bioanalytical chemistry 2009-09, Vol.395 (2), p.479-487
• Main Authors: Zhang, Shao-Wen, Xing, Jun, Cai, Ling-Shuang, Wu, Cai-Ying
• Format: Article
• Language: English
• Subjects: Adult; Analytical Chemistry; Biochemistry; Cancer; Characterization and Evaluation of Materials; Chemistry; Chemistry and Materials Science; Chromatographic methods and physical methods associated with chromatography; Chromatography, High Pressure Liquid; Deoxyguanosine - analogs & derivatives; Deoxyguanosine - urine; Enrichment; Equipment Design; Exact sciences and technology; Extraction; Food Science; Guanosine; Humans; Laboratory Medicine; Mass transfer; Middle Aged; Molecular Imprinting - economics; Molecular Imprinting - methods; Monitoring/Environmental Analysis; Neoplasms - metabolism; Original Paper; Other chromatographic methods; Patients; Selectivity; Sensitivity and Specificity; Signal to noise ratio; Solid Phase Microextraction - economics; Solid Phase Microextraction - instrumentation; Solid Phase Microextraction - methods; Urine; Young Adult
• ISSN: 1618-2642; 1618-2650
• DOI: 10.1007/s00216-009-2964-9

Urinary 8-hydroxy-2'-deoxyguanosine (8-OHdG) has been widely used as a biomarker of oxidative DNA damage. Measurements of 8-OHdG in urinary samples are challenging owing to the low level of 8-OHdG and the complex matrix. In this study, a novel molecularly imprinted polymer (MIP) monolithic column was synthesized with guanosine as a dummy template which was used as the medium for in-tube solid-phase microextraction (SPME). In-tube SPME coupled with HPLC/UV detection for extraction and determination of urinary 8-OHdG was developed. The synthesized MIP monolithic column exhibited high extraction efficiency owing to its greater phase ratio with convective mass transfer and inherent selectivity. The enrichment factor for 8-OHdG was found to be 76 and the limits of detection and quantification of the method for urinary samples were 3.2 nmol/L (signal-to-noise ratio 3) and 11 nmol/L (signal-to-noise ratio 10), respectively. The MIP's selectivity also made the sample preparation procedure and chromatographic separation much easier. The linear range of the proposed method was from 0.010 to 5.30 μmol/L (r = 0.9997), with a relative standard deviation of 1.1-6.8%, and the recovery for spiked urine samples was 84 ± 3%. The newly developed method was successfully applied to determine urinary samples of healthy volunteers, coking plant workers, and cancer patients. The 8-OHdG level in cancer patients was significantly higher than that in healthy people.