title: Development and evaluation of a hydrophilic interaction liquid chromatography-MS/MS method to quantify 19 nucleobases and nucleosides in rat plasma.
abstract: As essential endogenous compounds, nucleobases and nucleosides fulfill various functions in living organisms. This study presents the development and validation of a new hydrophilic interaction liquid chromatography tandem mass spectrometry method for simultaneous quantification of 19 nucleobases and nucleosides in rat plasma. For the sample preparation, 15 kinds of protein precipitants were evaluated according to the chromatographic profile and ion response of analytes. The optimization of chromatographic separation was respectively performed using reversed-phase liquid chromatography and hydrophilic interaction liquid chromatography mode; each separation mode included two test columns with different stationary phases. The chromatographic profile and parameters such as half-width (W1/2), capacity factor (K') and tailing factor (ft) were used to evaluate the separation efficiencies. Furthermore, the adopted composition of two mobile phase systems and the concentrations of the additives in the optimum buffer system were also investigated. The developed method was fully validated and successfully applied quantitatively to determine 19 nucleobases and nucleosides in plasma from normal and diabetic nephropathy (DN) rats. Significant differences between normal and DN rats were found in plasma levels of cytosine, xanthine, thymidine, adenosine, guanosine, inosine and 8-hydroxy-2'-deoxyguanosine. This information may provide a useful reference for the discovery of potential biomarkers of DN.
authors.affiliation: Jiangsu Key Laboratory of New Drug Research and Clinical Pharmacy, Xuzhou Medical University, Xuzhou, China.
authors.forename: Yan
authors.initials: Y
authors.lastname: Du
authors.affiliation: Jiangsu Key Laboratory of New Drug Research and Clinical Pharmacy, Xuzhou Medical University, Xuzhou, China.|Department of Pharmacy, Jiangsu Provincial Xuzhou Pharmaceutical Vocational College, Xuzhou, China.
authors.forename: Yin-Jie
authors.initials: YJ
authors.lastname: Li
authors.affiliation: Jiangsu Key Laboratory of New Drug Research and Clinical Pharmacy, Xuzhou Medical University, Xuzhou, China.
authors.forename: Xun-Xiu
authors.initials: XX
authors.lastname: Hu
authors.affiliation: Jiangsu Key Laboratory of New Drug Research and Clinical Pharmacy, Xuzhou Medical University, Xuzhou, China.
authors.forename: Xu
authors.initials: X
authors.lastname: Deng
authors.affiliation: Jiangsu Key Laboratory of New Drug Research and Clinical Pharmacy, Xuzhou Medical University, Xuzhou, China.
authors.forename: Zeng-Ting
authors.initials: ZT
authors.lastname: Qian
authors.affiliation: Jiangsu Key Laboratory of New Drug Research and Clinical Pharmacy, Xuzhou Medical University, Xuzhou, China.
authors.forename: Zheng
authors.initials: Z
authors.lastname: Li
authors.affiliation: Jiangsu Key Laboratory of New Drug Research and Clinical Pharmacy, Xuzhou Medical University, Xuzhou, China.|Department of Pharmaceutical Analysis, School of Pharmacy, Xuzhou Medical University, Xuzhou, China.
authors.forename: Meng-Zhe
authors.initials: MZ
authors.lastname: Guo
authors.affiliation: Jiangsu Key Laboratory of New Drug Research and Clinical Pharmacy, Xuzhou Medical University, Xuzhou, China.|Department of Pharmaceutical Analysis, School of Pharmacy, Xuzhou Medical University, Xuzhou, China.
authors.forename: Dao-Quan
authors.initials: DQ
authors.lastname: Tang
mesh_terms: D000080242:8-Hydroxy-2'-Deoxyguanosine; D000225:Adenine / Q000097:blood; D000818:Animals; D002853:Chromatography, Liquid / Q000379:methods*; D056148:Chromatography, Reverse-Phase / Q000379:methods; D003596:Cytosine / Q000097:blood; D003849:Deoxyguanosine / Q000031:analogs & derivatives / Q000097:blood; D003928:Diabetic Nephropathies / Q000097:blood; D057927:Hydrophobic and Hydrophilic Interactions; D009705:Nucleosides / Q000097:blood*; D011743:Pyrimidines / Q000097:blood; D017207:Rats, Sprague-Dawley; D015203:Reproducibility of Results; D053719:Tandem Mass Spectrometry / Q000379:methods* / Q000592:standards; D019820:Xanthine / Q000097:blood
chemical_list: D009705:Nucleosides; D011743:Pyrimidines; D019820:Xanthine; D000080242:8-Hydroxy-2'-Deoxyguanosine; D003596:Cytosine; D003849:Deoxyguanosine; D000225:Adenine
keywords: comparative evaluation; hydrophilic interaction liquid chromatography; nucleobases; nucleosides; quantitative determination