Targeted UPLC-MS/MS high-throughput metabolomics approach to assess the purine and pyrimidine metabolism.

Abstract

Purines and pyrimidines, the important components of DNA and RNA, are closely related to metabolic syndrome and disorder, such as renal disease, gout and diabetic nephropathy etc. Given the importance of the biological significance of purines and pyrimidines, it is necessary to further develop a rapid and sensitive method for practical detection of a large-scale analyses. In this study, based on 96-well solid phase extraction plates-ultra performance liquid chromatography-triple quadrupole mass spectrometry (SPE-UPLC-QqQ-MS/MS), a novel approach for simultaneous determination of 23 purines and pyrimidines in biological samples was developed. First, plasma samples were pretreated by SPE using 96-well plates, which lead to an automated, simplified and rapid sample preparation process. In the methodology development, a large-scale test was performed to evaluate the stability and reliability of the approach. Finally, the levels of purines and pyrimidines in the biological samples were analyzed by this strategy. Experimental results showed that lowest limit of quantification (LLOQ) range from 6.678 × 10-2 μg/mL to 4.275 × 10-6 μg/mL; intra- and inter-day precision are <15% for all analytes. The stability and maximal capability of a single analytical batch could be extended to at least 431 injections (about 70 h). Analysis time of a single run was controlled in 10 min. Under the optimized conditions, wide linear ranges and good correlation coefficients (R2 > 0.99) were acquired. The successful development of this method provides a feasible protocol for a large-scale metabolomics study and it also lays the foundation of quantitative analysis in endogenous analytes.

• Authors: Chuanxin Liu, Caiyun Gu, Wei Huang, Xue Sheng, Jun Du, Yubo Li
• Journal: Journal of chromatography. B, Analytical technologies in the biomedical and life sciences (J Chromatogr B Analyt Technol Biomed Life Sci) Vol. 1113 Pg. 98-106 (Apr 15 2019) ISSN: 1873-376X [Electronic] Netherlands
• PMID: 30925277 (Publication Type: Journal Article)
• Copyright: Copyright © 2019 Elsevier B.V. All rights reserved.

Chemical References

• Purines
• Pyrimidines

Topics

• Adult
• Animals
• Chromatography, High Pressure Liquid (methods)
• High-Throughput Screening Assays
• Humans
• Limit of Detection
• Linear Models
• Male
• Metabolomics (methods)
• Purines (blood, metabolism)
• Pyrimidines (blood, metabolism)
• Rats
• Rats, Wistar
• Reproducibility of Results
• Tandem Mass Spectrometry (methods)