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  3. High-throughput intracellular pteridinic profiling by liquid chromatography-quadrupole time-of-flight mass spectrometry

High-throughput intracellular pteridinic profiling by liquid chromatography-quadrupole time-of-flight mass spectrometry

  • Anal Chim Acta. 2015 Jan 1:853:442-450. doi: 10.1016/j.aca.2014.10.044.
Casey Burton 1,  Rui Weng 2,  Li Yang 2,  Yu Bai 3,  Huwei Liu 2,  Yinfa Ma 4
Affiliations

Affiliations

  • 1 Department of Chemistry and Center for Biomedical Science & Engineering, Missouri University of Science and Technology, 400 West 11th Street, Rolla, MO 65409, United States.
  • 2 Beijing National Laboratory for Molecular Sciences, Key Laboratory of Bioorganic Chemistry and Molecular Engineering of Ministry of Education, College of Chemistry and Molecular Engineering, Peking University, Beijing 100871, China.
  • 3 Beijing National Laboratory for Molecular Sciences, Key Laboratory of Bioorganic Chemistry and Molecular Engineering of Ministry of Education, College of Chemistry and Molecular Engineering, Peking University, Beijing 100871, China. Electronic address: yu.bai@pku.edu.cn.
  • 4 Department of Chemistry and Center for Biomedical Science & Engineering, Missouri University of Science and Technology, 400 West 11th Street, Rolla, MO 65409, United States. Electronic address: yinfa@mst.edu.
PMID: 25467489 DOI: 10.1016/j.aca.2014.10.044
Abstract

Pteridines are a diverse family of endogenous metabolites that may serve as useful diagnostic biomarkers for disease. While many preparative and analytical techniques have been described for analysis of selected pteridines in biological fluids, broad intracellular pteridine detection remains a significant analytical challenge. In this study, a novel, specific and sensitive extraction and high performance liquid chromatography-quadrupole time-of-flight mass spectrometry (HPLC-QTOF MS) method was developed to simultaneously quantify seven intracellular pteridines and monitor 18 additional, naturally-occurring intracellular pteridines. The newly developed method was validated through evaluation of spiked recoveries (84.5-109.4%), reproducibility (2.1-5.4% RSD), method detection limits (0.1-3.0 μg L(-1)) and limits of quantitation (0.1-1 μg L(-1)), and finally application to non-small cell lung Cancer A549 cells. Twenty-three pteridine derivatives were successfully detected from cell lysates with an average RSD of 12% among culture replicates. Quantified intracellular pteridine levels ranged from 1 to 1000 nM in good agreement with previous studies. Finally, this technique may be applied to cellular studies to generate new biological hypotheses concerning pteridine physiological and pathological functions as well as to discovery new pteridine-based biomarkers.

Keywords

A549; HPLC–QTOF MS; Pteridine extraction; Pteridines; Pteridinic profiling.

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