Simultaneous and high-throughput quantitation of urinary tetranor PGDM and tetranor PGEM by online SPE-LC-MS/MS as inflammatory biomarkers

Yizhong Zhang, Guodong Zhang, Philip A. Clarke, Jeffrey T. J. Huang, Eddie Takahashi, David Muirhead, Rick C. Steenwyk, Zhaosheng Lin (Lead / Corresponding author)

Research output: Contribution to journalArticle

19 Citations (Scopus)

Abstract

Quantitation of urinary tetranor PGDM or tetranor PGEM (tPGDM and tPGEM) in the past was performed separately using off-line SPE LC-MS/MS methods. The manual SPE procedure is generally time-consuming and cost-ineffective. In addition, simultaneous quantitation of tPGDM and tPGEM is favorable yet very challenging because of the similar chemical structures and identical MRM transitions. This work describes the development and validation of a high-throughput online SPE-LC-MS/MS method, allowing simultaneous and high-throughput measurement of tPGDM and tPGEM in human urine. The reportable range of the assay was 0.2-40 ng/ml for tPGDM and 0.5-100 ng/ml for tPGEM. Intra- and inter-assay precision and accuracy determined using quality control samples were all within acceptable ranges (% CV and % Bias < 15%). Tetranor PGDM was stable under all tested conditions while tPGEM was stable at 4 °C and after three F/T cycles but not stable at room temperature for 24 h (recovery below 80%). The assay was applied to measure urinary tPGDM and tPGEM among healthy volunteers, smokers and COPD patients. Significantly higher urinary levels of both tPGDM and tPGEM were observed in COPD patients than those of non-smoking healthy volunteers. These results demonstrated that the high-throughput online SPE-LC-MS/MS assay provides sensitive, reproducible and accurate measurement of urinary tPGDM and tPGEM as biomarkers for assessing inflammatory diseases such as COPD.

Original languageEnglish
Pages (from-to)705-11
Number of pages7
JournalJournal of Mass Spectrometry
Volume46
Issue number7
Early online date27 Jun 2011
DOIs
Publication statusPublished - Jul 2011

Keywords

  • Biomarkers/urine
  • High-Throughput Screening Assays/methods
  • Humans
  • Inflammation/urine
  • Least-Squares Analysis
  • Prostaglandin D2/analogs & derivatives
  • Prostaglandins/urine
  • Pulmonary Disease, Chronic Obstructive/urine
  • Reproducibility of Results
  • Sensitivity and Specificity
  • Smoking
  • Solid Phase Extraction/methods
  • Tandem Mass Spectrometry/methods

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