Tartrate inhibition of prostatic acid phosphatase improves seminal fluid metabolite stability

Roberts, Matthew J., Hattwell, Jake P. N., Chow, Clement W. K., Lavin, Martin, Pierens, Gregory K., Gardiner, Robert A. and Schirra, Horst Joachim (2016) Tartrate inhibition of prostatic acid phosphatase improves seminal fluid metabolite stability. Metabolomics, 12 11: 162.1-162.14. doi:10.1007/s11306-016-1102-x


Author Roberts, Matthew J.
Hattwell, Jake P. N.
Chow, Clement W. K.
Lavin, Martin
Pierens, Gregory K.
Gardiner, Robert A.
Schirra, Horst Joachim
Title Tartrate inhibition of prostatic acid phosphatase improves seminal fluid metabolite stability
Journal name Metabolomics   Check publisher's open access policy
ISSN 1573-3890
1573-3882
Publication date 2016-11-01
Year available 2016
Sub-type Article (original research)
DOI 10.1007/s11306-016-1102-x
Open Access Status Not yet assessed
Volume 12
Issue 11
Start page 162.1
End page 162.14
Total pages 14
Place of publication New York, NY, United States
Publisher Springer New York LLC
Collection year 2017
Language eng
Formatted abstract
Introduction: Human seminal fluid (hSF) has been suggested as a biofluid suitable to characterise male reproductive organ pathology with metabolomics. However, various enzymatic processes, including phosphorylcholine hydrolysis mediated by prostatic acid phosphatase (PAP), cause unwanted metabolite variation that may complicate metabolomic analysis of fresh hSF samples.

Objectives: To investigate the effects of PAP inhibition with tartrate.

Methods: Using NMR spectroscopy, the kinetics of phosphorylcholine to choline hydrolysis was characterized in hSF samples from three subjects at different temperatures and tartrate concentrations. Principal components analysis was used to characterise the effects of tartrate and temperature on personal differences in metabolite profiles. Potential effects of tartrate on RNA quantification were also determined.

Results: Metabolite profiles and the kinetics of phosphorylcholine degradation are reproducible in independent samples from three ostensibly normal subjects. Increasing concentrations of tartrate and refrigerated sample storage (279 K) resulted in greatly reduced reaction rates as judged by apparent rate constants. Multivariate statistical analysis showed that personal differences in metabolite profiles are not overshadowed by tartrate addition, which stabilises phosphorylcholine and choline concentrations. The tartrate signal also served as an internal concentration standard in the samples, allowing the determination of absolute metabolite concentrations in hSF. Furthermore, the presence of tartrate did not affect RNA expression analysis by qPCR.

Conclusion: Based on these results we recommend as standard protocol for the collection of hSF samples, that 10 mM tartrate are added immediately to samples, followed by sample storage/handling at 277 K until clinical processing within 6 h to remove/inactivate enzymes and isolate metabolite supernatant and other cellular fractions.
Keyword Enzyme inhibition
Metabolomics
Nuclear magnetic resonance (NMR)
Prostate cancer
Prostatic acid phosphatase
Seminal fluid
Q-Index Code C1
Q-Index Status Provisional Code
Institutional Status UQ

 
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