Quantification of L-ergothioneine in human plasma and erythrocytes by liquid chromatography-tandem mass spectrometry
Article first published online: 12 MAR 2013
Copyright © 2013 John Wiley & Sons, Ltd.
Journal of Mass Spectrometry
Volume 48, Issue 3, pages 406–412, March 2013
How to Cite
Wang, L.-Z., Thuya, W.-L., Toh, D. S.-L., Lie, M. G.-L., Lau, J.-Y. A., Kong, L.-R., Wan, S.-C., Chua, K.-N., Lee, E. J.-D. and Goh, B.-C. (2013), Quantification of L-ergothioneine in human plasma and erythrocytes by liquid chromatography-tandem mass spectrometry. J. Mass Spectrom., 48: 406–412. doi: 10.1002/jms.3150
- Issue published online: 27 FEB 2013
- Article first published online: 12 MAR 2013
- Manuscript Accepted: 27 NOV 2012
- Manuscript Revised: 24 NOV 2012
- Manuscript Received: 4 OCT 2012
A sensitive analytical method has been developed and validated for the quantification of L-ergothioneine in human plasma and erythrocytes by liquid chromatography-tandem mass spectrometry. A commercially available isotope-labeled L-ergothioneine-d9 is used as the internal standard. A simple protein precipitation with acetonitrile is utilized for bio-sample preparation prior to analysis. Chromatographic separation of L-ergothioneine is conducted using gradient elution on Alltime C18 (150 mm × 2.1 mm, 5 µ). The run time is 6 min at a constant flow rate of 0.45 ml/min. The mass spectrometer is operated under a positive electrospray ionization condition with multiple reaction monitoring mode. The mass transitions of L-ergothioneine and L-ergothioneine-d9 are m/z 230 > 127 and m/z 239 > 127, respectively. Excellent linearity [coefficient of determination (r2) ≥ 0.9998] can be achieved for L-ergothioneine quantification at the ranges of 10 to 10 000 ng/ml, with the intra-day and inter-day precisions at 0.9–3.9% and 1.3–5.7%, respectively, and the accuracies for all quality control samples between 94.5 and 101.0%. This validated analytical method is suitable for pharmacokinetic monitoring of L-ergothioneine in human and erythrocytes. Based on the determination of bio-samples from five healthy subjects, the mean concentrations of L-ergothioneine in plasma and erythrocytes are 107.4 ± 20.5 ng/ml and 1285.0 ± 1363.0 ng/ml, respectively. Copyright © 2013 John Wiley & Sons, Ltd.