Resolution, quantification and confirmation of betamethasone and dexamethasone in equine plasma by liquid chromatography/tandem mass spectrometry.
Abstract: This method describes the simultaneous separation, identification, quantification and confirmation of betamethasone (BTM) and dexamethasone (DXM) in equine plasma by liquid chromatography (LC) integrated with multidimensional tandem mass spectrometry. Analytes were directly extracted from equine plasma by methyl tert-butyl ether (MTBE). The residues were reconstituted with sample solvent. LC separation of the analytes was performed on a Hypercarb column using acetonitrile/water/formic acid (95:5:0.5, v/v/v) as the mobile phase. Sample screening, quantification and confirmation were performed in multiple reaction monitoring (MRM) mode. The method was linear over the concentration range of 0.1-75 ng/mL for both analytes. Limit of detection (LOD) was 50 pg/mL and that of quantification (LOQ) was 100 pg/mL for both analytes. The limit of confirmation (LOC) for the presence of BTM or DXM by MRM was 0.5 ng/mL. The intra-and inter-day precisions expressed as coefficient of variation (CV) for quantification of DXM and BTM from 0.1 to 50 ng/mL were less than 7% and the accuracy was in the range of 97-105%. This method is capable of distinguishing BTM from DXM when both analytes are simultaneously present in equine plasma. Measurement uncertainty for both analytes was estimated at less than 16%. The method is rapid, specific, selective, sensitive, simple and reliable. The importance of this method is its usefulness in directly identifying and differentiating BTM from DXM without derivatization.
Copyright 2005 John Wiley & Sons, Ltd.
Publication Date: 2005-02-17 PubMed ID: 15714597DOI: 10.1002/rcm.1851Google Scholar: Lookup
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- Comparative Study
- Evaluation Study
- Journal Article
- Research Support
- Non-U.S. Gov't
Summary
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The study presents a method for the simultaneous separation, quantification, and identification of betamethasone and dexamethasone in horse plasma using a specific type of mass spectrometry. This allows for the effective differentiation between the two substances when both are present.
Methodology
- The researchers utilized liquid chromatography (LC) combined with tandem mass spectrometry to carry out the separation, identification, quantification, and confirmation of betamethasone (BTM) and dexamethasone (DXM) in horse plasma.
- The substances were directly extracted from equine plasma using methyl tert-butyl ether (MTBE). After extraction, the residues were reconstituted with sample solvent.
- LC separation was performed using a Hypercarb column with a mobile phase made up of acetonitrile/water/formic acid in a particular volume ratio. Sample screening, quantification, and confirmation were performed using multiple reaction monitoring (MRM) mode.
Results
- The method was found to be linear across a concentration range of 0.1-75 ng/mL for both BTM and DXM.
- Further details were provided for the limits of detection (LOD), confirmation (LOC), and quantification (LOQ) for both analytes, contributing to the precision and accuracy of the process.
- The intra- and inter-day precision of the method demonstrated the reproducibility and reliability of the results, with a coefficient of variation (CV) less than 7% and accuracy within 97-105%.
Significance of Findings
- This method can effectively distinguish BTM from DXM when both substances are presented in horse plasma.
- Measurement uncertainty for both analytes was estimated to be less than 16%, indicating high reliability.
- The method proposed is quick, specific, selective, sensitive, simple, and thus reliable. This highlights the practicality and efficiency of directly detecting and differentiating BTM from DXM without requiring additional steps such as derivatization or conversion.
Cite This Article
APA
Luo Y, Uboh CE, Soma LR, Guan F, Rudy JA, Tsang DS.
(2005).
Resolution, quantification and confirmation of betamethasone and dexamethasone in equine plasma by liquid chromatography/tandem mass spectrometry.
Rapid Commun Mass Spectrom, 19(6), 825-832.
https://doi.org/10.1002/rcm.1851 Publication
Researcher Affiliations
- University of Pennsylvania, School of Veterinary Medicine, Department of Clinical Studies, New Bolton Center Campus, Kennett Square, PA 19348, USA.
MeSH Terms
- Animals
- Betamethasone / blood
- Chromatography, High Pressure Liquid / methods
- Chromatography, High Pressure Liquid / veterinary
- Dexamethasone / blood
- Doping in Sports / prevention & control
- Horses / blood
- Reproducibility of Results
- Sensitivity and Specificity
- Spectrometry, Mass, Electrospray Ionization / methods
- Spectrometry, Mass, Electrospray Ionization / veterinary
- Substance Abuse Detection / methods
- Substance Abuse Detection / veterinary
Citations
This article has been cited 4 times.- Chen JW, Uboh CE, Soma LR, You Y, Jiang Z, Li X, Guan F, Liu Y. Identification of sample donor by 24-plex short tandem repeat in a post-race equine plasma containing dexamethasone.. Springerplus 2014;3:94.
- Della Torre M, Hibbard JU, Jeong H, Fischer JH. Betamethasone in pregnancy: influence of maternal body weight and multiple gestation on pharmacokinetics.. Am J Obstet Gynecol 2010 Sep;203(3):254.e1-12.
- Samtani MN, Jusko WJ. Quantification of dexamethasone and corticosterone in rat biofluids and fetal tissue using highly sensitive analytical methods: assay validation and application to a pharmacokinetic study.. Biomed Chromatogr 2007 Jun;21(6):585-97.
- Samtani MN, Lohle M, Grant A, Nathanielsz PW, Jusko WJ. Betamethasone pharmacokinetics after two prodrug formulations in sheep: implications for antenatal corticosteroid use.. Drug Metab Dispos 2005 Aug;33(8):1124-30.
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