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Urinary myoglobin quantification by high-performance liquid chromatography: an alternative measurement for exercise-induced muscle damage
journal contribution
posted on 2015-12-15, 00:00 authored by Angus Lindsay, Sam Carr, Nick Draper, Steven P GiesegThis study investigated a means of quantifying urinary myoglobin using a novel reverse-phase high-performance liquid chromatography (RP-HPLC) method that is an alternative measure of exercise-induced muscle damage. It also investigated the effect of storage and alkalization on urinary myoglobin stability issues. An RP-HPLC method was validated by precision and repeatability experiments. Myoglobin stability was determined through spiked urine samples stored at various temperatures over an 8-week period using alkalization and dilution in a pH 7.0 buffer. The method was validated with urine collected from mixed martial arts fighters during a competition and training session. The method produced linearity from 5 to 1000 μg/ml (R(2) = 0.997), intra- and inter-assay coefficients of variation from 0.32 to 2.94%, and a lower detection limit of 0.2 μg/ml in the final dilution and 2 μg/ml in the original urine sample. Recovery ranged from 96.4 to 102.5%, myoglobin remained stable at 4 °C when diluted in a pH 7.0 buffer after 20 h, and a significant increase (P < 0.01) and an identifiable peak were observed following a mixed martial arts contest and training session. Storage length and conditions had significant effects (P < 0.05) on stability. The method's simplicity and noninvasive nature means it can be used as an alternative muscle damage assay following exercise and trauma.
History
Journal
Analytical biochemistryVolume
491Pagination
37 - 42Publisher
ElsevierLocation
Amsterdam, The NetherlandsPublisher DOI
eISSN
1096-0309Language
engPublication classification
C1 Refereed article in a scholarly journalCopyright notice
2015, Elsevier Inc.Usage metrics
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