Abstract
Measurement of 3-hydroxyisovaleric acid (3HIA) in human urine has been shown to be a useful indicator of biotin status for a variety of clinical situations, including pregnancy. The work described herein presents a novel UPLC-MS/MS method for accurate and precise quantitation of urinary 3HIA. This method utilizes sample preparation prior to quantitation that has been simplified compared to the previous GC-MS method. To demonstrate the suitability of the UPLC-MS/MS method for human bio-monitoring, this method was used to measure 3-HIA in 64 human urine samples from eight healthy adults in whom marginal biotin deficiency had been induced experimentally by egg white feeding. 3HIA was detected in all specimens; the mean concentration [±standard deviation (SD)] was 80.6 ± 51 μM prior to inducing biotin deficiency. Mean excretion rate for 3HIA (expressed per mol urinary creatinine) before beginning the biotin-deficient diet was 8.5 ± 3.2 mmol 3HIA per mol creatinine and the mean increased threefold with deficiency. These specimens had been previously analyzed by GC-MS; the two data sets showed strong linear relationship with a correlation coefficient of 0.97. These results provide evidence that this method is suitable for bio-monitoring of biotin status in larger populations.
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Acknowledgments
This work was supported by the following agencies: National Institutes of Health, grants R37 DK36823 (DMM), R37 DK36823-26S1 (DMM), and R01 DK79890-01S1 (DMM); Arkansas Biosciences Institute, Arkansas Tobacco Settlement Proceeds Act of 2000 (DMM and GB). The project described was supported by Award Number 1UL1RR029884 from the National Center for Research Resources. We thank Katie Estes and Anna Bogusiewicz, Ph.D., at UAMS for their technical assistance, and Marie Tippett at UAMS for editorial support.
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Horvath, T.D., Matthews, N.I., Stratton, S.L. et al. Measurement of 3-hydroxyisovaleric acid in urine from marginally biotin-deficient humans by UPLC-MS/MS. Anal Bioanal Chem 401, 2805–2810 (2011). https://doi.org/10.1007/s00216-011-5356-x
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DOI: https://doi.org/10.1007/s00216-011-5356-x