Abstract
Recently, it has become possible to examine metabolism abnormalities by detecting increases in specific acylcarnitines in blood tests of newborn babies using tandem mass spectrometer. However, acylcarnitine standard solutions with metrological traceability to the International System of Units (SI) for accurate calibration of tandem mass spectrometer are not yet available worldwide. In this study, we examined a quantitative 1H NMR procedure for obtaining accurate and SI-traceable purity evaluations of acylcarnitines having different numbers of carbon atoms as each raw material for their standard solutions. In particular, the solvent composition and measurement temperature were optimized to reduce the influence of water signal overlapping on analyte signals. It was found that, when the signal of that 1H which directly bound to the asymmetric carbon of the acylcarnitine is the target signal, it was possible to reduce overlapping of the water signal on the target signal by using deuterium oxide as a solvent. On the other hand, in the case of an acylcarnitine that was poorly soluble in deuterium oxide, it was possible to reduce overlapping of the water signal on the target signal by adding an appropriate amount of deuterium oxide to methanol-d 4 in which the acylcarnitine had high solubility. At this time, the optimum mixing volume ratio of methanol-d 4/deuterium would be 80:20. The overlapping of the water signal could be also reduced when the measurement temperature was 15 °C to 40 °C. When the measurement temperature was an around room temperature (in this study, 25 °C), fine shimming could be performed easily. Therefore, the optimum temperature would be 25 °C, because fine shimming was essential to quantify any signal area accurately. Finally, this study confirmed that accurate values with SI traceability could be obtained at about 1 % or less expanded uncertainty for five kinds of acylcarnitines.
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I am grateful to Dr. M. Numata for help to read and correct the English manuscript.
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Presented in part at the 54th Annual Meeting of the Nuclear Magnetic Resonance Society of Japan, Narashino, Japan, November 2015.
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Saito, N., Saito, T., Yamazaki, T. et al. Establishment of an analytical method for accurate purity evaluations of acylcarnitines by using quantitative 1H NMR spectroscopy. Accred Qual Assur 22, 171–178 (2017). https://doi.org/10.1007/s00769-017-1263-y
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DOI: https://doi.org/10.1007/s00769-017-1263-y