Abstract
A simple protocol for rapid quantitation of acylcarnitines in serum and whole blood has been developed using paper spray mass spectrometry. Dried serum and whole blood containing a mixture of ten acylcarnitines at various concentrations were analyzed as spots from paper directly without any sample pretreatment, separation, or derivatization. The composition of the spray solvent was found to be a critical factor: for serum samples, spray solvent of methanol/water/formic acid (80:20:0.1) gave the best signal intensity while for blood samples which contain more matrix components, acetonitrile/water (90:10) was a much more suitable spray solvent. For the paper type and size used, 0.5 μL of sample provided an optimal signal for both serum and whole blood samples. For quantitative profiling, the limits of quantitation obtained from both serum and blood were much lower than the clinically validated cutoff values for diagnosis of fatty acid oxidation disorders in newborn screening. Linearity (R 2 > 0.95) and reproducibility (RSD ∼10 %) were achieved in the concentration ranges from 100 nM to 5 μM for the C2 acylcarnitine, and for other acylcarnitines, these values were from 10 to 500 nM. Acylcarnitine profiles offer an effective demonstration of the fact that paper spray mass spectrometry is an appropriate, simple, rapid method with high sensitivity and high reproducibility applicable to newborn screening tests.
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Acknowledgments
This work was supported by the National Science Foundation (CHE 0847205 and CHE 0848650), National Science Foundation Instrumentation Development for Biological Research (DBI 0852740), National Center for Research Resources (5R21RR031246-02) and the National Institute of General Medical Sciences (8 R21 GM103454) from the National Institutes of Health, and the Alfred Mann Institute at Purdue University.
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Yang, Q., Manicke, N.E., Wang, H. et al. Direct and quantitative analysis of underivatized acylcarnitines in serum and whole blood using paper spray mass spectrometry. Anal Bioanal Chem 404, 1389–1397 (2012). https://doi.org/10.1007/s00216-012-6211-4
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DOI: https://doi.org/10.1007/s00216-012-6211-4