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Open AccessArticle

Profiling of Amino Acids in Urine Samples of Patients Suffering from Inflammatory Bowel Disease by Capillary Electrophoresis-Mass Spectrometry

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Department of Pharmaceutical Analysis and Nuclear Pharmacy, Faculty of Pharmacy, Comenius University in Bratislava, Odbojarov 10, SK-832 32 Bratislava, Slovak Republic
2
Toxicological and Antidoping Center, Faculty of Pharmacy, Comenius University in Bratislava, Odbojárov 10, SK-832 32 Bratislava, Slovak Republic
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Institute of Neuroimmunology, Slovak Academy of Science, Dubravska cesta 9, SK-845 10, Bratislava, Slovak Republic
4
SK-Lab s.r.o., Partizanska 15, SK-984 01, Lucenec, Slovak Republic
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Department of Gastroenterology, St Michael’s Hospital, Satinskeho 1, SK-811 08 Bratislava, Slovak Republic
*
Author to whom correspondence should be addressed.
Molecules 2019, 24(18), 3345; https://doi.org/10.3390/molecules24183345
Received: 13 August 2019 / Revised: 9 September 2019 / Accepted: 11 September 2019 / Published: 14 September 2019
(This article belongs to the Section Analytical Chemistry)
Urine represents a convenient biofluid for metabolomic studies due to its noninvasive collection and richness in metabolites. Here, amino acids are valuable biomarkers for their ability to reflect imbalances of different biochemical pathways. An impact of amino acids on pathology, prognosis and therapy of various diseases, including inflammatory bowel disease (IBD), is therefore the subject of current clinical research. This work is aimed to develop a capillary electrophoresis-tandem mass spectrometry (CE-MS/MS) method for the quantification of the 20 proteinogenic amino acids in human urine samples obtained from patients suffering from IBD and treated with thiopurines. The optimized CE-MS/MS method, with minimum sample preparation (just “dilute and shoot”), exhibited excellent linearity for all the analytes (coefficients of determination were higher than 0.99), with inter-day and intra-day precision yielding relative standard deviations in the range of 0.91–15.12% and with accuracy yielding relative errors in the range of 85.47–112.46%. Total analysis time, an important parameter for the sample throughput demanded in routine practice, was shorter in ca. 17% when compared to established CE-MS methods. Favorable performance of the proposed CE-MS/MS method was also confirmed by the comparison with corresponding ultra-high performance liquid chromatography-mass spectrometry (UHPLC-MS) method. Consistent data for the investigated amino acid metabolome were obtained using both methods. For the first time, the amino acid profiling by CE-MS approach was applied on the clinical IBD samples. Here, significant differences observed in the concentration levels of some amino acids between IBD patients undergoing thiopurine treatment and healthy volunteers could result from the simultaneous action of the disease and the corresponding therapy. These findings indicate that amino acids analysis could be a valuable tool for the study of mechanism of the IBD treatment by thiopurines. View Full-Text
Keywords: inflammatory bowel disease; amino acids profiling; clinical analysis; human urine; capillary zone electrophoresis; mass spectrometry inflammatory bowel disease; amino acids profiling; clinical analysis; human urine; capillary zone electrophoresis; mass spectrometry
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Piestansky, J.; Olesova, D.; Galba, J.; Marakova, K.; Parrak, V.; Secnik, P.; Secnik, P., Jr.; Kovacech, B.; Kovac, A.; Zelinkova, Z.; Mikus, P. Profiling of Amino Acids in Urine Samples of Patients Suffering from Inflammatory Bowel Disease by Capillary Electrophoresis-Mass Spectrometry. Molecules 2019, 24, 3345.

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