Measurement of very low-molecular weight metabolites by traveling wave ion mobility and its use in human urine samples

• Published in: Journal of pharmaceutical analysis Vol. 14; no. 5; pp. 100921 - 11
• Main Authors: Kurilung, Alongkorn, Limjiasahapong, Suphitcha, Kaewnarin, Khwanta, Wisanpitayakorn, Pattipong, Jariyasopit, Narumol, Wanichthanarak, Kwanjeera, Sartyoungkul, Sitanan, Wong, Stephen Choong Chee, Sathirapongsasuti, Nuankanya, Kitiyakara, Chagriya, Sirivatanauksorn, Yongyut, Khoomrung, Sakda
• Format: Journal Article
• Language: English
• Published: China Elsevier B.V 01.05.2024; Xi'an Jiaotong University, Journal of Pharmaceutical Analysis; Chakri Naruebodindra Medical Institute,Faculty of Medicine Ramathibodi Hospital,Mahidol University,Samut Prakan,10540,Thailand%Department of Medicine,Ramathibodi Hospital,Mahidol University,Bangkok,10400,Thailand%Siriraj Center of Research Excellent in Metabolomics and Systems Biology(SiCORE-MSB),Faculty of Medicine Siriraj Hospital,Mahidol University,Bangkok,10700,Thailand; Siriraj Metabolomics and Phenomics Center,Faculty of Medicine Siriraj Hospital,Mahidol University,Bangkok,10700,Thailand; Center of Excellence for Innovation in Chemistry(PERCH-CIC),Faculty of Science,Mahidol University,Bangkok,10400,Thailand; Department of Biochemistry,Faculty of Medicine Siriraj Hospital,Mahidol University,Bangkok,10700,Thailand; Siriraj Center of Research Excellent in Metabolomics and Systems Biology(SiCORE-MSB),Faculty of Medicine Siriraj Hospital,Mahidol University,Bangkok,10700,Thailand; SingHealth Duke-NUS Institute of Biodiversity Medicine,National Cancer Centre Singapore,168583,Singapore%Siriraj Center of Research Excellent in Metabolomics and Systems Biology(SiCORE-MSB),Faculty of Medicine Siriraj Hospital,Mahidol University,Bangkok,10700,Thailand; Department of Biomedical Informatics,University of Arkansas for Medical Sciences,Little Rock,AR,72205,USA%Siriraj Metabolomics and Phenomics Center,Faculty of Medicine Siriraj Hospital,Mahidol University,Bangkok,10700,Thailand%Siriraj Center of Research Excellent in Metabolomics and Systems Biology(SiCORE-MSB),Faculty of Medicine Siriraj Hospital,Mahidol University,Bangkok,10700,Thailand; Siriraj Metabolomics and Phenomics Center,Faculty of Medicine Siriraj Hospital,Mahidol University,Bangkok,10700,Thailand%Waters Pacific Pte.,Ltd.,Singapore,117528,Singapore%Program in Translational Medicine,Faculty of Medicine Ramathibodi Hospital,Mahidol University,Bangkok,10400,Thailand; Elsevier
• Subjects: Chromatography; Collision cross-section; Electric fields; Human urine; Liquid chromatography; Mass spectrometry; Mass spectroscopy; Metabolites; Metabolomics; Mobility; Molecular weight; Scientific imaging; Traveling wave ion mobility; Urine; Very low-molecular-weight metabolites; United States > US; Collision cross-section; Human urine; Very low-molecular-weight metabolites; Traveling wave ion mobility
• ISSN: 2095-1779; 2214-0883; 2214-0883
• DOI: 10.1016/j.jpha.2023.12.011

The collision cross-sections (CCS) measurement using ion mobility spectrometry (IMS) in combination with mass spectrometry (MS) offers a great opportunity to increase confidence in metabolite identification. However, owing to the lack of sensitivity and resolution, IMS has an analytical challenge in studying the CCS values of very low-molecular-weight metabolites (VLMs ≤ 250 Da). Here, we describe an analytical method using ultrahigh-performance liquid chromatography (UPLC) coupled to a traveling wave ion mobility-quadrupole-time-of-flight mass spectrometer optimized for the measurement of VLMs in human urine samples. The experimental CCS values, along with mass spectral properties, were reported for the 174 metabolites. The experimental data included the mass-to-charge ratio (m/z), retention time (RT), tandem MS (MS/MS) spectra, and CCS values. Among the studied metabolites, 263 traveling wave ion mobility spectrometry (TWIMS)-derived CCS values (TWCCSN2) were reported for the first time, and more than 70% of these were CCS values of VLMs. The TWCCSN2 values were highly repeatable, with inter-day variations of <1% relative standard deviation (RSD). The developed method revealed excellent TWCCSN2 accuracy with a CCS difference (ΔCCS) within ±2% of the reported drift tube IMS (DTIMS) and TWIMS CCS values. The complexity of the urine matrix did not affect the precision of the method, as evidenced by ΔCCS within ±1.92%. According to the Metabolomics Standards Initiative, 55 urinary metabolites were identified with a confidence level of 1. Among these 55 metabolites, 53 (96%) were VLMs. The larger number of confirmed compounds found in this study was a result of the addition of TWCCSN2 values, which clearly increased metabolite identification confidence. [Display omitted] •Reduce bias voltages and WH and increase WV enabled the measurement of VLMs by TWIMS.•A total of 263 TWCCSN2 were first reported.•An experimental IM-MS library of 174 standards with m/z, RT, MS/MS, and CCS was created.•The application of the IM-MS library successfully identified 55 metabolites in human urine.