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J Pharm Biomed Anal

Title:		Beyond the limit of assignment of metabolites using minimal serum samples and (1)H NMR spectroscopy with cross-validation by mass spectrometry
Author(s):		Gupta A; Kumar D;
Address:		"Centre of Biomedical Research, SGPGIMS Campus, Lucknow, India. Electronic address: ashishg24@yahoo.co.in. Centre of Biomedical Research, SGPGIMS Campus, Lucknow, India"
Journal Title:		J Pharm Biomed Anal
Year:		2018
Volume:		20180109
Issue:
Page Number:		356 - 364
DOI:		10.1016/j.jpba.2018.01.015
ISSN/ISBN:		1873-264X (Electronic) 0731-7085 (Linking)
Abstract:		"Identification of NMR-based metabolic indexes is limited by the deleterious effects of copious proteins and lipoproteins in the serum that accentuate the need for advance and high-throughput method. We tried to explore the use of a novel filtration (2KDa molecular weight cut-off) approach to remove the proteins from serum following use of less sample volume (only 150?ª+muL of filtered serum), combining an array of 1D/2D NMR experiments (at 800?ª+MHz spectrometer), spiking experiments with standard compounds, and validated by mass spectrometry. This novel method enabled the identification of a large number (n?ª+=?ª+73) of metabolites and their percentage of abundance in the present study cohort. Mass spectrometry further validates and confirms the presence of all these 73 metabolites using same filtered serum. This study reveals seven new metabolites (citrulline, inosine, taurine, trimethyl amine, methylmalonate, uracil, methanol) in filtered serum using 1D/2D NMR spectroscopy that were not observed in earlier available literature using protein precipitation approach. This novel method delineates volatile metabolites, nitrogenous bases and nucleosides that may provide a milestone for the identification of inborn error of metabolism, pathogenicity at molecular level, disease identification and prognosis, and forensic studies using minimal volume of filtered serum samples and NMR spectroscopy"
Keywords:		"Feasibility Studies Forensic Sciences/instrumentation/methods Humans Limit of Detection Mass Spectrometry/instrumentation/*methods Metabolism, Inborn Errors/blood/diagnosis/metabolism *Metabolome Metabolomics/instrumentation/*methods Micropore Filters Mol;"
Notes:		"MedlineGupta, Ashish Kumar, Deepak eng Comparative Study England 2018/02/08 J Pharm Biomed Anal. 2018 Mar 20; 151:356-364. doi: 10.1016/j.jpba.2018.01.015. Epub 2018 Jan 9"