biostudies-literatureLenco JNCI NIH HHS7https://www.ebi.ac.uk/biostudies/studies/S-EPMC3331852biostudies-literatureUnknown10(1)Naturally occurring native peptides provide important information about physiological states of an organism and its changes in disease conditions but protocols and methods for assessing their abundance are not well-developed. In this paper, we describe a simple procedure for the quantification of non-tryptic peptides in body fluids. The workflow includes an enrichment step followed by two-dimensional fractionation of native peptides and MS/MS data management facilitating the design and validation of LC- MRM MS assays. The added value of the workflow is demonstrated in the development of a triplex LC-MRM MS assay used for quantification of peptides potentially associated with the progression of liver disease to hepatocellular carcinoma.Proteome scienceMS/MS library facilitated MRM quantification of native peptides prepared by denaturing ultrafiltration.PMC3331852R01 CA135069Lenco JLan RGoldman REdwards NfalseMS/MS library facilitated MRM quantification of native peptides prepared by denaturing ultrafiltration.Naturally occurring native peptides provide important information about physiological states of an organism and its changes in disease conditions but protocols and methods for assessing their abundance are not well-developed. In this paper, we describe a simple procedure for the quantification of non-tryptic peptides in body fluids. The workflow includes an enrichment step followed by two-dimensional fractionation of native peptides and MS/MS data management facilitating the design and validation of LC- MRM MS assays. The added value of the workflow is demonstrated in the development of a triplex LC-MRM MS assay used for quantification of peptides potentially associated with the progression of liver disease to hepatocellular carcinoma.2012-01-01T00:00:00Z2012 Feb2025-04-04T09:12:35.771Z2019-03-27T00:52:31ZS-EPMC33318522230475610.1186/1477-5956-10-7