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. 2020 Feb 26;10:3506. doi: 10.1038/s41598-020-60526-0

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© The Author(s) 2020

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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Overview of GAGDoMa. (a) Workflow of GAGDoMa and structure of a xyloside-primed CS/DS chain including the cleavage sites of the bacterial depolymerizing enzymes; chondroitinase ABC (ChABC), chondroitinase AC (ChAC), and chondroitinase B (ChB). (b–d) Extracted ion chromatograms (XICs) at m/z 458.06 (dp2nSn, where n = 1, 2, 3…) after chondroitinase ABC (b), chondroitinase AC (c), and chondroitinase B (d) to demonstrate the separation of internal oligosaccharides using GAGDoMa. Additionally sulfated precursor ions, dp2nS(n + 1) (where n = 1*, 2*, 3*…), detected at m/z 458.06 owing to in-source sulfate loss, are indicated by an asterisk. (e) MS¹ spectra at 25.05–25.75 min displaying the dp2S2-related precursor ions including [M − 2H]^2- at m/z 268.50, [M − SO₃-H]^- at m/z 458.06, [M − H]^- at m/z 538.02, and [M+DBA-2H]^- at m/z 667.17. DBA, dibutylamine. (f) XIC at m/z 143.05, arising from the naphthyl fragment ion in xyloside-primed linkage region variants generated after chondroitinase B depolymerization displays the elution profile of linkage region variants using GAGDoMa.