In-depth characterization of 1,4-butanediol diglycidyl ether substituted hyaluronic acid hydrogels

Tianji Zhang; Siran Zhao; Yujuan Chen; Jiandong Wang; Wei Zhang; Jianjian Liu; Ying Kan; Jin-Ping Li; Xueping Guo; Hongmei Li

doi:10.1016/j.carbpol.2023.120611

In-depth characterization of 1,4-butanediol diglycidyl ether substituted hyaluronic acid hydrogels

Carbohydr Polym. 2023 May 1:307:120611. doi: 10.1016/j.carbpol.2023.120611. Epub 2023 Jan 31.

Authors

Tianji Zhang¹, Siran Zhao², Yujuan Chen³, Jiandong Wang², Wei Zhang², Jianjian Liu³, Ying Kan¹, Jin-Ping Li⁴, Xueping Guo⁵, Hongmei Li⁶

Affiliations

¹ Division of Chemistry and Analytical Science, National Institute of Metrology, Beijing, China; Key Laboratory of Chemical Metrology and Applications on Nutrition and Health for State Market Regulation, China.
² Division of Chemistry and Analytical Science, National Institute of Metrology, Beijing, China.
³ Bloomage Biotechnology Corporation Limited, Jinan, China.
⁴ Beijing Advanced Innovation Centre for Soft Matter Science and Engineering, Beijing University of Chemical Technology, Beijing, China; Department of Medical Biochemistry and Microbiology, University of Uppsala, Uppsala, Sweden. Electronic address: Jin-ping.Li@imbim.uu.se.
⁵ Bloomage Biotechnology Corporation Limited, Jinan, China. Electronic address: guoxp@bloomagebiotech.com.
⁶ Division of Chemistry and Analytical Science, National Institute of Metrology, Beijing, China; Key Laboratory of Chemical Metrology and Applications on Nutrition and Health for State Market Regulation, China. Electronic address: lihm@nim.ac.cn.

PMID: 36781273
DOI: 10.1016/j.carbpol.2023.120611

Abstract

BDDE substituted HA hydrogels remain the most commonly used HA product in the biomedical field. The physical and biochemical properties of the hydrogels are dependent on the degree of modification and substitution patterns/positions, thus, characterizing their fine structure is of great importance for quality assurance. In this study, we developed novel LC-MS methods for accurate determination of MoD as well as in-depth characterization of the linkage network. Fragments resulted from enzymatic depolymerization were resolved by a porous graphitic carbon column followed by online tandem-MS for determining the modification site/residue. With high-resolution separation, two types of previously unknown structures were detected in the cross-linked fragments of 2-B-2 and 4-B-2. Based on the feature of resistance to NaBH₄ reduction, these structures contain a GlcNAc residue modified at OH1. This special sugar unit likely derived from reducing end of the native polysaccharide could be a signature to discriminate subtle batch to batch variations.

Keywords: BDDE cross-linking; HPLC; Hyaluronic acid; Tandem mass spectrometry.