title: Determination of modification degree in BDDE-modified hyaluronic acid hydrogel by SEC/MS.
abstract: Determination of modification degree in BDDE-modified hyaluronic acid (HA) hydrogel is of particular interest. In this paper, three crosslinking parameters (degree of total modification, t-MOD; degree of cross-link modification, c-MOD; degree of pendent modification, p-MOD) are defined and determined by quantification of the modified fragments in hydrogel digestion by size exclusion chromatography combined with mass spectrometry (SEC-MS). The digestion products of a novel hyaluronidase HAase-B produced by Bacillus sp. A50 are studied and only a few modified fragments are identified by (1)H NMR and MS. As a result, Three HA hydrogels prepared in lab have different t-MOD, c-MOD and p-MOD, but the ratio of c-MOD to p-MOD result in the almost same value of 75%. Hydrogel products from Q-Med have nearly same t-MOD about 0.8% and c-MOD about 0.1%, the ratio of c-MOD to p-MOD is about 13%. Hydrogels from ANTEIS S.A all have much higher t-MOD values, the ratio of c-MOD and p-MOD is about 8%.
authors.affiliation: School of Pharmaceutical Science, Shandong University, Jinan, China.
authors.forename: Biao
authors.initials: B
authors.lastname: Yang
authors.affiliation: School of Pharmaceutical Science, Shandong University, Jinan, China; Bloomage Freda Biopharm Co., Ltd., Jinan, China. Electronic address: guoxp@bloomagefreda.com.
authors.forename: Xueping
authors.initials: X
authors.lastname: Guo
authors.affiliation: School of Pharmaceutical Science, Shandong University, Jinan, China.
authors.forename: Hengchang
authors.initials: H
authors.lastname: Zang
authors.affiliation: Bloomage Freda Biopharm Co., Ltd., Jinan, China.
authors.forename: Jianjian
authors.initials: J
authors.lastname: Liu
mesh_terms: D002072:Butylene Glycols / Q000737:chemistry*; D002850:Chromatography, Gel; D003432:Cross-Linking Reagents / Q000737:chemistry; D000067548:Dermal Fillers; D017666:Deuterium Oxide / Q000737:chemistry; D005782:Gels / Q000737:chemistry; D006820:Hyaluronic Acid / Q000737:chemistry*; D006821:Hyaluronoglucosaminidase / Q000378:metabolism; D020136:Hydrogel, Polyethylene Glycol Dimethacrylate / Q000737:chemistry*; D013058:Mass Spectrometry; D008970:Molecular Weight; D066244:Proton Magnetic Resonance Spectroscopy; D013997:Time Factors
chemical_list: D002072:Butylene Glycols; D003432:Cross-Linking Reagents; D000067548:Dermal Fillers; D005782:Gels; D020136:Hydrogel, Polyethylene Glycol Dimethacrylate; D006820:Hyaluronic Acid; C014376:1,4-bis(2,3-epoxypropoxy)butane; D006821:Hyaluronoglucosaminidase; D017666:Deuterium Oxide
keywords: (1)H NMR; Crosslinking parameters; HA hydrogel; Hyaluronic acid; Hyaluronidase; Modification degree; SEC–MS