Low molecular weight chondroitin sulfate (LMCS) has attention for enhanced bioavailability and bioactivity compared to native CS. We optimized H₂O₂/ ascorbic acid (Vc) degradation conditions to prepare LMCS from chicken, bovine, and shark cartilages. Degradation kinetics models and chemical composition data of LMCS showed the GlcA residues of chondroitin-4-sulfate (CSA) may be preferentially attacked. Nuclear magnetic resonance (NMR) spectroscopy and high-performance liquid chromatography-electrospray mass spectrometry (HPLC-MS) indicated that the CH of GlcA in CS was broken through a hydrogen abstraction reaction to break the β-(1 → 3) bond and form the hexendioic acid product. Standard density functional theory (DFT) calculations indicated that the energy required for the hydrogen abstraction from the C1-H bond in GlcA was lower than that of GalNAc. Molecular dynamics (MD) showed that CSA was more likely to interact with hydroxyl radicals (·OH) than non-sulfated chondroitin (CSO) and chondroitin-6-sulfate (CSC). These results provide guidance for producing LMCS.

与天然硫酸软骨素相比，低分子量硫酸软骨素 (LMCS) 因其生物利用度和生物活性的增强而受到关注。我们优化了 H ₂ O ₂ / 抗坏血酸 (Vc) 降解条件，从鸡、牛和鲨鱼软骨中制备 LMCS。LMCS 的降解动力学模型和化学成分数据表明，4-硫酸软骨素 (CSA) 的 GlcA 残基可能会优先受到攻击。核磁共振(NMR)光谱和高效液相色谱-电喷雾质谱(HPLC-MS)表明CS中GlcA的C H通过夺氢反应被打破，从而破坏了β-(1 → 3)键，形成己二酸产物。标准密度泛函理论（DFT）计算表明，GlcA中C1-H键夺氢所需的能量低于GalNAc。分子动力学（MD）表明，CSA 比非硫酸化软骨素（CSO）和 6-硫酸软骨素（CSC）更容易与羟基自由基（·OH）相互作用。这些结果为生产 LMCS 提供了指导。