Substituent distribution and clouding behavior of hydroxypropyl methyl cellulose analyzed using enzymatic degradation

The distribution of substituents along the polymer backbone will have a strong influence on the properties of modified cellulose. Endoglucanases were used to degrade three different batches of hydroxypropyl methyl cellulose (HPMC) derivatives with similar chemical properties. The phase separation of the HPMCs as a function of temperature, i.e., the clouding behavior, was analyzed prior to degradation. The total amount of unsubstituted glucose was determined using total acid hydrolysis followed by high-performance anion-exchange chromatography with pulsed amperometric detection (HPAEC-PAD). The products after enzymatic degradation were analyzed with size-exclusion chromatography with online multiangle light scattering and refractive index detection and also with reducing end determination. To further characterize the formed products, matrix-assisted laser desorption/ionization time-of-flight mass spectrometry was employed for analysis of short-chained oligosaccharides. The different endoglucanases showed varying degradation capability of HPMC derivatives, depending on structure of the active site. The investigated HPMCs had different susceptibility to degradation by the endoglucanases. The results showed a difference in substituent distribution between HPMC batches, which could explain the differing clouding behaviors. The batch with the lowest cloud point was shown to contain a higher number of non-degradable, highly substituted regions.