Establishment And Application Of A Novel SDS-PAGE Method To Characterise The Collagen Hydrolyses And Modified Gelatin Product Distributed Between10-90K Dalton

A novel SDS-PAGE method, which integrates the concentrated gel and separatedgel as a whole, is established in present study aiming to separate and characterise therod-like collagen hydrolyses. A series of CB-peptides are used as standard makers toevaluate the collagens whose molecular weight is distributed between10-90K Dalton.Some factors, such as the formulations of gel preparation, electrophoresis conditionswere co-related to the electrophoresis mobility and the separated sharpness of differentcomponents contained in collagen hydrolyses(CH). The established novel SDS-PAGEmethod is firstly and successfully used to characterise a good many of CH preparaed bydifferent raw materials(pigskin, bone, gelatin etc.) and various treated methods (acid,enzymatic and dialysed) as well. A series of scanned chromatograms of variouscomponents with molecular weight ranging10-90K Dalton is clearly demonstrated ononly one single gel plate in particular for the CH from pigskin treated by enzymaticmethod. This CH sample has components whose molecular weight are distributedbetween10-90K Dalton and separated with good definintion. It is great importance andsignificance to characterise CH whose molecular weight is smaller than90K Dalton.The established novel SDS-PAGE method is also employed successfully to estimatethe magnitude and depth of a side reaction of gelatin hydrolysis which is frequently metand accompanied with a gelatin-modified main reaction with some organic modifiers. Atypical example clearly demonstrated by this SDS-PAGE method that the gelatinhydrolysis side reaction percentage can be estimated and reached as high as20-40%depending on the reaction time when gelatin molecular is modified by esterified3,4-dihydroxy bensoyl chloride at pH=8.5as a main reaction to preparemodified gelatinused for making intestine-solubilised capsule.