title: Immobilised lipase for in vitro lipolysis experiments.
abstract: In vitro lipolysis experiments are used to assess digestion of lipid-based formulations, and probe solubilisation by colloidal phases during digestion. However, proteins and other biological components in the pancreatin often used as the lipase result in high-background scattering when interrogating structures using scattering approaches, complicating the resolution of colloidal structures. In this study, to circumvent this problem, a modified in vitro digestion model employing lipase immobilised on polymer beads, which allows for separation of the lipid digestion components during lipolysis, was investigated. Titration of the fatty acids released during digestion of medium chain triglycerides using pancreatin compared with immobilised lipase, combined with HPLC was used to follow the digestion, and small-angle X-ray scattering was used to determine colloidal structure formation. Digestion of medium chain triglycerides at the same nominal activity revealed that for the immobilised lipase, a longer digestion time was required to achieve the same extent of digestion. However, the same structural endpoint was observed, indicating that structure formation was not affected by the choice of lipase used. Lipolysis with immobilised lipase led to the reduction of parasitic scattering, resulting in clearer and more defined scattering from the structures generated by the lipolysis products.
authors: Drug Delivery, Disposition and Dynamics, Monash Institute of Pharmaceutical Sciences, Monash, Parkville, Victoria, 3052, Australia.
authors: Stephanie
authors: S
authors: Phan
authors: Stefan
authors: S
authors: Salentinig
authors: Adrian
authors: A
authors: Hawley
authors: Ben J
authors: BJ
authors: Boyd
mesh_terms: D002021:Buffers; D002210:Caprylates / Q000378:metabolism; D002851:Chromatography, High Pressure Liquid; D004800:Enzymes, Immobilized / Q000737:chemistry / Q000378:metabolism*; D005227:Fatty Acids / Q000378:metabolism*; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D008049:Lipase / Q000737:chemistry / Q000378:metabolism*; D008066:Lipolysis; D010194:Pancreatin / Q000737:chemistry / Q000378:metabolism; D011108:Polymers / Q000737:chemistry*; D011208:Powders; D053838:Scattering, Small Angle; D013696:Temperature; D014280:Triglycerides / Q000378:metabolism*; D014961:X-Ray Diffraction
chemical_list: D002021:Buffers; D002210:Caprylates; D004800:Enzymes, Immobilized; D005227:Fatty Acids; D011108:Polymers; D011208:Powders; D014280:Triglycerides; C003637:tricaprylin; D010194:Pancreatin; D008049:Lipase; C010800:tricaprin
keywords: colloid; emulsion; enzyme kinetics; immobilised lipase; in vitro lipolysis; lipids; structure; synchrotron small-angle X-ray scattering