ppt课件-methodsandmaterialsmicroscopicdrugdistributionstudies.pptVIP

Methods and Materials: Microscopic  Drug Distribution Studies Presented By: Rich Dominiak Laura Kuczynski John Roszko Microscopic Study Procedure 1st – 10 μm thin sections were obtained using a cryogenic microtome set at -25 °C with a microtome knife 2nd – 1 mL of embedding medium was poured onto the chucks 30 seconds later it became an opaque solid Procedure cont’d Next, about 3 mm x 1 mm x 1 mm of release matrix was taken from various location in the original 1 cm x 1 cm x 1 mm slab. These pieces of the matrix were then placed on the planed embedding medium, which was in contact to the planed surface. Procedure cont’d More hardening medium was added to the planned surface. Sections 10 μm in thickness were cut and stuck to the microtome knife They were then taken off of the knife with a glass slide at room temperature Final Matrix The final sections were 3 mm x 1 mm x 10 μm. The 1 mm represents the original depth of the matrix. Observation They were observed under scanning electron microscopy (SEM). Dried to enable high vacuum conditions for SEM. Drying Drying Procedure Water to 100% ethanol 100% ethanol to 100% amyl acetate Cooled to 4 °C and filled with liquid CO2 Exhausted the CO2 vapor while adding CO2 liquid Temperature and Pressure increased over 20 minutes to 40 °C and 55 atm, respectively (when amyl acetate was gone) Drying cont’d After 20 min it is assumed all amyl acetate is gone because all the CO2 is vapor Pressure and Temperature went back to ambient conditions Sample is ready for analysis Drug Distribution Studies Separating matrix into four sections Removed and frozen on dry ice to terminate drug release Serially cut with cryomicrotome into four sections for analysis The samples were set in 0.9% NaCl solution for 3 days The medium was then filtered and protein concentration was determined by UV spectroscopy at 220 nm Questions ? * * February 14, 2006 *