Millipore Technical Publications |
 | TN- TN1103EN00 | |
Automation of HDM-PAMPA using a MultiScreen Permeability Plate on a Hamilton MICROLAB® STAR Workstation | ||
| Lit No: | TN1103EN00 | |
| Year: | 2004 | |
| The HDM-PAMPA is a non-cell based assay designed to predict passive, transcellular permeability of drugs in early drug discovery1. The assay is carried out in a 96-well MultiScreen Permeability filter plate and measures the ability of compounds to diffuse from a donor to an acceptor compartment separated by a hexadecane/hexane artificial membrane on a polycarbonate membrane support. HDM-PAMPA can be used to determine the effect of pH on compound permeability by adjusting the pH of the solutions used in the analysis. The pH permeability profiles in the permeability assay are valuable in predicting gastrointestinal absorption of ionizable drugs. An artificial membrane is applied to the polycarbonate membrane in a 96-well filter plate (Donor plate). The Donor plate is filled with buffer solutions containing the compounds to be tested. The Donor plate is placed in a 96-well Acceptor plate filled with sufficient buffer to ensure liquid contact between the liquid in the Acceptor plate and the polycarbonate membrane. The Donor and Acceptor plates are incubated together for 5-7 hours after which time the Donor plate is removed from the Acceptor plate. Samples from the Acceptor and Donor plates are analyzed by LC/MS or transferred to a UV compatible 96 well plate and analyzed immediately in a UV/Vis spectrophotometer. At the end of the incubation time, the integrity of the artificial membrane layer can be measured using electrical resistance. Automation of HDM-PAMPA on the Hamilton MICROLAB STAR takes about 1 hour and 17 minutes (this does not included the 5 – 7 hours incubation). This time includes the formation of the artificial membrane, dilution of compounds from a mother plate, addition of compounds to the Donor plate and removal of samples for UV/Vis analysis from the Acceptor and Donor plates. |
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