Cassette Incubation Followed By Bioanalysis Using High-Resolution Ms For In Vitro Adme Screening Assays

Abstract

Cassette incubation with bioanalysis using HRMS is a feasible approach for high-throughput in vitro ADME assays evaluated in this study.

Evolution of Orbitrap Mass Spectrometry Instrumentation

“…The detection of accurate mass information for all ions in the full scan spectrum, providing a degree of specificity equal to most SRM-based assays, is the principal advantage of the HR/AM MS approach (75). This technique has been used in an in vitro absorption, distribution, metabolism, and excretion workflow of cassette incubation for as many as 32 compounds, followed by quantitative bioanalysis using full scan acquisition (64). Targeting of compounds can also now be achieved on the fly for a variety of analyte groups, including those containing metals (76) or those with distinct isotopic patterns such as sulfur-and bromine-containing (77,78).…”

“…A wide variety of compounds are now routinely analyzed by Orbitrap-based instruments, typically coupled with UHPLC. The role of Orbitrap-based analysis is growing rapidly in a variety of applications, including those of food safety and environmental analysis (51)(52)(53), metabolite and metabolomics analysis (54)(55)(56)(57)(58)(59)(60), clinical analysis (61)(62)(63), bioanalysis (64,65), doping control (66)(67)(68), and lipidomics (69)(70)(71)(72).…”

Development of Cassette PAMPA for Permeability Screening

“…6,7) In the current study we establish a cassette PAMPA assay that would be especially useful for screening the permeability of lipophilic compounds. A double-sink PAMPA consisting of a pH gradient (i.e., pH 6.5 and 7.4 for the donor and receiver compartments, respectively) and a lipophilic sink (i.e., a surfactant in the receiver solution) was utilized with cassette incubation of reference compounds in our studies.…”

Database of Optimized Proteomic Quantitative Methods for Human Drug Disposition‐Related Proteins for Applications in Physiologically Based Pharmacokinetic Modeling

“…Similarly, although mRNA expression is often used as a surrogate of protein expression, mRNA data do not always correlate well with protein expression and activity, particularly in human tissues due to the instability of mRNA and the time elapsed between tissue acquisition and storage . Therefore, quantification methods for important ADME proteins have been developed here using triple quadrupole MS instruments, which are commonly available in ADME laboratories both in industry and academia settings …”