title: Combined approach for high-throughput preparation and analysis of plasma samples from exposure studies.
abstract: In drug discovery today, drug exposure is determined in preclinical efficacy and safety studies and drug effects are related to measured concentrations rather than to the administered dose. This leads to a strong increase in the number of bioanalytical samples, demanding the development of higher throughput methods to cope with the increased workload. Here, a combined approach is described for the high-throughput preparation and liquid chromatography/tandem mass spectrometry (LC/MS/MS) analysis of drug levels in plasma samples from the preclinical efficacy and safety studies, i.e. exposure studies. Appropriate pharmacokinetic (PK) compartmental models were fitted to data from PK screening studies in the rat, which were subsequently used to simulate the expected plasma concentrations of the respective exposure studies. Information on the estimated drug concentrations was used to dilute the samples to appropriate concentration levels. A Tecan Genesis RSP liquid handling system was utilized to perform automated plasma sample preparation including serial dilution of standard solutions, dilution of plasma samples, addition of internal standard solution and precipitation with acetonitrile. This robotic sample preparation process permitted two studies of 1-96 samples each to be run simultaneously. To ensure the performance of this method the accuracy and precision for diazepam were examined. Two novel drugs were used to illustrate the suggested approach. In conclusion, our method for sample preparation of exposure samples, based on the combined use of PK simulations, a liquid handling system and a fast LC/MS/MS method, increased the throughput more than three times and minimized the errors, while maintaining the required accuracy and precision.
authors.affiliation: DMPK, AstraZeneca R&D Södertälje, S-151 85 Södertälje, Sweden.
authors.forename: Sveinn
authors.initials: S
authors.lastname: Briem
authors.forename: Stefan
authors.initials: S
authors.lastname: Martinsson
authors.forename: Tjerk
authors.initials: T
authors.lastname: Bueters
authors.forename: Erika
authors.initials: E
authors.lastname: Skoglund
mesh_terms: D000818:Animals; D014151:Anti-Anxiety Agents / Q000009:adverse effects / Q000097:blood / Q000493:pharmacokinetics; D002853:Chromatography, Liquid / Q000379:methods*; D003198:Computer Simulation; D003975:Diazepam / Q000009:adverse effects / Q000097:blood / Q000493:pharmacokinetics; D004353:Drug Evaluation, Preclinical; D064420:Drug-Related Side Effects and Adverse Reactions; D008297:Male; D015394:Molecular Structure; D004364:Pharmaceutical Preparations / Q000097:blood* / Q000737:chemistry; D051381:Rats; D017207:Rats, Sprague-Dawley; D012015:Reference Standards; D012371:Robotics; D053719:Tandem Mass Spectrometry / Q000379:methods*; D013997:Time Factors; D014859:Warfarin / Q000737:chemistry
chemical_list: D014151:Anti-Anxiety Agents; D004364:Pharmaceutical Preparations; D014859:Warfarin; D003975:Diazepam