title: Development and validation of an ultrafast chromatographic method for quantification of the immunosuppressant mycophenolic acid in canine, feline and human plasma.
abstract: Mycophenolic acid (MPA) is the active metabolite of the prodrug mycophenolate mofetil. In this study, we developed and validated a novel ultra-high performance liquid chromatography (UHPLC) method for the rapid quantification of MPA in plasma from dogs, cats and humans. Following the protein precipitation, calibration standards and quality controls were separated by UHPLC reversed-phase on a 1.5μm 2.1×100mmC18column and quantified using UV detection at 215nm. The procedure produced a linear curve (r2>0.997) over the concentration range 0.4-50μg/mL and exhibited a high degree of repeatability (CV% <11%). The limit of detection (LOD) and lower limit of quantitation (LLOQ) were 0.1 and ≤0.4μg/mL, respectively and the overall recovery was ≥87%. By combining isocratic conditions with a UHPLC column containing solid core particles, we were able to elute MPA and the internal standard (mycophenolic acid carboxybutoxy ether) within 3.0min. The short total run time makes this method ideal to study the disposition of MPA in large batches of plasma samples and/or monitor plasma drug concentrations, as recommended for patients that require optimized immunosuppression.
authors.affiliation: Program in Individualized Medicine, Department of Veterinary Clinical Sciences, College of Veterinary Medicine, Washington State University, Pullman 99164, WA, United States.
authors.forename: Sol-Maiam
authors.initials: SM
authors.lastname: Rivera Vélez
authors.affiliation: Program in Individualized Medicine, Department of Veterinary Clinical Sciences, College of Veterinary Medicine, Washington State University, Pullman 99164, WA, United States.
authors.forename: Alice
authors.initials: A
authors.lastname: Morassi
authors.affiliation: Program in Individualized Medicine, Department of Veterinary Clinical Sciences, College of Veterinary Medicine, Washington State University, Pullman 99164, WA, United States.
authors.forename: Michael H
authors.initials: MH
authors.lastname: Court
authors.affiliation: Program in Individualized Medicine, Department of Veterinary Clinical Sciences, College of Veterinary Medicine, Washington State University, Pullman 99164, WA, United States.
authors.forename: Jennifer E
authors.initials: JE
authors.lastname: Slovak
authors.affiliation: Program in Individualized Medicine, Department of Veterinary Clinical Sciences, College of Veterinary Medicine, Washington State University, Pullman 99164, WA, United States. Electronic address: nvillarino@vetmed.wsu.edu.
authors.forename: Nicolas F
authors.initials: NF
authors.lastname: Villarino
mesh_terms: D061605:Administration, Intravenous; D000284:Administration, Oral; D000818:Animals; D002415:Cats; D002626:Chemistry, Pharmaceutical / Q000379:methods* / Q000592:standards; D002851:Chromatography, High Pressure Liquid / Q000379:methods / Q000592:standards; D004285:Dogs; D006801:Humans; D007166:Immunosuppressive Agents / Q000008:administration & dosage / Q000032:analysis / Q000097:blood*; D009173:Mycophenolic Acid / Q000008:administration & dosage / Q000032:analysis / Q000097:blood*; D015203:Reproducibility of Results
chemical_list: D007166:Immunosuppressive Agents; D009173:Mycophenolic Acid
keywords: Cats; Dogs; Human; Mycophenolic acid; UHPLC