Free mycophenolic acid determination in human plasma ultrafiltrate by a validated liquid chromatography-tandem mass spectrometry method.
The aim of this study was to develop and validate fully the liquid chromatography-tandem mass spectrometry method for free mycophenolic acid (MPA) concentration measurements in plasma ultrafiltrate that will be reliable and simple in preparation with deuterated MPA (MPA-d3) chosen as an internal standard. The chromatographic separation was made with Zorbax Eclipse XDB-C18 column (4.6 × 150 mm) using a gradient of two solutions as a mobile phase: (A) water and (B) methanol, each containing 0.1% formic acid and 2.5 mm ammonium acetate. Satisfactory repeatability of retention times was achieved with average values of 7.54 ± 0.20 min and 7.50 ± 0.19 min for MPA and MPA-d3, respectively. The method was selective, with no carry-over or matrix effect observed. The analytical range was proven for MPA ultrafiltrate concentrations of 1-500 ng/mL. The accuracy and precision fell within the acceptance criteria for intraday (accuracy: 100.63-110.46%, imprecision: 6.23-7.76%), as well as interday assay (accuracy: 98.81-110.63%; imprecision: 5.36-10.22%). The method was used for free MPA determination in plasma samples from patients treated with mycophenolate mofetil. To the best of our knowledge this is the first liquid chromatography-tandem mass spectrometry method for free MPA monitoring using MPA-d3 that allows to measure plasma ultrafiltrate concentrations as low as 1 ng/mL.
Department of Drug Chemistry, Faculty of Pharmacy, Medical University of Warsaw, Banacha 1, 02-097, Warsaw, Poland.
Paulina
http://orcid.org/0000-0003-4622-8935
P
Łuszczyńska
Department of Drug Chemistry, Faculty of Pharmacy, Medical University of Warsaw, Banacha 1, 02-097, Warsaw, Poland.
Tomasz
T
Pawiński
Department of Drug Chemistry, Faculty of Pharmacy, Medical University of Warsaw, Banacha 1, 02-097, Warsaw, Poland.|Clinical Pharmacology Unit, Department of Medical Biology, (previous name: Department of Clinical Biochemistry), Institute of Cardiology, Alpejska 42, 04-628, Warsaw, Poland.
Paweł K
PK
Kunicki
Department of Drug Chemistry, Faculty of Pharmacy, Medical University of Warsaw, Banacha 1, 02-097, Warsaw, Poland.
Katarzyna
K
Sikorska
Department of Bioanalysis and Drugs Analysis, Faculty of Pharmacy, Medical University of Warsaw, Banacha 1, 02-097, Warsaw, Poland.
Ryszard
R
Marszałek
D002853:Chromatography, Liquid / Q000379:methods*; D004355:Drug Stability; D006801:Humans; D057230:Limit of Detection; D016014:Linear Models; D009173:Mycophenolic Acid / Q000097:blood* / Q000737:chemistry; D015203:Reproducibility of Results; D053719:Tandem Mass Spectrometry / Q000379:methods*; D014462:Ultrafiltration
D009173:Mycophenolic Acid
LC-MS/MS; MPA-d3; free fraction; mycophenolate mofetil; mycophenolic acid; validation