title: Pharmacokinetic application of a bio-analytical LC-MS method developed for 5-fluorouracil and methotrexate in mouse plasma, brain and urine.
abstract: In the past we have reported significant cognitive deficits in mice receiving 5-fluorouracil in combination with low-dose methotrexate. To explain such interactions, a pharmacokinetic study was designed. A sensitive bio-analytical method was therefore developed and validated for 5-fluorouracil and methotrexate in mouse plasma, brain and urine with liquid chromatography coupled to a single quadrupole mass spectrometer. Chromatographic separation was accomplished by Agilent® Zorbax® SB-C18 column, with isocratic elution (5 mM ammonium acetate and methanol, 70:30, %v/v) at a flow rate of 300 μL/min. The limit of quantitation for both drugs was 15.6 ng/mL (plasma and brain) and 78.1 ng/mL (urine), with interday and intraday precision and accuracy ≤15% and a total run time of 6 min. This bio-analytical method was used for the pharmacokinetic characterization of 5-fluorouracil and methotrexate in mouse plasma, brain and urine over a period of 24 h. This method allowed characterization of the brain concentrations of 5-fluorouracil over a period of 24 h.
authors.affiliation: Department of Pharmaceutical Sciences, Temple University School of Pharmacy, Philadelphia, PA 19140, USA.
authors.forename: Vaishnavi
authors.initials: V
authors.lastname: Ganti
authors.forename: Ellen A
authors.initials: EA
authors.lastname: Walker
authors.forename: Swati
authors.initials: S
authors.lastname: Nagar
mesh_terms: D000818:Animals; D001921:Brain / Q000378:metabolism; D001923:Brain Chemistry; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D004355:Drug Stability; D005472:Fluorouracil / Q000032:analysis / Q000097:blood / Q000737:chemistry / Q000493:pharmacokinetics*; D016014:Linear Models; D008297:Male; D013058:Mass Spectrometry / Q000379:methods*; D008727:Methotrexate / Q000032:analysis / Q000097:blood / Q000737:chemistry / Q000493:pharmacokinetics*; D051379:Mice; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity
chemical_list: D005472:Fluorouracil; D008727:Methotrexate
keywords: 5-fluorouracil; LC-MS; brain; methotrexate; pharmacokinetics