title: Liquid chromatography tandem mass spectrometry method for the quantitation of mycophenolate mofetil in human plasma: Application to a bioequivalence study and metabolite identification.
abstract: We established a sensitive, selective, and rapid analytical method for the quantitation and pharmacokinetic investigation of mycophenolate mofetil in human plasma. To our knowledge, this is the first method that characterizes presence of mycophenolate mofetil glucuronide in clinical samples through tandem mass spectrometry detection and resolves mycophenolate mofetil from its glucuronide metabolite. Liquid chromatography coupled to tandem mass spectrometry detection in positive ion mode was selected to provide optimal selectivity and sensitivity. Due to the ionizable characteristics of the mycophenolate mofetil, a mixed-mode cation-exchange disposable extraction cartridge was prudently chosen. The chromatographic separation was achieved on Luna(®) C18(2) (100×4.60 mm) column using mobile phase consisting of a mixture of 1±0.05 mM ammonium formate in water, titrated to pH 3.1±0.1 with formic acid, and methanol (20:80, v/v), at a flow rate of 0.7 mL/min. The detection was led at m/z ratios of 434.4→ 114.2 and 438.4→ 118.3, for mycophenolate mofetil and mycophenolate mofetil-D4, respectively. The developed method was linear between 40.2-4986.0 pg/mL. All validation parameters were within the defined limits. The validated method was then successfully applied for the evaluation of bioequivalence parameters of mycophenolate mofetil after an oral administration of 500 mg mycophenolate mofetil tablet to healthy male Indian volunteers.
authors.affiliation: Department of Clinical Pharmacology and Pharmacokinetics, Sun Pharmaceutical Industries Ltd, HSIIDC, Gurgaon, Haryana, India.
authors.forename: Pankaj
authors.initials: P
authors.lastname: Partani
authors.affiliation: Department of Pharmaceutical Sciences and Technology, Birla Institute of Technology, Mesra, Ranchi, Jharkhand, India.
authors.forename: Saurabh Manaswita
authors.initials: SM
authors.lastname: Verma
authors.affiliation: Department of Clinical Pharmacology and Pharmacokinetics, Sun Pharmaceutical Industries Ltd, HSIIDC, Gurgaon, Haryana, India.
authors.forename: Tausif
authors.initials: T
authors.lastname: Monif
mesh_terms: D000328:Adult; D002853:Chromatography, Liquid; D064368:Healthy Volunteers; D006801:Humans; D008297:Male; D015394:Molecular Structure; D009173:Mycophenolic Acid / Q000031:analogs & derivatives* / Q000097:blood / Q000378:metabolism / Q000493:pharmacokinetics; D053719:Tandem Mass Spectrometry; D013810:Therapeutic Equivalency
chemical_list: D009173:Mycophenolic Acid
keywords: Bioequivalence studies; Mycophenolate mofetil; Solid-phase extraction; Tandem mass spectrometry