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.

Assessing the Potential of Untargeted SWATH Mass Spectrometry-Based Metabolomics to Differentiate Closely Related Exposures in Observational Studies

“…As with EC-MPA, the drug is absorbed as MPA in case of the prodrug since “after oral administration, MMF can hardly be detected at any time in plasma because it is rapidly de-esterified in the stomach to produce MPA”, according to the recently published consensus report by the International Association of Therapeutic Drug Monitoring and Clinical Toxicology [ 25 ]. Comparable urinary metabolite profiles are thus expected in users of the two products, although possible biotransformation products of intact MMF [ 28 ] and/or the cleaved mofetil group [ 29 ] might be detected in urine.…”

The development of a population physiologically based pharmacokinetic model for mycophenolic mofetil and mycophenolic acid in humans using data from plasma, saliva, and kidney tissue

“…(b) 90% PI of modelpredicted and experimentally observed (Bullingham et al, 1996) tMPA, phenyl mycophenolic acid glucuronide concentration in plasma (tMPAG), and mycophenolate mofetile concentration in plasma (tMMF) after intravenous infusion of MMF. (c) 90% PI of model-predicted and experimentally observed (Partani et al, 2015) tMMF after single oral administration of MMF. (d) 90% PI of model-predicted and experimentally observed (Bullingham et al, 1996) tMPA after single oral administration of MMF.…”

In Vitro Predictive Dissolution Test Should Be Developed and Recommended as a Bioequivalence Standard for the Immediate-Release Solid Oral Dosage Forms of the Highly Variable Mycophenolate Mofetil

“…For example, Patel et al and Almeida et al, as in Table , recruited 126 and 103 healthy subjects for BE study under fasting conditions, respectively, due to the high WSV of MMF BE measures. However, studies conducted by Partani et al and Reigner et al failed to demonstrate BE, possibly due to insufficient numbers of subjects relative to the high WSV of a HV drug. Although successful BE test results were reported by Zhang et al and Estevez-Carrizo et al with only a few subjects, the two cases will not be discussed in this paper as the two generic MMF IR products have not been approved by FDA.…”

“…4 For example, Patel et al 15 and Almeida et al, 16 as in Table 2, recruited 126 and 103 healthy subjects for BE study under fasting conditions, respectively, due to the high WSV of MMF BE measures. However, studies conducted by Partani et al 17 and Reigner et al 18 failed to demonstrate BE, possibly due to insufficient numbers of subjects relative to the high WSV of a HV drug.…”