title: Development of an ion-pairing reversed-phase liquid chromatography method using a double detection analysis (UV and evaporative light scattering detection) to monitor the stability of Alimta(®)-pemetrexed preparations: identification and quantification of L-glutamic acid as a potential degradation product.
abstract: A new method based on high-performance liquid chromatography coupled to ultraviolet and evaporative light scattering detection (HPLC-UV-ELSD) was developed for the determination of L-glutamic acid, a potential degradation product of pemetrexed, and for the quantification of pemetrexed itself. This is an ion-pairing, reversed-phase method. The column was a Synergi MAX-RP C12 4 μm (150 mm x 4.6 mm). The mobile phase was 1 mM tridecafluoroheptanoic acid in aqueous solution and acetonitrile under gradient elution mode. L-Glutamic acid was detected by ELSD, and pemetrexed by UV at 254 nm. Good resolution was achieved between pemetrexed and L-glutamic acid. The HPLC method was validated according to SFSTP and ICH guidelines, and applied the accuracy profile procedure with a five-level validation experimental design. For pemetrexed, the decision criteria selected consisted of the acceptability limits (±3%) and the proportion of results within the calculated tolerance intervals (95%). In conclusion, the proposed analytical procedures were validated over the selected validation domains for L-glutamic acid (0.005-0.025 mg/mL) and pemetrexed (0.4-0.6 mg/mL) and shown to provide a very effective method.
authors.affiliation: Equipe 5, Génétique, Immunothérapie, Chimie et Cancer (GICC), UMR CNRS 6239, Laboratoire de Chimie organique, Faculté de Pharmacie, 31 Avenue Monge, Université de Tours, 37100 Tours, France; Unité de Pharmacie Clinique Oncologique, 2 Bd Tonnellé, 37044 Tours Cedex 9, France. renaud.respaud-2@etu.univ-tours.fr
authors.forename: Renaud
authors.initials: R
authors.lastname: Respaud
authors.forename: Jean-François
authors.initials: JF
authors.lastname: Tournamille
authors.forename: Cécile
authors.initials: C
authors.lastname: Croix
authors.forename: Hélène
authors.initials: H
authors.lastname: Laborie
authors.forename: Claire
authors.initials: C
authors.lastname: Elfakir
authors.forename: Marie-Claude
authors.initials: MC
authors.lastname: Viaud-Massuard
mesh_terms: D002138:Calibration; D002851:Chromatography, High Pressure Liquid; D056148:Chromatography, Reverse-Phase / Q000379:methods*; D004355:Drug Stability; D005971:Glutamates / Q000737:chemistry*; D018698:Glutamic Acid / Q000032:analysis; D006147:Guanine / Q000031:analogs & derivatives* / Q000737:chemistry; D007477:Ions; D008027:Light; D057230:Limit of Detection; D000068437:Pemetrexed; D015203:Reproducibility of Results; D012542:Scattering, Radiation; D012680:Sensitivity and Specificity; D014466:Ultraviolet Rays; D014835:Volatilization
chemical_list: D005971:Glutamates; D007477:Ions; D000068437:Pemetrexed; D018698:Glutamic Acid; D006147:Guanine