title: HPLC determination of methylphenidate and its metabolite, ritalinic acid, by high-performance liquid chromatography with peroxyoxalate chemiluminescence detection.
abstract: An HPLC-peroxyoxalate chemiluminescence (PO-CL) method for simultaneous determination of methylphenidate (MPH) and ritalinic acid (RA) was developed. The method was used to monitor MPH and RA after administration of MPH to rats. Deproteinized plasma spiked with 1-(3-trifluoromethylphenyl)piperazine (IS) was dried and labeled with 4-(N,N-dimethylaminosulfonyl)-7-fluoro-2,1,3-benzoxadiazole (DBD-F). The labeled sample was cleaned with two kinds of solid-phase extraction cartridge, and the DBD-labels were separated on an ODS column with gradient elution using a mixture of CH(3)CN and imidazole-HNO(3) buffer. Separation of MPH and RA can be achieved within 33 min. The LODs of MPH and RA at a signal-to-noise ratio of 3 were 2.2 and 0.4 ng mL(-1), respectively. Moreover, monitoring of MPH and RA after MPH administration (10 mg kg(-1)) to rat could be performed. The concentration of RA 480 min after administration was eight times higher than that of MPH. The proposed HPLC-PO-CL method was useful for determination of MPH and RA in rat plasma and was successfully used to monitor these substances after MPH administration.
authors.affiliation: Graduate School of Biomedical Sciences, Nagasaki University, 1-14 Bunkyo-machi, Nagasaki 852-8521, Japan.
authors.forename: Mitsuhiro
authors.initials: M
authors.lastname: Wada
authors.forename: Keisuke
authors.initials: K
authors.lastname: Abe
authors.forename: Rie
authors.initials: R
authors.lastname: Ikeda
authors.forename: Ruri
authors.initials: R
authors.lastname: Kikura-Hanajiri
authors.forename: Naotaka
authors.initials: N
authors.lastname: Kuroda
authors.forename: Kenichiro
authors.initials: K
authors.lastname: Nakashima
mesh_terms: D000818:Animals; D000697:Central Nervous System Stimulants / Q000097:blood* / Q000378:metabolism; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D008163:Luminescent Measurements / Q000295:instrumentation / Q000379:methods*; D008297:Male; D008774:Methylphenidate / Q000031:analogs & derivatives* / Q000097:blood* / Q000378:metabolism; D010070:Oxalates / Q000737:chemistry*; D051381:Rats; D017208:Rats, Wistar
chemical_list: D000697:Central Nervous System Stimulants; D010070:Oxalates; D008774:Methylphenidate; C044622:peroxyoxalate; C009189:ritalinic acid