title: High-performance liquid chromatographic determination of tiapride and its phase I metabolite in blood plasma using tandem UV photodiode-array and fluorescence detection.
abstract: New bioanalytical SPE-HPLC-PDA-FL method for the determination of the neuroleptic drug tiapride and its N-desethyl metabolite was developed, validated and applied to xenobiochemical and pharmacokinetic studies in humans and animals. The sample preparation process involved solid-phase extraction of diluted plasma spiked with sulpiride (an internal standard) using SPE cartridges DSC-PH Supelco, USA. Chromatographic separation of the extracts was performed on a Discovery HS F5 250 mm × 4 mm (Supelco) column containing pentafluorophenylpropylsilyl silica gel. Mobile phase (acetonitrile-0.01 M phosphate buffer pH=3, flow rate 1 ml min(-1)) in the gradient mode was employed in the HPLC analysis. Tandem UV photodiode-array→fluorescence detection was used for the determination of the analytes. Low concentrations of tiapride and N-desethyl tiapride were determined using a more selective fluorescence detector (λ(exc.)/λ(emiss.)=232 nm/334 nm), high concentrations (500-6000 pmol ml(-1)) using a UV PDA detector at 212 nm with a linear response. Each HPLC run lasted 15 min. Lower limits of quantification (LLOQ) for tiapride (N-desethyl tiapride) were found to be 8.24 pmol ml(-1) (10.11 pmol ml(-1)). The recoveries of tiapride ranged from 89.3 to 94.3%, 81.7 to 86.8% for internal standard sulpiride and 90.9 to 91.8% for N-desethyl tiapride.
authors.affiliation: Institute of Experimental Biopharmaceutics, Joint Research Center of PROMEDCS Praha as and Academy of Sciences of the Czech Republic, Hradec Králové, Czech Republic. nobilis@uebf.cas.cz
authors.forename: Milan
authors.initials: M
authors.lastname: Nobilis
authors.forename: Zuzana
authors.initials: Z
authors.lastname: Vybíralová
authors.forename: Barbora
authors.initials: B
authors.lastname: Szotáková
authors.forename: Květa
authors.initials: K
authors.lastname: Sládková
authors.forename: Martin
authors.initials: M
authors.lastname: Kuneš
authors.forename: Zbyněk
authors.initials: Z
authors.lastname: Svoboda
mesh_terms: D000818:Animals; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D006801:Humans; D057230:Limit of Detection; D016014:Linear Models; D008297:Male; D008862:Microsomes, Liver / Q000378:metabolism; D051381:Rats; D015203:Reproducibility of Results; D052616:Solid Phase Extraction; D013050:Spectrometry, Fluorescence / Q000379:methods*; D013056:Spectrophotometry, Ultraviolet / Q000379:methods*; D013469:Sulpiride / Q000097:blood; D013976:Tiapamil Hydrochloride / Q000031:analogs & derivatives* / Q000097:blood* / Q000493:pharmacokinetics; D055815:Young Adult
chemical_list: D013469:Sulpiride; D013976:Tiapamil Hydrochloride