title: Determination of a novel calmodulin antagonist, 3-[2-[4-(3-chloro-2-methylphenyl)-1-piperazinyl]ethyl]-5,6-dimethoxy-1- (4-imidazolylmethyl)-1H-indazole dihydrochloride 3.5 hydrate, DY-9760e, in human plasma using solid-phase extraction and high-performance liquid chromatography with fluorescence detection.
abstract: A high-performance liquid chromatographic method for the determination of a novel calmodulin antagonist, 3-[2-[4-(3-chloro-2-methylphenyl)-1-piperazinyl]ethyl]-5,6-dimethoxy-1-( 4-imidazolylmethyl)-1H-indazole dihydrochloride 3.5 hydrate, DY-9760e and its major metabolite, 3-[2-[4-(3-chloro-2-methylphenyl)-1-piperazinyl]ethyl]-5,6-dimethoxyi ndazole, DY-9836 in human plasma has been developed. DY-9760e, DY-9836 and the internal standard (I.S.) were extracted from plasma by means of an Isolute C18 (EC) column. The extracts were chromatographed on a reversed-phase TSK-gel ODS-80Ts column using 0.1 M acetate buffer (pH 5)-CH3CN (65:35, v/v) as the mobile phase at a flow-rate of 1.0 ml/min. Fluorescence detection at an excitation wavelength of 303 nm and an emission wavelength of 347 nm resulted in a limit of quantitation of 1.000 ng/ml for plasma. The method showed satisfactory sensitivity, precision, accuracy, recovery and selectivity. Stability studies showed that DY-9760 and DY-9836 were stable in plasma up to at least eight weeks at -80 degrees C.
authors.affiliation: Drug Metabolism and Analytical Chemistry Research Laboratory, Daiichi Pharmaceutical Co. Ltd., Tokyo, Japan.
authors.forename: S
authors.initials: S
authors.lastname: Tachibana
authors.forename: A
authors.initials: A
authors.lastname: Fukano
authors.forename: K
authors.initials: K
authors.lastname: Sudo
authors.forename: M
authors.initials: M
authors.lastname: Tanaka
mesh_terms: D002021:Buffers; D002147:Calmodulin / Q000037:antagonists & inhibitors*; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D004355:Drug Stability; D006801:Humans; D006863:Hydrogen-Ion Concentration; D007191:Indazoles / Q000097:blood*; D011786:Quality Control; D012680:Sensitivity and Specificity; D012996:Solutions; D013050:Spectrometry, Fluorescence
chemical_list: C109602:3-(2-(4-(3-chloro-2-methylphenyl)1-piperazinyl)ethyl)5,6-dimethoxy-1-(4-imidazolylmethyl)-1H-indazol dihydrochloride 3.5 hydrate; D002021:Buffers; D002147:Calmodulin; D007191:Indazoles; D012996:Solutions