Analysis of 3-(2-(ethylamino)propyl)-1,2,3,4-tetrahydro-5H(1)benzopyrano (3,4-c)pyridin-5-one in plasma by liquid chromatographic column switching after derivatizing the secondary amine with fluorescein-6-isothiocyanate.
A high-performance liquid chromatographic (HPLC) assay has been developed to quantify the bronchodilator 3-(2-ethylamino)propyl)-1,2,3,4-tetrahydro-5H(1)benzopyrano(3,4-c)pyridi n-5-one (CI-923) (1) in human plasma. The drug and internal standard (2) were extracted from plasma (pH 10.5) with ether, and the secondary amines were derivatized with fluorescein-6-isothiocyanate (F6ITC) (3). The derivatives were isolated from the mixture by on-line HPLC column switching and were quantified with fluorometric detection (Ex: 484 nm; Em: 518 nm). Linearity was observed over the range 0.5-50.0 ng/mL of 1, and the recovery of drug from plasma was greater than 86%. The precision of the method was 3.7-5.2% RSD based on the analysis of seeded control pools containing 0.8, 20.0 and 40.0 ng/mL of 1. The accuracy of the method was +/- 3.3% relative error for the same three pools.
D001993:Bronchodilator Agents / Q000097:blood*; D055598:Chemical Phenomena; D002621:Chemistry; D002851:Chromatography, High Pressure Liquid; D002853:Chromatography, Liquid; D003374:Coumarins / Q000097:blood*; D016650:Fluorescein-5-isothiocyanate; D005452:Fluoresceins; D006801:Humans; D013861:Thiocyanates
D001993:Bronchodilator Agents; D003374:Coumarins; D005452:Fluoresceins; D013861:Thiocyanates; D016650:Fluorescein-5-isothiocyanate; C047548:CI 923