title: Determination of A 3,4-diaminopyridine in plasma by liquid chromatography with electrochemical detection using solid-phase extraction.
abstract: In order to quantify a small amount of a drug, 3,4-diaminopyridine (3,4-DAP), in animal plasma samples, an analytical method was developed. It involved an extraction of 3,4-DAP and phenylephrine, used as internal standard (IS), from plasma with solid-phase extraction (SPE) on C18 cartridges. This analytical method is a hyphenated technique based on high-performance liquid chromatography with electrochemical detection (HPLC-EC) whose purpose is to obtain first a sensitive method and second a satisfying separation between 3,4-DAP and phenylephrine. The analytical method is accurate, specific, and linear between 10 and 500 g of 3,4-DAP per litre. The recovery of 3,4-DAP is estimated at 70.8% with a 95% confidence interval of (66.0 -75.6%). Intermediate precision was evaluated on three quality control samples; the intra-day precision was estimated at 13.5, 9.1, 7.8% and the inter-day precision at 17.9, 8.4, 9.3%. The limit of quantification of the method was evaluated at 10 g l-1. First toxicokinetic parameters determined on dogs plasma samples after one 3,4-DAP oral administration of 1 mg kg-1 were: Cmax=395.7 microg l-1; Tmax =15 min; t1/2=113.6 min; Clearance/F=16.8 ml kg-1 min-1 and Vd/F=2.7 l kg -1.
authors.affiliation: Analytical Development Department, 75005 Paris, France. sophie.goulay@eps.ap-hop-paris.fr
authors.forename: S
authors.initials: S
authors.lastname: Goulay-Dufaÿ
authors.forename: B
authors.initials: B
authors.lastname: Do
authors.forename: M D
authors.initials: MD
authors.lastname: Le Hoang
authors.forename: J A
authors.initials: JA
authors.lastname: Raust
authors.forename: H
authors.initials: H
authors.lastname: Graffard
authors.forename: F
authors.initials: F
authors.lastname: Guyon
authors.forename: D
authors.initials: D
authors.lastname: Pradeau
mesh_terms: D015761:4-Aminopyridine / Q000031:analogs & derivatives* / Q000097:blood*; D000077770:Amifampridine; D000818:Animals; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D004285:Dogs; D004563:Electrochemistry / Q000379:methods*; D005260:Female; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity
chemical_list: D015761:4-Aminopyridine; D000077770:Amifampridine