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Journal Article
Research Support, U.S. Gov't, P.H.S.
Rapid quantification of the non-competitive NMDA antagonist MK-801 in canine cerebrospinal fluid and plasma by capillary gas chromatography-nitrogen phosphorus detection.
Journal of Pharmaceutical and Biomedical Analysis 2003 Februrary 27
A facile and sensitive method utilizing solid-phase cartridge extraction and capillary gas chromatography (GC) with nitrogen phosphorus detection was validated for the determination of MK-801 [(+)-5-methyl-10,11-dihydro-5H-dibenzo(a,d)cyclo-hepten-5,10-imine maleate], a non-competitive NMDA receptor antagonist, in dog cerebrospinal fluid (CSF) and plasma. Clonidine hydrochloride was used as the internal standard (ISTD), after evaluation of several ISTD candidates. Separations were performed with an intermediate polarity fused silica capillary column, yielding typical retention times of 3.20 min for MK-801 and 4.90 min for ISTD. Plasma and CSF samples were extracted with 100 mg Bond Elut C(18) TCA Copyright cartridges to yield methanolic eluates that were evaporatively enriched before reconstitution in anhydrous ethanol prior to injection. The standard curve was validated from 1 to 100,000 ng/ml for CSF, and from 0.1 to 1,000 ng/ml for plasma. Chromatograms from naive plasma and CSF exhibited no endogenous interfering peaks. The efficiency of extraction recovery was >94%, and the intra-assay and inter-assay precision was within 9% relative standard deviation (%R.S.D.) for both fluids. MK-801 and ISTD were stable in the injection solvent at 22 degrees C for at least 48 h. The assay was applied to the toxocologic study of intrathecal MK-801 administration in the dog.
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