High-performance liquid chromatographic method for the quantification of anthranilic and 3-hydroxyanthranilic acid in rat brain dialysate.
Anthranilic acid (ANA) and 3-hydroxyanthranilic acid (3-HANA) have attracted considerable attention as two of the L-tryptophan kynurenine pathway metabolites in the central nervous system. In this study, a highly sensitive and accurate method for the quantification of ANA and 3-HANA has been developed using reversed-phase high performance liquid chromatography (HPLC) with fluorimetric detection. The HPLC assay was carried out using a C(18) column (5 microm, 250 x 4.6 mm i.d.). The mobile phase consisted of a mixture of 25 mM sodium/acetic acid buffer (pH 5.5) and methanol (90:10 v/v). Fluorimetric detection at lambda(ex)=316 nm and lambda(em)=420 nm was used. The assay was applied to the measurement of ANA and 3-HANA acid in rat brain dialysate following administration of L-tryptophan or L-kynurenine. 3-HANA and ANA levels were progressively increased during 90 min following administration of L-tryptophan, then decreased progressively to basal levels. 3-HANA levels were significantly higher than ANA levels after L-kynurenine administration. These findings suggest that the assay developed should provide an improved means for investigation of neurobiology of kynurenine pathway.
Cannazza
G
G
Dipartimento di Scienze Farmaceutiche, Università degli Studi di Modena e Reggio Emilia, Via Campi 183, 41100 Modena, Italy. pippi@unimo.it
Baraldi
M
M
Braghiroli
D
D
Tait
A
A
Parenti
C
C
D015095:3-Hydroxyanthranilic Acid / Q000032:analysis* / Q000378:metabolism; D000818:Animals; D001921:Brain / Q000378:metabolism*; D002851:Chromatography, High Pressure Liquid / Q000379:methods; D008297:Male; D017551:Microdialysis / Q000379:methods*; D051381:Rats; D017207:Rats, Sprague-Dawley; D062367:ortho-Aminobenzoates / Q000032:analysis* / Q000378:metabolism
D062367:ortho-Aminobenzoates; C031385:anthranilic acid; D015095:3-Hydroxyanthranilic Acid