Novel immunoaffinity extraction for liquid chromatographic determination of major metabolites of 4-acetoxy-2-(4-methylphenyl)benzothiazole in plasma.
Group extraction of the metabolites of 4-acetoxy-2-(4-methylphenyl)benzothiazole has been achieved through the use of an immunoaffinity adsorbent. The antisera elicited from an immunogen, 4-hydroxy-2-(4-formylphenyl)benzothiazole O-carboxymethyloxime-bovine serum albumin conjugate, were characterized to have a broad affinity spectrum for major metabolites oxidized at the 4-methyl group of the benzene moiety. One millilitre of the immunoaffinity adsorbent prepared by immobilization of antibodies (12.5 mg ml-1) was capable of retaining up to 4 micrograms of benzothiazoles. The adsorbates were recovered quantitatively by elution with 90% (v/v) methanol without any interfering peaks on the high-performance liquid chromatogram. The peak-height ratio of each metabolite to an internal standard was plotted against the concentration of the former substance; good linearity was observed in the range of 10-500 ng ml-1.
Miyairi
S
S
Pharmaceutical Institute, Tohoku University, Sendai, Japan.
Shimada
H
H
Awata
N
N
Goto
J
J
Nambara
T
T
D000818:Animals; D000918:Antibody Specificity; D052160:Benzothiazoles; D002417:Cattle; D002851:Chromatography, High Pressure Liquid; D006241:Haptens; D007106:Immune Sera; D007163:Immunosorbent Techniques; D012710:Serum Albumin, Bovine; D013844:Thiazoles / Q000097:blood*
D052160:Benzothiazoles; D006241:Haptens; D007106:Immune Sera; D013844:Thiazoles; C088347:4-acetoxy-2-(4-methylphenyl)benzothiazole; D012710:Serum Albumin, Bovine