Enantioselective immunoaffinity extraction for simultaneous determination of optically active bufuralol and its metabolites in human plasma by HPLC.
A combined method of immunoaffinity extraction with high-performance liquid chromatography has been developed for the enantioselective determination of bufuralol and its metabolites in human plasma. The antibodies having high affinity toward the asymmetric center at the C-1 position of bufuralol and its 1'-oxidized metabolites and low affinity to their antipodes were elicited by immunization of rabbits with immunogens, (1R)- and (1S)-1'-oxobufuralol O-carboxymethyloxime-bovine serum albumin conjugates, respectively. 0.5 ml Of the immunoaffinity adsorbent (7.6 mg.ml-1 for anti-(1S)-antibody and 28.8 mg.ml-1 for anti-(1R)-antibody) prepared by immobilization of an antibody was capable of retaining up to 1 microgram of (R)- and (S)-bufuralol and up to 500 ng of other metabolites. The adsorbates were recovered quantitatively by elution with methanol-10 mM ammonium acetate buffer (pH 5) (95:5, v/v) without any interfering peaks on the high-performance liquid chromatogram. The proposed method was evaluated to be useful for the simultaneous determination of optically active bufuralol and its metabolite in plasma with acceptable recovery and precision.
Ikegawa
S
S
Faculty of Pharmaceutical Sciences, Tohoku University, Aobayama, Sendai, Japan.
Matsuura
K
K
Sato
T
T
Isriyanthi
N M
NM
Niwa
T
T
Miyairi
S
S
Takashina
H
H
Kawashima
Y
Y
Goto
J
J
D000319:Adrenergic beta-Antagonists / Q000097:blood* / Q000276:immunology; D002846:Chromatography, Affinity / Q000379:methods*; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D004983:Ethanolamines / Q000097:blood* / Q000276:immunology; D006801:Humans; D007106:Immune Sera; D015203:Reproducibility of Results; D013237:Stereoisomerism
D000319:Adrenergic beta-Antagonists; D004983:Ethanolamines; D007106:Immune Sera; C010831:bufuralol