Immunoaffinity extraction for liquid chromatographic determination of equilin and its metabolites in plasma.
Immunoaffinity extraction for the high-performance liquid chromatographic determination of equilin and its metabolites in plasma has been achieved. The antibody raised against an equilin 3-O-carboxymethyl ether-bovine serum albumin conjugate was characterized as having a high affinity for equilin and equilenin. One mL of the immunoaffinity adsorbent prepared by immobilization of an antibody was capable of retaining up to 1 microgram of equilin and equilenin, to 100 ng of other metabolites including 2-methoxylated and 17 beta-reduced compounds, and to 0.3 micrograms of glucuronic acid conjugates at C-3. The adsorbates were recovered qualitatively by elution with 90% aqueous (v/v) methanol without any interfering peaks on the chromatogram.
Ikegawa
S
S
Faculty of Pharmaceutical Sciences, Tohoku University, Sendai, Japan.
Itoh
M
M
Murao
N
N
Kijima
H
H
Suzuki
M
M
Fujiyama
T
T
Goto
J
J
D002846:Chromatography, Affinity; D004857:Equilin / Q000097:blood*; D006801:Humans; D007120:Immunochemistry; D007124:Immunoenzyme Techniques; D010084:Oxidation-Reduction; D012710:Serum Albumin, Bovine; D013056:Spectrophotometry, Ultraviolet; D000519:alpha-Galactosidase / Q000737:chemistry
D004857:Equilin; D012710:Serum Albumin, Bovine; D000519:alpha-Galactosidase