Immunoaffinity Extraction for Liquid Chromatographic Determination of Equilin and its Metabolites in Plasma

✍ Scribed by Shigeo Ikegawa; Manabu Itoh; Naoaki Murao; Hiromi Kijima; Mio Suzuki; Tomoharu Fujiyama; Junichi Goto; Masahiko Tohma

• Publisher: John Wiley and Sons
• Year: 1996
• Tongue: English
• Weight: 432 KB
• Volume: 10
• Category: Article
• ISSN: 0269-3879
• DOI: 10.1002/(sici)1099-0801(199603)10:2<73::aid-bmc561>3.0.co;2-5

✦ Synopsis

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 3-0-carboxymethyl etherbovine 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 pg of equilin and equilenin, to 100 ng of other metabolites including 2-methoxylated and l'lpreduced compounds, and to 0.3 pg of glucuronic acid conjugates at C-3. The adsorbates were recovered quantitatively by elution with 90% aqueous ( v h ) methanol without any interfering peaks on the chromatogram. R p H , RpB-OH DHEqu : R'=H, Rz-P-OH N -H y d r o x y s u c c i n i m i d y l 2-MeOEqu : RlzOCH3, Rz=O I-McODHEqu : R1=OCH,, Rz=P-OH 0 figure 1. Structures of equilin and related compounds.