Ikegawa S, Matsuura K, Sato T, Isriyanthi N M, Niwa T, Miyairi S, Takashina H, Kawashima Y, Goto J
Faculty of Pharmaceutical Sciences, Tohoku University, Aobayama, Sendai, Japan.
J Pharm Biomed Anal. 1998 May;17(1):1-9. doi: 10.1016/s0731-7085(97)00147-7.
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.
已开发出一种免疫亲和萃取与高效液相色谱联用的方法，用于对人血浆中布呋洛尔及其代谢物进行对映体选择性测定。通过分别用免疫原（1R）-和（1S）-1'-氧代布呋洛尔O-羧甲基肟-牛血清白蛋白偶联物免疫兔子，制备出了对布呋洛尔C-1位不对称中心及其1'-氧化代谢物具有高亲和力、对其对映体具有低亲和力的抗体。将抗体固定化制备的0.5 ml免疫亲和吸附剂（抗（1S）-抗体为7.6 mg·ml-1，抗（1R）-抗体为28.8 mg·ml-1）能够保留多达1微克的（R）-和（S）-布呋洛尔以及多达500纳克的其他代谢物。用甲醇-10 mM醋酸铵缓冲液（pH 5）（95:5，v/v）洗脱可定量回收吸附物，高效液相色谱图上无任何干扰峰。经评估，所提出的方法可用于同时测定血浆中具有光学活性的布呋洛尔及其代谢物，回收率和精密度均可接受。
