title: Peroxyoxalate chemiluminescence detection for the highly sensitive determination of fluorescence-labeled chlorpheniramine with Suzuki coupling reaction.
abstract: A sensitive and selective high performance liquid chromatography-peroxyoxalate chemiluminescence (PO-CL) method has been developed for the simultaneous determination of chlorpheniramine (CPA) and monodesmethyl chlorpheniramine (MDCPA) in human serum. The method combines fluorescent labeling with 4-(4,5-diphenyl-1H-imidazole-2-yl)phenyl boronic acid using Suzuki coupling reaction with PO-CL detection. CPA and MDCPA were extracted from human serum by liquid-liquid extraction with n-hexane. Excess labeling reagent, which interfered with trace level determination of analytes, was removed by solid-phase extraction using a C18 cartridge. Separation of derivatives of both analytes was achieved isocratically on a silica column with a mixture of acetonitrile and 60 mM imidazole-HNO(3) buffer (pH 7.2; 85:15, v/v) containing 0.015% triethylamine. The proposed method exhibited a good linearity with a correlation coefficient of 0.999 for CPA and MDCPA within the concentration range of 0.5-100 ng/mL. The limits of detection (S/N = 3) were 0.14 and 0.16 ng/mL for CPA and MDCPA, respectively. Using the proposed method, CPA could be selectively determined in human serum after oral administration.
authors.affiliation: Graduate School of Biomedical Sciences, Course of Pharmaceutical Sciences, Nagasaki University, 1-14 Bunkyo-machi, Nagasaki, 852-8521, Japan.
authors.forename: Lawrence Asamoah
authors.initials: LA
authors.lastname: Adutwum
authors.forename: Naoya
authors.initials: N
authors.lastname: Kishikawa
authors.forename: Kaname
authors.initials: K
authors.lastname: Ohyama
authors.forename: Shiro
authors.initials: S
authors.lastname: Harada
authors.forename: Kenichiro
authors.initials: K
authors.lastname: Nakashima
authors.forename: Naotaka
authors.initials: N
authors.lastname: Kuroda
mesh_terms: D002744:Chlorpheniramine / Q000031:analogs & derivatives* / Q000097:blood* / Q000737:chemistry; D002851:Chromatography, High Pressure Liquid / Q000379:methods; D005453:Fluorescence; D005456:Fluorescent Dyes / Q000737:chemistry; D006801:Humans; D008163:Luminescent Measurements / Q000379:methods*; D010070:Oxalates / Q000737:chemistry; D012680:Sensitivity and Specificity
chemical_list: D005456:Fluorescent Dyes; D010070:Oxalates; D002744:Chlorpheniramine; C044622:peroxyoxalate