Determination of haloperidol and reduced haloperidol in human serum by liquid chromatography after fluorescence labeling based on the Suzuki coupling reaction

• Published in: Analytical and bioanalytical chemistry 2006-10, Vol.386 (3), p.719-724
• Main Authors: Kishikawa, Naoya, Hamachi, Chiyuki, Imamura, Yoshihiro, Ohba, Yoshihito, Nakashima, Kenichiro, Tagawa, Yashuhiro, Kuroda, Naotaka
• Format: Article
• Language: English
• Subjects: Boron Compounds - chemistry; Chromatography, High Pressure Liquid - methods; Drug Monitoring; Female; Fluorescence; Fluorescence derivatization; Fluorescent arylboronic acid; haloperidol; Haloperidol - analogs & derivatives; Haloperidol - blood; Haloperidol - chemistry; Humans; Male; Molecular Structure; Oxidation-Reduction; Reduced haloperidol; Reference Values; Reproducibility of Results; Schizophrenia - blood; Sensitivity and Specificity; Suzuki coupling reaction; Therapeutic drug monitoring
• ISSN: 1618-2642; 1618-2650
• DOI: 10.1007/s00216-006-0755-0

A simultaneous method for the determination of haloperidol (HP) and its metabolite, reduced haloperidol (RHP), in human serum was developed by means of high-performance liquid chromatography (HPLC) with fluorescence detection. Suzuki coupling reaction with a fluorescent arylboronic acid, 4-(4,5-diphenyl-1H-imidazol-2-yl)phenylboronic acid (DPA), was employed to convert HP and RHP into highly fluorescent compounds. HP and RHP were extracted from human serum by liquid-liquid extraction with a mixture of n-hexane and isoamyl alcohol (99:1, v/v) and subsequently labeled by reaction with DPA. Separation of DPA derivatives of HP and RHP was performed on a silica column with a mixture of acetonitrile and H₂O (90:10, v/v) containing triethylamine and acetic acid as a mobile phase. The proposed method allowed sensitive detection of HP and RHP in human serum with a detection limit (at a signal to noise ratio of 3) of 0.22 and 0.20 ng/mL, respectively. The applicability of the method for therapeutic drug monitoring (TDM) was demonstrated by analyzing human serum samples from schizophrenic patients receiving HP.