title: Simultaneous determination of dibucaine and naphazoline in human serum by monolithic silica spin column extraction and liquid chromatography-mass spectrometry.
abstract: A simple, sensitive, and specific liquid chromatography-mass spectrometry (LC-MS) method for simultaneous determination of dibucaine and naphazoline from serum was developed and validated. The extraction procedure was performed using a monolithic silica spin column. Chromatographic separation of dibucaine and naphazoline was achieved on a C(18) reverse phase column with a mobile phase gradient (mobile phase A: 10 mM ammonium formate and mobile phase B: acetonitrile) at a flow rate of 0.2 mL/min. LC-MS was operated under the selective ion monitoring mode using the electrospray ionization technique in the positive mode. The retention times for naphazoline, dibucaine, and the internal standard (IS) were 6.7, 7.8, and 8.0 min, respectively. A linear graph was obtained for dibucaine and naphazoline with correlation coefficients >0.998 for all analytes by this method. The limit of quantification of dibucaine and naphazoline was 10 and 25 ng/mL, respectively. The mean recoveries were greater than 70%. Both compounds were stable under conditions of short-term storage, long-term storage as well as after freeze-thaw cycles. Monolithic spin column extraction and LC-MS analysis enabled the separation of dibucaine and naphazoline within 20 min.
authors.affiliation: Department of Emergency and Critical Care Medicine, Tokai University School of Medicine, Isehara, Kanagawa 259-1143, Japan. saito@is.icc.u-tokai.ac.jp
authors.forename: Takeshi
authors.initials: T
authors.lastname: Saito
authors.forename: Seiji
authors.initials: S
authors.lastname: Morita
authors.forename: Izumi
authors.initials: I
authors.lastname: Kishiyama
authors.forename: Shota
authors.initials: S
authors.lastname: Miyazaki
authors.forename: Akihiro
authors.initials: A
authors.lastname: Nakamoto
authors.forename: Manami
authors.initials: M
authors.lastname: Nishida
authors.forename: Akira
authors.initials: A
authors.lastname: Namera
authors.forename: Masataka
authors.initials: M
authors.lastname: Nagao
authors.forename: Sadaki
authors.initials: S
authors.lastname: Inokuchi
mesh_terms: D000368:Aged; D000369:Aged, 80 and over; D002138:Calibration; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D003992:Dibucaine / Q000097:blood*; D006801:Humans; D008297:Male; D009278:Naphazoline / Q000097:blood*; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D021241:Spectrometry, Mass, Electrospray Ionization / Q000379:methods*
chemical_list: D009278:Naphazoline; D003992:Dibucaine