title: Screening procedure for the analysis of distigmine bromide in serum by high-performance liquid chromatography-electrospray ionization mass spectrometry.
abstract: A screening procedure was developed for the identification and quantification of distigmine bromide in serum samples by using liquid chromatography (LC)-electrospray ionization (ESI)-mass spectrometry (MS). In this method, distigmine bromide was analyzed in 0.5 mL serum by using pancuronium bromide as the internal standard, and gradient elution was performed using a reversed-phase column and a mixture of 10 mM-ammonium formate and methanol as the mobile phase. A highly sensitive assay could be performed with simple solid phase extraction using a cation exchange cartridge column by carrying out selected ion monitoring analysis in the positive ion detection mode. The procedure was validated in terms of linearity (0.997<r(2)<0.999 for concentrations ranging from 5 to 250 ng/mL), extraction recovery (83.0% to 89.3%, n=5), and detection limit (S/N ratio, >3 at 2.5 ng/mL). The inter- and intra-day precisions (coefficient of variation; CV%) were <8.5% and < 9.7%, respectively. The analytes were evaluated for stability and were found to be stable in serum for 1 week at 4 degrees C and 4 weeks at -30 degrees C, and successfully applied to in the analysis of two overdose cases. This method is sensitive and useful for the detection, quantification, and confirmation of distigmine bromide in serum.
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: Fumiko
authors.initials: F
authors.lastname: Satoh
authors.forename: Kozo
authors.initials: K
authors.lastname: Tamura
authors.forename: Hiroyuki
authors.initials: H
authors.lastname: Otsuka
authors.forename: Shigeaki
authors.initials: S
authors.lastname: Inoue
authors.forename: Isotoshi
authors.initials: I
authors.lastname: Yamamoto
authors.forename: Sadaki
authors.initials: S
authors.lastname: Inokuchi
mesh_terms: D000328:Adult; D000368:Aged; D002138:Calibration; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D005260:Female; D006801:Humans; D008297:Male; D011726:Pyridinium Compounds / Q000097:blood*; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D021241:Spectrometry, Mass, Electrospray Ionization / Q000379:methods*
chemical_list: D011726:Pyridinium Compounds; C084645:distigmine