title: Sensitive isotope dilution liquid chromatography/tandem mass spectrometry method for quantitative analysis of bumetanide in serum and brain tissue.
abstract: We have developed and validated a simple and sensitive stable isotope dilution liquid chromatography/tandem mass spectrometric (LC-MS/MS) method for the quantification of bumetanide in human serum. Samples were prepared with a simple acetonitrile based protein precipitation. The supernatant was then analyzed directly using LC-MS/MS. Chromatographic separation was achieved on a C18 reversed phase column using a methanol and water gradient. The detection was performed in selected reaction monitoring (SRM) mode via a positive electrospray ionization (ESI) interface. The method had a lower limit of quantification (LLOQ) of 1 ng/mL, linearity up to 1250 ng/mL, intra- and inter-day precision less than 10%, and accuracy within ±10%. This method was also demonstrated to be suitable for the analysis of bumetanide in rat serum and brain tissue. Bumetanide concentrations in rat serum and brain were determined for samples collected at several intervals following intraperitoneal (i.p.) injection of bumetanide, and were used to calculate bumetanide permeability through the blood-brain barrier.
authors.affiliation: Division of Genetics, Children's Hospital Boston, Harvard Medical School, Boston, MA, USA.
authors.forename: Yijun
authors.initials: Y
authors.lastname: Li
authors.forename: Ryan
authors.initials: R
authors.lastname: Cleary
authors.forename: Mark
authors.initials: M
authors.lastname: Kellogg
authors.forename: Janet S
authors.initials: JS
authors.lastname: Soul
authors.forename: Gerard T
authors.initials: GT
authors.lastname: Berry
authors.forename: Frances E
authors.initials: FE
authors.lastname: Jensen
mesh_terms: D000097:Acetonitriles; D000818:Animals; D001921:Brain / Q000378:metabolism; D001923:Brain Chemistry; D002034:Bumetanide / Q000032:analysis* / Q000097:blood / Q000493:pharmacokinetics; D002853:Chromatography, Liquid / Q000379:methods*; D006801:Humans; D007553:Isotope Labeling; D016018:Least-Squares Analysis; D051381:Rats; D020318:Rats, Long-Evans; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D053719:Tandem Mass Spectrometry / Q000379:methods*
chemical_list: D000097:Acetonitriles; D002034:Bumetanide; C032159:acetonitrile