title: Development and validation of a rapid and sensitive assay for the determination of anisodamine in 50 μL of beagle dog plasma by LC-MS/MS.
abstract: A simple, rapid, high-throughput, and highly sensitive LC-MS/MS was developed to determine anisodamine in a small volume (50 μL) of beagle dog plasma using atropine sulfate as the internal standard. The analyte and internal standard were isolated from 50 μL plasma samples after a one-step protein precipitation using Sirocco 96-well protein precipitation filtration plates. The separation was accomplished on a Hanbon Hedera CN column (100 × 4.6 mm, 5 μm) and the run time was 4 min. A Micromass Quatro Ultima mass spectrometer equipped with an ESI source was operated in the multiple reaction monitoring mode with the precursor-to-product ion transitions m/z 306.0→140.0 (anisodamine) and 290.0→123.9 (atropine) used for quantitation. The method was sensitive with a low LOQ of 0.05 ng/mL, and good linearity in the range 0.05-50 ng/mL for anisodamine (r(2) ≥ 0.995). All the validation data, such as accuracy, intra- and interrun precision, were within the required limits. The method was successfully applied to the pharmacokinetic study of anisodamine hydrochloride injection in beagle dogs.
authors.affiliation: Department of Pharmaceutical Analysis, School of Pharmacy, Second Military Medical University, Shanghai, P. R. China; College of Pharmacy, Fujian University of Traditional Chinese Medicine, Fuzhou, Fujian, P. R. China; Shanghai Key Laboratory for Pharmaceutical Metabolite Research, Shanghai, P. R. China.
authors.forename: Wenxue
authors.initials: W
authors.lastname: Li
authors.forename: Jun
authors.initials: J
authors.lastname: Wen
authors.forename: Jingyu
authors.initials: J
authors.lastname: He
authors.forename: Di
authors.initials: D
authors.lastname: Cao
authors.forename: Fanlu
authors.initials: F
authors.lastname: Sun
authors.forename: Jinying
authors.initials: J
authors.lastname: Li
authors.forename: Guorong
authors.initials: G
authors.lastname: Fan
mesh_terms: D000818:Animals; D002851:Chromatography, High Pressure Liquid / Q000295:instrumentation / Q000379:methods*; D004285:Dogs; D015394:Molecular Structure; D012991:Solanaceous Alkaloids / Q000097:blood* / Q000737:chemistry; D053719:Tandem Mass Spectrometry / Q000295:instrumentation / Q000379:methods*
chemical_list: D012991:Solanaceous Alkaloids; C003922:anisodamine
keywords: Anisodamine; Dog plasma; LC-MS/MS; Pharmacokinetics; Tropane alkaloids