title: Selective reaction monitoring of negative electrospray ionization acetate adduct ions for the bioanalysis of dapagliflozin in clinical studies.
abstract: Dapagliflozin (Farxiga), alone, or in the fixed dose combination with metformin (Xigduo), is an orally active, highly selective, reversible inhibitor of sodium-glucose cotransporter type 2 (SGLT2) that is marketed in United States, Europe, and many other countries for the treatment of type 2 diabetes mellitus. Here we report a liquid chromatography-tandem mass spectrometry (LC-MS/MS) bioanalytical assay of dapagliflozin in human plasma. A lower limit of quantitation (LLOQ) at 0.2 ng/mL with 50 μL of plasma was obtained, which reflects a 5-fold improvement of the overall assay sensitivity in comparison to the previous most sensitive assay using the same mass spectrometry instrumentation. In this new assay, acetate adduct ions in negative electrospray ionization mode were used as the precursor ions for selective reaction monitoring (SRM) detection. Sample preparation procedures and LC conditions were further developed to enhance the column life span and achieve the separation of dapagliflozin from potential interferences, especially its epimers. The assay also quantifies dapagliflozin's major systemic circulating glucuronide metabolite, BMS-801576, concentrations in human plasma. The assay was successfully transferred to contract research organizations (CROs), validated, and implemented for the sample analysis of pediatric and other critical clinical studies. This assay can be widely used for bioanalytical support of future clinical studies for the newly approved drug Farxiga or any combination therapy containing dapagliflozin.
authors: †Research and Development, Bristol-Myers Squibb, Princeton, New Jersey 08543, United States.
authors: Qin C
authors: QC
authors: Ji
authors: †Research and Development, Bristol-Myers Squibb, Princeton, New Jersey 08543, United States.
authors: Xiaohui
authors: X
authors: Xu
authors: ‡PPD, Richmond, Virginia 23230, United States.
authors: Eric
authors: E
authors: Ma
authors: †Research and Development, Bristol-Myers Squibb, Princeton, New Jersey 08543, United States.
authors: Jane
authors: J
authors: Liu
authors: †Research and Development, Bristol-Myers Squibb, Princeton, New Jersey 08543, United States.
authors: Shenita
authors: S
authors: Basdeo
authors: †Research and Development, Bristol-Myers Squibb, Princeton, New Jersey 08543, United States.
authors: Guowen
authors: G
authors: Liu
authors: ‡PPD, Richmond, Virginia 23230, United States.
authors: William
authors: W
authors: Mylott
authors: †Research and Development, Bristol-Myers Squibb, Princeton, New Jersey 08543, United States.
authors: David W
authors: DW
authors: Boulton
authors: †Research and Development, Bristol-Myers Squibb, Princeton, New Jersey 08543, United States.
authors: Jim X
authors: JX
authors: Shen
authors: †Research and Development, Bristol-Myers Squibb, Princeton, New Jersey 08543, United States.
authors: Bruce
authors: B
authors: Stouffer
authors: †Research and Development, Bristol-Myers Squibb, Princeton, New Jersey 08543, United States.
authors: Anne-Françoise
authors: AF
authors: Aubry
authors: †Research and Development, Bristol-Myers Squibb, Princeton, New Jersey 08543, United States.
authors: Mark E
authors: ME
authors: Arnold
mesh_terms: D000085:Acetates / Q000737:chemistry*; D053000:Analytic Sample Preparation Methods; D001559:Benzhydryl Compounds / Q000097:blood* / Q000737:chemistry*; D001774:Blood Chemical Analysis / Q000379:methods*; D005960:Glucosides / Q000097:blood* / Q000737:chemistry*; D006801:Humans; D015203:Reproducibility of Results; D021241:Spectrometry, Mass, Electrospray Ionization / Q000379:methods*; D053719:Tandem Mass Spectrometry
chemical_list: D000085:Acetates; D001559:Benzhydryl Compounds; D005960:Glucosides; C529054:dapagliflozin