title: The development and validation of a turbulent flow-liquid chromatography-tandem mass spectrometric method for the simultaneous quantification of citalopram, sertraline, bupropion and hydroxybupropion in serum.
abstract: OBJECTIVE
  Depression is a rapidly growing issue in the United States. There are many drug classes that may be used to treat depression, including the selective serotonin-reuptake inhibitors (SSRIs) citalopram (Celexa®) and sertraline (Zoloft®), as well as the aminoketone bupropion (Wellbutrin®). However, therapeutic efficacy and treatment success is often variable, requiring changes in dosing regimens or drug selection. Methods for drug quantification can become important tools in the assessment of drug efficacy to optimize treatment regimens. Here, we present a turbulent flow-liquid chromatography-tandem mass spectrometric (TFC-MS/MS) method for the robust, simultaneous quantification of citalopram, sertraline, bupropion and its active metabolite, hydroxybupropion (OH-bupropion).
  
  
  METHODS
  Serum spiked with the aforementioned antidepressants, along with their corresponding isotopically labeled internal standards was subjected to protein precipitation. Samples were injected onto a TFC column for on-line solid phase extraction and a Hypersil Gold C18 column for chromatographic separation. Detection was achieved using a TSQ Vantage mass spectrometer. Assay validation followed FDA bioanalytical guidelines.
  
  
  RESULTS
  The analytical measuring range for all analytes spanned from 5 to 1000ng/mL. Intra- and inter-assay precision across four quality control levels were ≤9.2% and ≤14.8%, respectively. A comparison to other LC-MS/MS methods resulted in a strong correlation with correlation coefficients ranging from 0.9929 to 0.9971. Carryover, stability, recovery, matrix effects, extraction and processing efficiency were also deemed acceptable in accordance with FDA recommendations.
  
  
  CONCLUSIONS
  The development and validation of this TFC-MS/MS method allow for the robust and high-throughput quantification of commonly prescribed antidepressants.
authors.affiliation: Department of Pathology, Johns Hopkins University School of Medicine, Baltimore, MD, USA.
authors.forename: Athena K
authors.initials: AK
authors.lastname: Petrides
authors.affiliation: Department of Pathology, Johns Hopkins University School of Medicine, Baltimore, MD, USA.
authors.forename: Joshua
authors.initials: J
authors.lastname: Moskowitz
authors.affiliation: ARUP Laboratories, University of Utah School of Medicine, Salt Lake City, UT, USA.
authors.forename: Kamisha L
authors.initials: KL
authors.lastname: Johnson-Davis
authors.affiliation: Department of Laboratory Medicine and Pathology, Mayo Clinic College of Medicine, Mayo Clinic, Rochester MN, USA.
authors.forename: Paul J
authors.initials: PJ
authors.lastname: Jannetto
authors.affiliation: Department of Laboratory Medicine and Pathology, Mayo Clinic College of Medicine, Mayo Clinic, Rochester MN, USA.
authors.forename: Loralie J
authors.initials: LJ
authors.lastname: Langman
authors.affiliation: Department of Pathology, Johns Hopkins University School of Medicine, Baltimore, MD, USA.
authors.forename: William
authors.initials: W
authors.lastname: Clarke
authors.affiliation: Department of Pathology, Johns Hopkins University School of Medicine, Baltimore, MD, USA; Department of Medicine, Johns Hopkins University School of Medicine, Baltimore, MD, USA. Electronic address: mmarzin1@jhmi.edu.
authors.forename: Mark A
authors.initials: MA
authors.lastname: Marzinke
mesh_terms: D000928:Antidepressive Agents / Q000097:blood / Q000302:isolation & purification; D016642:Bupropion / Q000031:analogs & derivatives* / Q000097:blood*; D002853:Chromatography, Liquid / Q000379:methods*; D015283:Citalopram / Q000097:blood*; D004355:Drug Stability; D006801:Humans; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D020280:Sertraline / Q000097:blood*; D052616:Solid Phase Extraction; D053719:Tandem Mass Spectrometry / Q000379:methods*
chemical_list: D000928:Antidepressive Agents; C580748:hydroxybupropion; D016642:Bupropion; D015283:Citalopram; D020280:Sertraline
keywords: Antidepressants; Method validation; TFC–MS/MS