title: A simple dried blood spot method for therapeutic drug monitoring of the tricyclic antidepressants amitriptyline, nortriptyline, imipramine, clomipramine, and their active metabolites using LC-MS/MS.
abstract: Therapeutic drug monitoring (TDM) of tricyclic antidepressants (TCAs) is considered useful in patients with major depressive disorder, since these drugs display large individual differences in clearance, and the therapeutic windows of these drugs are relatively small. We developed an assay for determination of amitriptyline (ATP), nortriptyline (NTP), imipramine (IMP), desipramine (DSP) clomipramine (CMP) and desmethyl-clomipramine (DCMP) in dried blood spots (DBS). A fast and robust LC-MS/MS method was developed and analytically validated for simultaneous determination of ATP, NTP, IMP, DSP, CMP, and DCMP in DBS. Six mm circles were punched out from DBS collected on Whatman DMPK-C paper and mixed with acetonitrile: methanol 1:3 containing the internal standard. The extract was analyzed by LC-MS/MS. Total LC-MS/MS runtime was 4.8 min. The assay was linear in the range 20-500 µg/L for all compounds. Overall-assay accuracy and precision were<20% for the lower limit of quantification (LLOQ), except for CMP (CV=22.3%), and <15% at other concentrations. The initial LLOQ was 20 µg/L however for CMP and DMCP it was increased to 40 µg/L. The blood volume per spot did not influence the results, but a low hematocrit (≤ 30%) was associated with a >15% negative bias for all compounds. Punching at the perimeter of the blood spot instead of the center was associated with a positive bias. A good correlation was found between patients plasma and DBS samples of ATP, NTP and DMCP, but not for CMP. In addition, proportional differences were found. This LC-MS/MS method was analytically validated for determination of TCAs in DBS. Future validation will focus on the clinical application of the method.
authors.affiliation: Dept. of Pharmacy, section Pharmacotherapy and Pharmaceutical Care, University of Groningen, Antonius Deusinglaan 1, 9713 AV Groningen, the Netherlands. Electronic address: e.j.j.berm@rug.nl.
authors.forename: E J J
authors.initials: EJJ
authors.lastname: Berm
authors.affiliation: Laboratory for Drug Analysis &Toxicology, Diaconessen Hospital Meppel& Bethesda Hospital, Hoogeveenseweg 38, 7943 KA, Meppel, the Netherlands.
authors.forename: J
authors.initials: J
authors.lastname: Paardekooper
authors.affiliation: Laboratory for Drug Analysis &Toxicology, Diaconessen Hospital Meppel& Bethesda Hospital, Hoogeveenseweg 38, 7943 KA, Meppel, the Netherlands.
authors.forename: E
authors.initials: E
authors.lastname: Brummel-Mulder
authors.affiliation: Dept. of Pharmacy, section Pharmaco Epidemiology and Pharmaco Economics, University of Groningen, Antonius Deusinglaan 1, 9713 AV Groningen, the Netherlands.
authors.forename: E
authors.initials: E
authors.lastname: Hak
authors.affiliation: Dept. of Pharmacy, section Pharmacotherapy and Pharmaceutical Care, University of Groningen, Antonius Deusinglaan 1, 9713 AV Groningen, the Netherlands; Dept. of Clinical Pharmacy and Pharmacology, University Medical Center Groningen, P.O. Box 30.001, 9700RB, Groningen, the Netherlands.
authors.forename: B
authors.initials: B
authors.lastname: Wilffert
authors.affiliation: Laboratory for Drug Analysis &Toxicology, Diaconessen Hospital Meppel& Bethesda Hospital, Hoogeveenseweg 38, 7943 KA, Meppel, the Netherlands. Electronic address: maring@noorderboog.nl.
authors.forename: J G
authors.initials: JG
authors.lastname: Maring
mesh_terms: D000639:Amitriptyline / Q000008:administration & dosage / Q000097:blood*; D000929:Antidepressive Agents, Tricyclic / Q000008:administration & dosage / Q000097:blood*; D001711:Biotransformation; D002138:Calibration; D002853:Chromatography, Liquid; D002997:Clomipramine / Q000008:administration & dosage / Q000097:blood*; D003865:Major Depressive Disorder / Q000097:blood* / Q000188:drug therapy; D059788:Dried Blood Spot Testing; D016903:Drug Monitoring / Q000379:methods; D006400:Hematocrit; D006801:Humans; D007099:Imipramine / Q000008:administration & dosage / Q000097:blood*; D057230:Limit of Detection; D009661:Nortriptyline / Q000008:administration & dosage / Q000097:blood*; D015203:Reproducibility of Results; D053719:Tandem Mass Spectrometry
chemical_list: D000929:Antidepressive Agents, Tricyclic; D000639:Amitriptyline; D009661:Nortriptyline; D002997:Clomipramine; D007099:Imipramine
keywords: LC-MS/MS; Therapeutic drug monitoring; dried blood spot; tricyclic antidepressants