Stereoselective determination of citalopram and desmethylcitalopram in human plasma and breast milk by liquid chromatography tandem mass spectrometry.
A high performance liquid chromatography (HPLC) tandem mass spectrometry (MS/MS) method was developed for the simultaneous, stereoselective quantification of the antidepressant citalopram and its active metabolite desmethylcitalopram in human plasma and breast milk. Sample preparation was performed by a two-step approach, including generic protein precipitation with acetonitrile followed by solid phase extraction. Enantiospecific separation of analytes was achieved on a Phenomenex®Lux Cellulose-2 column (4.6mm×150mm; 5μm), using reversed phase chromatography conditions characterized by a gradient elution of ammonium acetate buffer (pH 9.0; 20mM) and acetonitrile at a flow rate of 0.6ml/min. The compounds were detected by a tandem quadrupole mass spectrometer equipped with an electrospray ionization source and operating in multiple reaction monitoring mode. The method was fully validated in both biological fluids over a large concentration range of 0.1-100ng/ml for S-(+)- and R-(-)-citalopram, and 0.3-100ng/ml for S-(+)- and R-(-)-desmethylcitalopram. Trueness (90.0-113.3% and 97.1-103.6%), repeatability (0.9-15.9% and 0.9-8.4%) and intermediate precision (1.3-17.8% and 0.9-9.6%) in plasma and breast milk, respectively, meet international guidelines for method validation. Internal standard-normalized matrix effects ranged between 99 and 101% and 98-105%, respectively. The accuracy profiles (total error of trueness and precision) were mostly within the acceptance limits for biological samples defined as ±30%. The method was successfully applied to patient samples in a clinical trial setting.
School of Pharmaceutical Sciences, University of Geneva and University of Lausanne, Geneva, Switzerland.
Etienne
E
Weisskopf
School of Pharmaceutical Sciences, University of Geneva and University of Lausanne, Geneva, Switzerland; Division of Clinical Pharmacology, Lausanne University Hospital, Lausanne, Switzerland.
Alice
A
Panchaud
Department of Neonatology, Hospices Civils de Lyon, Lyon, France.
Kim An
KA
Nguyen
EPICIME-CIC 1407, Inserm, Service of Clinical Pharmacology, CHU-Lyon, Bron, France.
Delphine
D
Grosjean
Department of Neonatology, Maternité Régionale, Université de Lorraine, Nancy, France.
Jean-Michel
JM
Hascoët
School of Pharmaceutical Sciences, University of Geneva and University of Lausanne, Geneva, Switzerland; Division of Clinical Pharmacology, Lausanne University Hospital, Lausanne, Switzerland.
Chantal
C
Csajka
School of Pharmaceutical Sciences, University of Geneva and University of Lausanne, Geneva, Switzerland; Unit of Pharmacogenetics and Clinical Psychopharmacology, Centre for Psychiatric Neuroscience, Department of Psychiatry, Lausanne University Hospital, Prilly, Switzerland.
Chin B
CB
Eap
Unit of Pharmacogenetics and Clinical Psychopharmacology, Centre for Psychiatric Neuroscience, Department of Psychiatry, Lausanne University Hospital, Prilly, Switzerland. Electronic address: nicolas.ansermot@chuv.ch.
Nicolas
N
Ansermot
D002851:Chromatography, High Pressure Liquid / Q000379:methods; D056148:Chromatography, Reverse-Phase / Q000379:methods; D015283:Citalopram / Q000031:analogs & derivatives* / Q000097:blood* / Q000378:metabolism*; D006801:Humans; D008895:Milk, Human / Q000378:metabolism*; D015203:Reproducibility of Results; D052616:Solid Phase Extraction; D021241:Spectrometry, Mass, Electrospray Ionization / Q000379:methods; D013237:Stereoisomerism; D053719:Tandem Mass Spectrometry / Q000379:methods
D015283:Citalopram; C046304:desmethylcitalopram
Breast milk; Citalopram; Enantiomer; LC–MS/MS; Plasma; Quantification