title: Quantification of trabectedin in human plasma: validation of a high-performance liquid chromatography-mass spectrometry method and its application in a clinical pharmacokinetic study.
abstract: A rapid, sensitive and specific HPLC-MS/MS method was developed and validated for the quantification of trabectedin in human plasma after using deuterated trabectedin as Internal Standard (IS). After the addition of ammonium sulphate, the analyte was extracted from human plasma with acidified methanol (0.1 M HCl). Chromatographic separation was done on an Accucore XL C₁₈ column (4 μm; 50 mm × 2.1 mm) using a Mobile Phase (MP) consisting of CH₃COONH₄ 10 mM, pH 6.8 (MP A) and CH₃OH (MP B). The mass spectrometer worked with electrospray ionization in positive ion mode and Selected Reaction Monitoring (SRM), using target ions at [M-H₂O+H]⁺ m/z 744.4 for trabectedin and [M-H₂O+H]⁺m/z 747.5 for the IS. The standard curve was linear (R² ≥ 0.9955) over the concentration range 0.025-1.0 ng/ml and had good back-calculated accuracy and precision. The intra- and inter-day precision and accuracy determined on three quality control samples (0.04, 0.08 and 0.80 ng/ml) were <9.9% and between 98.3% and 105.3%, respectively. The extraction recovery was >81% and the lower limit of quantification 0.025 ng/ml. The method was successfully applied to study trabectedin pharmacokinetics in a patient with a liposarcoma who received 1.3 mg/m² as a 24 h continuous i.v. infusion.
authors.affiliation: IRCCS-Istituto di Ricerche Farmacologiche "Mario Negri", Laboratory of Cancer Pharmacology, Department of Oncology, Via G. La Masa 19, 20156 Milan, Italy.
authors.forename: Monique
authors.initials: M
authors.lastname: Zangarini
authors.affiliation: IRCCS-Istituto di Ricerche Farmacologiche "Mario Negri", Laboratory of Cancer Pharmacology, Department of Oncology, Via G. La Masa 19, 20156 Milan, Italy.
authors.forename: Laura
authors.initials: L
authors.lastname: Ceriani
authors.affiliation: IRCCS-Istituto di Ricerche Farmacologiche "Mario Negri", Laboratory of Cancer Pharmacology, Department of Oncology, Via G. La Masa 19, 20156 Milan, Italy.
authors.forename: Federica
authors.initials: F
authors.lastname: Sala
authors.affiliation: Centro di Riferimento Oncologico (CRO), National Cancer Institute, Aviano (PN), Italy.
authors.forename: Elena
authors.initials: E
authors.lastname: Marangon
authors.affiliation: IRCCS-Istituto di Ricerche Farmacologiche "Mario Negri", Department of Environmental Health, Via G. La Masa 19, 20156 Milan, Italy.
authors.forename: Renzo
authors.initials: R
authors.lastname: Bagnati
authors.affiliation: IRCCS-Istituto di Ricerche Farmacologiche "Mario Negri", Laboratory of Cancer Pharmacology, Department of Oncology, Via G. La Masa 19, 20156 Milan, Italy.
authors.forename: Maurizio
authors.initials: M
authors.lastname: D'Incalci
authors.affiliation: Oncology, SS Antonio e Biagio General Hospital, Via Venezia 16, Alessandria, Italy.
authors.forename: Federica
authors.initials: F
authors.lastname: Grosso
authors.affiliation: IRCCS-Istituto di Ricerche Farmacologiche "Mario Negri", Laboratory of Cancer Pharmacology, Department of Oncology, Via G. La Masa 19, 20156 Milan, Italy. Electronic address: massimo.zucchetti@marionegri.it.
authors.forename: Massimo
authors.initials: M
authors.lastname: Zucchetti
mesh_terms: D018906:Antineoplastic Agents, Alkylating / Q000097:blood*; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D004149:Dioxoles / Q000097:blood* / Q000493:pharmacokinetics; D006801:Humans; D057230:Limit of Detection; D053719:Tandem Mass Spectrometry / Q000379:methods*; D044005:Tetrahydroisoquinolines / Q000097:blood* / Q000493:pharmacokinetics; D000077606:Trabectedin
chemical_list: D018906:Antineoplastic Agents, Alkylating; D004149:Dioxoles; D044005:Tetrahydroisoquinolines; D000077606:Trabectedin
keywords: HPLC–MS/MS; Human plasma; Liposarcoma; Pharmacokinetics; Trabectedin