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  • Quantification of trabectedin in human plasma: validation of a high-performance liquid chromatography-mass spectrometry method and its application in a clinical pharmacokinetic study.

Quantification of trabectedin in human plasma: validation of a high-performance liquid chromatography-mass spectrometry method and its application in a clinical pharmacokinetic study.

Journal of pharmaceutical and biomedical analysis (2014-03-26)
Monique Zangarini, Laura Ceriani, Federica Sala, Elena Marangon, Renzo Bagnati, Maurizio D'Incalci, Federica Grosso, Massimo Zucchetti
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.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
Methanol, suitable for HPLC, ≥99.9%
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Methanol, anhydrous, 99.8%
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Methanol, HPLC Plus, ≥99.9%
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Ammonium acetate, suitable for mass spectrometry (MS), LiChropur, eluent additive for LC-MS
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Ammonium sulfate, ACS reagent, ≥99.0%
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Ammonium acetate, reagent grade, ≥98%
USP
Methyl alcohol, United States Pharmacopeia (USP) Reference Standard
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Ammonium acetate, ACS reagent, ≥97%
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Ammonium acetate, ≥99.99% trace metals basis
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Methanol, Pharmaceutical Secondary Standard; Certified Reference Material
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Ammonium sulfate, BioXtra, ≥99.0%
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Ammonium acetate, BioUltra, for molecular biology, ≥99.0%
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Methanol, BioReagent, ≥99.93%
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Methanol, ACS spectrophotometric grade, ≥99.9%
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Ammonium acetate solution, BioUltra, for molecular biology, ~5 M in H2O
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Ammonium acetate, BioXtra, ≥98%
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Ammonium acetate, 99.999% trace metals basis
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Methanol, ACS reagent, ≥99.8%
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Methanol, puriss., meets analytical specification of Ph Eur, ≥99.7% (GC)