title: Bio-analytical method based on MALDI-MS analysis for the quantification of CIGB-300 anti-tumor peptide in human plasma.
abstract: A fully validated bio-analytical method based on Matrix-Assisted-Laser-Desorption/Ionization-Time of Flight Mass Spectrometry was developed for quantitation in human plasma of the anti-tumor peptide CIGB-300. An analog of this peptide acetylated at the N-terminal, was used as internal standard for absolute quantitation. Acid treatment allowed efficient precipitation of plasma proteins as well as high recovery (approximately 80%) of the intact peptide. No other chromatographic step was required for sample processing before MALDI-MS analysis. Spectra were acquired in linear positive ion mode to ensure maximum sensitivity. The lower limit of quantitation was established at 0.5 μg/mL, which is equivalent to 160 fmol peptide. The calibration curve was linear from 0.5 to 7.5 μg/mL, with R(2)>0.98, and permitted quantitation of highly concentrated samples evaluated by dilution integrity testing. All parameters assessed for five validation batches met the FDA guidelines for industry. The method was successfully applied to analysis of clinical samples obtained in a phase I clinical trial following intravenous administration of CIGB-300 at a dose of 1.6 mg/kg body weight. With the exception of Cmax and AUC, pharmacokinetic parameters were similar for ELISA and MALDI-MS methods.
authors.affiliation: Chemical Synthesis Department, Center for Genetic Engineering and Biotechnology, Havana, Cuba.
authors.forename: Ania
authors.initials: A
authors.lastname: Cabrales-Rico
authors.affiliation: Departament de Ciències Experimentals i de la Salut, UPF/PRBB, Barcelona, Spain.
authors.forename: Beatriz G
authors.initials: BG
authors.lastname: de la Torre
authors.affiliation: Chemical Synthesis Department, Center for Genetic Engineering and Biotechnology, Havana, Cuba.
authors.forename: Hilda E
authors.initials: HE
authors.lastname: Garay
authors.affiliation: Mass Spectrometry Department, Center for Molecular Immunology, Havana, Cuba.
authors.forename: Yoan J
authors.initials: YJ
authors.lastname: Machado
authors.affiliation: Mass Spectrometry Department, Center for Molecular Immunology, Havana, Cuba.
authors.forename: Jose A
authors.initials: JA
authors.lastname: Gómez
authors.affiliation: Mass Spectrometry Department, Center for Molecular Immunology, Havana, Cuba.
authors.forename: Enrique
authors.initials: E
authors.lastname: Audain
authors.affiliation: Mass Spectrometry Department, Center for Molecular Immunology, Havana, Cuba.
authors.forename: Orlando
authors.initials: O
authors.lastname: Morales
authors.affiliation: Mass Spectrometry Laboratory, Proteomics Department, CIGB, Havana, Cuba.
authors.forename: Vladimir
authors.initials: V
authors.lastname: Besada
authors.affiliation: Quality Assurance Unit, CIGB, Havana, Cuba.
authors.forename: Jose Luis
authors.initials: JL
authors.lastname: Marcelo
authors.affiliation: Pharmaceutical Research Division, CIGB, Havana, Cuba.
authors.forename: Vilcy
authors.initials: V
authors.lastname: Reyes
authors.affiliation: Pharmaceutical Research Division, CIGB, Havana, Cuba.
authors.forename: Yasser
authors.initials: Y
authors.lastname: Perera
authors.affiliation: Pharmaceutical Research Division, CIGB, Havana, Cuba.
authors.forename: Silvio E
authors.initials: SE
authors.lastname: Perea
authors.affiliation: Chemical Synthesis Department, Center for Genetic Engineering and Biotechnology, Havana, Cuba.
authors.forename: Osvaldo
authors.initials: O
authors.lastname: Reyes
authors.affiliation: Mass Spectrometry Laboratory, Proteomics Department, CIGB, Havana, Cuba. Electronic address: luis.javier@cigb.edu.cu.
authors.forename: Luis Javier
authors.initials: LJ
authors.lastname: González
mesh_terms: D000107:Acetylation; D000970:Antineoplastic Agents / Q000097:blood* / Q000737:chemistry; D002986:Clinical Trials as Topic; D006801:Humans; D007275:Injections, Intravenous; D057230:Limit of Detection; D009369:Neoplasms / Q000097:blood / Q000188:drug therapy; D010456:Peptides, Cyclic / Q000097:blood* / Q000737:chemistry; D012015:Reference Standards; D015203:Reproducibility of Results; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization / Q000295:instrumentation / Q000379:methods*
chemical_list: D000970:Antineoplastic Agents; D010456:Peptides, Cyclic; C532510:CIGB-300
keywords: Anti-tumor peptide; Bio-analytical method; Full validation; Human plasma; MALDI-TOF