title: Validation of a stability-indicating RP-LC method for the assessment of recombinant human interleukin-11 and its correlation with bioassay.
abstract: A stability-indicating reversed-phase liquid chromatography (RP-LC) method was validated for the assessment of recombinant human interleukin-11 (rhIL-11), based on the ICH guidelines. The method was carried out on a Jupiter C(4) column (250 mm × 4.6 mm i.d.), maintained at 25°C. The mobile phase A consisted of 0.1% trifluoroacetic acid (TFA) and the mobile phase B was acetonitrile with 0.1% TFA, run at a flow rate of 1 mL/min, and using a photodiode array (PDA) detection at 214 nm. Separation was obtained with a retention time of 27.6 min, and was linear over the concentration range of 1-200 µg/mL (r(2) = 0.9995). Specificity was established in degradation studies, which also showed that there was no interference of the excipients. The accuracy was 100.22% with bias lower than 1.25%. Moreover, the in vitro cytotoxicity test of the degraded products showed non-significant differences (p > 0.05). The method was applied to the assessment of rhIL-11 and related proteins in biopharmaceutical dosage forms, and the results were correlated to those of a bioassay.
authors.affiliation: Department of Industrial Pharmacy, Federal University of Santa Maria, Santa Maria-RS, Brazil.
authors.forename: Ricardo Bizogne
authors.initials: RB
authors.lastname: Souto
authors.forename: Fernanda Pavani
authors.initials: FP
authors.lastname: Stamm
authors.forename: Maria Teresa de Carvalho Pinto
authors.initials: MT
authors.lastname: Ribela
authors.forename: Paolo
authors.initials: P
authors.lastname: Bartolini
authors.forename: Guilherme Zanini
authors.initials: GZ
authors.lastname: Calegari
authors.forename: Sérgio Luiz
authors.initials: SL
authors.lastname: Dalmora
mesh_terms: D000818:Animals; D001681:Biological Assay; D002460:Cell Line; D049109:Cell Proliferation / Q000187:drug effects; D056148:Chromatography, Reverse-Phase / Q000379:methods*; D006801:Humans; D017370:Interleukin-11 / Q000032:analysis / Q000302:isolation & purification* / Q000494:pharmacology* / Q000633:toxicity; D057230:Limit of Detection; D011786:Quality Control; D011994:Recombinant Proteins / Q000032:analysis / Q000302:isolation & purification* / Q000494:pharmacology* / Q000633:toxicity
chemical_list: D017370:Interleukin-11; D011994:Recombinant Proteins