Quality by design approach for development and validation of a RP-HPLC method for simultaneous determination of co-administered levetiracetam and pyridoxine HCl in prepared tablets

• Published in: Microchemical journal 2018-12, Vol.143, p.55-63
• Main Authors: Hashem, Hisham, El-Sayed, Heba M.
• Format: Article
• Language: English
• Subjects: HPLC-DAD; Levetiracetam; Prepared tablets; Pyridoxine HCl; QbD
• ISSN: 0026-265X; 1095-9149
• DOI: 10.1016/j.microc.2018.07.031

A new simple, rapid and sensitive reversed-phase high-performance liquid chromatography (RP-HPLC) method was developed by employing Quality by Design (QbD) approach for determination of levetiracetam and pyridoxine HCl in prepared tablets. Levetiracetam and pyridoxine HCl are co-administered because treatment with the antiepileptic levetiracetam leads to a deficiency in pyridoxine HCl. Fractional factorial design (FFD) was applied for screening of the four independent factors: pH of the aqueous part of the mobile phase, flow rate, injection volume and % of the organic modifier. Analysis of variance (ANOVA) confirmed that the four factors were significant. Optimization of the chromatographic conditions was performed using central composite design (CCD). The Analysis was achieved on BDS Hypersil C8 (250 × 4.6 mm, 5 μm) column applying an isocratic mobile phase containing MeOH and 25 mM KH2PO4 buffer pH 3 (38.4:61.6, v/v) at 0.8 mL/min flow rate with UV detection at 214 nm and 5 μL injection volume. The proposed method was validated according to ICH guidelines. Under optimized conditions, linear ranges of 1.56–100 μg/mL and 0.39–100 μg/mL were obtained for levetiracetam and pyridoxine HCl, respectively, with correlation coefficients (R2) of 0.999. The average recoveries ranged from 95.46 to 101.14%. The inter- and intra-day precisions were lower than 1%. The proposed method showed good predictability and robustness. [Display omitted] •Levetiracetam and pyridoxine HCl were simultaneously determined by a RP-HPLC method.•Critical factors were determined, then optimized by response surface methodology.•The suggested models relating the responses and critical factors were significant.•Method predictability was confirmed, and the design space (robustness zone) was identified.•The developed method was applied to analysis of both analytes in prepared tablets.