Development and validation of method for glucosaminylmuramyl dipeptide determining in aqueous solutions by high-performance liquid chromatography [Разработка и валидация методики определения глюкозаминилмурамилдипептида в водных растворах методом высокоэффективной жидкостной хроматографии]

Introduction. For the determination of peptide-based drugs the reverse phase HPLC method with a gradient elution mode is widely used. Among the published works no validated methods suitable for the study of new dosage forms of glucosaminyl muramyl dipeptide (GMDP) were found. Due to the significant drawbacks of gradient elution, it is recommended to avoid the use of a gradient when developing methods for HPLC analysis, preferring isocratic elution to it. The aim of the study is the development and validation of a method for the quantitative determination of glucosaminyl muramyl dipeptide in aqueous solutions using the isocratic elution regimen, which can be used in the development of new dosage forms of GMDP. Material and methods. Quantitative determination of GMDP in solutions was carried out by reverse phase HPLC in isocratic mode on a Mediterranea Sea18 15 × 0.4 cm, 5 μm column using an eluent of the composition acetonitrile: 25 mM phosphate buffer solution (3:97), pH 7.3; a flow rate of 0.7 ml/min; a detector wavelength of 200 nm; and a column thermostat temperature of 25 °C. Results. The developed technique was validated by the following characteristics: specificity, linearity, correctness, repeatability (convergence), intermediate (laboratory) precision, analytical area, suitability of the chromatographic system. The technique has been successfully tested on laboratory samples of transdermal therapeutic systems containing GMDP. Conclusion. A method has been developed for the quantitative determination of GMDP in aqueous solutions by reverse phase HPLC with isocratic elution. The suitability of the method is confirmed by the compliance of the main validation characteristics with the acceptance criteria. The confirmed analytical range of the method was 0.5–50 μg/ml. The technique can be used in the analytical part of preclinical studies of new dosage forms of GMDP. © 2020 Meditsina Publishers. All rights reserved.