Development and validation of analytical methods for characterization and quantification of ertapenem sodium in lyophilized powder for injectable solution

Author(s)

Tahisa Marcela Pedroso

• Document type: Doctoral Thesis
• Press: Araraquara. 2017-10-11.
• Institution: Universidade Estadual Paulista (Unesp). Faculdade de Ciências Farmacêuticas. Araraquara
• Defense date: 2017-09-15
• Advisor: Hérida Regina Nunes Salgado

Abstract

Ertapenem sodium is an antimicrobial from the carbapenems class, commercially available in lyophilized powder form for injectable solution. It exhibits action against Gram-negative, Gram-positive, aerobic and anaerobic bacteria. The ertapenem sodium was characterized for its physical appearance, solubility, moisture content, melting point, by spectroscopic and chromatographic methods. Seven quantitative methods have been developed and validated. The method by spectroscopy on the ultraviolet region was developed using purified water as solvent and obtained linear regression of y = 0.0219x - 0.0017, R2 = 0.9999. For the method for spectroscopy in the infrared region, KBr pellets were prepared and measured, and the linear regression was y = 0.021 + 0.5141x, R2 = 0.9993. Among the chromatographic methods, a hydrophilic interaction liquid chromatography method was developed; reversed-phase highperformance liquid chromatography, and supercritical fluid chromatography. The hydrophilic interaction liquid chromatography was developed using as mobile phase A: acetonitrile and B: water (88:12 v/v) 0.1% formic acid pH 2.5, in isocratic mode, linear regression was y = 29928x - 547879, R2 = 0.9994. Whereas the method by reversed-phase high-performance liquid chromatography to the mobile phase was used A: water and B: ethanol (80:20 v/v) with 0.1% formic acid pH 2.5 in isocratic mode, linear regression was y = 23043x - 38525, and R2 = 0.9999. Regarding the supercritical fluid chromatography method the mobile phase consists of A: carbon dioxide (CO2) and B: a ternary mixture of 70: 25: 5 methanol: ethanol: water v/v/v, in the gradient mode. For this method the back pressure regulator was set to 175 Bars. The linear regression was y = 7426.6 x + 51489, R2 0.9996. The capillary electrophoresis method was developed using potassium phosphate buffer 10 mM pH 7 and 15 kV voltage, the linear regression was y = 2281.7x - 24495 and R2 = 0.9994. In addition to the analytical methods, a microbiological method by turbidimetry was developed, capable of determining the mean percentage of ertapenem sodium powder lyophilized for injection solution, before the microorganism Staphylococcus aureus ATCC 25923. The microbiological method was evaluated by the 3 x 3 test described in the Brazilian Pharmacopoeia. The microbiological method by turbidimetry obtained a linear regression of y = -1.140 Ln (x) + 3.6706, R2 = 0.9996 for the standard and y = -1.148 Ln (x) + 3.6942, R2 = 0.9998 for the sample. Finally, this work contemplates interchangeable methods for the evaluation of ertapenem sodium that contribute to improve the quality control of commercially available ertapenem sodium. (AU)

• FAPESP's process: 13/12959-0 - DEVELOPMENT AND VALIDATION OF ANALYTICAL METHODS FOR CHARACTERIZATION AND QUANTIFICATION OF ERTAPENEM SODIUM IN LYOPHILIZED POWDER FOR INJECTABLE SOLUTION
• Grantee: Tahisa Marcela Pedroso
• Support type: Scholarships in Brazil - Doctorate