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| Author(s): |
Danilo Fernando Rodrigues
Total Authors: 1
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| Document type: | Doctoral Thesis |
| Institution: | Universidade Estadual Paulista (Unesp) |
| Defense date: | 2017-05-19 |
| Advisor: | Hérida Regina Nunes Salgado |
| Abstract | |
The emergence of resistant microorganisms to a large number of clinically approved antimicrobials has been increasingly used, which limits the options for the treatment of bacterial infections. As a strategy, drugs with high antimicrobial activities are in evidence. Stands out a class of antimicrobial, the cephalosporins, having as fourth generation cefepime (CEF), a semi- synthetic product which has activity against various Gram-positive bacteria (e.g. oxacillin resistant Staphylococcus aureus) and Gram-negative (e.g. Pseudomonas aeruginosa) aerobic. Its mechanism of action is similar to other β-lactams, that mean, inhibits bacterial cell wall synthesis by binding to one or more penicillin-binding proteins - PBPs. In this context, innovative research involving analytical methods present fundamental importance and it is highly relevant to optimize its analysis in routine pharmaceutical industry and ensure quality of product already marketed. Qualitative analyzes were developed and used during work in order to promote the identification of the drug. In this research were also developed and validated methods for the quantification of CEF in lyophilised powder for solution for injection. Among them, microbiological test by turbidimetric method was validated using Staphylococcus aureus ATCC 6538 strain, being linear in the range from 36.0 to 81.0 μg.mL-1, which has potency of 102.46% and 99.92% accuracy. The method using high performance liquid chromatography (HPLC) was performed using as mobile phase only purified water and ethanol (45:55, v/v) at a wavelength () of 258 nm, showed the linear range in 10 to 70 μg.mL-1 and retention time (tR) equal to 4.99 minutes. The determination shown by HPLC was 106.31% and the accuracy obtained a value of 100.08%. Assays by spectrophotometry with absorption in the regions of ultraviolet (UV) and of visible (VIS) had their validated parameters to quantification CEF using purified water as solvent in the wavelengths of 258 and 740 nm, respectively. The method by UV showed linearity in the range of 10 to 20 μg.mL-1, with content of 103.81% and accuracy of 100.80%. The VIS method was developed on miniaturized scale, reducing the use of materials and solvents, as well as the time of prepare of the analyzes. The linearity of the VIS method was presented in the range of 15 to 40 μg.mL-1 and the content value obtained was of 104.26%. Lastly, another method was developed and validated by spectrophotometry with absorption in the region of infrared (IR), which did not use organic solvents for the analysis, important feature in reducing residues. The CEF content obtained by the IR method was 103.86%, within the linear range shown in concentrations of 0.2 to 0.6 mg/pellet of potassium bromide (KBr) and with value resulting of accuracy of 100.83%. Analysis of variance (ANOVA) were performed for validated methods, proving the safety and reliability of the results obtained. Analyses of mass spectrometry were performed to detect possible products after CEF degradation. The accelerated and long term stability studies of CEF in lyophilized powder also were performed, decreasing the percentages of the contents, but in according with the limit described in the literature. Thus, all the methods developed and validated in this work are turned on green chemistry and recommended for the quality control of CEF in pharmaceutical formulation. (AU) | |
| FAPESP's process: | 13/20017-5 - Chemical-pharmaceutical analysis of Cefepime hydrochloride in lyophilized powder for injectable solution |
| Grantee: | Danilo Fernando Rodrigues |
| Support Opportunities: | Scholarships in Brazil - Doctorate |