Desenvolvimento e validação de métodos analíticos para quantificação de vancomicina pó liofilizado para solução injetável
Carregando...
Data
2020-02-27
Autores
Nascimento, Patrícia Aleixa do
Título da Revista
ISSN da Revista
Título de Volume
Editor
Universidade Estadual Paulista (Unesp)
Resumo
Os antibióticos glicopeptídicos são uma classe de antimicrobianos que se apresenta como uma importante alternativa para o tratamento de infecções causadas por algumas cepas bacterianas resistentes. Estudos presentes na literatura relacionadosà vancomicina (glicopeptídeo de primeira geração) e métodos analíticos apresentam propostas eficazes para sua quantificação, entretanto, observa-se um predomínio no uso de solventes tóxicos, que contribuem para a geração de resíduos, que podem ser prejudiciais ao meio ambiente, ao operador e reduzir a vida média dos materiais utilizados nas análises. Desta forma, foram propostos quatro novos métodos analíticos para a análise quantitativa de vancomicina em pó liofilizado para solução injetável, por meio de cromatografia líquida de alta eficiência (CLAE), espectrofotometria região do ultravioleta (UV) e na região do infravermelho (IV) e análise microbiológica por turbidimetria. A proposta deste trabalho foi desenvolver e validar métodos de análise, objetivando métodos inovadores, rápidos e com uso de solventes de baixa toxicidade, minimizando, assim, as ações tóxicas aos operadores bem como resíduos ao meio ambiente, seguindo os princípios da Química Analítica Verde. As análises qualitativas foram realizadas para o peso médio, faixa de fusão, cromatografia em camada delgada, UV, IV, CLAE e turbidimetria, possibilitando a identificação deste fármaco na forma de pó liofilizado. Para a análise quantitativa, foram desenvolvidos e validados métodos para a quantificação de vancomicina em pó liofilizado por espectrofotometria na região do UV(linear entre 50 e 150 µg/mL, preciso com desvio padrão relativo – DPR – de até 1,90 %, exato com recuperação de 101,10 %, seletivo perante degradação ácida, alcalina, neutra, oxidativa e fotolítica, robusto perante modificações na comprimento de onda, volume da cubeta, filtração, marca do fosfato monobásico e dibásico de potássio, tempo de ultrassom e fonte de água), IV (linear entre 1,0 e 2,0 mg/10 mg, preciso com DPR de até 3,12 %, exato com recuperação de 99,37 %, seletivo perante a sobreposição dos espectro de vancomicina amostra e referência, robusto perante modificações na força de compressão), CLAE (linear entre 40 e 140 µg/mL, preciso com DPR de até 2,35 %, exato com recuperação de 100,19 %, seletivo perante degradação ácida, alcalina neutra, oxidativa e fotolítica, robusto perante modificações no comprimento de onda, concentração de ácido na fase móvel, marca de etanol, proporção da fase móvel, tempo de ultrassom e fonte de água) e turbidimetria (linear entre 30 e 50,7 µg/mL, preciso com DPR de até 4,42 %, exato com recuperação de 100,71 %, seletivo perante a comparação das absorbâncias de vancomicina amostra e referência, robusto perante modificações no comprimento de onda de leitura e temperatura de incubação),atendendo os limites propostos pela legislação para cada parâmetro de validação. Além disso, estes quatro métodos foram comparados estatisticamente, mostrandoque as técnicas CLAE e UV são intercambiáveis entre si (tcalculado 0,049 < ttabelado 2,77), devendo sempre estar associadas ao método microbiológico por ser um fármaco antimicrobiano. Assim, o trabalho traz métodos ecologicamente sustentáveis para a análise farmacêutica da vancomicina em produto farmacêutico.
The glycopeptide antibiotics are a class of antimicrobial agents that is an important alternative for the treatment of infections caused by some resistant bacterial strains. Studies found in literature relating vancomycin (first generation glycopeptide) and analytical methods show effective proposals for its quantification, however, there is a prevalence in the use of toxic solvents, that contributes to the generation of waste, that can be harmful to the environment, to the operator and reduce the half-life of the materials used in the analysis. In this way, it was proposed four new analytical methods for the quantitative analysis of vancomycin lyophilized powder for injectable solution, using High Performance Liquid Chromatography (HPLC), spectrophotometry in ultraviolet region (UV), in infrared region (IR) and the microbiological analysis using turbidimetry. The proposal of this work was to develop and validate analytical methods, aiming innovative and quickly methods, with the use of organic solvents of low toxicity, minimizing, in this way, the toxic action against the operators, as well as, the generation of waste to the environment, following the Green Analytical Chemistry principles. The qualitative analysis was performed by average weight, melting point, thin layer chromatography, UV, IR, HPLC and turbidimetry, making possible the identification of this drug in dosage form. For quantitative analysis, it was developed and validated analytical methods for quantification of vancomycin using UV (linear between50 to 150 µg/mL, precisewith relative standard deviation – RSD – up to 1.90 %, accuratewith mean recovery of 101.10 %, selectiveunder acid, alkaline, aqueous, oxidative and photolitic degradation, robustwhen variations were performed in wavelength, cubette volumn, filtration, monobasic and dibassic phosphate brand, ultrasound time and source of water), IR (linear between 1.0 to 2.0 mg/150 mg, precisewith RSD up to 3.12 %, accurate with mean recovery of 99.37 %, selectivewhen vancomycin sample and reference spectra were overlapped, robustwhen variations were performed in strength of compression), HPLC(linear between 40 to 140 µg/mL, precisewith RSD up to 2.35%, accurate with mean recovery of 100,19 %, selective under acid, alkaline, aqueous, oxidative and photolytic degradation, robustwhen variations were performed in wavelength, acid concentration in mobile phase, ethanol brand, mobile phase proportion, ultrasound time and source of wwater) andbioassay turbidimetry (linear between 30 and 50.7 µg/mL, precise with RSD up to 4.42%, accurate ith mean recovery of 100.71 %, selectivewhen vancomycin sample and reference absorbance values were compared, robust when variations were performed in wavelength and temperature of incubation), in accordance with the legislation stipulated limits for each validation parameter. Moreover, these four analytical methods were statistically compared and proved that the HPLC and UV methods are interchangeable (tcalculated 0.049 < ttabulated 2.77), and must always be apllied in association with the microbiological method, because this is an antimicrobial drug. In this way, this work proposes ecologically correct and sustainable methods for pharmaceutical analysis of vancomycin dosage form.
The glycopeptide antibiotics are a class of antimicrobial agents that is an important alternative for the treatment of infections caused by some resistant bacterial strains. Studies found in literature relating vancomycin (first generation glycopeptide) and analytical methods show effective proposals for its quantification, however, there is a prevalence in the use of toxic solvents, that contributes to the generation of waste, that can be harmful to the environment, to the operator and reduce the half-life of the materials used in the analysis. In this way, it was proposed four new analytical methods for the quantitative analysis of vancomycin lyophilized powder for injectable solution, using High Performance Liquid Chromatography (HPLC), spectrophotometry in ultraviolet region (UV), in infrared region (IR) and the microbiological analysis using turbidimetry. The proposal of this work was to develop and validate analytical methods, aiming innovative and quickly methods, with the use of organic solvents of low toxicity, minimizing, in this way, the toxic action against the operators, as well as, the generation of waste to the environment, following the Green Analytical Chemistry principles. The qualitative analysis was performed by average weight, melting point, thin layer chromatography, UV, IR, HPLC and turbidimetry, making possible the identification of this drug in dosage form. For quantitative analysis, it was developed and validated analytical methods for quantification of vancomycin using UV (linear between50 to 150 µg/mL, precisewith relative standard deviation – RSD – up to 1.90 %, accuratewith mean recovery of 101.10 %, selectiveunder acid, alkaline, aqueous, oxidative and photolitic degradation, robustwhen variations were performed in wavelength, cubette volumn, filtration, monobasic and dibassic phosphate brand, ultrasound time and source of water), IR (linear between 1.0 to 2.0 mg/150 mg, precisewith RSD up to 3.12 %, accurate with mean recovery of 99.37 %, selectivewhen vancomycin sample and reference spectra were overlapped, robustwhen variations were performed in strength of compression), HPLC(linear between 40 to 140 µg/mL, precisewith RSD up to 2.35%, accurate with mean recovery of 100,19 %, selective under acid, alkaline, aqueous, oxidative and photolytic degradation, robustwhen variations were performed in wavelength, acid concentration in mobile phase, ethanol brand, mobile phase proportion, ultrasound time and source of wwater) andbioassay turbidimetry (linear between 30 and 50.7 µg/mL, precise with RSD up to 4.42%, accurate ith mean recovery of 100.71 %, selectivewhen vancomycin sample and reference absorbance values were compared, robust when variations were performed in wavelength and temperature of incubation), in accordance with the legislation stipulated limits for each validation parameter. Moreover, these four analytical methods were statistically compared and proved that the HPLC and UV methods are interchangeable (tcalculated 0.049 < ttabulated 2.77), and must always be apllied in association with the microbiological method, because this is an antimicrobial drug. In this way, this work proposes ecologically correct and sustainable methods for pharmaceutical analysis of vancomycin dosage form.
Descrição
Palavras-chave
Vancomicina, Química Analítica Verde, Controle de Qualidade, Validação, CLAE, Espectrofotometria, Turbidimetria