Development and validation of a microbiological assay for determination of chlorhexidine digluconate in aqueous solution

Chlorhexidine (CHX) is a broad-spectrum antiseptic that is used in many topical pharmaceutical formulations. Because there is no official microbiological assay reported in the literature that is used to quantify CHX, this paper reports the development and validation of a simple, sensitive, accurate and reproducible agar diffusion method for the dosage of chlorhexidine digluconate (CHX-D) in an aqueous solution. The assay is based on the inhibitory effect of CHX-D upon the strain of Staphylococcus aureus ATCC 25923, which is used as the test microorganism. The design 3x3 parallel-line model was used. The results were treated statistically by analysis of variance (ANOVA), and they were excellent in terms of linearity (r = 0.9999), presenting a significant regression between the zone diameter of growth inhibition and the logarithm of the concentration within the range of 0.5 to 4.5%. The results obtained were precise, having relative standard deviations (RSD) for intra-day and inter-day precision of 2.03% and 2.94%, respectively. The accuracy was 99.03%. The method proved to be very useful and appropriate for the microbiological dosage of CHX-D in pharmaceutical formulations; it might also be used for routine drug analysis during quality control in pharmaceutical industries.Clorexidina (CHX) é um antisséptico com amplo espectro de ação utilizada em muitos tipos de preparações farmacêuticas para uso tópico. Uma vez que não há na literatura ensaio microbiológico oficial para quantificar a clorexidina, este trabalho objetivou o desenvolvimento e validação de um ensaio microbiológico simples, sensível, exato e reprodutível, por difusão em ágar, para doseamento de digliconato de clorexidina (CHX-D) em solução aquosa. O ensaio é baseado no efeito da inibição de Staphylococcus aureus ATCC 25923, utilizado como microorganismo teste, pela CHX-D. Utilizou-se o delineamento 3x3. Os resultados foram verificados estatisticamente pela análise de variância (ANOVA) e apresentaram excelente linearidade (r = 0,9999), demonstrando que o método segue o modelo linear com regressão significativa entre o diâmetro da zona de inibição e o lagaritmo da concentração no intervalo de 0,5 a 4,5%. Os resultados obtidos foram precisos apresentando desvio padrão relativo (DPR) para precisão intra-dia de 2,03% e DPR para precisão inter-dias de 2,94%. A exatidão foi 99,03%. O método provou ser muito útil e apropriado para doseamento microbiológico da CHX-D em formas farmacêuticas e pode ser empregado para análise desta substância no controle de qualidade em indústrias farmacêuticas

Development and validation of a microbiological assay for determination of chlorhexidine digluconate in aqueous solution

Chlorhexidine (CHX) is a broad-spectrum antiseptic that is used in many topical pharmaceutical formulations. Because there is no official microbiological assay reported in the literature that is used to quantify CHX, this paper reports the development and validation of a simple, sensitive, accurate and reproducible agar diffusion method for the dosage of chlorhexidine digluconate (CHX-D) in an aqueous solution. The assay is based on the inhibitory effect of CHX-D upon the strain of Staphylococcus aureus ATCC 25923, which is used as the test microorganism. The design 3x3 parallel-line model was used. The results were treated statistically by analysis of variance (ANOVA), and they were excellent in terms of linearity (r = 0.9999), presenting a significant regression between the zone diameter of growth inhibition and the logarithm of the concentration within the range of 0.5 to 4.5%. The results obtained were precise, having relative standard deviations (RSD) for intra-day and inter-day precision of 2.03% and 2.94%, respectively. The accuracy was 99.03%. The method proved to be very useful and appropriate for the microbiological dosage of CHX-D in pharmaceutical formulations; it might also be used for routine drug analysis during quality control in pharmaceutical industries

Análise químico-farmacêutica e estudo de estabilidade e de dissolução de cápsulas de flucloxacilina sódica

A flucloxacilina (FLU) é uma isoxazolilpenicilina semissintética, classificada como penicilina resistente à penicilinase estafilocócica. É ativa contra bactérias Gram-positivas, mas não é ativa contra Staphylococcus aureus resistentes à meticilina. A FLU é indicada para tratamento de infecções respiratórias, das articulações, síndrome do choque tóxico, pele, tecidos moles, endocardite e osteomielite, podendo ser administrada por via oral, intramuscular e endovenosa. A literatura apresenta alguns métodos de análise para FLU em fluidos biológicos, seus metabólitos e formas farmacêuticas. Neste trabalho foram desenvolvidos e validados métodos para análise qualitativa e quantitativa de FLU sódica em cápsula, bem como estudos de estabilidade para este fármaco. Para análise qualitativa, foram realizados os métodos de análise térmica, cromatografia em camada delgada, espectrofotometria nas regiões do infravermelho (IV) e ultravioleta (UV) e cromatografia líquida de alta eficiência (CLAE), possibilitando a identificação da amostra. Para a quantificação do fármaco foram desenvolvidos e validados os métodos: (1) ensaios microbiológicos por métodos de difusão em ágar e turbidimétrico, utilizando o microorganismo Staphylococcus aureus ATCC 25923 e ATCC 25923 IAL 1606, com faixa linear de 1,5 a 6,0 μg/mL e 4,0 a 16,0 μg/mL, respectivamente; exatidão de 98,98 e 99,73% e potência de 94,32 e 96,25%, respectivamente; (2) método espectrofotométrico na região do UV a 274 nm, utilizando água como solvente, com faixa linear em concentração de 50 a 100 μg/mL, exatidão de 100,09% e teor de 106,59%; (3) CLAE indicativo de estabilidade em comprimento de onda de 225 nm, fase móvel composta por ácido fosfórico (0,025 M), pH 5,5 e metanol (40:60 v/v) e o tempo de retenção médio foi de 4,5 minutos; a faixa linear foi de...Flucloxacillin (FLU) is a semi-synthetic isoxazolilpenicillin, classified as resistant penicillin to penicillinase staphylococal. It is active against Gram-positive, but not against methicillin resistant Staphylococcus aureus. FLU is indicated to treat respiratory infections, joints, toxic shock syndrome, skin, soft tissue, endocarditis and osteomyelitis and it can be administered by mouth, intramuscular and intravenous via. The literature describes some methods to analyse FLU in biologic fluids, its metabolites and pharmaceutical forms. In this work, methodologies were developed and validated to qualitative and quantitative analysis of FLU in capsules and also, stability studies were carried out. To qualitative analysis, thermal analysis, infrared, ultraviolet spectrophotometry, thin layer chromatography and high performance liquid chromatographic methods were studied, allowing the identification of sample. To drug quantification, the following methods were validated: (1) Microbiological assays by agar diffusion and turbidimetric methods, using Staphylococcus aureus ATCC 25923 and ATCC 25923 IAL 1606 organisms, with linear range from 1.5 to 6.0 μg/mL and 4.0 to 16.0 μg/mL, accuracy of 98.98 and 99.73% and potency of 94.32 and 96.25%, respectively; (2) ultraviolet spectrophotometric method at 274 nm, using water as solvent; with concentration linear range from 50 to 100 μg/mL, accuracy of 100.09% and quantitation of 106.59%; (3) HPLC stability-indicating, using detection wavelength at 225 nm, mobile phase consisted of fosforic acid (0.025 M) pH 5.5 methanol (40:60, v/v), with retention time of 4.5 minutes; linear range was from 20 to 80 μg/mL, with accuracy of 100.06% and quantitation of 94.07% were obtained; (4) capillary electrophoresis stability-indicating method at 210 nm, eletrolite consisted of borate buffer solution (50 mM) and... (Complete abstract click electronic access below)Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP

Desenvolvimento e controle de qualidade de formulação cosmética contendo digluconato de clorexidina

Os sabonetes são preparações destinadas à higiene e conhecidos há mais de 4000 anos. São constituídos por sais de ácidos graxos com propriedades detergentes, resultantes da saponificação entre ácidos graxos superiores e seus glicerídeos à custa de um material alcalino. Podem ser incorporados com diversas substâncias que possuem algum efeito terapêutico ou preventivo sobre a pele, por facilitarem o contato de tais substâncias com o tecido alvo. Porém, isto nem sempre é possível. Por tratar-se de um tensoativo aniônico carboxilado e ter elevada alcalinidade, muitos dos materiais ou ativos a serem incorporados a este tipo de preparação podem apresentar instabilidade. Com o advento dos detergentes sintéticos tal situação pode ser contornada, e apesar de constituírem-se também em tensoativos aniônicos, e por terem origem em ácidos fortes, permitem o preparo de sabonetes líquidos não alcalinos e apresentam maior compatibilidade frente a diversos ativos. Entre as diversas possibilidades de ativos, os agentes antimicrobianos, merecem atenção especial, uma vez que encontra nos sabonetes um excelente veículo para desempenhar sua função na assepsia da pele. A clorexidina é um anti-séptico de amplo espectro de ação, sendo ativo frente a bactérias Gram-positivas e Gram-negativas, fungos, leveduras e vírus. Atua sobre a membrana celular causando perda de material intracelular, tais como ácido nucléico e potássio, inibição respiratória e coagulação citoplasmática. Pode apresentar-se na forma de diversos sais como acetato, cloridrato, gluconato e digluconato. Neste trabalho foram estudadas formulações contendo solução de digluconato de clorexidina. A solução amostra utilizada para preparação dos sabonetes foi avaliada quanto a alguns parâmetros como pH, densidade relativa, ponto de fusão, cromatografia em camada delgada, espectroscopia...The soaps are preparation for hygiene and they are well known for 4000 years ago. They are constituited by salts of fatty acids with detergent properties, resulting from saponification of fatty acids superiorities and glycerides using an alkaline material. They may be incorporated with several substances that have a preventive or therapeutic effect on the skin, by facilitating the contact of such substances with the target tissue. However, this is not always possible. The soaps are anionic surfactant and carboxylic and they have high alkalinity, many of the materials or activities to be incorporated into this type of preparation may be instable. With the advent of synthetic detergents such situation can be avoided, and despite of they be also anionic surfactants, and are based on strong acids, allow the preparation of liquid soaps and alkali not have greater compatibility front of several assets. Among the various possibilities of active, antimicrobial agents deserve special attention, since the soap is an excellent vehicle to perform its function in the asepsis of the skin. The chlorhexidine is an antiseptic of wide spectrum of action, being active against the bacteria Gram-positive and Gram-negative, fungi, yeasts and viruses. Chlorhexidine is an excellent antiseptic of wide spectrum of action, being active against the bacteria Gram-positive and Gram-negative, fungi, yeasts and viruses. Acts on the cell membrane causing loss of intracellular material, such as nucleic acid and potassium, respiration inhibition and cytoplasmic coagulation. The salts of chlorhexidine can be acetate, hydrochloride, gluconate and digluconate. In this work formulation were studied with solution of chlorhexidine digluconate. The sample solution employed to prepare the liquid soaps was evaluated for some parameters such as pH, relative density, melting point, thin-layer chromatography... (Complete abstract click electronic access below)Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES

Development and validation of a microbiological assay for determination of chlorhexidine digluconate in aqueous solution

Chlorhexidine (CHX) is a broad-spectrum antiseptic that is used in many topical pharmaceutical formulations. Because there is no official microbiological assay reported in the literature that is used to quantify CHX, this paper reports the development and validation of a simple, sensitive, accurate and reproducible agar diffusion method for the dosage of chlorhexidine digluconate (CHX-D) in an aqueous solution. The assay is based on the inhibitory effect of CHX-D upon the strain of Staphylococcus aureus ATCC 25923, which is used as the test microorganism. The design 3x3 parallel-line model was used. The results were treated statistically by analysis of variance (ANOVA), and they were excellent in terms of linearity (r = 0.9999), presenting a significant regression between the zone diameter of growth inhibition and the logarithm of the concentration within the range of 0.5 to 4.5%. The results obtained were precise, having relative standard deviations (RSD) for intra-day and inter-day precision of 2.03% and 2.94%, respectively. The accuracy was 99.03%. The method proved to be very useful and appropriate for the microbiological dosage of CHX-D in pharmaceutical formulations; it might also be used for routine drug analysis during quality control in pharmaceutical industries.Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES

Corrositex®, BCOP and HET-CAM as alternative methods to animal experimentation

Tests in animals are used as models in toxicological and investigative studies. However, such tests have been considered inhumane because they can cause pain and suffering to experimental animals, while these methods can often be subjective. Protests calling for animal protection have questioned the effectiveness of in vivo tests and suggest the introduction of alternative, in vitro methods. International organizations, such as the Interagency Coordinating Committee on the Validation of Alternative Methods (ICCVAM), the National Institute of Health (NIH), the Organization for Economic Co-operation and Development (OECD), that regulate and develop new alternative animal models, have indicated the running of preliminary assays and execution of sequential tests, which consider physical-chemical properties and data of in vitro assays, before performing in vivo studies. Towards this background, the objective of the present article was to select promising alternative methods such as Corrositex®, BCOP and HET-CAM, intended to refine or replace the use of animals and reduce their suffering