A fully validated microbiological assay to evaluate the potency of ceftriaxone sodium

Abstract

Ceftriaxone (CFTX) sodium is a third-generation, broad-spectrum cephalosporin that is resistant to betalactamases. An alternative bioassay for the assessment of the potency of this drug in pharmaceutical formulations has not been previously reported. Thus, this paper reports the development and full validation of a 3 x 3 agar diffusion bioassay using a cylinder-plate method to quantify CFTX sodium in pharmaceutical samples. The strain Staphylococcus aureus ATCC 6538P was used as the test microorganism, and the results of the proposed bioassay displayed high linearity, precision, accuracy, specificity and robustness. All potency results were statistically analyzed using an analysis of variance (ANOVA) and were found to be linear (r=0.99999) in the range of 16-64 μg/mL, accurate (100.5%), and precise [repeatability: relative standard deviation (RSD)=1.4%; intermediate precision: between-day RSD=2.1% and between-analyst RSD=2.5%]. The specificity of the bioassay was determined by evaluating a degraded sample (50 ºC) at 0, 24 and 48 hours as compared against the results from the pharmacopeial liquid chromatography method for CFTX. The results validated the proposed microbiological assay, which allows reliable quantitation of CFTX in pharmaceutical samples. Moreover, it is a useful, simple and low-cost alternative method for monitoring the quality of this medicine.Uniterms: Ceftriaxone/agar diffusion assay. Ceftriaxone/quality control. Ceftriaxone/microbiological assay. Drugs/quality control.A ceftriaxona sódica é uma cefalosporina de terceira geração de uso parenteral, com amplo espectro de atividade e resistente a b-lactamases. Este estudo apresenta o desenvolvimento e validação de um bioensaio por difusão em ágar usando o método de cilindros em placas para determinação da potência deste antibiótico. A validação desenvolvida apresentou bons resultados em termos de linearidade, precisão, exatidão, especificidade e robustez. Empregou-se o Staphylococcus aureus ATCC 6538P como micro-organismo teste. Os resultados dos ensaios foram tratados estatisticamente utilizando-se análise de variância (ANOVA). O método apresentou linearidade (r=0,99999) na faixa de doses selecionada (16-64 µg/mL), precisão (repetibilidade: DPR=1,4%; precisão intermediária: inter-dias DPR=2,1% e inter-analistas: DPR=2,5%) e exatidão de 100,5%. A especificidade do bioensaio foi avaliada através da análise comparativa, por cromatografia líquida de alta eficiência, de amostras degradadas a 50 ºC nos tempos zero, 24 e 48 h. Os resultados encontrados demonstraram a validade do bioensaio proposto, o qual permite a quantificação confiável de ceftriaxona sódica em produtos farmacêuticos comerciais. Por ser metodologia simples e de baixo custo constitui-se em alternativa para a análise de rotina do controle de qualidade de medicamentos.Unitermos: Ceftriaxona/ensaio de difusão em ágar. Ceftriaxona/controle de qualidade. Ceftriaxona/ ensaio microbiológico. Fármacos/controle de qualidade.

Effect of quercetin on the disposition kinetics of ciprofloxacin

“…The microbiological assay employed in the present study to measure the concentrations of ciprofloxacin in plasma, was also used previously by many authors to measure the concentrations of fluoroquinolones including ciprofloxacin in the plasma/serum of animals (Haritova et al, 2006;Akhtar et al, 2007;Raina et al, 2008;Muhammad and Muhammad, 2009). It was found that bioassay method correlates well with HPLC studies (Zuluaga et al, 2009, Abdelaziz et al, 2011, Manfio et al, 2013.…”

Biotransformation of Pharmaceuticals by Comamonas and Aeromonas Species

“…Biotransformation of antibiotics by resistant isolates under optimum culture conditions with time was evaluated by a microbiological assay (62). For this purpose, fractions of culture medium (2ml) were collected during growth curve analysis at different time intervals (3, 24, and 96 h).…”

A Critical Review of Analytical Methods for Determination of Ceftriaxone Sodium

“…[2][3][4][5] Ceftriaxone sodium is a third-generation semi-synthetic cephalosporin, derived from a fermentation product, for parenteral use, being this group of extreme importance because they are able to overcome the blood-brain barrier, since previous generations do not have this capacity. [1,6,7] It acts on Gram-positive and Gram-negative bacteria, the activity against Gram-positive is noticeably smaller in relation to the first-generation cephalosporins. Its major activity is related to Gram-negative bacteria, and it is also capable of acting against Gram-negative bacilli resistant to the firstand second-generation cephalosporins.…”

“…Its major activity is related to Gram-negative bacteria, and it is also capable of acting against Gram-negative bacilli resistant to the firstand second-generation cephalosporins. [2,7,8,9] Ceftriaxone is presented in the form of single-ingredient preparations in the USA, United Kingdom, Japan and Canada as Rocephin TM ; in Brazil as Rocefin TM , Ceftriax TM , Celltriaxon TM , Keftron TM , Triaxin TM , Amplospec TM , Ceftriona TM , Triaxton TM ; in China as Ansailong TM , Cefin TM , Dezhi TM , Likang Kesong TM , Livzonphin TM , Locekin TM , Oframax TM , Rocephin TM , Xianqin TM ; in Germany as Cefotrix TM , Rocephin TM ; in Italy as Axobat TM , Bixon TM , Cefrag TM , Davixon TM , Daytrix TM , Deixim TM , Diaxone TM , Eftry TM , Eraxitron TM , Fidato TM , Frieng TM , Kocefan TM , Monoxar TM , Nilson TM , Panatrix TM , Pantoxon TM , Ragex TM , Rocefin TM , Setriox TM , Sirtap TM , Valexime TM . [10][11][12] It is indicated for cases of septicemia, meningitis, disseminated Lyme borreliosis (early and late stages of the disease) (Lyme disease), intra-abdominal infections (peritonitis, gastrointestinal and biliary tract infections), bone, joint, soft tissue, skin and wound infections, infections in immunocompromised patients, kidney and urinary tract infections; infections of the respiratory tract, particularly pneumonia andotolaryngological infections, genital infections, including gonorrhea, perioperative prophylaxis of infections.…”