ABSTRACT
A UV spectrophotometric method was developed for determination of ceftiofur sodium in the drug substance and sterile powder for injection. The method validation, which yielded good results, included evaluation of the range, linearity, intra- and interday precision, accuracy, recovery, specificity, robustness, LOQ, and LOD. The UV spectrophotometric determinations were performed at 292 nm. Good linearity was obtained between 2.5 and 20.0 microg/mL. A prospective validation showed that the method is linear (r = 0.9999) and precise, with RSD values of 0.3% for product A and 0.4% for product B. The intra- and interday precision values were < 2% for all samples analyzed. Comparison of UV spectrophotometry and LC by analysis of variance and Student's t-test showed no significant difference between methodologies. Moreover, the accuracy and precision obtained with the UV method correlated well with the values obtained with the LC method, and this correlation suggests that UV spectrophotometric analysis can be an inexpensive, reliable, and less time-consuming alternative to chromatographic analysis. The results demonstrated the validity of the proposed method as a simple and useful alternative for the determination of ceftiofur in routine QC analyses.
Subject(s)
Anti-Bacterial Agents/analysis , Cephalosporins/analysis , Powders , Reference Standards , Reproducibility of Results , Solutions , Spectrophotometry, UltravioletABSTRACT
Ceftiofur sodium is a third-generation broad-spectrum cephalosporin antibiotic, formulated as an intramuscular injection, that is approved for use in pigs, cattle, poultry, and dogs. The present work reports a method to quantify ceftiofur in powder for injection by comparing the cylinder plate assay and the liquid chromatographic (LC) method. The assay is based on the inhibitory effect of ceftiofur upon the strain of Micrococcus luteus ATCC 10240 used as the test microorganism. Ceftiofur sodium at concentrations ranging from 2.0 to 8.0 microg/mL can be measured in powder for injection. A prospective validation showed that the method is linear (r2 = 0.9998), with precise relative standard deviation (RSD) of 0.8% for product A (Excenel; Pharmacia and Upjohn Co., Kalamazoo, MI) and of 0.6% for product B (Topcef; Eurofarma Lab. Ltda, São Paulo, Brazil), with intermediate precision; between-day RSD = 1.0 and 1.1%, between-analyst RSD = 0.8 and 0.8% for products A and B, respectively and accurately. The comparison between bioassay and LC by analysis of variance and Student's t-test showed no significant difference among methodologies. The results demonstrated the validity of the proposed bioassay that is simple and a useful alternative methodology for analysis of ceftiofur in routine quality control.
Subject(s)
Anti-Bacterial Agents/analysis , Cephalosporins/analysis , Biological Assay , Chromatography, Liquid , Indicators and Reagents , Micrococcus luteus/drug effects , Powders , Reproducibility of Results , SolutionsABSTRACT
Cefepime is a new parenteral cephalosporin that has been described as a fourth-generation, broad-spectrum antibiotic. This paper reports the development and in-house validation of an agar diffusion bioassay using a cylinder-plate method for the determination of cefepime in powder for injection. The validation performed yielded good results in terms of linearity, precision, accuracy, and robustness. The assay is based on the inhibitory effect of cefepime upon the strain of Micrococcus luteus ATCC 10240 used as the test microorganism. The results of assays were treated statistically by analysis of variance (ANOVA) and were found to be linear (r = 0.99993) in the selected range of 8.0-32.0 microg/mL; precise [repeatability: relative standard deviation (RSD) = 1.39%, intermediate precision: between-day RSD = 1.77%, and between-analyst RSD = 1.97%] and accurate. Comparison of bioassay and liquid chromatography by ANOVA showed no significant difference between methodologies. The results demonstrated the validity of the proposed bioassay, which is a simple and useful alternative methodology for cefepime determination in routine quality control.
Subject(s)
Biological Assay/methods , Cephalosporins/analysis , Analysis of Variance , Anti-Bacterial Agents/administration & dosage , Anti-Bacterial Agents/analysis , Anti-Bacterial Agents/pharmacology , Anti-Bacterial Agents/standards , Biological Assay/standards , Biological Assay/statistics & numerical data , Cefepime , Cephalosporins/administration & dosage , Cephalosporins/pharmacology , Cephalosporins/standards , Chromatography, Liquid , Injections , Mass Spectrometry , Micrococcus luteus/drug effects , Powders , Reference Standards , Reproducibility of ResultsABSTRACT
A simple, sensitive and specific agar diffusion bioassay for the antibacterial enrofloxacin was developed. Using a strain of Staphylococcus aureus ATCC 6538P as the test organism, enrofloxacin at concentrations ranging from 3.2 to 12.8 microg ml(-1) could be measured in injection. A prospective validation of the method showed that method was linear (r = 0.99998), precise (R.S.D. = 0.27) and accurate (it measured the added quantities). The method shows results that confirm its precision, not differing significantly the other method described in the literature. We conclude that microbiological assay is satisfactory for quantitation of in vitro antibacterial activity of enrofloxacin.
Subject(s)
Anti-Bacterial Agents/pharmacology , Colony Count, Microbial/methods , Fluoroquinolones/pharmacology , Quinolones/pharmacology , Staphylococcus aureus/drug effects , Anti-Bacterial Agents/standards , Enrofloxacin , Fluoroquinolones/standards , Injections , Quinolones/standards , Reference Standards , Reproducibility of Results , Sensitivity and Specificity , Staphylococcus aureus/growth & developmentABSTRACT
A high-performance liquid chromatography isocratic procedure was developed for the assay of cefetamet pivoxil hydrochloride in drug substance and powder for oral suspension. The method validation yielded good results and included the range, linearity, precision intra- inter-day, accuracy, specificity, LOD and LOQ values. The chromatographic system consisted of a C(18) absorbosphere column (150 x 4.6 mm i.d., 5 microm particle size), a mobile phase composed of water-acetonitrile-methanol-phosphate buffer, pH 3.5 (50:35:10:5, v/v), flow rate of 1.5 ml min(-1) and UV detection at 254 nm. The relative standard deviation varied between 0.03 and 1.76%, and accuracy of 100.09% was found. Calibration curve was linear from 30.0-80.0 microg ml(-1); its correlation coefficient was 0.99989.
Subject(s)
Ceftizoxime/analogs & derivatives , Ceftizoxime/analysis , Administration, Oral , Ceftizoxime/chemistry , Chemistry, Pharmaceutical , Chromatography, Liquid/methods , PowdersABSTRACT
A simple, rapid and sensitive high-performance liquid chromatographic (HPLC) method was developed for the assay of enrofloxacin in raw material and injection. The validation method yielded good results and included the range, linearity, precision, accuracy, specificity, recovery, limit of detection (LOD) and limit quantification (LOQ) values. The HPLC separation was carried out by reversed phase chromatography on a C-18 absorbosphere column (150 x 4.6 mm i.d. 5 microm particle size) with a phase composed of sodium acetate (pH 4.7; 0.1 M): acetonitrile (60:40, v/v; pH 5.0), pumped isocratically at a flow rate of 1.5 ml min(-1). The effluent was monitored at 278 nm with the eluting solvent. The calibration graph for enrofloxacin was linear from 10.0 to 80.0 microg ml(-1).
