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Clavulanic acid (CA) is an important secondary metabolite commercially produced by cultivation of Streptomyces clavuligerus (Sc). It is a potent inhibitor of bacterial β-lactamases. In this work, a specific and improved high performance liquid chromatography (HPLC) method, using a C-18 reversed phase column, diode array detector and gradient elution for CA quantification in fermentation broths of Sc, was developed and successfully validated. Samples were imidazole-derivatized for the purpose of creating a stable chromophore (clavulanate-imidazole). The calibration curve was linear over a typical range of CA concentration between 0.2 and 400mg/L. The detection and quantification limits were 0.01 and 0.02mg/L, respectively. The precision of the method was evaluated for CA spiked into production media and a recovery of 103.8%, on average, was obtained. The clavulanate-imidazole complex was not stable when the samples were not cooled during the analysis. The recovery rate was 39.3% on average. This assay was successfully tested for CA quantification in samples from Sc fermentation, using both, a chemically defined and a complex medium., (Copyright © 2015 Elsevier B.V. All rights reserved.)

This article deals with the simultaneous determination of three dissolution profiles in the same pharmaceutical formulation. The officially proposed procedure from the pharmacopoeias is adapted to the FIA methodology to obtain the officially recommended profile or "global profile", and two "individual" profiles, corresponding to dissolution rate of two different active principles present in the formulation; both drugs have overlapped UV-vis spectra. The simultaneous determination of several profiles is based on the derivative spectra and the zero crossing mathematical procedure for the "individual" profiles of an active principle; the "global" profile of the formulation is obtained from the order zero derivative. The empirical profiles were adjusted by regression analysis using the three-parameter (Higuchi equation) plot method which was selected as the most suitable. The analytical errors when the concentration of one drug is very small or very high are also checked.

A micellar electrokinetic chromatography method for simultaneous assay of ticarcillin and clavulanic acid in Timentin i.v. injection preparation was developed. This method ensures excellent separation of both components of Timentin preparation. The validation of the method was performed, and specificity, reproducibility, precision and accuracy were confirmed. The detection and quantitative limits for Timentin were established in the concentrations 0.04 and 0.08 mg/ml, respectively. The elaborated technique was compared with two methods routinely used-UV and high performance liquid chromatography (HPLC). The obtained results and their statistical analysis proved the same precision of all methods, however, no significant differences were observed between CE and HPLC.

Two almost independent potentiometric methods were implemented in a multi-task flow system based on the concept of sequential injection analysis for the analytical control of potassium clavulanate in pharmaceutical formulations. In addition to in-line preparation of the solutions submitted to the analysis, the system simultaneously detected the clavulanate anion and the potassium cation using two potentiometric detectors selective to each of the species referred. For the two determinations, the system developed presented an analytical range of 2 mM-0.1 M. Relative standard deviations of 0.5 and 0.6%, respectively, for clavulanate and potassium, were calculated from the signals obtained for 10 consecutive injections of a solution of 5x10(-3) M potassium clavulanate. The frequency of sampling was 53 samples h(-1). The quality of results supplied was evaluated by comparison with the reference procedure revealing relative errors less than 2% for the two determinations. The simultaneous attainment of two measurements permitted the standardisation of results in real time, the detection of faults in the procedure and monitoring the chemical stability of clavulanate.

A rapid, capillary electrophoresis method was evaluated for determination of amoxicillin and clavulanic acid in Augmentin as well as ampicillin and sulbactam in Unasyn preparations for injections. Phosphate-borate buffer at pH 8.66 containing 14.4% sodium dodecyl sulfate was used as a mobile phase. The method was validated. Reproducibility, precision, accuracy and assay linearity in concentration of amoxicillin 0.05-3.03 mg/ml and ampicillin 0.05-3.08 mg/ml, as well as clavulanic acid 0.02-2.02 mg/ml and sulbactam 0.05-2.08 mg/ml were established. This new method is fast, inexpensive and limits consumption of organic solvents when compared with alternative high performance liquid chromatography (HPLC) method, used for drug analysis. Statistical analysis by Student's t-test showed no significant differences between the results obtained by the two methods t(calculated) 0.32 and 1.69 for amoxicillin and clavulanic acid and 0.67 and 1.93 for ampicillin and sulbactam were smaller than t(tabulated).

A simple and sensitive liquid chromatographic method with electrochemical (EC) detection is described for the quantitative determination of amoxycillin and clavulanic acid in pharmaceutical dosage forms. Sample components were separated by a reversed phase C18 column, using a mixture of methanol-phosphate buffer (pH 3.2-3.4) (5:95, v/v) as the mobile phase. Detection of antibiotics was performed amperometrically by applying a potential of +1.25 V. High linearity over a concentration range of 15.625-500 ng was demonstrated for amoxycillin (r=0.9999) and clavulanic acid (r=0.9979). Detection limits were 0.8 ng ml(-1) for amoxycillin and 15 ng ml(-1) for clavulanic acid. This method was found to be convenient and reproducible for analysis of these two components in oral suspensions and tablets and might be useful for other pharmaceutical dosage forms.

The effects of gamma irradiation on the stability of potassium clavulanate, amoxicillin sodium and their combination were investigated. A decrease in purity and increase in degradation products up to 30 days after the irradiation were evaluated by reversed phase HPLC. The comparison between unirradiated and irradiated amoxicillin sodium, performed within 24 h following the irradiation process, showed no significant increase in the pre-existing impurities and no evidence of newly induced degradation products. On the contrary, an appreciable increase in the content of some impurities was evidenced 30 days after the irradiation. The chromatographic profile of irradiated potassium clavulanate showed the appearance of one unidentified new product and a slight increase of one pre-existing impurity. No further change in the impurity content was noted 30 days after the irradiation. The amoxicillin sodium-potassium clavulanate combination underwent the same kind of radiation induced degradation as the single compounds.