Degradation of cefradine in alga-containing water environment: a mechanism and kinetic study.

Large quantities of antibiotics are manufactured, used, and eventually discharged into alga-containing water environment as prototypes, by-products, or transformation products. Different activities of Chlamydomonas reinhardtii toward cefradine (CFD) were studied, and the results indicated that CFD is resistant (removal rate of 5.45-14.72%) in simulated natural water environment. Cefradine was mainly removed by hydrolysis, adsorption, desorption, photodecarboxylation, and photoisomerization. The effects of C. reinhardtii density, initial solution pH, and different light sources on CFD removal efficiency were investigated. The optimum conditions occurred at a density of algae 10 × 104 cells/mL, a solution pH of 9.0, and the ultraviolet (UV) light. Additionally, the removal kinetics under 16 different conditions was explored. The results showed that the removal of CFD fits well with a pseudo-first-order kinetic, and the half-life times are from 0.8 to 261.6 days. This study summarizes the CFD removal mechanisms in alga-containing water environment, highlights the important role played by light irradiation in eliminating CFD, and obtains the important kinetic data on CFD removal.

A comparative study of different aspects in manipulating ratio spectra used for the analysis of cefradine in the presence of its alkaline degradation product.

Six stability-indicating UV-spectrophotometric methods manipulating ratio spectra were utilized for the analysis of cefradine in presence of its alkaline degradate. These methods are different forms of transformations; ratio difference, mean centering, derivative ratio using numerical differentiation, derivative ratio using Savitsky-Golay filter, continuous wavelet transform and derivative continuous wavelet transform. Water was used as a solvent and the linearity ranges were 6-26 µg/mL. Determination of accuracy and precision for the suggested procedures were executed. Assessment of specificity was run through analyzing laboratory prepared mixtures containing cefradine and its alkaline degradate. The suggested methods were useful for cefradine estimation in tablets. Statistically, the outputs obtained from the recommended and published methods reveal no significant differences.

Study on the interaction mechanism between cefradine and Chlamydomonas reinhardtii in water solutions under dark condition.

Our research investigated the hormesis effect of cefradine on the specific growth rates (µ) of single-celled algae (Chlamydomonas reinhardtii) from aqueous solutions. We found the specific growth rate of C. reinhardtii slightly increased with cefradine concentrations within the range 0.5-10 mg/L. Effects of algae density, initial solution pH, and temperature on the adsorption batch assays were investigated. The optimum conditions for cefradine adsorption occurred at a density of 5 × 106 algae cells/mL, a solution pH of 7.0, and a temperature of 25.0 °C. A Box-Behnken design was employed to evaluate correlations between influential factors and cefradine adsorption. The results showed a significant interaction between algae density and temperature. The maximum removal rate could reach 50.13% under the optimal conditions. Additionally, the adsorption mechanisms were explored through Langmuir and Freundlich isotherm equations, adsorption kinetics, and thermodynamics. The results suggested that the adsorption process was monolayer, spontaneous, and endothermic with an increase in randomness at the algae-solution interface, which followed a pseudo-second-order model. All the data indicated that the alga performed a better removal capacity in the antibiotic-containing wastewater treatment process. This study lays the groundwork for a better understanding of the interaction mechanism between cefradine and Chlamydomonas reinhardtii in water solutions under dark condition.

Detection of the iron complexes with hydrolysis products of cephalexin and cefradine upon high-performance liquid chromatography/electrospray ionization mass spectrometry analysis.

RATIONALE: Cephalosporins (e.g. cephalexin, cefradine) are a major group of widely used ß-lactam antibiotics. Hydrolysis of the ß-lactam ring is an important reaction (often undesired) which leads to deactivation of ß-lactams. To the best of our knowledge there is no electrospray ionization mass spectrometry (ESI-MS) data reported concerning the products of hydrolysis of cephalosporins. METHODS: The hydrolysis of cephalexin and cefradine was performed in aqueous NaOH solutions. After the process the solutions were analyzed by high-performance liquid chromatography (HPLC)/ESI-MS. The elemental compositions of the ions discussed were confirmed by the accurate mass measurements on a quadrupole time-of-flight (QTOF) mass spectrometer. RESULTS: Unexpectedly, complexes between the hydrolysis products of cephalexin and cefradine (CFLh and CFRh ) and iron cation were detected upon HPLC/ESI-MS analysis, namely the ions [(CFLh -H)2 +Fe]+ and [(CFRh -H)2 +Fe]+ , although iron was not added to the analyzed solutions or to the mobile phase. These ions were found to be very stable in the gas phase. CONCLUSIONS: The detection of the complexes between the hydrolysis products of cephalosporins and iron may have a positive impact on the sensitivity and specificity of HPLC/ESI-MS analyses of the hydrolysis products of some cephalosporins.

In situ degradation of antibiotic residues in medical intravenous infusion bottles using high energy electron beam irradiation.

This study reported an immediate approach for the degradation of three antibiotic (amoxicillin, ofloxacin, and cefradine) residues in medical intravenous infusion bottles (MIIBs) using high energy electron beam (HEEB) irradiation. The effects of irradiation doses, initial concentrations, initial pH, and scavengers of active radicals on the degradation of three antibiotic residues (ARs) were investigated, and the results displayed that 97.02%, 97.61% and 96.87% of amoxicillin, ofloxacin, and cefradine residues could be degraded in situ through HEEB irradiation respectively. Fourier transform infrared spectroscopy (FTIR) and high performance liquid chromatography-mass spectrometry (HPLC-MS) analysis demonstrated that ARs were mainly decomposed into inorganic ions and alkanes. Typically, the detailed degradation mechanism of ARs was also investigated, and the dominant active particle inducing the degradation of antibiotics during the HEEB irradiation process was demonstrated to be hydroxyl radical.

Sensitive chemiluminescence determination of thirteen cephalosporin antibiotics with luminol-copper(II) reaction.

A new chemiluminescence reaction, the luminol-Cu(2+) reaction, was investigated for the determination of thirteen (13) cephalosporin antibiotics, namely cefalexin, cefadroxil, cefradine, cefazolin sodium, cefaclor, cefuroxime axetil, cefotaxime sodium, cefoperazone sodium, ceftriaxone sodium, ceftazidime, cefetamet pivoxil hydrochloride, cefixime, and cefpodoxime. It was found that, without adding any special oxidant, strong chemiluminescent (CL) signal could be produced from the reaction of the alkaline luminol with the above-mentioned antibiotics in the presence of Cu(2+). The experimental conditions for the reaction were carefully optimized with flow-injection mode. The detection limits are 0.3 ng/mL cefalexin, 3 ng/mL cefadroxil, 0.3 ng/mL cefradine, 0.02 µg/mL cefazolin sodium, 0.8 ng/mL cefaclor, 0.02 µg/mL cefuroxime axetil, 5 ng/mL cefotaxime sodium, 0.02 µg/mL cefoperazone sodium, 0.8 ng/mL ceftriaxone sodium, 1 ng/mL ceftazidime, 0.08 ng/mL cefetamet pivoxil hydrochloride, 0.8 ng/mL cefixime, and 2 ng/mL cefpodoxime. The proposed method was validated by direct application to commercial formulations and spiked milk samples containing cefradine. A possible reaction mechanism is also discussed.

Biotic and abiotic degradation of four cephalosporin antibiotics in a lake surface water and sediment.

Cephalosporins are widely used veterinary and human antibiotics, but their environmental fate and impacts are still unclear. We studied degradation of four cephalosporins (cefradine, cefuroxime, ceftriaxone, and cefepime) from each generation in the surface water and sediment of Lake Xuanwu, China. The four cephalosporins degraded abiotically in the surface water in the dark with half-lives of 2.7-18.7d, which were almost the same as that in sterilized surface water. Under exposure to simulated sunlight, the half-lives of the cephalosporins decreased significantly to 2.2-5.0d, with the maximal decrease for ceftriaxone from 18.7d in the dark to 4.1d under the light exposure. Effects of dissolved organic matter (DOM) and nitrate on photodegradation of the cephalosporins were compound-specific. While DOM (5 mg L(-1)) stimulated the photodegradation of only cefradine (by 9%) and cefepime (by 34%), nitrate (10 microM) had effects only on cefepime (stimulation by 13%). Elimination rates of the cephalosporins in oxic sediment (half-lives of 0.8-3.1d) were higher than in anoxic sediment (half-lives of 1.1-4.1d), mainly attributed to biodegradation. The data indicate that abiotic hydrolysis (for cefradine, cefuroxime, and cefepime) and direct photolysis (for ceftriaxone) were the primary processes for elimination of the cephalosporins in the surface water of the lake, whereas biodegradation was responsible for the elimination of the cephalosporins in the sediment. Further studies are needed on chemical structure, toxicity, and persistence of transformation products of the cephalosporins in the environment.

Colorimetric detection of cephradine in pharmaceutical formulations via fluorosurfactant-capped gold nanoparticles.

The aggregation of gold nanoparticles (GNPs) capped with nonionic fluorosurfactant (FSN) could be induced rapidly and selectively by cephradine degradation products, but not by cephradine and other excipients in pharmaceutical formulations. A new detection method for cephradine has been developed based on the cephradine degradation products-induced aggregation of the GNPs. The present approach offers various advantages, such as simplicity and high selectivity. Under optimum conditions, the lowest detectable concentration of cephradine through this approach (S/N=3) is 0.8 microg mL(-1). The calibration curve was linear over the range of 2.0-10.0 microg mL(-1) for the detection of cephradine. The recoveries of cephradine were found to fall in the range between 97% and 105%. We have validated the applicability of our method through the analyses of cephradine in pharmaceutical formulations. Good agreements were obtained for the determination of cephradine between the present approach and official method.

Chemiluminescence flow-injection analysis of beta-lactam antibiotics using the luminol-permanganate reaction.

A new flow injection chemiluminescence (CL) method has been developed for the determination of six beta-lactam antibiotics, including amoxicillin, cefadroxil, cefoperazone sodium, cefazolin sodium, cefradine and ceftriaxone sodium. When the antibiotic was injected into a stream of KMnO4 with alkaline luminol, a strong CL signal was produced. The method allows the measurements of 0.1-50.0 mg/L amoxicillin, 0.1-80.0 mg/L cefadroxil, 1.0-30.0 mg/L cefoperazone sodium, 1.0-30.0 mg/L cefazolin sodium, 3.0-50.0 mg/L cefradine and 3.0-50.0 mg/L ceftriaxone sodium. The detection limits are 0.05 mg/L for amoxycillin, 0.05 mg/L for cefadroxil, 0.4 mg/L for cefoperazonum sodium, 0.4 mg/L for cefazolin sodium, 0.8 mg/L for cefradine and 0.8 mg/L for ceftriaxone sodium. The relative standard deviations in 11 repeated measurements are 0.6%, 0.8%, 1.5%, 1.2%, 0.4% and 0.3% for 3.0 mg/L amoxicillin, 1.0 mg/L cefadroxil, 10.0 mg/L cefoperazone sodium, 10.0 mg/L cefazolin sodium, 10.0 mg/L cefradine and 10.0 mg/L ceftriaxone sodium, respectively. The method was successfully applied to the determination of amoxicillin in pharmaceutical preparations. A possible CL reaction mechanism is also discussed.

Spectrofluorometric determination of alpha-aminocephalosporins in biological fluids and pharmaceutical preparations.

A selective and highly sensitive fluorometric method was developed for the determination of four alpha-aminocephalosporins, namely cefaclor, cefadroxil, cephalexin and cephradine. The method involves the reaction of the target compounds with fluorescamine at a specific pH, ranging from 7.8 to 8.4. The produced derivatives exhibit maximum fluorescence intensities at 472-478 nm after excitation at 370-372 nm. The method is highly specific because other alpha-aminocephalosporins whose alpha amino group was blocked do not react similarly and hence do not interfere. At the specific pH range of the reaction where no degradation may occur with that medium the proposed method can be utilised as a stability-indicating assay. The different experimental parameters affecting the derivatisation reaction were carefully studied and incorporated into the procedure. Under the described conditions, the proposed method is linear over the concentration range of 0.05(-1) microg/ml(-1) for both cefaclor and cephalexin, and 0.05-0.65 and 0.025-0.5 microg/ml(-1) for cefadroxil and cepharadine, respectively and the coefficients of determination were greater than 0.999 (n = 3). The recoveries of the title compounds from spiked serum ranged from 88.6 to 89.7% and from spiked urine from 92.2 to 93.3% with a limit of quantitation (LOQ) of 25-50 ng/ml(-1) and limit of detection (LOD) of 5 ng/ml(-1) (S/N = 2) for all drugs. The mechanism of the fluorometric reaction is proposed and the advantages of the proposed method are discussed.

[Studies on the simultaneous measurement of several cephalosporins by RP-HPLC (I)].

This paper reported the research on the simultaneous separation and determination of six cephalosporins by RP-HPLC. Six cephalosporins are cefalcor, cefalexin, cefradine, cefadroxil, cefominox and cefoxitin. The analytical conditions for this method were as follows: a Hypersil ODS C18(200 mm x 4.6 mm i.d., 5 microns), detection wavelength: 254 nm; a mobile phase solution of 50 mmol/L monopotassium phosphate (pH 3.4)-acetonitrile (87.5:12.5) and DAD detector. The flow rate was 1.0 mL/min. The calibration curves of the six compounds were linear, the correlation coefficients were 0.9951 for cefominox, 0.9999 for the others, the range were 164 ng-16.4 micrograms for cefominox, 99 ng-9.934 micrograms for cefadroxil, 104 ng-10.358 micrograms for cefalcor, 122 ng-12.224 micrograms for cefalexin, 107 ng-10.702 micrograms for cefradine and 115 ng-11.506 micrograms for cefoxitin. The recovery rates were 103.5% for cefominox, 99.3% for cefadroxil, 101.4% for cefalcor, 101.5% for cefalexin, 98.7% for cefradine and 97.6% for cefoxitin. Six cephalosporins were all stable in 50 mmol/L monopotassium phosphate (pH 3.4-4.6). When preparations of these cephalosporins were determined, it is indicated there were no difference between the results by using this method and the pharmacopoeia methods. The total separation time of these cephalosporins was within fifteen minutes. This method is simple, sensitive, rapid and accurate.

The analysis of aqueous humor constituents using capillary zone electrophoresis.

One of the difficulties encountered in the study of aqueous humor is the relatively small volume generally available for analysis. The purpose of this study was to investigate the potential use of capillary zone electrophoresis (CE) for the analysis of nanolitre quantities of this fluid. Twelve samples of aqueous humor were obtained from patients undergoing cataract surgery and a further three samples were from non cataract post mortem subjects within 6 hr of death. CE was carried out in an uncoated fused silica glass capillary, 75 mu internal diameter and 100 cm long using a run buffer of 40 mM borate pH 9.4 containing 0.4 g l-1 methylcellulose. Detection of the separated zones was by ultra violet absorption at 200 nm. Preliminary identification of peaks was achieved by enzymatic hydrolysis and spiking with purified analytes. A number of very well resolved peaks were obtained from both cataract and post mortem samples using nanolitre quantities of unmodified fluid. Additional peaks were noted in the post mortem samples, most of which were likely to be due to a partial breakdown of the blood aqueous humor barrier. The profiles obtained were not significantly affected by various drugs routinely administered during cataract surgery. This preliminary study has demonstrated the potential value of CE in the analysis of aqueous humor in health and disease.

Isolation and structural elucidation of an impurity of cefradine.

An impurity of unknown identity was isolated from commercial cefradine by liquid chromatography on poly (styrene-divinylbenzene) with HOAc (0.01 M)-CH3CN (94:6, v/v) as the mobile phase. The structure was elucidated as 4',5'-dihydrocefradine using nuclear magnetic resonance spectroscopy (NMR) and mass spectrometry (MS). The structure was confirmed by comparison with the chromatographic retention characteristics and photodiode-array detected ultraviolet spectrum of the synthetic compound and with its infrared, NMR and MS spectra. The presence of 4',5'-dihydrocefradine in cefradine has not been described previously.

A comparative study of LC methods for analysis of cefradine.

A comparative study of two isocratic liquid chromatographic methods for the analysis of cefradine is described. The first method is prescribed by the European Pharmacopoeia for the assay of cefradine, using classical alkyl bonded phase (C18) as the stationary phase. Poor reproducibility of the selectivity towards cefradine and its related substances was observed when this method was used and none of the C18 columns examined was able to separate cefradine completely from its potential related substances under the prescribed LC conditions. On the other hand, the second method, which uses poly(styrene-divinylbenzene) as the stationary phase, shows good selectivity even when using columns from different manufacturers and of different age. Four bulk samples of cefradine were analysed following both methods and the results were compared.

Quantitative analysis of cefradine by liquid chromatography on poly(styrene-divinylbenzene).

A method is described for isocratic analysis of cefradine by liquid chromatography on a poly(styrene-divinylbenzene) column (PLRP-S, 250 x 4.6 mm i.d.) at 50 degrees C. Cefradine is separated from its related substances using a mobile phase of acetonitrile-0.02 M sodium 1-octanesulphonate-0.2 M phosphoric acid-water (14.5:10:5:up to 100, v/v/v/v). The flow rate was 1.0 ml min-1 and UV-detection was performed at 254 nm. The method was employed for the quantitative analysis of reference substances, bulk samples and pharmaceutical dosage forms.

Cephradine (Velosef) penetration of mandibular bone.

The concentration of cephradine in serum and mandibular bone was assayed in 28 patients undergoing 3rd molar surgery following a single 1 g intravenous injection. Serum and cortical bone samples taken simultaneously, contained mean cephradine concentrations of 42.11 micrograms/ml and 2.61 micrograms/g respectively. These results, when compared with those reported for other bony sites including the femoral head and knee, show a reduced bone penetration with a bone-to-serum ratio of approximately 0.06:1.

Qualitative and quantitative analysis of cephradine in biological materials by high performance liquid chromatography.

A reversed phase high performance liquid chromatographic method was developed for the determination of cephradine, one of the commonly used antibiotics, in biological materials. Mimic samples for stomach contents, miso soup, were applied to high performance liquid chromatography (HPLC) after centrifugation and purification by Sep-Pak C18 cartridge treatment. Serum samples deproteinized or urine samples diluted were directly injected into the HPLC. The recoveries of cephradine from these materials were 95-97% and the detection limit was 0.01 microgram/injection. This method was applied to the analysis of cephradine in stomach contents obtained by autopsy. After purification by the cartridge treatment, cephradine in the sample was identified and determined by HPLC and further confirmed by thin-layer chromatography (TLC) and mass spectrometry (MS).