Search

Miniaturization of liquid-phase extraction is an increasingly emerging field of sample preparation aiming to reduce the solvent consumption and to protect the environment. In this paper, a homogeneous liquid phase microextraction using a hydrophilic extraction media in combination with a salt as phase-forming agent was developed for the determination of four fluoroquinolones (ciprofloxacin, norfloxacin, moxifloxacin and prulifloxacin) in human urine. Important parameters affecting the extraction performance including sample volume, organic solvent volume, salt concentration etc were systematically investigated and optimized. Multivariate experimental designs namely Plackett-Burman and Box-Behnken designs were sequentially applied for factor screening and optimization of the method parameters. The method was fully validated using the “total error” approach. Accuracy profiles – a graphical decision-making tool – were constructed using the results of the validation procedures. The β-expectation tolerance intervals did not exceed the acceptance criteria of ± 15%, meaning that 95% of future results will be included in the defined bias limits. The relative bias ranged between ─ 2.8 to 7.7 % for all analytes, while the RSD values for repeatability and intermediate precision were less 6.6%. The achieved limits of detection (LOD) were ranged between 3 and 11 ng mL−1 while the lower limits of quantitation (LLOQ) were established as 10 ng mL−1 for ciprofloxacin and norfloxacin and 100 ng mL−1 for moxifloxacin and prulifloxacin, respectively. The ruggedness of the microextraction procedure was assessed through Monte-Carlo simulations and capability analysis. The proposed approach improves the procedures proposed to date for the bioanalysis of fluoroquinolones in terms of efficiency, reduction of the sample volume and extraction time.

Based on the thiourea quenching of the chemiluminescence of Tb3+/ prulifloxacin (PUFX) sensitized KMnO4-Na2SO3 system, a convenient and rapid chemiluminescence method for the determination of thiourea was proposed. The reaction between KMnO4 and Na2SO3 brought only weak chemiluminescence, but the chemiluminescence increased sharply in the presence of sensitizer Tb3+/ PUFX. Addition of thiourea can prevent the reaction between KMnO4 and Na2SO3, thus the chemiluminescence intensity was significantly decreased. Under the optimum conditions, the calibration graphs for thiourea were linear in the range of 1.0 × 10-7 to 4.0 × 10-5 mol•L-1. The limit of detection was 6.4 × 10-8 mol•L-1. The method was applied satisfactorily to the determination of thiourea in tap water, lake water and rice noodles and the spiked recoveries were between 104.7 ~ 113.4%. The possible mechanism of sensitization and quenching was also proposed.

This study provides an insight into the prevalence of (fluoro)quinolones (FQs) and their specific quinolone qnrS resistance gene in the Avon river catchment area receiving treated wastewater from 5 wastewater treatment plants (WWTPs), serving 1.5 million people and accounting for 75% of inhabitants living in the catchment area in the South West of England.. Ofloxacin, ciprofloxacin, nalidixic acid and norfloxacin were found to be ubiquitous with daily loads reaching a few hundred g/day in wastewater influent and tens of g/day in receiving waters. This was in contrast to other FQs analysed: flumequine, nadifloxacin, lomefloxacin, ulifloxacin, prulifloxacin, besifloxacin and moxifloxacin, which were hardly quantified. Enantiomeric profiling revealed that ofloxacin was enriched with the S-(-)-enantiomer, likely deriving from its prescription as the more potent enantiomerically pure levofloxacin, alongside racemic ofloxacin. While ofloxacin's enantiomeric fraction (EF) remained constant, high stereoselectivity was observed in the case of its metabolite ofloxacin-N-oxide. The removal efficiency of quinolones during wastewater treatment at 5 WWTPs utilising either trickling filters (TF) or activated sludge (AS), was compound and wastewater treatment process dependent, with AS providing better efficiency than TF. The qnrS resistance gene was ubiquitous in wastewater. Its removal was WWTP treatment process dependent with TF performing best and resulting in significant removal of the gene (from 28 to 75%). AS underperformed with only 9% removal in the case of activated sludge and actual increase in the gene copy number within sequencing batch reactors (SBRs). Interestingly, the data suggests that higher removal of antibiotics could be linked with high prevalence of the gene (SBR and WWTP E) and vice versa, low removal of antibiotic is correlated with lower prevalence of the gene in wastewater effluent (TF, WWTP B and D). This is especially prominent in the case of ofloxacin and could indicate that AS might be facilitating antimicrobial resistance (AMR) prevalence to higher extent than TF. Wastewater-based epidemiology (WBE) was also applied to monitor any potential misuse (e.g. direct disposal) of FQs in the catchment. In most cases higher use of antibiotics with respect to official statistics (i.e. ciprofloxacin, ofloxacin) was observed, which suggests that FQs management practice require further attention.

Antibiotics are the most consumed therapeutic classes worldwide and are released to the environment in their original form as well as potentially active metabolites and/or degradation products. Consequences of the occurrence of these compounds in the environment are primarily related to bacterial resistance development. This work presents a validated analytical method based on solid phase extraction (SPE) using HLB cartridges, followed by liquid chromatography-tandem mass spectrometry (LC-MS/MS) for quantification of seven different fluoroquinolone antibiotics, namely ciprofloxacin (CPF), enrofloxacin (ENR), lomefloxacin (LOM), norfloxacin (NOR), ofloxacin (OFL), prulifloxacin (PLF) and moxifloxacin (MOX) and its application to detect the target compounds in influents and effluents of wastewater treatment plants (WWTP). Linearity was established through calibration curves in solvent and matrix match using internal calibration method in the range of 50–1300 ng L−1 and all the fluoroquinolones showed good linear fit (r2 ≥ 0.991). Accuracy ranged between 80.3 and 92.9%, precision was comprised between 7.2 and 14.6%, and 10.7 and 18.1% for intra- and inter-batch determinations, respectively. Method detection and quantification limits ranged from 6.7 to 59.0 ng L−1 and 22.3–196.6 ng L−1, respectively. Influents and effluents of fifteen WWTPs of North of Portugal were analyzed. OFL was the fluoroquinolone found at the highest concentration, up to 4587.0 ng L−1 and 987.9 ng L−1, in influent and effluent, respectively. NOR and PLF were not detected. The present works characterizes WWTP influents and effluents regarding seven different fluoroquinolone antibiotics in fifteen Portuguese northern WWTPs with diverse operational features and treatment procedures.

A novel HPLC method for the simultaneous determination of Prulifloxacin, Ulifloxacin, its process impurities in a tablets formulation and Enrofloxacin, used as Internal Standard, is developed. The separation was successfully carried out with a new stationary phase, HILIC, under isocratic elution mode using ammonium acetate buffer (5 mM, pH 5.8) and acetonitrile (12:88, v/v) at a flow rate of 1.0 mL/min. Column was thermostated at 25 °C (±1 °C) and 20 μL were injected for the analysis. Calibration curves were linear in the investigated range with correlation coefficient better than 0.9880, while the limit of quantifications ranged from 0.25 to 5 μg/mL, depending from the analyte. The within and between batch precision (RSD%) values ranged from 0.11% to 13.9% while within and between batch trueness (bias%) values ranged from 14.0% to -11.3%. This method for the direct determination and quantification of process impurities in pharmaceutical formulations is suitable for routine analyses in quality control laboratories and was applied to evaluate for the first time, the presence and the quantities of cited analytes in commercially available formulation.

This experiment was designed to investigate correlation among 28-days repeated oral dose toxicity, toxicokinetics and tissue distribution data of ulifloxacin (active metabolite of prulifloxacin) in Wistar albino rats. Prulifloxacin was administered for 28-days in rats at 0, 100, 200, 400mg/kg/day followed by 14-days recovery period. Simultaneously different toxicokinetic parameters and tissue distributions of ulifloxacin was examined by LC-MS/MS method. Plasma levels and tissue concentrations of ulifloxacin were increased with dose-related manner. Ulifloxacin was also distributed to many tissues, and concentration in lungs nearly equivalent to the plasma concentration. Based on these results it was concluded that long-term repeated dose of prulifloxacin may produce different blood parameters abnormality, liver damage, stomach ulcer, joint damage and dysfunction of lungs in rats which relates to high tissue distribution and accumulation of ulifloxacin in these tissues. These findings help in management of prulifloxacin induced adverse effects by appropriate dose selection in clinical practice.

A rapid, specific and novel gradient LC-MS method has been developed and validated for the identification and characterization of stressed degradation products (DPs) of prulifloxacin (PF) using liquid chromatography combined with quadrupole time-of-flight electrospray ionization tandem mass spectrometry (LC/Q-TOF-ESI-MS/MS). PF was subjected to hydrolytic (acidic, alkaline and neutral), oxidation, photolytic and thermal stress, as per ICH guidelines Q1A (R2). The drug showed extensive degradation in hydrolytic and oxidative, while it was stable to thermal and photolytic stress conditions. In total, 13 DPs were formed and the chromatographic separation of drug and its DPs was achieved on a C-18 column (4.6 × 250 mm, 5 μm) using gradient elution method. All the DPs have been identified and characterized using MS(n) experiments and accurate mass measurements. The LC-MS method was validated with respect to specificity, linearity, accuracy, precision and robustness.

Prulifloxacin is a fluoroquinolone antibiotic that has been approved in several European countries for the treatment of lower urinary tract infections and exacerbations of chronic bronchitis. In this review, PubMed and Scopus databases were searched for potential uses of prulifloxacin beyond respiratory and urinary tract infections. Nine individual articles (eight randomised controlled trials and one cohort study) were regarded as eligible for inclusion in the review. Three of the studies were double-blinded, whilst six were open-label trials. Three studies referred to the treatment of patients with chronic bacterial prostatitis (CBP), one to prophylaxis of patients undergoing transrectal prostate biopsy, one to prophylaxis of women undergoing surgical abortion, two to patients with traveller's diarrhoea, one to diabetic patients with soft tissue infections or osteomyelitis, and one to improving tolerance of Bacillus Calmette-Guérin (BCG) instillations in patients with bladder cancer. Regarding CBP, prulifloxacin was non-inferior to its comparators, with a trend towards better microbiological outcomes at follow-up. Regarding traveller's diarrhoea, prulifloxacin resulted in better clinical and microbiological outcomes compared with placebo. Finally, prulifloxacin decreased the adverse events associated with BCG instillations in patients with bladder cancer, without affecting cancer recurrence rates. In summary, prulifloxacin appears to be a promising agent for the treatment of bacterial prostatitis and traveller's diarrhoea.

Prulifloxacin is a kind of new oral taking antibiotic of fluoroquinolone. Conjugation reaction of prulifloxacin with trypsin in Britton–Robinson buffer solution of pH 7.96 was analyzed by UV–vis spectrophotometry and fluorescence spectrometry. The intrinsic fluorescence of trypsin was strongly quenched by prulifloxacin. This effect was rationalized in terms of a static quenching procedure. The binding parameters have been evaluated by fluorescence quenching methods. Negative values Δ G 0 for the formation of prulifloxacin–trypsin complex implied that both hydrogen bonds and hydrophobic interactions might play a significant role in prulifloxacin binding to trypsin. The binding distance deduced from the efficiency of energy transfer was 0.84 nm for prulifloxacin–trypsin. Furthermore, association constants and binding mechanism were successfully derived from the fluorescence spectra. UV–vis detections supported a change in the secondary structure of proteins caused by the interaction of prulifloxacin with trypsin.

The interaction between prulifloxacin, a kind of new oral taking antibiotic and pepsin, a kind of enzyme in the stomach has been investigated in vitro under a simulated physiological condition by different spectroscopic methods. The intrinsic fluorescence of pepsin was strongly quenched by prulifloxacin. This effect was rationalized in terms of a static quenching procedure. The binding parameters have been evaluated by fluorescence quenching methods. The negative value of ΔG0 reveals that the binding process is a spontaneous process. The binding distance R between donor (pepsin) and acceptor (prulifloxacin) was obtained according to the Forster's resonance energy transfer theory and found to be 0.95 nm. The results obtained herein will be of biological significance in pharmacology and clinical medicine.

We report the results of a prospective randomised study to evaluate the therapeutic effect of Serenoa repens, Urtica dioica (ProstaMEV), quercitin and curcumin (FlogMEV) extracts associated with prulifloxacin in patients affected by chronic bacterial prostatitis (CBP). From a whole population of 284 patients, 143 patients affected by CBP [National Institutes of Health (NIH) class II prostatitis] were enrolled. All patients received prulifloxacin 600 mg daily for 14 days, in accordance with antibiogram results. Patients were split into two groups: Group A received prulifloxacin associated with ProstaMEV and FlogMEV; Group B received only antibiotic therapy. Microbiological and clinical efficacies were tested by two follow-up visits at 1 month and 6 months, respectively. Quality of life (QoL) was measured using the NIH Chronic Prostatitis Symptom Index (CPSI) and International Prostatic Symptom Score (IPSS) questionnaires. Group A comprised 106 patients and Group B comprised 37 patients. One month after treatment, 89.6% of patients who had received prulifloxacin associated with ProstaMEV and FlogMEV did not report any symptoms related to CBP, whilst only 27% of patients who received antibiotic therapy alone were recurrence-free (P < 0.0001). Significant differences were found between groups in terms of symptoms and QoL (P < 0.0001 for both). Six months after treatment, no patients in Group A had recurrence of disease whilst two patients in Group B did. Questionnaire results demonstrated statistically significant differences between groups (all P < 0.001). The association of S. repens, U. dioica (ProstaMEV), quercitin and curcumin (FlogMEV) extracts is able to improve the clinical efficacy of prulifloxacin in patients affected by CBP.

A capillary electrophoresis (CE)–chemiluminescence (CL) method for determining norfloxacin (NFLX) and prulifloxacin (PFLX) was developed based on the enhanced CL intensity of the cerium(IV)–sulfite–fluoroquinolone (FQ) reaction sensitized by terbium(III). The separation was conducted in buffer composed of 20 mM sodium citrate, 4 mM citric acid and 10 mM sodium sulfite at pH 6.1. The CL reagent solution consisted of 2 mM cerium(IV), 4 mM terbium(III) and 1.1 mM hydrochloric acid. NFLX and PFLX were baseline separated within 11 min with detection limits (S/N = 3) of 0.057 and 0.084 μg mL −1 , respectively. The maximum intra- and inter-day relative standard deviations (R.S.D.s) of migration time of the analytes were less than 4.0% and 4.2%, respectively. The proposed method was applied to detect NFLX and PFLX in fortified urine sample and the results were comparable to high-performance liquid chromatography (HPLC)–UV method. Moreover, the high selectivity of the CL detection and the high-separation efficiency of CE render the method the potential of quick analyzing fluoroquinolones in real complex matrix.

A rapid and simple method for determining two fluoroquinolones (FQNs), namely Ciprofloxacin and Ulifloxacin, this being the last active metabolite of Prulifloxacin, in aqueous human humor (AHH) has been developed and validated. The calibration data resulted linearly correlated in the 4-500 ng/mL concentration range with 8 ng/mL lower limit of quantification (LLOQ) for Ciprofloxacin, and 5-600 ng/mL concentration range with 6 ng/mL LOD for Ulifloxacin. The proposed analytic procedure has been validated by testing quality control sample (QCS) of AHH probed with the two FQNs at 10, 50, 500, and 1000 ng/mL concentration values. Validation of the method has been checked by accuracy and precision data of intra-day and long-term experiments. The two FQN concentrations have been measured by HPLC technique with UV detection at 278-nm wavelength for the AHH of patients to whom were supplied oral doses of FQNs (500 mg) twice in a day, within 1-24h before the surgery intervention of cataract. The average concentration of Ciprofloxacin resulted 186 ng/mL and that of Ulifloxacin 78 ng/mL. The nice quality of the proposed analytic procedure means that it may be suitable for in vivo studies of pharmacokinetics regarding these substances in the AHH medium.

Quinolones, an expanding class of clinically established potent antibiotics, is not freely soluble in water which prevents the design of liquid dosage forms and restricts their use in topical applications. In the present study we investigated the potential of polyamidoamine (PAMAM) dendrimers as drug carriers of quinolones (nadifloxacin and prulifloxacin) by aqueous solubility and antibacterial activity studies. Results showed that the aqueous solubility of nadifloxacin and prulifloxacin was significantly increased by PAMAM dendrimers. Microbiology studies showed that nadifloxacin and prulifloxacin still exhibit their strong antimicrobial activities in the presence of dendrimers. These studies indicated that PAMAM dendrimers might be considered as biocompatible carriers of quinolones under suitable conditions.

Foreign body-associated infectious disease is currently one of the most problematic hospital-acquired infections. Patients with placement of urinary catheters are especially susceptible to such infection, that is biofilm infection. In this study, we focused on the therapeutic efficacy of prulifloxacin (PUFX) against Pseudomonas aeruginosa OP 14-210, isolated from a patient with complicated urinary tract infection. This microbe formed a biofilm on the surface of a polyethylene tube (PT) placed in a rat bladder without surgical manipulation. In addition, we attempted to eradicate the biofilm by treatment with a combination of PUFX and fosfomycin (FOM). A single oral administration of PUFX at a dose of 20 mg/kg was effective against P. aeruginosa as a biofilm, yielding a significant reduction in CFU per PT of approximately 1 log(10) CFU/PT compared with that in untreated controls. A similar therapeutic effect was also observed in levofloxacin-treated rats, and albeit slightly weaker, in ciprofloxacin-treated animals as well. Because 3 days' consecutive treatment with each fluoroquinolone did not further decrease the viable cell counts on the PT, we tested the efficacy of combining PUFX and FOM. These two drugs, administered once a day for 3 days, at doses of 20 and 100 mg/kg, respectively, resulted in significant decreases of viable cell counts on the PT of more than 1.5 log(10) CFU/PT compared with PUFX alone (P0.05). As seen by scanning electron microscopy, destruction and disappearance of multilayer biofilms occurred after treatment with this drug combination. Such combination therapy with PUFX and FOM may be advantageous for treating biofilm-related infectious diseases.

Prulifloxacin, the prodrug of ulifloxacin, is a broad-spectrum oral fluoroquinolone antibacterial agent. After absorption, prulifloxacin is metabolized by esterases to ulifloxacin. The drug has a long elimination half-life, allowing once-daily administration. In vitro, ulifloxacin is generally more active than other fluoroquinolones against a variety of clinical isolates of Gram-negative bacteria, including community and nosocomial isolates of Escherichia coli, Klebsiella spp., Proteus, Providencia and Morganella spp., Moraxella catarrhalis, Haemophilus spp., and Pseudomonas aeruginosa. Gram-positive organisms, including methicillin- or oxacillin-susceptible Staphylococcus aureus, Enterococcus spp. and Streptococcus pneumoniae, are susceptible to ulifloxacin. In well-designed clinical trials, prulifloxacin showed good clinical and bacteriological efficacy (similar to that of ciprofloxacin) and was generally well tolerated, demonstrating a similar tolerability profile to that of ciprofloxacin. In conclusion, the in vitro inhibitory and bactericidal activities exhibited by ulifloxacin and the favorable characteristics shown by its prodrug (prulifloxacin) in clinical trials, particularly indicate this drug for the treatment of lung and urinary infections.

Minimal inhibitory concentrations (MICs) of prulifloxacin were evaluated in comparison with ciprofloxacin, levofloxacin and moxifloxacin against a large collection ( N =300) of Pseudomonas aeruginosa strains characterised according to the CLSI/NCCLS microdilution method. Additional in vitro tests (time–kill curves and mutant prevention concentration (MPC) determinations) were carried out. Assuming a susceptibility breakpoint for prulifloxacin identical to that of ciprofloxacin, the new fluoroquinolone emerged as the most potent antibiotic (72% of susceptible strains versus 65%, 61% and 23% for ciprofloxacin, levofloxacin and moxifloxacin, respectively). Time–kill tests at 4× MIC confirmed the pronounced bactericidal potency of the drug against P. aeruginosa . Amongst the members of the fluoroquinolone class assessed, prulifloxacin produced the lowest MPC values (≤4mg/L). Our in vitro results indicate that prulifloxacin represents the most powerful antipseudomonal drug available today.

Fluoroquinolones has been rarely associated with exacerbation of myasthenia gravis (MG). We present a case of MG following a treatment with prulifloxacin, a new broad-spectrum oral fluoroquinolone. Fluoroquinolones of any generation may interfere with neuromuscular transmission and should be avoided in patients with MG.

We report the results of a prospective randomised study to evaluate the therapeutic effect of Serenoa repens, Urtica dioica (ProstaMEV®), quercitin and curcumin (FlogMEV®) extracts associated with prulifloxacin in patients affected by chronic bacterial prostatitis (CBP). From a whole population of 284 patients, 143 patients affected by CBP [National Institutes of Health (NIH) class II prostatitis] were enrolled. All patients received prulifloxacin 600 mg daily for 14 days, in accordance with antibiogram results. Patients were split into two groups: Group A received prulifloxacin associated with ProstaMEV® and FlogMEV ® ; Group B received only antibiotic therapy. Microbiological and clinical efficacies were tested by two follow-up visits at 1 month and 6 months, respectively. Quality of life (QoL) was measured using the NIH Chronic Prostatitis Symptom Index (CPSI) and International Prostatic Symptom Score (IPSS) questionnaires. Group A comprised 106 patients and Group B comprised 37 patients. One month after treatment, 89.6% of patients who had received prulifloxacin associated with ProstaMEV® and FlogMEV® did not report any symptoms related to CBP, whilst only 27% of patients who received antibiotic therapy alone were recurrence-free (P < 0.0001). Significant differences were found between groups in terms of symptoms and QoL (P < 0.0001 for both). Six months after treatment, no patients in Group A had recurrence of disease whilst two patients in Group B did. Questionnaire results demonstrated statistically significant differences between groups (all P < 0.001). The association of S. repens, U. dioica (ProstaMEV®), quercitin and curcumin (FlogMEV ® ) extracts is able to improve the clinical efficacy of prulifloxacin in patients affected by CBP.