Quantitative determination of the ß-methyl carbapenem doripenem in powder for injection by a stability-indicating capillary zone electrophoresis method.

A capillary zone electrophoresis method for quantitative determination of doripenem in synthetic matrix was developed. The stability-indicating capability was performed applying stress testing protocols. The selected analytical conditions include 100 mM sodium borate buffer (pH 8.0) as run electrolyte, voltage of +15 kV, hydrodynamic injection of 5s (50 mBar), detection at 298 nm and temperature of analysis of 25 degrees C. The electrophoretic separation was carried out in a fused silica capillary (effective length 40 cm, 50 µm i.d.), using procainamide hydrochloride as internal standard. The proposed method showed quickness and reproducibility, with an analytical run in a total time of 5 min. The percentage of drug amount estimated was 101.33% (RSD = 0.80), with satisfactory intra-day and inter-day precision. In the recovery test, the method was found to be reliable and accurate in the drug quantitation (mean recovery = 101.86%). The robustness was performed applying the Plackett-Burman experimental design which confirmed the assay reliability. Based on results from forced degradation study, the stability-indicating capability was established, being observed a major degradation in alkaline, photolytic and thermal conditions. In comparison to HPLC method previously developed, the proposed capillary electrophoresis assay is statistically equivalent.

Capillary zone electrophoresis for determination of vildagliptin (a DPP-4 inhibitor) in pharmaceutical formulation and comparative study with HPLC.

A stability-indicating capillary zone electrophoresis (CZE) method was validated for the determination of vildagliptin (VLG) in pharmaceutical dosage forms using ranitidine hydrochloride (RH) as internal standard. The CZE method was carried out in a fused silica capillary (64.5 cm total length and 56.0 cm effective length, 50 microm i.d.) by applying a potential of 25 kV (positive polarity), hydrodynamic injection by 50 mbar for 5 s and the temperature of the capillary cartridge was 25 degreesC. The selected background electrolyte (BGE) consisted of 25 mM potassium phosphate (pH 8.0) with UV/PDA detection at 207 nm. The electrophoretic separation was obtained within 6 min and was linear in the range of 50-200 microg/mL (r= 0.9994). The specificity and the stability-indicating capability were demonstrated through degradation studies, which also showed that there was no interference of the formulation excipients. The method was validated in accordance to ICH guidelines acceptance criteria for specificity, linearity, precision, accuracy, robustness and system suitability. The proposed method was compared with HPLC method previously validated for this drug, and statistical analysis showed no significant difference between the methods.

Levodopa microparticles for pulmonary delivery: photodegradation kinetics and LC stability-indicating method.

Levodopa, (S)-2-amino-3-(3,4-dihydroxyphenyl) propanoic acid, is still considered the gold standard treatment for Parkinson's disease. However, oral levodopa shows poor pharmacokinetics and its efficacy becomes problematic with the progression of the disease. Pulmonary delivery using the association of the polymers: chitosan, hyaluronic acid and HPMC, represents a novel approach to overcome this problem. A stability-indicating liquid chromatography method for the quantitative determination of levodopa microparticles for pulmonary delivery was developed as well as its photodegradation kinetics in solution. The developed and validated method was applied for the analyses of the novel formulation as well as for protocols of stability studies.

Photodegradation kinetics, cytotoxicity assay and determination by stability-indicating HPLC method of mianserin hydrochloride.

A stability-indicating HPLC method for the determination of mianserin hydrochloride in coated tablets was developed and validated. Also, drug photodegradation kinetics and cytotoxicity were determined. Chromatographic analyses were performed in an Ace RP-18 octadecyl silane column (250 mm x 4.6 mm i.d., particle size 5 microm) maintained at ambient temperature (25 degrees C). The mobile phase was composed of methanol, 50 mM monobasic potassium phosphate buffer and 0.3% triethylamine solution adjusted to pH 7.0 with phosphoric acid 10% (85:15, v/v) in isocratic mode at a flow rate of 1.0 mL x min(-1). The performed degradation conditions were: acid and basic media with HCl 1.0 M and NaOH 1.0 M, respectively, oxidation with H2O2 3% and the exposure to UV-C light. No interference in the mianserin hydrochloride elution was verified by degradation products formed. Linearity was assessed and ANOVA showed non-significant linearity deviation (p > 0.05). Adequate results were obtained for repeatability, intermediate precision, accuracy and robustness. The photodegradation kinetics of mianserin hydrochloride was evaluated in methanol. The degradation of mianserin could be better described as zero order kinetic (r = 0.9982). The UV-C degraded samples of mianserin hydrochloride were also studied in order to determine the preliminary cytotoxicity in vitro against mononuclear cells.

Microparticles containing lemongrass volatile oil: preparation, characterization and thermal stability.

Lemongrass volatile oil (LVO) is an important ingredient in cosmetics, presenting antimicrobial properties, in particular antifungal activity, and it is a promising raw material for the development of pharmaceutical products. However, its volatility and susceptibility to degradation are the major drawbacks for the use of Cymbopogon citratus oil in pharmaceutical compounding. Thus, the aim of this work was to develop and to characterize microparticles containing this oil viewing the stabilization of LVO. Two techniques of preparation were evaluated; spray drying and precipitation, and two encapsulation materials, beta-cyclodextrin (beta-CD) and hydroxypropyl-beta-cyclodextrin (HP-beta-CD) were tested. The microparticles were characterized in terms of content of water, yield, percentage of inclusion, infrared spectroscopy. Morphology was evaluated by scanning electronic microscopy. Studies of stability were also conducted. The content of citral (neral and geranial), major component of the oil, present in microparticles was assayed by a validated HPLC method. The percentage of inclusion of LVO into the microparticles was 56-60% and 26-29% using beta-CD and HP-beta-CD, respectively. The results showed that the use of the beta-CD as encapsulant material was more efficient. Additionally, an increased inclusion of lemongrass oil was observed with the precipitation technique.

Identification and stability of a new bichalcone in Achyrocline satureioides spray dried powder.

A new chemical structure, the 4,2',4",2'''-tetrahydroxy-6',6'''-dimethoxy-4'-O-4'''- bichalcone, named achyrobichalcone was isolated and identified from an Achyrocline satureioides spray-dried powder (SDP80). The thermal and photo stability of this new compound as well as that of the main polyphenols present in the spray dried powder, quercetin, luteolin, 3-O-metylquercetin and the corresponding kinetics of degradation are reported. In the long-term testing (30 +/- 2 degrees C/75 +/- 5% RH, 12 months), the total polyphenols contained in SDP80 demonstrated to be stable, remaining higher than 90% after a 12 month exposure. The photo stability testing revealed that all polyphenols were stable for 48 h when SDP80 was conditioned in amber or transparent flasks and exposed to UV-C radiation (light express LE UV, 254 nm, 30W). In contrast, when unprotected, the polyphenols demonstrated to be sensitive to both, thermal stress testing (80 +/- 2 degrees C), for 14 days and to UV-C radiation. Luteolin showed to be the most stable against UVC light and 3-O-methylquercetin against temperature. The achyrobichalcone demonstrated to be the more unstable against both, temperature and light. The kinetics of polyphenol thermal degradation (80 +/- 2 degrees C, 49 days) and photodegradation (UV-C radiation, 96 h) followed, 2nd and 1st order reaction, respectively.

Increasing sodium pantoprazole photostability by microencapsulation: effect of the polymer and the preparation technique.

Pantoprazole sodium is a proton pump inhibitor, used in acid-related disorders, like peptic ulcers and gastroesophageal reflux. This drug is unstable in acid solution and in the presence of salts. The aim of this work was to study the photostability under UVC radiation of pantoprazole and to determine its kinetics. A methanol solution and the solid pantoprazole were evaluated by HPLC within 120 min and 10 days, respectively. The work was also dedicated to evaluate and compare the ability of microencapsulation in stabilizing pantoprazole after UVC radiation. Pantoprazole-loaded microparticles prepared by emulsification/solvent evaporation or spray drying were compared. Pantoprazole was encapsulated using Eudragit S100 or its blend with poly(epsilon-caprolactone) or HPMC. In methanol solution, pantoprazole was completely degraded after 120 min and presented zero-order kinetics with t1/2 of 6.48 min. In the solid form, after 10 days, pantoprazole concentration was reduced to 27% following zero-order kinetic. The microparticles prepared only with Eudragit S100 demonstrated an increase of the drug photostability. After 10 days of irradiation, 56 and 44% of the drug was stable when encapsulated by emulsification/solvent evaporation and spray drying, respectively. The use of polymer blends did not improve the pantoprazole photostability.

Thermal and photo stability of phenolic constituents of an Achyrocline satureioides spray-dried powder.

Thermal and the photo stabilities of an Achyrocline satureioides powder (SDP40) were evaluated in particular concerning the total polyphenol content as well as the main identified constituents quercetin, luteolin, 3-O-methylquercetin and caffeic acid. SDP40 presented good stability for nine months under normal storage conditions of 25 degrees C temperature and 60% relative humidity (RH). In accelerated term testing, 50 degrees C temperature and 90% RH and also in stress testing, 80 degrees C, caffeic acid and a non-identified constituent P3 were the most instable constituents. Luteolin and 3-O-methylquercetin were the most stable constituents. Quercetin presented an unusual behavior, improving its concentration after 1 month at 50 degrees C or 2 days at 80 degrees C exposition, followed by a decrease in its concentration. The hypothesis that this observation is related to the simultaneous decreasing of a non-identified peak P3 or to the hydrolysis of a non-identified precursor as a quercetin heteroside is being investigated. The SDP40 presented good stability against UV-C light when conditioned in amber or transparent containers, but it suffered degradation when stored in open-dishes. In summary, the total polyphenol content remains within acceptable limits of 10% under normal storage conditions for nine months. However, the LC polyphenol analysis demonstrated that the behavior of individual constituents has still to be enlightened.

Antioxidant, a pro-oxidant and cytotoxic effects of Achyrocline satureioides extracts.

In this study we compared the antioxidant properties of five different extracts of different composition obtained from Achyrocline satureioides' inflorescences (Compositae), a widely used Brazilian folk medicinal herb. All of the extracts presented significant antioxidant potential identified by TRAP assay, which increased in the presence of human plasma. Characterization of the content of flavonoids in each extract showed that the FDP80 (ethanol 80%) and FFr (enriched flavonoid fraction) extracts contained a higher content of flavonoids. Cytotoxicity of the extracts as determined in Sertoli cell culture showed that FDP80 and FFr were highly toxic at most concentrations tested. The extracts induced a significant increase in lipid peroxidation levels in Sertoli cells. These results suggest that medicinal herb extracts that contain higher flavonoid concentrations and shows higher antioxidant protection in vitro might not always produce the greatest benefit.

Determination of dexamethasone acetate in cream by HPLC.

The aim of this research was to validate a high performance liquid chromatographic method for the quantitative determination of dexamethasone acetate contained in cream preparation. A MetaSil octadecyl silane (250 x 4.6 mm, 5 microm) column, a methanol: water (65:35; v/v) mobile phase (1.0 ml min(-1)) and an UV detector (set at 254 nm) were used to evaluate the parameters: linearity, precision, accuracy, specificity, as well as, quantitation and detection limits. The calibration curve showed a correlation coefficient of 0.9999. The precision was demonstrated by the relative standard deviation (RSD) of 0.53. The recovery test resulted in an average of 97.85%, what confirmed the accuracy of the method. The quantitation and detection limits determined were 1.41 and 0.47 microg ml(-1), respectively. The specificity test showed there was no interference in the drug peak.

Microbiological assay for azithromycin in pharmaceutical formulations.

The validation of a microbiological assay, applying the cylinder-plate method, for the determination of the antibiotic azithromycin is described. Using a strain of Micrococcus luteus ATCC 9341 as the test organism, azithromycin at concentrations ranging from 0.1 to 0.4 microgml(-1) could be measured in capsules and suspensions. A prospective validation of the method showed that it was linear (r=0.998), precise (RSD=1.40-capsules; RSD=1.19-powder for suspension and RSD=1.73-oral suspension) and accurate (it measured the added quantities). We conclude that the microbiological assay is satisfactory for quantitation of in vitro antibacterial activity of azithromycin.

LC determination of flavonoids: separation of quercetin, luteolin and 3-O-methylquercetin in Achyrocline satureioides preparations.

The pharmacological activities of the flavonoids show the interest in quantifying these constituents in phytopharmaceutical preparations, as well as in the validation of the analytical methodologies. LC methods have been reported to quantify isolated flavonoids or these compounds in complex biological matrices, such as herbal raw materials and extractive preparations. This work was designed, therefore, to develop an LC system to separate quercetin, luteolin and 3-O-methylquercetin and to quantify them in extractive solutions from Achyrocline satureioides. The main validation parameters of the method are also determined. The method showed linearity for quercetin and luteolin in the range 1-10 microg/ml. The aqueous and ethanol 80% extractive solutions showed linear response in the range 2.5-20 microl/ml and ethanol 40% extractive solution in the range 2.5-10 microl/ml. Precision and accuracy were determined for ethanol 80% extractive solution, in the concentration of 10 microl/ml. The LC method showed an excellent performance in separating the flavonoids quercetin, luteolin and 3-O-methylquercetin in A. satureioides extracts, since the presence of interference has been previously evaluated.

Ethnopharmacological studies of antimicrobial remedies in the south of Brazil.

This study reports the antimicrobial evaluation of the species most commonly used in Rio Grande do Sul (RS), the southernmost state of Brazil, for treating conditions likely to be associated with microorganisms. A four-stage process of documentation and evaluation was conducted: (a). review of RS ethnobotanical studies; (b). analysis of traditional uses; (c). literature survey on phytochemical and pharmacological data; (d). microbiological screening of selected plants. From the 149 species initially identified, 49 were cited as being used for microbial associated conditions in at least two other regions in RS, and 18 were further selected for screening. The crude methanol extract of these 18 plants were evaluated against seven microorganisms using the diffusion agar test. Extracts from Chaptalia nutans, Cordia monosperma, Echinodorus grandiflorus, Eugenia uniflora, Leonurus sibiricus, Luehea divaricata, Malva sylvestris, Ocotea odorifera, Parapiptadenia rigida, Pluchea sagittalis, Psidium cattleyanum and Senna neglecta were active against at least one microorganism. Although preliminary, these results are useful for rationalizing the use of medicinal plants in established systems of traditional medicine in primary health care.

Influence of penetration enhancers and molecular weight in antifungals permeation through bovine hoof membranes and prediction of efficacy in human nails.

This work aimed to evaluate the effect of different substances on the permeation of geraniol through bovine hoof membranes. Different penetration enhancers were able to increase the permeability up to 25 times compared to control. It was demonstrated that acetilcysteine in association with ascorbic acid increased the permeation, even in acid formulations. In addition, some antifungal drugs were incorporated into a gel formulation of HPMC containing acetylcysteine 5% and ascorbic acid 0.2% and then the permeation coefficient through bovine hoof membranes was evaluated. The relationship between permeability and molecular weight was established for fluconazole, miconazole, terbinafine, butenafine, geraniol and nerol. Geraniol and nerol, the antifungals with lower molecular weight, had the better permeability results. Permeability coefficients for nail plates were estimated and geraniol demonstrated similar or even better efficacy index values against T. rubrum, T. menthagrophytes and M. canis compared with terbinafine and miconazole.

Degradation kinetics of telithromycin determined by HPLC method.

The degradation kinetics of the antibiotic telithromycin using a stability-indicating high-performance liquid chromatography (HPLC) method is demonstrated. The photodegradation is performed by UVC lamp-254 nm (15W), installed in a chamber internally coated with mirrors, where telithromycin solutions prepared from coated tablets are placed in quartz cells. To promote oxidation, the reaction between the telithromycin solution and 3% hydrogen peroxide solution is carried out. The kinetics parameters of order of reaction and the rate constants of the degradation are determined for both conditions. The degradation process of telithromycin can be described by first-order kinetics under both experimental conditions used in this study. The results reveal the photo and oxidation lability of the drug and confirm the reliability of HPLC method for telithromycin in the presence of its degradation products.

LC: analysis of photodegradation kinetics of nitazoxanide in pharmaceutical formulations.

Nitazoxanide is a new broad-spectrum, antiparasitic drug agent. The photodegradation of nitazoxanide was studied in order to investigate the degradation kinetics of this drug. The analyses of the degraded samples were performed by a stability-indicating liquid chromatographic method. The degradation was carried out in acetonitrile with coated tablets or oral suspension powder in quartz cells under UVC light at 254 nm. The kinetics parameters, such as order of reaction, rate constants, half-life (t(1/2)), and the time when 90% of the drug original concentration was left, were determined. The photodegradation of nitazoxanide for both pharmaceutical formulations in acetonitrile solution shows a zero-order kinetics under the applied experimental conditions. The obtained results confirm the reliability of the chromatographic method for determining the kinetics run of nitazoxanide in the presence of its degradation products. The present study reveals the photolability of the drug in solution. Thus, appropriated photoprotection is recommended during the manipulation of the drug.

Influence of excipients and technological process on anti-inflammatory activity of quercetin and Achyrocline satureioides (Lam.) D.C. extracts by oral route.

The flavonoids quercetin, 3-O-methylquercetin and luteolin play an important role in the anti-inflammatory activity of Achyrocline satureioides ethanol extracts when administered intraperitoneally. The present work describes the oral anti-inflammatory effect of quercetin and A. satureioides extracts and the role played by the solvent concentration, adjuvant and drying processes of freeze-drying (FD) or spray-drying (SD) on the effect. The best anti-edema effect was observed with 250 mg/kg body wt of the freeze-dried powder (FDP), prepared with 40% (v/v) ethanol (FDP40). In contrast, 250 mg/kg body wt of FDP80, prepared with ethanol 80% (ES80), did not significantly inhibit the carrageenan-induced rat paw edema. However, when ES80 was freeze-dried in the presence of polysorbate 80 (FDP80-P80) or spray-dried in the presence of colloidal silicon dioxide (CSD) and P80 (SDP80), both dried extracts became more active. Quercetin suspension in saline did not inhibit paw edema, but the mixture of quercetin with polysorbate 80 was effective in edema inhibition by the oral route. Aqueous extract (ESAQ), freeze-dried (FDPAQ, FDPAQ-P80) or spray-dried (SDPAQ) did not exhibit the edema-inhibition effect. Taken together, the results point to the following order of efficacy (at 4 h, for example): FDP40 > indomethacin > SDP40 > SDP80 = FDP80-80 > Quercetin-P80. Additionally, the FDP40, SDP40 (prepared from 40% v/v ethanol added of CSD) and SDP80 reduced the total leukocyte and polymorphonuclear cell migration in the pleural cavity.

Antimicrobial activity of some Hypericum species.

The crude methanolic extracts of six species of Hypericum [H. caprifoliatum Cham. & Schlecht., H. carinatum Griseb., H. connatum Lam., H. ternum A. St. Hil., H. myrianthum Cham. & Schlecht. and H. polyanthemum Klotzsch ex Reichardt] growing in southern Brazil were analyzed for antimicrobial activity against several microorganisms (bacteria and fungi). The most active plant was H. caprifoliatum, which showed activity against Staphylococcus aureus. Only H. polyanthemum and H. ternum extracts were active against Bacillus subtilis. None of the crude methanolic extracts showed activity against S. epidermidis, Escherichia coli or Saccharomyces cerevisiae. Extracts from these species were evaluated chemically and tannin, flavonoid and phenolic acids were the prominent compounds. The plants contained quercitrin, hyperoside (except H. connatum) and, less frequently, isoquercitrin and chlorogenic acid. In contrast to H. perforatum, which has high concentrations of rutin, these species do not produce this flavonoid or it appears as traces. The tannin concentration varied between 5.1 and 16.7% in H. myrianthum and H. ternum, respectively.

Antiinflammatory investigation of some species of Mikania.

Mikania laevigata Schultz Bip. ex Baker, M. involucrata Hook. et Arn. and M. hirsutissima DC. (Asteraceae), commonly occurring in the southern Brazilian State of Rio Grande do Sul, were submitted to biological tests to evaluate their potential antiinflammatory activity. Decoctions from the leaves and stems were analysed by the induced rat paw oedema and pleurisy models. The animals were treated orally with different decoction doses. In the induced rat paw oedema test, the animals treated with leaf decoctions from M. laevigata (200 mg/ kg) and M. involucrata (50 mg/ kg) presented an oedema inhibition of 81.56% and 81.67%, respectively, 3 h after the administration of the phlogistic agent. Leaf decoctions from M. hirsutissima (400 mg/ kg) did not show such an activity. Stem decoctions displayed lower antiinflammatory activity when compared with the same doses and response time of the leaf decoctions for all analysed species. In the pleurisy assay, leaf decoctions from M. laevigata (400 mg/ kg) and M. involucrata (200 mg/ kg) inhibited leukocyte migration to the pleural exudate by 28.26% and 54.35%, respectively.

Análise térmica de esparfloxacino / Thermal analysis of sparfloxacin

Foi realizada a análise térmica de esparfloxacino, fluorquinolona de terceira geração que possui potente atividade contra bactérias Gram-positivas, como Streptococcus pneumoniae e Staphylococcus aureus inclusive cepas meticilina resistentes (MRSA), bactérias Gram-negativas, anaeróbios, Legionella spp., Mycoplasma spp., Chlamydia spp. e Mycobacterium spp. Nas curvas DTA observa-se pico endotérmico de fusão na temperatura de 276,5ºC. A curva DTA, em ar sintético, apresenta dois picos exotérmicos (341,6 e 579,2ºC), atribuídos à decomposição do composto. A curva TG permite observar a perda de massa total em duas etapas, entre as temperaturas de 285,5 e 645,3ºC. A curva DTA, em atmosfera de nitrogênio, apresenta pico exotérmico de decomposição na temperatura de 340,0ºC e na curva TG, a perda de massa inicia-se na temperatura de 254,4º