RESUMEN
The imbalance in skin microbiota is characterized by an increased number of pathogens in respect to commensal microorganisms. Starting from a skin microbiota collection, the aim of this work was to evaluate the possible role of Pomegranate (Punica granatum L.) Peel Extract (PPE) in restoring the skin microbiota balance acting on Staphylococcus spp. PPE was extracted following green methodology by using n-butane and the Dimethyl Ether (DME) solvents and analyzed for phytochemical composition and antimicrobial activity. The PPE antimicrobial action was evaluated against Gram +, Gram - bacteria and yeast reference strains and the most effective extract was tested against the main skin microbiota isolated strains. PPE extracted with DME showed the best antimicrobial action with MICs ranging from 1 to 128 mg/mL; the main active compounds were Catechin, Quercetin, Vanillic acid and Gallic acid. The PPE in DME anti-adhesive effect was examined against S. epidermidis and S. aureus mono and dual-species biofilm formation by biomass quantification and CFU/mL determination. The extract toxicity was evaluated by using Galleria mellonella larvae in vivo model. The extract displayed a significant anti-adhesive activity with a remarkable species-specific action at 4 and 8 mg/mL against S. epidermidis and S. aureus mono and dual-species biofilms. PPE in DME could represent an eco-sustainable non-toxic strategy to affect the Staphylococcal skin colonization in a species-specific way. The innovation of this work is represented by the reuse of food waste to balance skin microbiota.
Asunto(s)
Biopelículas , Pruebas de Sensibilidad Microbiana , Microbiota , Extractos Vegetales , Granada (Fruta) , Piel , Staphylococcus aureus , Staphylococcus epidermidis , Staphylococcus epidermidis/efectos de los fármacos , Granada (Fruta)/química , Piel/microbiología , Piel/efectos de los fármacos , Extractos Vegetales/farmacología , Extractos Vegetales/química , Microbiota/efectos de los fármacos , Staphylococcus aureus/efectos de los fármacos , Biopelículas/efectos de los fármacos , Humanos , Animales , Antibacterianos/farmacología , Frutas/microbiología , Frutas/químicaRESUMEN
The increased risk of illness and disability is related to the age inevitable biological changes. Oxidative stress is a proposed mechanism for many age-related diseases. The crucial importance of polyphenol pharmacophore for aging process is largely described thanks to its effects on concentrations of reactive oxygen species. Resveratrol (3,5,4'-trihydroxy-trans-stilbene, RSV) plays a critical role in slowing the aging process but has a poor bioavailabity after oral intake. In this present work, a series of RSV derivatives was designed, synthesized, and evaluated as potential antioxidant agents. These derivatives contain substituents with different electronic and steric properties in different positions of aromatic rings. This kind of substituents affects the activity and the bioavailability of these compounds compared with RSV used as reference compound. Studies of Log P values demonstrated that the introduction of halogens gives the optimum lipophilicity to be considered promising active agents. Among them, compound 6 showed the higher antioxidant activity than RSV. The presence of trifluoromethyl group together with a chlorine atom increased the antioxidant activity compared to RSV.
Asunto(s)
Técnicas de Química Sintética , Estilbenos/síntesis química , Estilbenos/farmacología , Animales , Línea Celular , Halogenación , Humanos , Ratones , Modelos Teóricos , Estructura Molecular , Estilbenos/químicaRESUMEN
INTRODUCTION: The analysis of plant and herbal samples is a challenging task for analytical chemists due to the complexity of the matrix combined with the low concentration of analytes. In recent years different liquid-phase microextraction (LPME) techniques coupled with a variety of analytical equipment have been developed for the determination of both organic and inorganic analytes. OBJECTIVE: Over the past few years, the number of research papers in this field has shown a markedly growing tendency. Therefore, the purpose of this review paper is to summarise and critically evaluate research articles focused on the application of LPME techniques for the analysis of plant and herbal samples. RESULTS: Due to the complex nature of the samples, the direct application of LPME techniques to the analysis of plants has not often been done. LPME techniques as well as their modalities have been commonly applied in combination with other pretreatment techniques, including a solid-liquid extraction technique supported by mechanical agitation or auxiliary energies for plant analysis. Applications and the most important parameters are summarised in the tables. CONCLUSION: This review summarises the application of the LPME procedure and shows the major benefits of LPME, such as the low volume of solvents used, high enrichment factor, simplicity of operation and wide selection of applicable detection techniques. We can expect further development of microextraction analytical methods that focus on direct sample analysis with the application of green extraction solvents while fully automating procedures for the analysis of plant materials.
Asunto(s)
Microextracción en Fase Líquida , Plantas , SolventesRESUMEN
Phenolic compounds are secondary metabolites that are found ubiquitously in plants, fruits, and vegetables. Many studies have shown that regular consumption of these compounds could have a positive effect on our health. The aim of this study was to compare the phytochemical contents of the water extracts from three different plants used as folk remedies in Turkey: Aesculus hippocastanum, Olea europaea, and Hypericum perforatum. A liquid chromatography-electrospray tandem mass spectrometry (LC-ESI-MS/MS) analysis was performed to explore the phenolic profiles. The biological activities of these extracts were also evaluated in terms of their antioxidant activities (2,2-diphenyl-1-picrylhydrazyl DPPH, 2,2'-azino-bis-(3-ethylbenzothiazoline-6-sulfonic acid ABTS, Ferric Reducing Antioxidant Power Assay FRAP, cupric ion reducing antioxidant capacity CUPRAC, ß-carotene, phosphomolybdenum, and metal chelating) and enzyme inhibitory properties (against acetylcholinesterase, butyrylcholinesterase, and tyrosinase). The aqueous extract of H. perforatum showed the highest levels of total phenolic, flavonoid, and saponin contents. Protocatechuic acid, vanillic acid, verbascoside, hesperidin, hyperoside, apigenin 7-hexosides, and quercetin were the most common compounds found in this species. The results confirm that A. hippocastanum, O. europaea, and H. perforatum represent a potential source of natural-derived molecules with positive properties that could be used as valid starting point for new food supplements, and drugs in the pharmaceutical, cosmetic, and food industries.
Asunto(s)
Aesculus/enzimología , Hypericum/enzimología , Medicina Tradicional , Olea/enzimología , Extractos Vegetales/química , Extractos Vegetales/farmacología , Antioxidantes/química , Antioxidantes/farmacología , Flavonoides , Fenoles , Fitoquímicos/química , Fitoquímicos/farmacología , Saponinas , TurquíaRESUMEN
In this paper a dispersive magnetic-solid phase extraction (MSPE) using a graphene nanocomposite (rG/Fe3O4) followed by ultra high performance liquid chromatography with photodiode array detection has been developed for the simultaneous analysis of new class of oral anticoagulants (NOAs) in human plasma. The performance of the nanocomposite graphene@Fe3O4 on the magnetic solid phase extraction of apixaban, rivaroxaban and dabigatran has been optimized using a Box-Behnken design of experiment. The amount of graphene nanocomposite, the sample pH and the adsorption time were the investigated parameters as a function of the extraction recovery. The analytical method was fully validated based on linearity, limit of detection (LOD), limit of detection (LOQ), inter- and intra-day precision and trueness, and extraction yield. Under optimal condition, excellent linearity (R2 > 0.9987) over the range (0.001-5.0⯵g/mL), limit of detection (0.003⯵g/mL), precision (0.81-8.97% RSD) and trueness (-5 to 9 % BIAS%) were observed for the target drugs. The average extraction recovery under optimal from plasma samples ranged between 96.6-98.6% for apixaban, rivaroxaban and dabigatran and the internal standard. The proposed method was developed, validated and successfully applied to the measurement of these NOAs in patients. The new approach offers an attractive alternative for the simultaneous analysis of the selected NOAs from plasma samples, providing several advantages including fewer sample preparation steps, ease of performance, and higher recoveries compared to traditional methodologies.
Asunto(s)
Anticoagulantes/análisis , Cromatografía Líquida de Alta Presión/métodos , Grafito/química , Plasma/química , Extracción en Fase Sólida/métodos , Dabigatrán/análisis , Humanos , Límite de Detección , Nanopartículas de Magnetita/química , Nanocompuestos/química , Pirazoles/análisis , Piridonas/análisis , Rivaroxabán/análisisRESUMEN
The Lamiaceae family comprises many flowering plants classified into about 236 genera. The genus Ziziphora is one of the well-known genera of this family and its species are important in different fields of pharmaceutical, chemical, traditional, and folk medicines. The phytochemicals present in Ziziphora include monoterpenic essential oils, triterpenes, and phenolic substances. The aim of this paper was to study the phytochemical profile of Ziziphora taurica subsp. taurica and compare and evaluate the biological activities of its ethyl acetate (ZTT-EtOAc), methanolic (ZTT-MeOH), and aqueous (ZTT-W) extracts based on their enzyme inhibition and antioxidant capacities. Determination of total phenolic (TPC) and total flavonoid (TFC) contents as well as biological activities were determined using spectrophotometric procedures. Subsequently, the individual phenolic compounds were detected by liquid chromatography-electrospray ionization-tandem mass spectrometry (LC-ESI-MS/MS). In total, twenty-two different phenolic compounds were identified, including apigenin, ferulic acid, and luteolin which were the most common. ZTT-MeOH extract showed the best antioxidant activity, whereas ZTT-EtOAc extract was the most effective against tyrosinase and α-amylase. Ziziphora taurica subsp. taurica represents a potential source of natural compounds with positive effects on human health.
Asunto(s)
Antioxidantes/farmacología , Inhibidores Enzimáticos/farmacología , Flavonoides/farmacología , Lamiaceae/química , Fenoles/farmacología , Fitoquímicos/farmacología , Antioxidantes/análisis , Benzotiazoles/antagonistas & inhibidores , Compuestos de Bifenilo/antagonistas & inhibidores , Inhibidores Enzimáticos/análisis , Flavonoides/análisis , Humanos , Estructura Molecular , Monofenol Monooxigenasa/antagonistas & inhibidores , Monofenol Monooxigenasa/metabolismo , Fenoles/análisis , Fitoquímicos/análisis , Picratos/antagonistas & inhibidores , Extractos Vegetales/química , Ácidos Sulfónicos/antagonistas & inhibidores , alfa-Amilasas/antagonistas & inhibidores , alfa-Amilasas/metabolismoRESUMEN
In the present work we analyzed the hydrophobicity and hydrophilicity properties of several non-steroidal anti-inflammatory drugs (NSAIDs) by investigating the structural changes of the dynamic hydrogen bond network in order to predict the extraction recovery of NSAIDs from biological fluids set by solid phase extraction (SPE). This work allows investigating the relationship between theoretical descriptors and experimental data using a parameter free method with a strong correlation (Pearson correlation 0.95, p-value 0.0003). The identification and quantification of analytes in human plasma were carried out by high performance liquid chromatography coupled with photodiode array detection (HPLC-PDA) using a Kinetex Evo C18 (150â¯xâ¯4.6â¯mm I.D) protected by a guard column and a mixture of acetonitrile and 10â¯mM phosphate buffer (pH 2.5) (50:50, v/v) as mobile phase at isocratic conditions. Accuracy (BIAS%) ranged within -2.33% andâ¯+â¯8.05% while precision (RSD%) was less than 5.73%.The mean extraction recovery of the carprofen (IS) was 84.1% and the recovery of NSAIDs from human plasma ranged between 81.9% to 86.6%. LODs and LOQs for all the investigated NSAIDs were 0.003 and 0.01⯵g/mL, respectively. The method was validated according to the ICH guide line in the range 0.010-20.0⯵g/mL.
Asunto(s)
Antiinflamatorios no Esteroideos/química , Análisis Químico de la Sangre/métodos , Cromatografía de Fase Inversa , Extracción en Fase Sólida , Antiinflamatorios no Esteroideos/sangre , Cromatografía Líquida de Alta Presión , Humanos , Límite de Detección , Plasma/química , Reproducibilidad de los ResultadosRESUMEN
A simple and rapid ultra-high-performance liquid chromatographic (UHPLC) for the simultaneous determination of meropenem and ciprofloxacin in human plasma was developed and validated. All of the analytes were separated in <5 min. A solid-phase extraction method was applied from sample preparation. Analytical separation was performed on a Poroshell SB C18 column (50 × 2.1 mm, 2.7 µm particle size) with photodiode array (PDA) detection. Meropenem and ciprofloxacin were determined at wavelengths of 300 and 277 nm, respectively. The mobile phase was a mixture of acetonitrile-10 mm ammonium acetate-methanol in gradient elution. The method has been validated for both drugs in gastric surgery for cancer patients. The method showed good linearity with correlation coefficients, r2 = 0.994 for the two drugs, as well as high precision (RSD < 10.5% in each case); accuracy ranged from -5.8 to +6.0%. The limit of quantitation of the two drugs was established at 0.02 and 0.01 µg/mL, respectively. Meropenem, ciprofloxacin and the internal standard were extracted from human plasma with a mean recovery ranging from 92.5 to 98.6%. The method was applied to quantify the drugs dosage in complicated gastric surgery patients.
Asunto(s)
Antibacterianos/sangre , Cromatografía Líquida de Alta Presión/métodos , Ciprofloxacina/sangre , Meropenem/sangre , Extracción en Fase Sólida/métodos , Antibacterianos/farmacocinética , Antibacterianos/uso terapéutico , Profilaxis Antibiótica , Ciprofloxacina/farmacocinética , Ciprofloxacina/uso terapéutico , Infección Hospitalaria/tratamiento farmacológico , Infección Hospitalaria/prevención & control , Humanos , Límite de Detección , Modelos Lineales , Meropenem/farmacocinética , Meropenem/uso terapéutico , Reproducibilidad de los Resultados , Neoplasias Gástricas/cirugíaRESUMEN
Owing to their pronounced polarity, hydrophilic interaction liquid chromatography (HILIC) can be considered as the elective choice for the LC analysis of aminoglycoside (AG) antibiotics. In the present work, a gradient program was optimized for the first time with a diol-type stationary phase and an evaporative light scattering detector (ELSD), thus allowing the almost complete separation of the nine analysed AGs: spectinomycin, dihydrostreptomycin, streptomycin A, gentamicin C1, amikacin, kanamycin A, paromomycin, apramycin and neomycin. In the optimized analysis conditions, analyte retention was found to be governed by a multimodal mechanism encompassing electrostatic, partitioning and hydrophilic interactions. However, the gradient mode of elution complicated a deep understanding of the influence of each contribution on the retention behaviour. The developed HILIC-ELSD method was applied for the analysis of commercial tablets containing neomycin co-formulated with the polypeptide antibiotic bacitracin. The method was fully validated according to the guidelines enshrined in the International Conference on Harmonization (ICH). The use of the diol-type stationary phase was well suited for implementing a successful 2D-HPLC system. Indeed, in order to cope with the absence of chemoselectivity for the couples amikacin/kanamycin and paromomycin/apramycin, a successful 2D-HPLC method was implemented with the "heart-cut" approach and the use of either heptafluorobutyric (for the former) or perfluorooctanoic acid (for the latter) as the ion-pair reagent in the second RP-LC dimension.
Asunto(s)
Aminoglicósidos/química , Aminoglicósidos/aislamiento & purificación , Antibacterianos/química , Antibacterianos/aislamiento & purificación , Cromatografía Liquida , Interacciones Hidrofóbicas e HidrofílicasRESUMEN
A novel, rapid, simple graphene/Fe3O4 based dispersive magnetic solid phase extraction was developed for the simultaneous separation/preconcentration and determination of non steroidal anti-inflammatory drugs with ultra high performance liquid chromatography coupled with photodiode array detection. Several parameters influencing the extraction efficiency of the investigated analytes such as the extraction time, the amount of graphene/Fe3O4, the sample pH, the ionic strength and the elution solvent were evaluated and optimized. Under optimal conditions, the linearity was in the range of 0.002-25 µg/mL for furprofen, diclofenac and ketoprofen, 0.003-25 for flurbiprofen, naproxen and fenbufen, 0.004-25 for indoprofen with a good coefficient of determination (R2> 0.9991) for each analyte. The inter-and- intra day accuracy (BIAS%) for human plasma and urine ranged between -7.15% to 6.20% and -5.17% to 4.87%, respectively.The precision (RSD%) in human plasma and urine was less than 8.67% and 8.92%, respectively. The proposed method was applied to the determination of NSAIDs in human plasma and urine.
Asunto(s)
Antiinflamatorios no Esteroideos/sangre , Antiinflamatorios no Esteroideos/orina , Grafito/química , Magnetismo/métodos , Nanopartículas de Magnetita/química , Extracción en Fase Sólida/métodos , Cromatografía Líquida de Alta Presión/métodos , Humanos , Límite de DetecciónRESUMEN
An ultra high-performance liquid chromatographic (UHPLC) method with PDA detection was developed and validated for the simultaneous quantification of metronidazole, meropenem, ciprofloxacin, linezolid and piperacillin in human plasma and applied to patients suffering from hospital acquired pneumonia (HAP). The method uses an air assisted dispersive liquid-liquid microextraction for sample preparation. All parameters in the extraction step, including selection of extractant, amount of extractant, ionic strength, pH, and extraction cycles, were investigated and optimized. Chromatography was carried out using a Poroshell 120 SB C18 (50â¯×â¯2.1â¯mm I.D. 2.6⯵m particle size) column and a gradient mobile phase consisting of ammonium acetate buffer (10â¯mM, pH 4.0) (eluent A); and a mixture of acetonitrile-methanol in a ratio (80/20)(eluent B). Ulifloxacin was used as internal standard. The method demonstrated good linearity with correlation coefficients, r2â¯>â¯0.9995 for the drugs, as well as high precision (RSD%â¯≤â¯9.87%), accuracy ranged from -8.14% to +8.98. The enrichment factor (EF) obtained ranged within 87 and 121. During the validation, the concentrations of the analytes were found to be stable after 3 freeze-thaw cycles and for at least 24â¯h after extraction. Subsequently, this method was used to quantify the drugs in patients with HAP in order to establish if the dosage regimen given was sufficient to eradicate the infection at the target site.
Asunto(s)
Antibacterianos/sangre , Cromatografía Líquida de Alta Presión/métodos , Monitoreo de Drogas/métodos , Microextracción en Fase Líquida/métodos , Neumonía/tratamiento farmacológico , Antibacterianos/normas , Antibacterianos/uso terapéutico , Cromatografía Líquida de Alta Presión/instrumentación , Monitoreo de Drogas/instrumentación , Monitoreo de Drogas/normas , Fluoroquinolonas/sangre , Fluoroquinolonas/uso terapéutico , Humanos , Enfermedad Iatrogénica , Límite de Detección , Microextracción en Fase Líquida/instrumentación , Piperazinas/sangre , Piperazinas/uso terapéutico , Neumonía/sangre , Estándares de ReferenciaRESUMEN
A green dispersive liquid-liquid microextraction (DLLME) using deep eutectic solvent (DES) as the extracting solvent has been developed and applied for the simultaneous quantification of ferulic acid, umbelliferone, boropinic acid, 7-isopentenyloxycoumarin, 4'-geranyloxyferulic acid (GOFA), and auraptene in some vegetable oils using ultra high performance liquid chromatography (UHPLC) with photodiode array detection (PDA). All parameters in the extraction step, including selection and loading of both extracting and dispersing solvents, amount of both extractant and disperser solvent were investigated and optimized. PhAA/TMG DES achieved higher recovery and enrichment factor compared to other DESs. The validated method showed good linearity with correlation coefficients, r2>0.9990 for all the analytes. Furthermore, this is the first time that eco-friendly solvents are used for the extraction of oxyprenylated phenylpropanoids and the corresponding extract analyzed with ultra high performance liquid chromatography with photodiode array detection.
Asunto(s)
Arachis/química , Helianthus/química , Microextracción en Fase Líquida/métodos , Olea/química , Extractos Vegetales/aislamiento & purificación , Aceites de Plantas/química , Zea mays/química , Cromatografía Líquida de Alta Presión/métodos , Ácidos Cumáricos/análisis , Ácidos Cumáricos/aislamiento & purificación , Cumarinas/análisis , Cumarinas/aislamiento & purificación , Extractos Vegetales/análisis , Umbeliferonas/análisis , Umbeliferonas/aislamiento & purificaciónRESUMEN
An ultra high-performance liquid chromatographic (UHPLC) method with PDA detection was developed and validated for the simultaneous quantification of meropenem, linezolid, and levofloxacin in human plasma and applied in human plasma of critical care patients. A semi-automated microextraction by packed sorbent (MEPS) for sample preparation was used. All parameters in the extraction step (pH, sample volume, sample dilution and number of aspiration - ejection cycles) and in the desorption step (percentage of acetonitrile in the solvent of elution and number of aspirations of elution solvent through the device) were statistically significant when the recovery was used as response. The method showed good linearity with correlation coefficients, r2>0.9991 for the three drugs, as well as high precision (RSD%<10.83% in each case). Accuracy ranged from -7.8% to +6.7%. The limit of quantification of the three drugs was established at 0.01µg/mL for linezolid and levofloxacin and 0.02µg/mL for meropenem. Linezolid, meropenem, levofloxacin and the internal standard were extracted from human plasma with a mean recovery ranged from 92.4% to 97.4%. During validation, the concentration of meropenem, linezolid and levofloxacin was found to be stable after 3 freeze-thaw cycles and for at least 24h after extraction. This method will be subsequently used to quantify the drugs in patients to establish if the dosage regimen given is sufficient to eradicate the infection at the target site.
Asunto(s)
Cromatografía Líquida de Alta Presión , Automatización , Cuidados Críticos , Humanos , Levofloxacino , Límite de Detección , Linezolid , Meropenem , Microextracción en Fase Sólida , TienamicinasRESUMEN
This study developed a high performance liquid chromatography (HPLC) method involving dried blood spotting (DBS) as a sampling method for the therapeutic drug monitoring of antimicrobial combination therapy with linezolid and ciprofloxacin. DBS for standards, quality control samples, and patient samples was excised and then extracted using a mixture of methanol/water/formic acid 80:20:0.1 (v/v/v), respectively. Linezolid (LZD) and ciprofloxacin (CPR) were measured by HPLC using a Kinetex EVO C18 (100 × 4.6 mm I.D. 2.6 µm particle size). Mobile phase consisted of 10 mM ammonium acetate (A) and a mixture of acetonitrile and methanol (B), both containing 0.1% triethylamine, in gradient elution. Detection was carried out at 251 nm for linezolid (LZD) and 277 for ciprofloxacin (CPR). Ulifloxacin was used as an internal standard. The internal standard, LZD, and CPR were eluted in 7.90, 7.18, and 8.89 min, respectively. Total run-time was 20 min. Calibration curves were constructed in the range of 0.05-30 µg/mL for LZD and 0.02-10 µg/mL for CPR, respectively. The intra- and inter-day precision (RSD values) did not exceed 9.43%, the intra- and inter-day accuracy (accuracy %) ranged between 96.2 and 106.2%. Haematocrit (Hct) effects were investigated to obtain a linear correlation between haematocrit values and volume of blood. Copyright © 2017 John Wiley & Sons, Ltd.
Asunto(s)
Antibacterianos/sangre , Cromatografía Líquida de Alta Presión/métodos , Ciprofloxacina/sangre , Pruebas con Sangre Seca/métodos , Monitoreo de Drogas/métodos , Linezolid/sangre , Infección Hospitalaria/sangre , Infección Hospitalaria/tratamiento farmacológico , Humanos , Límite de Detección , Neumonía/sangre , Neumonía/tratamiento farmacológicoRESUMEN
An ultra high-performance liquid chromatographic (UHPLC) method with PDA detection was developed and validated for the simultaneous quantification of linezolid and ciprofloxacin in human plasma and applied in hospital acquired pneumonia patients (HAP). The method uses a semi-automated microextraction by packed sorbent for sample preparation. All parameters in the extraction step (pH, sample volume, sample dilution and number of aspiration - ejection cycles) and in the desorption step (percentage of acetonitrile in the solvent of elution and number of aspirations of elution solvent through the device) were statistically significant when the recovery was used as response. The method showed good linearity with correlation coefficients, r2>0.9995 for the two drugs, as well as high precision (RSD%<9.77% in each case), accuracy ranged from -6.2% to +8.2. The limit of quantification of the two drugs was established at 0.01 and 0.02µg/mL for ciprofloxacin and linezolid, respectively. Linezolid, ciprofloxacin and internal standard were extracted from human plasma with a mean recovery ranging from 92.4% to 97.4%. During validation, the concentrations of linezolid and ciprofloxacin were found to be stable after 3 freeze-thaw cycles and for at least 24h after extraction. This method will subsequently be used to quantify the drugs dosage in patients with HAP to establish if the dosage regimen given is sufficient to eradicate the infection at the target site.
Asunto(s)
Cromatografía Líquida de Alta Presión/instrumentación , Ciprofloxacina/sangre , Ciprofloxacina/aislamiento & purificación , Linezolid/sangre , Linezolid/aislamiento & purificación , Neumonía/sangre , Microextracción en Fase Sólida/métodos , Infección Hospitalaria/sangre , Equipos y Suministros Eléctricos , Humanos , Límite de Detección , Factores de TiempoRESUMEN
Oxyprenylated natural products were shown to exert in vitro and in vivo remarkable anti-cancer and anti-inflammatory effects. This paper describes a rapid, selective, and sensitive HPLC method with fluorescence detection for determination of 4'-geranyloxyferulic acid (GOFA) and its conjugate with l-nitroarginine methyl ester (GOFA-L-NAME) in mononuclear cells. Analytes were extracted from cells using methanol and eluted on a GraceSmart RP18 analytical column (250×4.6mm i.d., 5µm particle size) kept at 25°C. A mixture of formic acid 1% in water (A) and methanol (B) were used as mobile phase, at a flow-rate of 1.2mL/min in gradient elution. A fluorescence detector (excitation/emission wavelength of 319/398nm for GOFA and GOFA-L-NAME), was used for the two analytes. Calibration curves of GOFA and GOFA-L-NAME were linear over the concentration range of 1.0-50µg/mL, with correlation coefficients (r2)≥0.9995. Intra- and inter-assay precision do not exceed 6.8%. The accuracy was from 94% to 105% for quality control samples (2.0, 25.0 and 40µg/mL). The mean (RSD%) extraction recoveries (n=5) for GOFA and GOFA-L-NAME from spiked cells at 2.0, 25.0 and 40.0µg/mL were 92.4±1.5%, 94.7±0.9% and 93.8±1.1%, for GOFA and 95.3±1.2%, 94.8±1.0% and 93.9±1.3%, for GOFA-L-NAME. The limits of detection and quantification were 0.3µg/mL and 1.0µg/mL for GOFA and GOFA-L-NAME. This method was successfully applied to measure GOFA and GOFA-L-NAME concentrations in a mononuclear cells.
Asunto(s)
Células Cultivadas/química , Ácidos Cumáricos/análisis , Nitroarginina/análogos & derivados , Calibración , Humanos , Límite de Detección , Nitroarginina/análisis , Reproducibilidad de los Resultados , Células U937RESUMEN
A novel sensitive analytical method based on the use of a semi-automatic microextraction by packed sorbents (MEPS) techniques combined with ultra high-performance liquid chromatography (UHPLC) with PDA detection has been developed and validate for the analysis of ulifloxacin, the active metabolite of prulifloxacin using danofloxacin as internal standard in human plasma and urine. Different experimental parameters were optimized and validated according to international guidelines. Complete separation of the analytes was achieved with a Waters BEH C18 (50×2.1mm I.D., 1.7µm particle size) analytical column, a mixture of 10mM ammonium acetate (pH 3.0) (A) with and acetonitrile (B) both containing 1% triethylamine were used as mobile phase, at a flow rate of 0.6mL/min in gradient elution, and detection wavelength of 272nm. This method is linear in concentration range of 0.02-10.0µg/mL for plasma and urine, respectively. The limit of quantitation was 20ng/mL for the two fluids. The recoveries of the method were 95% for ulifloxacin in human plasma and urine and 95.5% for the internal standard. Intra- and inter- assay precision and accuracy for ulifloxacin were lower than 10% at all tested concentrations. The proposed method was successfully applied to measure plasma and urine concentrations of ulifloxacin in patients suffering from Peripheral Arterial Disease and for pharmacokinetics study.
Asunto(s)
Antibacterianos/análisis , Fluoroquinolonas/análisis , Enfermedad Arterial Periférica/metabolismo , Piperazinas/análisis , Antibacterianos/farmacocinética , Calibración , Cromatografía Líquida de Alta Presión , Fluoroquinolonas/farmacocinética , Humanos , Límite de Detección , Piperazinas/farmacocinética , Estándares de Referencia , Reproducibilidad de los Resultados , Microextracción en Fase SólidaRESUMEN
A procedure based on microextraction by packed sorbent (MEPS) followed by ultra-high performance liquid chromatography (UHPLC) with photodiode array (PDA) detection has been developed for the analysis of seven selected non steroidal anti-inflammatory drugs (NSAIDs) in human dialysates. The influence on MEPS efficiency of pH of the sample, pH of the washing solvent and methanol content in the hydro-alcoholic elution mixture has been investigated by response surface methodology based on a Box-Behnken design of experiments. Among the above factors, pH of sample is the variable that mostly influences MEPS recovery. UHPLC separation of the NSAIDs was completed within less than 4min under isocratic elution conditions on a Fortis SpeedCore C18 column (150×4.6mm I.D., 2.6µm) using acetonitrile-phosphate buffer as the mobile phase. Calibration curves of the NSAIDs were linear over the concentration range 0.025-15µg/mL, with correlation coefficients≥0.998. Intra- and inter-assay relative standard deviations were <8% and recovery values ranged from 94% to 100% for the quality control samples. The results reveal that the developed MEPS/PDA-UHPLC method exhibits a good accuracy and precision and is well suited for the rapid analysis of human dialysate from patients treated with the selected NSAIDs.
Asunto(s)
Antiinflamatorios no Esteroideos/aislamiento & purificación , Cromatografía Líquida de Alta Presión/métodos , Diálisis , Concentración de Iones de Hidrógeno , Reproducibilidad de los Resultados , Sensibilidad y Especificidad , Propiedades de SuperficieRESUMEN
A procedure based on solid-phase extraction (SPE) followed by high performance liquid chromatography (HPLC) with PDA detection has been developed for the analysis of multiple drugs in rat plasma. The analytes evaluated were ulifloxacin, fenbufen and felbinac. Eight different solid phase extraction cartridges were tested to evaluate their applicability for the isolation of drugs from rat plasma. Comparison were recovery of different drugs and reproducibility. The samples were analyzed by HPLC using a Kinetex C18 EVO column and acetonitrile-10mM ammonium acetate-methanol as the mobile phase under gradient elution conditions. SPE combined with HPLC-PDA allowed the determination of drugs over a linear range of 0.05-15 µg/mL for ulifloxacin while 0.5-50 µg/mL for felbinac and fenbufen, with limit of detection at 0.05 for ulifloxacin and 0.5 for felbinac and fenbufen. Bond Elut Plexa sorbent was found to provide the most effective clean-up, removing the greatest amount of interfering substance and simultaneously ensuring analyte recoveries higher than 93.54% with relative standard deviation (RSD) <10%. The method was applied with good accuracy and precision in the determination of ulifloxacin, fenbufen and felbinac in rat plasma obtained from rats treated with selected drugs. This method permits its application to pharmacokinetic and pharmacodynamic studies of these analytes and will facilitate detailed investigations on the interactions between new fluoroquinolones and fenbufen.
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Cromatografía Líquida de Alta Presión/métodos , Fluoroquinolonas/sangre , Fenilacetatos/sangre , Fenilbutiratos/sangre , Piperazinas/sangre , Plasma/química , Extracción en Fase Sólida/métodos , Animales , Fluoroquinolonas/química , Límite de Detección , Fenilacetatos/química , Fenilbutiratos/química , Piperazinas/química , Ratas , Reproducibilidad de los ResultadosRESUMEN
Several studies have shown that xanthones obtained from Garcinia Mangostana (GM) have remarkable biological activities. α-mangostin (α-MG) is the main constituent of the fruit hull of the GM. Several findings have suggested that SIRT-1, a nuclear histone deacetylase, could influence cellular function by the inhibition of NF-kB signaling. ROS can inhibit SIRT-1 activity by initiating oxidative modifications on its cysteine residues, and suppression of SIRT-1 enhances the NF-κB signaling resulting in inflammatory responses. The goals of the present study were to evaluate the quantity of α-MG in the methanolic extract of GM (Vithagroup Spa) and to investigate the activity of this xanthone in U937 cell line and in human monocytes from responsive to inflammatory insult analyzing the possible changes on the activation of SIRT-1 protein via NF-Kb. Cells were treated with the methanolic extract of GM and/or LPS. The chromatographic separation of α-MG was performed by an HPLC analysis. EX 527, a specific SIRT-1 inhibitor, was used to determine if SIRT-1/NfkB signaling pathway might be involved in α-MG action on cells. Our results show that α-MG inhibits p65 acetylation and down-regulates the pro-inflammatory gene products as COX-2, iNOS via SIRT-1 activation. Cells treated with EX 527 showed an up-regulation of NFkB acetylation and an over expression of inducible enzymes and their product of catalysis (NO and PGE2). These results suggest that α-MG may be useful for the development of alternative pharmacological strategies aimed at reducing the inflammatory process. J. Cell. Physiol. 231: 2439-2451, 2016. © 2016 Wiley Periodicals, Inc.