RESUMEN
BACKGROUND: When performing therapeutic drug monitoring (TDM) for flucloxacillin, it is advised to measure the unbound, not the total, flucloxacillin concentration. To be able to accurately quantify unbound flucloxacillin concentrations, a reliable analytical method is indispensable. OBJECTIVE: To determine the influence of temperature and pH of the sample during ultrafiltration on the measured unbound fraction of flucloxacillin. MATERIALS AND METHODS: We performed three different experiments. In a single laboratory experiment, we investigated the influence of ultrafiltration temperature (10°C, room temperature and 37°C) on the measured unbound fraction of flucloxacillin for three concentration levels. In a multiple laboratory experiment, the results of eight laboratories participating in an international quality control programme measuring unbound flucloxacillin concentrations were analysed. In the third experiment, patient samples were ultrafiltrated using four different conditions: (i) physiological pH and room temperature; (ii) unadjusted pH (pH 9 after freezing) and room temperature; (iii) physiological pH and 37°C and (iv) unadjusted pH and 37°C. RESULTS: For all experiments, measurement of samples that were ultrafiltrated at room temperature resulted in a substantially lower unbound fraction compared to samples that were ultrafiltrated at 37°C. Adjusting the pH to physiological pH only had a minimal impact on the measured unbound fraction. CONCLUSIONS: On the basis of these findings and considering the need for fast, simple and reproducible sample pretreatment for TDM purposes, we conclude that ultrafiltration of flucloxacillin should be performed at physiological temperature (37°C), but adjustment of pH does not seem to be necessary.
Asunto(s)
Antibacterianos , Monitoreo de Drogas , Floxacilina , Temperatura , Ultrafiltración , Floxacilina/farmacocinética , Ultrafiltración/métodos , Humanos , Antibacterianos/farmacocinética , Monitoreo de Drogas/métodos , Concentración de Iones de HidrógenoRESUMEN
Acute decompensated heart failure and fluid overload are the most common causes of hospitalization in heart failure patients, and often, they contribute to disease progression. Initial treatment encompasses intravenous diuretics although there might be a percentual of patients refractory to this pharmacological approach. New technologies have been developed to perform extracorporeal ultrafiltration in fluid overloaded patients. Current equipment allows to perform ultrafiltration in most hospital and acute care settings. Extracorporeal ultrafiltration is then prescribed and conducted by specialized teams, and fluid removal is planned to restore a status of hydration close to normal. Recent clinical trials and European and North American practice guidelines suggest that ultrafiltration is indicated for patients with refractory congestion not responding to medical therapy. Close interaction between nephrologists and cardiologists may be the key to a collaborative therapeutic effort in heart failure patients. Further studies are today suggesting that wearable technologies might become available soon to treat patients in ambulatory and de-hospitalized settings. These new technologies may help to cope with the increasing demand for the care of chronic heart failure patients. Herein, we provide a state-of-the-art review on extracorporeal ultrafiltration and describe the steps in the development of a new miniaturized system for ultrafiltration, called AD1 (Artificial Diuresis).
Asunto(s)
Insuficiencia Cardíaca , Ultrafiltración , Humanos , Insuficiencia Cardíaca/terapia , Ultrafiltración/métodos , Ultrafiltración/instrumentación , Miniaturización , Diseño de Equipo , Hemofiltración/instrumentación , Hemofiltración/métodosRESUMEN
Third-spacing of fluid is a common complication in hospitalized patients with decompensated cirrhosis. In addition to ascites, patients with advanced cirrhosis may develop significant peripheral edema, which may limit mobility and exacerbate debility and muscle wasting. Concomitant kidney failure and cardiac dysfunction may lead to worsening hypervolemia, which may ultimately result in pulmonary edema and respiratory compromise. Diuretic use in such patients may be limited by kidney dysfunction and electrolyte abnormalities, including hyponatremia and hypokalemia. A slow, continuous form of ultrafiltration known as aquapheresis is a method of extracorporeal fluid removal whereby a pump generates a transmembrane pressure that forces an isotonic ultrafiltrate across a semipermeable membrane. This leads to removal of an ultrafiltrate that is isotonic to blood without the need for dialysate or replacement fluid as is necessary in other forms of continuous kidney replacement therapy. This technique has been utilized in other conditions including acute decompensated heart failure, with trials showing mixed, but generally favorable results. Herein, we present a series of our own experience using aquapheresis among patients with cirrhosis, review the literature regarding its use in other hypervolemic states, and discuss how we may apply lessons learned from use of aquapheresis in heart failure to patients with end-stage liver disease.
Asunto(s)
Enfermedad Hepática en Estado Terminal , Insuficiencia Cardíaca , Insuficiencia Renal , Humanos , Ultrafiltración/métodos , Enfermedad Hepática en Estado Terminal/complicaciones , Cirrosis Hepática/complicaciones , Cirrosis Hepática/terapia , Insuficiencia Cardíaca/complicaciones , Insuficiencia Cardíaca/terapia , Insuficiencia Renal/complicacionesRESUMEN
Target-based drug discovery technology based on cell membrane targets has gained significant traction and has been steadily advancing. However, current methods still face certain limitations that need to be addressed. One of the challenges is the laborious preparation process of screening materials, which can be time-consuming and resource-intensive. Additionally, there is a potential issue of non-specific adsorption caused by carrier materials, which can result in false-positive results and compromise the accuracy of the screening process. To address these challenges, this paper proposes a target-based cell membrane affinity ultrafiltration technology for active ingredient discovery in natural products. In this technique, the cell membranes of human lung adenocarcinoma epithelial cells (A549) with a high expression of epidermal growth factor receptor (EGFR) were incubated with candidate drugs and then transferred to an ultrafiltration tube. Through centrifugation, components that interacted with EGFR were retained in the ultrafiltration tube as "EGFR-ligand" complex, while the components that did not interact with EGFR were separated. After thorough washing and eluting, the components interacting with EGFR were dissociated and further identified using LC-MS, enabling the discovery of bioactive compounds. Moreover, the target-based cell membrane affinity ultrafiltration technology exhibited commendable binding capacity and selectivity. Ultimately, this technology successfully screened and identified two major components from the Curcumae Rhizoma-Sparganii Rhizoma (CS) herb pair extracts, which were further validated for their potential anti-tumor activity through pharmacological experiments. By eliminating the need for laborious preparation of screening materials and the potential non-specific adsorption caused by carriers, the development of target-based cell membrane affinity ultrafiltration technology provides a simplified approach and method for bioactive compounds discovery in natural sources.
Asunto(s)
Productos Biológicos , Ultrafiltración , Humanos , Ultrafiltración/métodos , Productos Biológicos/farmacología , Tecnología , Receptores ErbB , Membrana CelularRESUMEN
BACKGROUND: The standard rate of sodium removal in adult anuric patients on continuous ambulatory peritoneal dialysis (CAPD) is 7.5 g/L of ultrafiltration volume (UFV). Although automated PD (APD) is widely used in pediatric patients, no attempt has yet been made to estimate sodium removal in APD. METHODS: The present, retrospective cohort study included pediatric patients with APD who were managed at Tokyo Metropolitan Children's Medical Center between July 2010 and November 2017. The patients underwent a peritoneal equilibrium test (PET) at our hospital. Sodium removal per UFV was calculated by peritoneal function and dwell time using samples from patients on APD with 1- and 2-h dwell effluent within three months of PET and 4- and 10-h dwell effluent at PET. RESULTS: In total, 217 samples from 18 patients were included, with 63, 81, and 73 of the samples corresponding to the High [H], High-average [HA], and Low-average [LA] PET category, respectively. Sodium removal per UFV (g/L in salt equivalent) for dwell times of one, two, four, and ten hours was 5.2, 8.8, 8.0, and 11.5 for PET [H], 5.3, 5.8, 5.6, and 8.1 for PET [HA], and 4.6, 5.1, 5.1, and 7.1 for PET [LA], respectively. CONCLUSIONS: Sodium removal per UFV in pediatric APD was less than the standard adult CAPD and tended to be lower with shorter dwell times, leading to sodium accumulation. Therefore, salt intake should be restricted in combination with one or more long daytime dwells, especially in anuric patients.
Asunto(s)
Sodio , Ultrafiltración , Humanos , Masculino , Estudios Retrospectivos , Femenino , Niño , Adolescente , Sodio/análisis , Ultrafiltración/métodos , Preescolar , Diálisis Peritoneal/métodos , Diálisis Peritoneal Ambulatoria Continua/métodos , Fallo Renal Crónico/terapia , Anuria/terapiaRESUMEN
The current study had conducted the life cycle analysis (LCA) to assess the environmental impact of microalgal wastewater treatment via an integrated membrane bioreactor. The functional unit selected for this analysis was 1 kg of treated microalgal wastewater with contaminants eliminated by ultrafiltration membrane fabricated from recycled polyethylene terephthalate waste. Meanwhile, the applied system boundary in this study was distinguished based on two scenarios, namely, cradle-to-gate encompassed wastewater treatment only and cradle-to-cradle which included the reutilization of treated wastewater to cultivate microalgae again. The environmental impacts and hotspots associated with the different stages of the wastewater treatment process had clearly elucidated that membrane treatment had ensued the highest impact, followed by microalgal harvesting, and finally cultivation. Among the environmental impact categories, water-related impact was found to be prominent in the following series: freshwater ecotoxicity, freshwater eutrophication and marine ecotoxicity. Notably, the key performance indicator of all environmental impact, i.e., the global warming potential was found to be very much lower at 2.94 × 10-4 kg CO2 eq as opposed to other literatures reported on the LCA of wastewater treatments using membranes. Overall, this study had proffered insights into the environmental impact of microalgal wastewater treatment and its stimulus for sustainable wastewater management. The findings of this study can be instrumental in making informed decision for optimizing microalgal wastewater treatment and reutilization assisted by membrane technology with an ultimate goal of enhancing sustainability.
Asunto(s)
Membranas Artificiales , Microalgas , Tereftalatos Polietilenos , Ultrafiltración , Aguas Residuales , Tereftalatos Polietilenos/química , Microalgas/crecimiento & desarrollo , Ultrafiltración/métodos , Aguas Residuales/química , Aguas Residuales/análisis , Eliminación de Residuos Líquidos/métodos , Ambiente , Reactores Biológicos , ReciclajeRESUMEN
Ultrafiltration (UF) is a highly efficient technique for algal-rich water purification, but it is heavily contaminated due to the complex water characteristics. To solve this problem, potassium permanganate (KMnO4) oxidation enhanced with sodium sulfite (Na2SO3) was proposed as a pretreatment means. The results showed that the end-normalized flux was elevated from 0.10 to 0.91, and the reversible fouling resistance was reduced by 99.95%. The membrane fouling mechanism also changed obviously, without the generation of cake filtration. Regarding the properties of algal-rich water, the zeta potential was decreased from -29.50 to -5.87 mV after KMnO4/Na2SO3 pretreatment, suggesting that the electrostatic repulsion was significantly reduced. Meanwhile, the fluorescent components in algal-rich water were significantly eliminated, and the removal of dissolved organic carbon was increased to 67.46%. In the KMnO4/Na2SO3 process, reactive manganese species (i.e., Mn(V), Mn(III) and MnO2) and reactive oxygen species (i.e., SO4â¢- and â¢OH) played major roles in purifying algal-rich water. Specifically, SO4â¢-, â¢OH, Mn(V) and Mn(III) could effectively oxidize algal pollutants. Simultaneously, the in-situ adsorption and coagulation of MnO2 could accelerate the formation of flocs by decreasing the electrostatic repulsion between cells, and protect the algal cells from being excessive oxidized. Overall, the KMnO4/Na2SO3 process showed significant potential for membrane fouling alleviation in purifying algal-rich water.
Asunto(s)
Permanganato de Potasio , Especies Reactivas de Oxígeno , Sulfitos , Purificación del Agua , Permanganato de Potasio/química , Purificación del Agua/métodos , Sulfitos/química , Especies Reactivas de Oxígeno/metabolismo , Membranas Artificiales , Manganeso/química , Contaminantes Químicos del Agua/análisis , Contaminantes Químicos del Agua/química , Ultrafiltración/métodos , Oxidación-ReducciónRESUMEN
The study investigated the influence of additives on the fabrication of mixed matrix membranes comprising polyethersulfone (PES), with a specific focus on hydrophilicity, flux, morphology, and antifouling properties. Carboxymethyl modified ß-cyclodextrin (CMß-CD) was used to enhance the dispersion and hydrophilicity of graphene oxide (GO), leading to the formation of a hydrophilic and stable composite nanoparticle (CMCD@GO). The hydrophilicity (WCA <51.5°) and water flux (32.6 L m-2.h-1) of the modified PES membranes (MCDGO-x) were improved by the incorporation of CMCD@GO nanoparticles, while that of PES membrane was 79.7° and 10.6 L m-2.h-1. The rate of backscattered light intensity (ΔBS) of MCDGO-x suspensions remains stable, suggesting stable dispersion of CMCD@GO in organic solvents. Compared to the bare PES membrane, the MCDGO-x membrane exhibits a thinner active layer and a finger-like structure. The MCDGO-x membrane exhibited excellent naphthenic acids (NAs) rejection (>93.2%) due to reduced roughness and higher hydrophilicity, while the GO-modified PES membrane (MGO-5) exhibited lower NAs rejection (87.2%). Furthermore, the MCDGO-5 membrane showed higher flux recovery ratio (FRR) of 79.3% compared to MGO-5 membrane (68.5%) after three cycles, indicating the antifouling performance of MCDGO-x for NAs was significantly improved. The combination of CMß-CD and GO enhance the flux and antifouling properties of PES ultrafiltration membranes, suggesting significant potential for applications in the purification of oil sands process water and the treatment of oily wastewater.
Asunto(s)
Incrustaciones Biológicas , Grafito , Membranas Artificiales , Ultrafiltración , Purificación del Agua , beta-Ciclodextrinas , beta-Ciclodextrinas/química , Grafito/química , Purificación del Agua/métodos , Ultrafiltración/métodos , Incrustaciones Biológicas/prevención & control , Interacciones Hidrofóbicas e Hidrofílicas , Sulfonas/química , PolímerosRESUMEN
Meconopsis integrifolia (Maxim.) Franch. is used extensively in traditional Tibetan medicine for its potent anti-inflammatory properties. In this study, six cyclooxygenase-2 (COX-2) inhibitors were purified from M. integrifolia using high-speed counter-current chromatography guided by ultrafiltration liquid chromatography (ultrafiltration-LC). First, ultrafiltration-LC was performed to profile the COX-2 inhibitors in M. integrifolia. The reflux extraction conditions were further optimized using response surface methodology, and the results showed that the targeted COX-2 inhibitors could be well enriched under the optimized extraction conditions. Then the six target COX-2 inhibitors were separated by high-speed countercurrent chromatography with a solvent system composed of ethyl acetate/n-butanol/water (4:1:4, v/v/v. Finally, the six COX-2 inhibitors, including 21.2 mg of 8-hydroxyluteolin 7-sophoroside, 29.6 mg of 8-hydroxyluteolin 7-[6'''-acetylallosyl-(1â2)-glucoside], 42.5 mg of Sinocrassoside D3, 54.1 mg of Hypolaetin 7-[6'''-acetylallosyll-(lâ2)-3''-acetylglucoside, 30.6 mg of Hypolaetin 7-[6'''-acetylallosyll-(lâ2)-6''-acetylglucoside and 17.8 mg of Hypolaetin were obtained from 500 mg of sample. Their structures were elucidated by 1 H-NMR spectroscopy. This study reveals that ultrafiltration-LC combined with high-speed counter-current chromatography is a robust and efficient strategy for target-guided isolation and purification of bioactive molecules. It also enhances the scientific understanding of the anti-inflammatory properties of M. integrifolia but also paves the way for its further medicinal applications.
Asunto(s)
Distribución en Contracorriente , Inhibidores de la Ciclooxigenasa 2 , Papaveraceae , Distribución en Contracorriente/métodos , Inhibidores de la Ciclooxigenasa 2/farmacología , Ultrafiltración/métodos , Cromatografía Líquida de Alta Presión/métodos , Cromatografía LiquidaRESUMEN
BACKGROUND: The evaluation and management of fluid balance are key challenges in critical care patients who require renal replacement therapies because cumulative fluid balance is an independent factor that increases morbidity and mortality in different clinical scenarios. SUMMARY: One of the strategies when fluid overload is refractory to diuretics is extracorporeal fluid removal (i.e., net ultrafiltration [UFNET] during kidney replacement therapy). However, problems with UFNET without individualized assessment are cardiovascular events and intradialytic hypotension, events that contribute to decreasing organ perfusion and sympathetic stress. Therefore, we must consider and try to predict the best timing for the start of ultrafiltration and find the point where the patient is most tolerant to ultrafiltration, making a simile to the concept of fluid tolerance. KEY MESSAGES: UFNET is a continuous and dynamic process, going through moments of tolerance and intolerance to ultrafiltration; as nephrologists, we must take the necessary measures to move through this period.
Asunto(s)
Ultrafiltración , Humanos , Ultrafiltración/métodos , Terapia de Reemplazo Renal/métodos , Equilibrio Hidroelectrolítico , FenotipoRESUMEN
INTRODUCTION: Continuous monitoring of relative blood volume (percentage BV) in hemodialysis (HD) is critical for determining dry weight and preventing intradialytic hypotension. However, the cause of the BV variation remains unknown. This research aimed to examine factors that influence the percentage BV. METHODS: We devised a formula based on coefficients ("a," "τ," and "b") to predict changes in percentage BV. "a" denotes a significant decrease in percentage BV in the early stages of HD. "τ" represents the transition from early to late phase of HD. "b" denotes the slope of the decrease in percentage BV in the late phase of HD. We measured the percentage BV in 18 patients with end-stage renal disease. The coefficients were estimated by fitting experimental data from patients using a least squares optimization algorithm. A correlation analysis of these parameters and patient predialysis data was performed. RESULTS: Ultrafiltration rate (UFR) was found to be negatively correlated with "b" (r = -0.851, p < 0.01). However, UFR was not significantly related to "a." Predialysis serum total protein level was negatively correlated with "a" (r = -0.531, p = 0.042). Predialysis serum albumin and predialysis sodium were not significantly correlated with "a" and "τ." Plasma osmolarity did not have a significant relationship with "a" and "τ." DISCUSSION/CONCLUSION: UFR influenced the decrease in percentage BV in the late phase but did not influence the decrease of percentage BV in the early phase. "a" was associated with predialysis serum total protein level but not with plasma osmolality or predialysis sodium. This implies that colloid oncotic pressure is important for plasma refilling immediately after dialysis begins. During the change of percentage BV, the decrease in the early phase of dialysis was not related to UFR, but related to other parameters, especially predialysis total protein level. A decrease in the late phase of dialysis is related to UFR.
Asunto(s)
Volumen Sanguíneo , Fallo Renal Crónico , Diálisis Renal , Humanos , Masculino , Femenino , Persona de Mediana Edad , Diálisis Renal/métodos , Diálisis Renal/efectos adversos , Fallo Renal Crónico/terapia , Fallo Renal Crónico/sangre , Anciano , Ultrafiltración/métodos , AdultoRESUMEN
Our research focuses on developing environmentally friendly biodegradable ultrafiltration (UF) membranes for small-scale water purification in areas lacking infrastructure or during emergencies. To address biofouling challenges without resorting to harmful chemicals, we incorporate bio-based extracts, such as methyl gallate from A. occidentale leaves, a Malaysian ulam herb, known for its quorum sensing inhibition (QSI) properties. The methyl gallate enriched extract was purified by solvent partitioning and integrated into cellulose-based UF membranes (0 to 7.5% w w-1) through phase inversion technique. The resulting membranes exhibited enhanced anti-organic fouling and anti-biofouling properties, with flux recovery ratio (FRR) of 87.84 ± 2.00% against bovine serum albumin and FRRs of 76.67 ± 1.89% and 69.57 ± 1.77% against E. coli and S. aureus, respectively. The CA/MG-5 membrane showed a 224% improvement in pure water flux (PWF) compared to the neat CA membrane. Our innovative approach significantly improves PWF, presenting an environmentally friendly method for biofouling prevention in UF membrane applications.
Asunto(s)
Anacardium , Incrustaciones Biológicas , Escherichia coli , Membranas Artificiales , Extractos Vegetales , Ultrafiltración , Purificación del Agua , Incrustaciones Biológicas/prevención & control , Ultrafiltración/métodos , Extractos Vegetales/farmacología , Extractos Vegetales/química , Escherichia coli/efectos de los fármacos , Anacardium/química , Purificación del Agua/métodos , Staphylococcus aureus/efectos de los fármacos , Ácido Gálico/análogos & derivados , Ácido Gálico/farmacología , Ácido Gálico/química , Albúmina Sérica Bovina/químicaRESUMEN
BACKGROUND: Considering no previous research into the utilization of ascending/descending ultrafiltration and linear sodium profiles in improving blood pressure among hemodialysis patients, the present study aimed to explore the effect of the A/D-UF along with linear sodium profiles on HD patients with hypotension. METHODS: Applying a crossover design, this clinical trial was fulfilled between December 2022 and June 2023 on 20 patients undergoing HD, randomized into two groups, each one receiving two intervention protocols, viz., (a) an intervention protocol in which the liquid sodium in the dialysis solution was linear and the UF profiling was A/D, and (b) a routine protocol or HD, wherein both liquid sodium and UF in the dialysis solution remained constant. The HD patients' BP was then checked and recorded at six intervals, namely, before HD, one, two, three, and four hours after it, and following its completion, within each session. The data were further statistically analyzed using the IBM SPSS Statistics 20 and the related tests. RESULTS: In total, 20 patients, including 12 men (60%) and 8 women (40%), with the mean age of 58.00 ± 14.54 on HD for an average of 54 months, were recruited in this study. No statistically significant difference was observed in the mean systolic and diastolic BP levels in the group receiving the A/D-UF profile all through the desired hours (p > 0.05), indicating that the patients did not face many changes in these two numbers during HD. Our cross-over clinical trial demonstrated a statistically significant reduction in symptomatic IDH episodes from 55 to 15% with the application of the A/D-UF profile (p < 0.05). CONCLUSION: The study demonstrated that the A/D-UF profile could contribute to the stability of blood pressure levels among HD patients, with no significant fluctuations observed during treatment sessions. TRIAL REGISTRATION: This study was registered in the Iranian Registry of Clinical Trials (no. IRCT20180429039463N5) on 07/01/2023.
Asunto(s)
Hipotensión , Ultrafiltración , Adulto , Anciano , Femenino , Humanos , Masculino , Persona de Mediana Edad , Presión Sanguínea , Estudios Cruzados , Soluciones para Diálisis , Hipotensión/etiología , Irán , Diálisis Renal/métodos , Sodio , Ultrafiltración/métodosRESUMEN
Red phycoerythrin (R-PE) is a highly valuable protein found in an edible seaweed, Pyropia yezoensis. It is used extensively in biotechnological applications due to its strong fluorescence and stability in diverse environments. However, the current methods for extracting and purifying R-PE are costly and unsustainable. The aim of the present study was to enhance the financial viability of the process by improving the extraction and purification of R-PE from dried P. yezoensis and to further enhance R-PE value by incorporating it into a tandem dye for molecular biology applications. A combination of ultrafiltration, ion exchange chromatography, and gel filtration yielded concentrated (1 mg·mL-1) R-PE at 99% purity. Using purified PE and Cyanine5 (Cy5), an organic tandem dye, phycoerythrin-Cy5 (PE-Cy5), was subsequently established. In comparison to a commercially available tandem dye, PE-Cy5 exhibited 202.3% stronger fluorescence, rendering it suitable for imaging and analyzes that require high sensitivity, enhanced signal-to-noise ratio, broad dynamic range, or shorter exposure times to minimize potential damage to samples. The techno-economic analysis confirmed the financial feasibility of the innovative technique for the extraction and purification of R-PE and PE-Cy5 production.
Asunto(s)
Carbocianinas , Ficoeritrina , Ficoeritrina/química , Ficoeritrina/aislamiento & purificación , Carbocianinas/química , Algas Marinas/química , Colorantes Fluorescentes/química , Cromatografía por Intercambio Iónico/métodos , Cromatografía en Gel/métodos , Ultrafiltración/métodos , Rhodophyta/química , Pigmentos Biológicos/aislamiento & purificación , Pigmentos Biológicos/química , Algas Comestibles , PorphyraRESUMEN
INTRODUCTION: Pancreatic lipase is one of the most important key targets in the treatment of obesity. Inhibition of pancreatic lipase can effectively reduce lipid absorption and treat obesity and other related metabolic disorders. OBJECTIVES: The goal of this study is the efficient screening of pancreatic lipase inhibitors in the root and rhizome of Rheum palmatum using affinity ultrafiltration-high-performance liquid chromatography (AUF-HPLC) combined with high-resolution inhibition profiling (HRIP). METHODS: Potential pancreatic lipase ligands and pancreatic lipase inhibitors in ethyl acetate fraction of R. palmatum were screened using AUF-HPLC and HRIP, respectively. All screened compounds were identified by HPLC- quadrupole time-of-flight (Q-TOF)/MS. Eight compounds were screened out by both AUF-HPLC and HRIP, and six compounds were screened out by either AUF-HPLC or HRIP alone. The pancreatic lipase inhibitory activities of all screened compounds were verified by enzyme inhibition assay and molecular docking. RESULTS: Five new potent pancreatic lipase inhibitors were discovered, namely procyanidin B5 3,3'-di-O-gallate (IC50 = 0.06 ± 0.01 µM), 1,6-di-O-galloyl-2-O-cinnamoyl-ß-D-glucoside (IC50 = 12.83 ± 0.67 µM), 1-O-(1,3,5-trihydroxy)phenyl-2-O-galloyl-6-O-cinnamoyl-ß-D-glucoside (IC50 = 17.84 ± 1.33 µM), 1,2-di-O-galloyl-6-O-cinnamoyl-ß-D-glucoside (IC50 = 18.39 ± 1.52 µM), and 4-(4'-hydroxyphenyl)-2-butanone-4'-O-ß-D-(2"-O-galloyl-6"-O-cinnamoyl)-glucoside (IC50 = 2.91 ± 0.40 µM). It was found that procyanidin B5 3,3'-di-O-gallate showed higher pancreatic lipase inhibitory activity than the positive control orlistat (IC50 = 0.12 ± 0.02 µM). CONCLUSION: The combination of affinity ultrafiltration-high-performance liquid chromatography (AUF-HPLC) and high-resolution inhibition profiling (HRIP) could reduce the risk of false-negative screening and missed screening and could achieve more efficient screening of bioactive compounds in complex natural products.
Asunto(s)
Rheum , Rheum/química , Cromatografía Líquida de Alta Presión/métodos , Ultrafiltración/métodos , Simulación del Acoplamiento Molecular , Glucósidos , Lipasa , Obesidad , Inhibidores Enzimáticos/farmacologíaRESUMEN
INTRODUCTION: Polygonum amplexicaule D. Don var. sinense Forb (PAF), a medicinal plant, has the effect of promoting blood circulation and removing blood stasis. However, the active compounds and targets of its anticoagulant effect are still unclear. OBJECTIVES: This study aims to establish an effective reversely thrombin-targeted screening method for anticoagulant active components in PAF by affinity ultrafiltration (AUF) coupled with ultrahigh-performance liquid chromatography-quadrupole time-of-flight mass spectroscopy (UPLC-Q-TOF-MS). METHODS: Different polar parts of PAF were screened for potential thrombin ligands by AUF-HPLC and identified by UPLC-Q-TOF-MS. After studying the affinity between ligands and thrombin by molecular docking, the antithrombotic activity of ligands was detected in vivo by zebrafish thrombus model, and in vitro by chromogenic substrate method. The mechanism of such ligands on thrombin was further studied by coagulation factor assay. RESULTS: Eleven potential thrombin ligands from PAF were screened by the AUF-UPLC-Q-TOF-MS method, and two compounds (butyl gallate and ß-sitosterol) with significant anticoagulant activity were discovered via in vitro and in vivo activity testing. CONCLUSION: A method system based on AUF-UPLC-Q-TOF-MS, molecular docking and in vivo and in vitro experiments also provided a powerful tool for further exploration of anticoagulant active components in PAF.
Asunto(s)
Anticoagulantes , Simulación del Acoplamiento Molecular , Polygonum , Trombina , Ultrafiltración , Pez Cebra , Polygonum/química , Cromatografía Líquida de Alta Presión/métodos , Anticoagulantes/farmacología , Anticoagulantes/química , Ultrafiltración/métodos , Animales , Trombina/metabolismo , Espectrometría de Masas/métodos , LigandosRESUMEN
Astragalus membranaceus is a traditional Chinese medicine with multiple pharmacological activities. Modern pharmacological research has found that Astragalus membranaceus extract has an inhibitory effect on α-glucosidase, however, which component can inhibit the activity of α-glucosidase and its degree of inhibition are unknown. To address this issue, this study used affinity ultrafiltration screening combined with UPLC-ESI-Orbitrap-MS technology to screen α-glucosidase inhibitors in Astragalus membranaceus. Using affinity ultrafiltration technology, we obtained the active components, and using UPLC-ESI-Orbitrap-MS technology, we quickly analyzed and identified them. As a result, a total of 8 ingredients were selected as α-glucosidase inhibitors.
Asunto(s)
Astragalus propinquus , Inhibidores de Glicósido Hidrolasas , Espectrometría de Masa por Ionización de Electrospray , Ultrafiltración , Inhibidores de Glicósido Hidrolasas/farmacología , Inhibidores de Glicósido Hidrolasas/química , Ultrafiltración/métodos , Cromatografía Líquida de Alta Presión/métodos , Estructura Molecular , Espectrometría de Masa por Ionización de Electrospray/métodos , Medicamentos Herbarios Chinos/química , Medicamentos Herbarios Chinos/farmacología , alfa-Glucosidasas/metabolismoRESUMEN
Phenolic compounds from a hydroalcoholic extract of wet olive pomace were purified and concentrated by an integrated membrane process in organic media. First, UF010104 (Solsep BV) and UP005 (Microdyn Nadir) membranes were tested to be implemented in the ultrafiltration stage, with the aim of purifying the extract and obtaining a permeate enriched in phenolic compounds. Despite the high flux observed with the UF010104 membrane (20.4 ± 0.7 L·h-1·m-2, at 2 bar), the UP005 membrane was selected because of a more suitable selectivity. Even though some secoiridoids were rejected, the permeate stream obtained with this membrane contained high concentrations of valuable simple phenols and phenolic acids, whereas sugars and macromolecules were retained. Then, the ultrafiltration permeate was subjected to a nanofiltration step employing an NF270 membrane (DuPont) for a further purification and fractionation of the phenolic compounds. The permeate flux was 50.2 ± 0.2 L·h-1·m-2, working at 15 bar. Hydroxytyrosol and some phenolic acids (such as vanillic acid, caffeic acid, and ferulic acid) were recovered in the permeate, which was later concentrated by reverse osmosis employing an NF90 membrane. The permeate flux obtained with this membrane was 15.3 ± 0.3 L·h-1·m-2. The concentrated phenolic mixture that was obtained may have important applications as a powerful antioxidant and for the prevention of diabetes and neurodegenerative diseases.
Asunto(s)
Membranas Artificiales , Olea , Fenoles , Ultrafiltración , Olea/química , Ultrafiltración/métodos , Fenoles/aislamiento & purificación , Fenoles/química , Fenoles/análisis , Ósmosis , Solventes/química , Extractos Vegetales/químicaRESUMEN
Membrane fouling induced by natural organic matter (NOM) has seriously affected the further extensive application of ultrafiltration (UF). Herein, a simple, green and robust vacuum ultraviolet (VUV) technology was adopted as pretreatment before UF and ultraviolet (UV) technology was used for comparison. The results showed that control effect of VUV pretreatment on membrane fouling was better than that of UV pretreatment, as evidenced by the increase of normalized flux from 0.27 to 0.38 and 0.73 after 30 min UV or VUV pretreatment, respectively. This is related to the fact that VUV pretreatment exhibited stronger NOM degradation ability than UV pretreatment owing to the formation of HOâ¢. The steady-state concentration of HO⢠was calculated as 3.04 × 10-13 M and the cumulative exposure of HO⢠reached 5.52 × 10-10 M s after 30 min of VUV irradiation. And the second-order rate constant between NOM and HO⢠was determined as 1.36 × 104 L mg-1 s-1. Furthermore, fluorescence EEM could be applied to predict membrane fouling induced by humic-enriched water. Standard blocking and cake filtration were major fouling mechanisms. Moreover, extension of UV pretreatment time increased the disinfection by-products (DBPs) formation, the DBPs concentration was enhanced from 322.36 to 1187.80 µg/L after 210 min pretreatment. However, VUV pretreatment for 150 min reduced DBPs content to 282.57 µg/L, and DBPs content continued to decrease with the extension of pretreatment time, revealing that VUV pretreatment achieved effective control of DBPs. The variation trend of cytotoxicity and health risk of DBPs was similar to that of DBPs concentration. In summary, VUV pretreatment exhibited excellent effect on membrane fouling alleviation, NOM degradation and DBPs control under a certain pretreatment time.
Asunto(s)
Desinfección , Purificación del Agua , Purificación del Agua/métodos , Ultrafiltración/métodos , Vacio , Rayos Ultravioleta , Membranas ArtificialesRESUMEN
Traditional Chinese medicine (TCM) is often composed of a variety of natural medicines. Its composition is complex, and many of its components can not be analyzed and identified. The first step in the rational application of TCM is to successfully separate the effective components which is also a great inspiration for the development of new drugs. Among the many separation technologies of TCM, the traditional heating concentration separation technology has high energy consumption and low efficiency. As a new separation technology, membrane separation technology has the characteristics of simple operation, high efficiency, environment-friendly and so on. The separation effect of high molecular weight difference solution is better. The applications of several main membrane separation technologies such as microfiltration, nanofiltration, ultrafiltration and reverse osmosis are reviewed, the methods of restoring membrane flux after membrane fouling are discussed, and their large-scale industrial applications in the future are prospected and summarized.