RESUMEN
BACKGROUND: Free tissue transfer to cover complex wounds with exposed critical structures results in donor-site morbidity. Perfusion decellularization and recellularization of vascularized composite tissues is an active area of research to fabricate complex constructs without a donor site. Sodium dodecyl sulfate (SDS)-based protocols remain the predominant choice for decellularization despite the deleterious effects on tissue ultrastructure and capillary networks. We aimed to develop an automated decellularization process and compare different SDS perfusion times to optimize the protocol. METHODS: A three-dimensional-printed closed-system bioreactor capable of continuously perfusing fluid through the vasculature was used for decellularization. The artery and vein of rat epigastric fasciocutaneous free flaps were cannulated and connected to the bioreactor. Protocols had varying durations of 1% SDS solution (3, 5, and 10 days) followed by 1 day of 1% Triton X-100 and 1 day of 1x phosphate-buffered saline. The residual DNA was quantified. Microarchitecture of the constructs was assessed with histology, and the vascular network was visualized for qualitative assessment. RESULTS: The structural integrity and the microarchitecture of the extracellular matrix was preserved in the 3- and 5-day SDS perfusion groups; however, the subcutaneous tissue of the 10-day protocol lost its structure. Collagen and elastin structures of the pedicle vessels were not compromised by the decellularization process. Five-day SDS exposure group had the least residual DNA content (p < 0.001). Across all protocols, skin consistently had twice as much residual DNA over the subcutaneous tissues. CONCLUSION: A compact and integrated bioreactor can automate decellularization of free flaps to bioengineer regenerative constructs for future use in reconstruction of complex defects. A decellularization protocol with 5 days of 1% SDS exposure was the most successful to keep the residual DNA content at a minimum while preserving the structural integrity of the tissues.
Asunto(s)
Colgajos Tisulares Libres , Ratas , Animales , Dodecil Sulfato de Sodio/farmacología , Dodecil Sulfato de Sodio/análisis , Dodecil Sulfato de Sodio/química , Roedores , Matriz Extracelular/química , Matriz Extracelular/ultraestructura , ADN/análisis , ADN/farmacología , Ingeniería de Tejidos/métodos , Andamios del TejidoRESUMEN
In this study, the caprine pancreas has been presented as an alternative to the porcine organ for pancreatic xenotransplantation with lesser risk factors. The obtained caprine pancreas underwent a systematic cycle of detergent perfusion for decellularization. It was perfused using anionic (0.5% w/v sodium dodecyl sulfate) as well as non-ionic (0.1% v/v triton X-100, t-octyl phenoxy polyethoxy ethanol) detergents and washed intermittently with 1XPBS supplemented with 0.1% v/v antibiotic and nucleases in a gravitation-driven set-up. After 48 h, a white decellularized pancreas was obtained, and its extracellular matrix (ECM) content was examined for scaffold-like properties. The ECM content was assessed for removal of cellular content, and nuclear material was evaluated with temporal H&E staining. Quantified DNA was found to be present in a negligible amount in the resultant decellularized pancreas tissue (DPT), thus prohibiting it from triggering any immunogenicity. Collagen and fibronectin were confirmed to be preserved upon trichrome and immunohistochemical staining, respectively. SEM and AFM images reveal interconnected collagen fibril networks in the DPT, confirming that collagen was unaffected. sGAG was visualized using Prussian blue staining and quantified with DMMB assay, where DPT has effectively retained this ECM component. Uniaxial tensile analysis revealed that DPT possesses better elasticity than NPT (native pancreatic tissue). Physical parameters like tensile strength, stiffness, biodegradation, and swelling index were retained in the DPT with negligible loss. The cytocompatibility analysis of DPT has shown no cytotoxic effect for up to 72 h on normal insulin-producing cells (MIN-6) and cancerous glioblastoma (LN229) cells in vitro. The scaffold was recellularized using isolated mouse islets, which have established in vitro cell proliferation for up to 9 days. The scaffold received at the end of the decellularization cycle was found to be non-toxic to the cells, retained biological and physical properties of the native ECM, suitable for recellularization, and can be used as a safer and better alternative as a transplantable organ from a xenogeneic source.
Asunto(s)
Detergentes , Insulinas , Animales , Antibacterianos/farmacología , Colágeno/metabolismo , ADN/metabolismo , Matriz Extracelular Descelularizada , Detergentes/química , Detergentes/metabolismo , Detergentes/farmacología , Etanol/farmacología , Matriz Extracelular/metabolismo , Fibronectinas/metabolismo , Cabras , Insulinas/análisis , Insulinas/metabolismo , Insulinas/farmacología , Ratones , Octoxinol/análisis , Octoxinol/metabolismo , Octoxinol/farmacología , Páncreas , Estudios Prospectivos , Dodecil Sulfato de Sodio/análisis , Dodecil Sulfato de Sodio/metabolismo , Dodecil Sulfato de Sodio/farmacología , Porcinos , Ingeniería de Tejidos/métodos , Andamios del Tejido/químicaRESUMEN
BACKGROUND: Hempseed meal, a by-product of the hempseed oil processing stream, is a potential alternative source for food proteins. Efficient extraction of proteins from hempseed meal is challenging owing to differences in the structure and solubility of various protein fractions present in the seed. In the present study, protein was extracted from hempseed meal using four different solvents, including aqueous NaOH, KOH, NaHCO3 and NaCl, at four different concentrations with the aim of improving the recovery of protein fractions rich in essential amino acids. RESULTS: Extraction using alkaline solvents provided superior protein recovery (60-78%) compared with NaCl solution and control extractions (20-48% and 21%, respectively). The concentration of alkali or salt (0.25-1 mol L-1 ) had a minor but significant impact on the yield. Amino acid composition analysis revealed that hempseed meal contains 24% (54.5 ± 0.19 mg g-1 ) essential amino acids of total amino acids, and extraction with NaOH, KOH, NaHCO3 or NaCl did not improve the selective extraction of essential amino acids compared to control experiments. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) analysis allowed the identification of edestin and albumin in the extracts obtained with NaHCO3 and NaCl solvents, with results further showing that the type of extraction solvent influences protein extraction selectivity. CONCLUSION: Although alkali solvents provide superior extraction yields, extraction with water resulted in extracts containing the highest proportion of proteins bearing essential amino acids. According to the results of SDS-PAGE, extraction using alkali solvents induced protein crosslinking. © 2022 The Authors. Journal of The Science of Food and Agriculture published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.
Asunto(s)
Semillas , Cloruro de Sodio , Albúminas/química , Aminoácidos/análisis , Aminoácidos Esenciales/análisis , Cannabis , Extractos Vegetales , Semillas/química , Cloruro de Sodio/análisis , Dodecil Sulfato de Sodio/análisis , Hidróxido de Sodio , Solventes/química , Agua/análisisRESUMEN
The method of colloid titration with poly(diallyldimethylammonium) chloride has been improved to detect the endpoint with an off-vessel light reflectance sensor. The digital color sensor used measures light reflectance by means of light guides, with no immersion into the reaction solution. In such a method, the optical signal is free of disturbances caused by sticky flocs in the solution. The improved automatic titration set was applied for the determination of sodium laureth sulfate (SLES) in industrial batches and commercial personal care products. The sample color and opacity do not disturb the SLES quantification. When the SLES content lies in the range from 5% to 9%, the optimal sample weight is from 6 g to 3 g.
Asunto(s)
Coloides/química , Cosméticos/química , Luz , Dodecil Sulfato de Sodio/análogos & derivados , Procesamiento de Imagen Asistido por Computador , Estándares de Referencia , Dodecil Sulfato de Sodio/análisis , Soluciones , Difracción de Rayos XRESUMEN
A microfluidic, label-free optical sensor for water pollutants, which is based on a packed micro-column of microspheres with refractive index similar to that of water, is presented. The perfluoropolyether microspheres are synthetized by membrane emulsification followed by UV irradiation. The microfluidic channel hosting the packed column is transparent when filled with pure water as a consequence of refractive index matching, whereas it scatters light in presence of compounds with lipophilic moieties that spontaneously adsorb on the fluorinated microspheres. The device is characterized by investigating the response to cationic and anionic surfactants. Both the signal growth rate and the recovery rate measured during washing with water depend on the type and concentration of the compounds. The cationic surfactants tested display a larger signal increase, linearly scaling with concentration. A limit of detection of 1 µM is obtained in the current configuration. The water index-matched microspheres enable to access an additional analytical parameter, that is the propagation velocity of the scattering signal along the column. This parameter is also found to scale linearly with concentration, hence providing a complementary analytical tool sensitive to the adhesion kinetics.
Asunto(s)
Técnicas Analíticas Microfluídicas/métodos , Microesferas , Compuestos de Amonio Cuaternario/análisis , Dodecil Sulfato de Sodio/análisis , Tensoactivos/análisis , Contaminantes Químicos del Agua/análisis , Adsorción , Polímeros de Fluorocarbono/química , Dispositivos Laboratorio en un Chip , Técnicas Analíticas Microfluídicas/instrumentación , Compuestos de Amonio Cuaternario/química , Refractometría , Dodecil Sulfato de Sodio/química , Tensoactivos/química , Contaminantes Químicos del Agua/químicaRESUMEN
BACKGROUND: The adulteration of milk by hazardous chemicals like surfactants has recently increased. It conceals the quality of the product to gain profit. As milk and milk-based products are consumed by many people, novel analytical procedures are needed to detect these adulterants. This study focused on Fourier-transform infrared (FTIR) spectroscopy equipped with an attenuated total reflection (ATR) accessory, and near-infrared (NIR) spectroscopy for the determination of milk-surfactant adulteration using a genetic algorithm (GA) coupled with multivariate methods. The model surfactant was sodium dodecyl sulfate (SDS), and its concentration varied from 1.94-19.4 gkg-1 in adulterated samples. RESULTS: Prominent peaks in the spectral range of 5500-6400 cm-1 , 1160-1260 cm-1 and 1049-1080 cm-1 may correspond to the sulfonate group in SDS. A genetic algorithm could significantly reduce the number of variables to almost one third by selecting the specific wavenumber region. Principal component analysis (PCA) for ATR and NIR data indicated separate clusters of samples in terms of the concentration level of SDS (P ≤ 0.05). Partial least squares regression (PLSR) was used to determine the maximum R2 value for ATR and NIR data for calibration, cross-validation and prediction, which were 0.980, 0.972, 0.980, and 0.970, 0.937, and 0.956 respectively. The results showed apparent differences between unadulterated and adulterated samples using partial least squares-discriminant analysis (PLS-DA), which was validated by the permutation test. CONCLUSION: The results clearly show the successful application of the proposed methods with multivariate analysis in the selection of variables, classification, clustering, and identification of the adulterant in amounts as low as 1.94 gkg-1 in milk. © 2020 Society of Chemical Industry.
Asunto(s)
Contaminación de Alimentos/análisis , Leche/química , Espectroscopía Infrarroja por Transformada de Fourier/métodos , Tensoactivos/análisis , Algoritmos , Animales , Bovinos , Análisis Discriminante , Análisis de los Mínimos Cuadrados , Dodecil Sulfato de Sodio/análisisRESUMEN
There are increasing concerns about the danger that water-borne pathogens and pollutants pose to the public. Of particular importance are those that disrupt the plasma membrane, since loss of membrane integrity can lead to cell death. Currently, quantitative assays to detect membrane-disrupting (lytic) agents are done offsite, leading to long turnaround times and high costs, while existing colorimetric point-of-need solutions often sacrifice sensitivity. Thus, portable and highly sensitive solutions are needed to detect lytic agents for health and environmental monitoring. Here, a lipid-based electrochemical sensing platform is introduced to rapidly detect membrane-disrupting agents. The platform combines benchtop fabricated microstructured electrodes (MSEs) with lipid membranes. The sensing mechanism of the lipid-based platform relies on stacked lipid membranes serving as passivating layers that when disrupted generate electrochemical signals proportional to the membrane damage. The MSE topography, membrane casting and annealing conditions were optimized to yield the most reproducible and sensitive devices. We used the sensors to detect membrane-disrupting agents sodium dodecyl sulfate and Polymyxin-B within minutes and with limits of detection in the ppm regime. This study introduces a platform with potential for the integration of complex membranes on MSEs towards the goal of developing Membrane-on-Chip sensing devices.
Asunto(s)
Técnicas Biosensibles/instrumentación , Técnicas Electroquímicas/instrumentación , Polimixina B/análisis , Dodecil Sulfato de Sodio/análisis , Membrana Celular/efectos de los fármacos , Colorimetría , Monitoreo del Ambiente , Dispositivos Laboratorio en un Chip , Límite de Detección , Polimixina B/farmacología , Dodecil Sulfato de Sodio/farmacologíaRESUMEN
Dissolved organic matter (DOM) is a common solute in snow and ice at Earth's surface. Its effects on reaction kinetics in ice and at air-ice interfaces can be large, but are currently difficult to quantify. We used Raman microscopy to characterize the surface and bulk of frozen aqueous solutions containing humic acid, sodium dodecyl sulfate (SDS), and citric acid at a range of concentrations and temperatures. The surface-active species (humic acid and SDS) were distributed differently than citric acid. Humic acid and SDS are almost completely excluded to the air-ice interface during freezing, where they form a film that coats the surface nearly completely. A liquid layer that coats the majority of the surface was observed at all humic acid and SDS concentrations. Citric acid, which is smaller and less surface active, is excluded to liquid channels at the air-ice interface and within the ice bulk, as has previously been reported for ionic solutes such as sodium chloride. Incomplete surface wetting was observed at all citric acid concentrations and at all temperatures (up to -5 °C). Citric acid appears to be solvated in frozen samples, but SDS and humic acid do not. These results will improve our understanding of the effects of organic solutes on environmental and atmospheric chemistry within ice and at air-ice interfaces.
Asunto(s)
Aire/análisis , Sustancias Húmicas/análisis , Cubierta de Hielo/química , Ácido Cítrico/análisis , Frío , Congelación , Cinética , Modelos Teóricos , Dodecil Sulfato de Sodio/análisis , Solubilidad , Soluciones , Espectrometría Raman , Tensoactivos/análisisRESUMEN
In this study, flaxseed mucilage (FSM) has been used as a green coagulant in the pretreatment stage of a combined process for the removal of an anionic surfactant, sodium dodecyl sulfate (SDS). In the post-treatment stage, heterogeneous photo-Fenton-like oxidation using MnFe2O4 nanocatalyst was applied to remove the remained SDS. Using response surface methodological approach, optimum condition in the coagulation process was obtained at pH 7.0, FSM dose of 100â¯mgâ¯L-1 and 30â¯min. In the photo-Fenton oxidation process, complete SDS removal was achieved using 76â¯mg of the nanocatalyst, 1.07â¯mL of H2O2 at 17â¯min. Application of the combined process on the real wastewater samples indicates that the proposed method can be used effectively for the treatment of industrial effluents containing surfactants.
Asunto(s)
Lino/metabolismo , Residuos Industriales/análisis , Dodecil Sulfato de Sodio/análisis , Tensoactivos/análisis , Eliminación de Residuos Líquidos/métodos , Contaminantes Químicos del Agua/análisis , Compuestos Férricos/química , Peróxido de Hidrógeno/química , Compuestos de Manganeso/química , Nanopartículas del Metal/química , Oxidación-Reducción , Óxidos/química , Dodecil Sulfato de Sodio/química , Tensoactivos/química , Aguas Residuales/químicaRESUMEN
Sodium dodecyl sulfate (SDS) is a commonly used surfactant in protein solubilization and also during the polysaccharide purification. A GC-MS method has been developed to quantitate residual SDS in meningococcal polysaccharide serogroups A,C,W,Y and X circumventing the need of spectroscopic assays and HPLC based methods which are either unstable or requires the confirmation by MS. The developed method is based on quantitative conversion of SDS to 1-dodecanol at elevated temperature. Meningococcal polysaccharides and SDS standards were treated with methanolic-HCl and extracted in n-Hexane. The conversion of SDS to 1-dodecanol was confirmed by mass spectra and separation was achieved using a DB-5ms column. The mass spectral analysis of 1-dodecanol showed characteristic ions at m/z 168, 140 and 125. The GC-MS method validation performed on intermediate and purified meningococcal polysaccharides showed linearity with r2â¯>â¯0.99 over the concentration range of 2.5-200⯵g/ml with LOD and LOQ of 1.27 and 3.85 respectively. The method was found to be precise, robust and accurate with spike recovery ranging 83-117%. The GC-MS method can be used in the quantitation of residual SDS during polysaccharide purification and provides valuable information about consistency of polysaccharide manufacturing process for development of pentavalent meningococcal conjugate vaccine.
Asunto(s)
Contaminación de Medicamentos , Cromatografía de Gases y Espectrometría de Masas , Vacunas Meningococicas/análisis , Neisseria meningitidis/química , Polisacáridos Bacterianos/análisis , Dodecil Sulfato de Sodio/análisis , Vacunas Meningococicas/química , Polisacáridos Bacterianos/química , Dodecil Sulfato de Sodio/químicaRESUMEN
As a commonly used surfactant, sodium dodecyl sulfate (SDS) usually coexists with inorganic anions in the industrial wastewater. These anions have a significant influence on SDS removal, indirectly threatening the environment. It is important to understand the relationship between the adsorption of SDS and inorganic anions. In this study, calcium-based layered double hydroxide (CaAl-LDH-Cl) as an efficient adsorbent was synthesized for investigating the effect of SO42- on SDS removal. The SDS adsorption capacities were enhanced to 3.21 and 4.21 mmol g-1 in the presence of SO42- with low/high SDS concentration, respectively. The phenomenon and mechanism were confirmed by X-ray diffraction (XRD), Fourier transform infrared (FTIR) spectroscopy, and Scanning electron microscopy (SEM). Anionic exchange played a dominant role in the adsorption of SDS onto CaAl-LDH-Cl at DS-/SO42- < 2, while both anion exchange and precipitation occurred when DS-/SO42- exceeded 2. Moreover, the thermal analysis (TG-DTA) was employed to further reveal the interaction mechanism. The results showed the highest total mass loss and the lowest loss temperature of interlayer water in the sulfate coexist system, confirming the enhancement of SDS adsorption amount in the presence of SO42-.
Asunto(s)
Hidróxido de Calcio/química , Dodecil Sulfato de Sodio/análisis , Sulfatos/química , Tensoactivos/análisis , Contaminantes Químicos del Agua/análisis , Purificación del Agua/métodos , Adsorción , Concentración de Iones de Hidrógeno , Cinética , Propiedades de Superficie , Aguas Residuales/químicaRESUMEN
Biomedical devices are complex products requiring numerous assembly steps along the industrial process chain, which can carry the potential of surface contamination. Cleanliness has to be analytically assessed with respect to ensuring safety and efficacy. Although several analytical techniques are routinely employed for such evaluation, a reliable analysis chain that guarantees metrological traceability and quantification capability is desirable. This calls for analytical tools that are cascaded in a sensible way to immediately identify and localize possible contamination, both qualitatively and quantitatively. In this systematic inter-comparative approach, we produced and characterized sodium dodecyl sulfate (SDS) films mimicking contamination on inorganic and organic substrates, with potential use as reference materials for ambient techniques, i.e., ambient mass spectrometry (AMS), infrared and Raman spectroscopy, to reliably determine amounts of contamination. Non-invasive and complementary vibrational spectroscopy techniques offer a priori chemical identification with integrated chemical imaging tools to follow the contaminant distribution, even on devices with complex geometry. AMS also provides fingerprint outputs for a fast qualitative identification of surface contaminations to be used at the end of the traceability chain due to its ablative effect on the sample. To absolutely determine the mass of SDS, the vacuum-based reference-free technique X-ray fluorescence was employed for calibration. Convex hip liners were deliberately contaminated with SDS to emulate real biomedical devices with an industrially relevant substance. Implementation of the aforementioned analytical techniques is discussed with respect to combining multimodal technical setups to decrease uncertainties that may arise if a single technique approach is adopted. Graphical abstract á .
Asunto(s)
Dodecil Sulfato de Sodio/análisis , Análisis Espectral/métodos , Vacio , Humanos , Estándares de Referencia , Dodecil Sulfato de Sodio/normas , Propiedades de SuperficieRESUMEN
The objectives of this study were to explore sodium dodecyl sulfate (SDS) and Soluplus on the crystallization inhibition and dissolution of felodipine (FLDP) extrudates by bottom-up and top-down approaches. FLDP extrudates with Soluplus and SDS were prepared by hot melt extrusion, and characterized by polarized light microscopy, differential scanning calorimetry, and fourier transform infrared spectroscopy. Results indicated that Soluplus inhibited FLDP crystallization, and the whole amorphous solid dispersions (ASDs) were binary FLDP-Soluplus (1:3) and ternary FLDP-Soluplus-SDS (1:2:0.15â¼0.3 and 1:3:0.2â¼0.4) extrudates. Internal SDS (5%-10%) decreased glass transition temperatures of FLDP-Soluplus-SDS ternary ASDs without presenting molecular interactions with FLDP or Soluplus. The enhanced dissolution rate of binary or ternary Soluplus-rich ASDs in the nonsink condition of 0.05% SDS was achieved. Bottom-up approach indicated that Soluplus was a much stronger crystal inhibitor to the supersaturated FLDP in solutions than SDS. Top-down approach demonstrated that SDS enhanced the dissolution of Soluplus-rich ASDs via wettability and complexation with Soluplus to accelerate the medium uptake and erosion kinetics of extrudates, but induced FLDP recrystallization and resulted in incomplete dissolution of FLDP-rich extrudates. In conclusion, top-down approach is a promising strategy to explore the mechanisms of ASDs' dissolution, and small amount of SDS enhances the dissolution rate of polymer-rich ASDs in the nonsink condition.
Asunto(s)
Química Farmacéutica/métodos , Felodipino/química , Polietilenglicoles/química , Polivinilos/química , Dodecil Sulfato de Sodio/química , Rastreo Diferencial de Calorimetría/métodos , Cristalización/métodos , Felodipino/análisis , Felodipino/farmacocinética , Polietilenglicoles/análisis , Polietilenglicoles/farmacocinética , Polivinilos/análisis , Polivinilos/farmacocinética , Dodecil Sulfato de Sodio/análisis , Dodecil Sulfato de Sodio/farmacocinética , Solubilidad , Espectroscopía Infrarroja por Transformada de Fourier/métodosRESUMEN
The aim of this study was to evaluate in vitro and in vivo the effect of sodium dodecyl sulfate (SDS) on the caprine lentivirus (CLV) in colostrum and milk. This was performed to develop a practical and efficient method of blocking the lactogenic transmission of the virus. In the in vitro experiment, colostrum and milk were treated with 0.25%; 0.50% and 1% SDS. Then, somatic cells of colostrum and milk were submitted to co-culture with caprine synovial membrane cells (CSM). In the in vivo test, goats were fed with colostrum and milk provided from CLV-positive goats treated with SDS in the same concentrations used in the in vitro experiment. Animals were tested by nested polymerase chain reaction (nPCR) and Western blot (WB) assays. In the in vitro experiment, inhibitory activity against CLV without inactivation occurred in colostrum with all SDS concentrations. However, concentrations of 0.25 and 0.5% SDS presented only inhibitory activity against CLV in milk cells, and 1% concentration provided inactivation of the virus. In the in vivo tests, none of the three concentrations of SDS was effective in inactivating LVC in colostrum or goat milk, which was confirmed by seroconversion and presence of proviral DNA in animals afterwards.(AU)
O objetivo da pesquisa foi avaliar in vitro e in vivo o efeito do dodecil sulfato de sódio (SDS) sobre o lentivírus caprino (LVC) no colostro e no leite, a fim de desenvolver um método prático e eficiente no bloqueio da via de transmissão lactogênica do vírus. No experimento in vitro, o colostro e o leite de cabras positivas foram tratados com SDS a 0,25%, 0,50% e 1,0%. Em seguida, as células somáticas do colostro e do leite foram obtidas e direcionadas ao cocultivo com células de membrana sinovial caprina (MSC). No teste in vivo, os cabritos foram alimentados com colostro e leite providos de cabras positivas para LVC, tratados com SDS nas mesmas concentrações usadas no teste in vitro. Os animais foram acompanhados pelos testes de reação em cadeia da polimerase nested (nPCR) e western blot (WB). Nos resultados in vitro, no colostro, observou-se que, em todas as concentrações de SDS, ocorreu uma atividade inibitória contra o LVC, sem a inativação. Em relação às células do leite, o SDS apresentou, nas concentrações de 0,25 e 0,5%, atividade inibitória contra o LVC, e na concentração de 1%, houve inativação viral. Nos testes in vivo, as três concentrações de SDS testadas não foram efetivas na inativação do LVC no colostro e no leite caprino, o que se comprovou pela soroconversão e pela presença de DNA proviral nos animais.(AU)
Asunto(s)
Animales , Femenino , Embarazo , Calostro/química , Lentivirus Ovinos-Caprinos , Dodecil Sulfato de Sodio/análisisRESUMEN
We report the fabrication of a 3D-printed water-heated cuvette that fits into a standard UV visible spectrophotometer. Full 3D-printable designs are provided and 3D-printing conditions have been optimised to provide options to print the cuvette in either acrylonitrile butadiene styrene or polylactic acid polymers, extending the range of solvents that are compatible with the design. We demonstrate the efficacy of the cuvette by determining the critical micelle concentration of sodium dodecyl sulphate at 40 °C, the molar extinction coefficients of cobalt nitrate and dsDNA and by reproducing the thermochromic UV visible spectrum of a mixture of cobalt chloride, water and propan-2-ol.
Asunto(s)
Resinas Acrílicas/química , Butadienos/química , Poliésteres/química , Poliestirenos/química , Impresión Tridimensional , ADN/análisis , Dodecil Sulfato de Sodio/análisis , Espectrofotometría Ultravioleta/instrumentación , Espectrofotometría Ultravioleta/métodosRESUMEN
It is still a challenge for clinical diagnostics to design non-toxic and selective sensors for a specific analyte in blood serum, which contains a large number of proteins, small molecules, and ions. Herein, H1, a synthetic hybrid of green fluorescent protein (GFP) chromophore and peptide for human serum albumin (HSA) detection via obvious fluorescence enhancement was reported. The fluorescence response was caused by the non-covalent binding between H1 and the subdomain IIA cavity of HSA, and this specific binding behavior can be confirmed by the decrease of fluorescence intensity of the sole tryptophan residue (Trp-214) located in the same subdomain. H1 showed remarkable sensitivity and selectivity toward HSA without interference from other proteins. A good linear relationship between the fluorescence intensity of H1 and HSA concentration from 0 to 67.0µgmL(-1) was obtained with the detection limit reaching as low as 198.6ngL(-1). Moreover, H1 was successfully utilized to detect trace HSA in healthy human urine (HU) and human blood serum (HBS), respectively. The resulting H1/HSA system also showed a satisfactory sensing ability toward anionic surfactant sodium dodecyl sulfate (SDS) through decreased fluorescence intensity with the detection limit of 3.48ngL(-1).
Asunto(s)
Albuminuria/orina , Técnicas Biosensibles/métodos , Proteínas Fluorescentes Verdes/química , Péptidos/química , Albúmina Sérica/análisis , Espectrometría de Fluorescencia/métodos , Animales , Bovinos , Humanos , Límite de Detección , Albúmina Sérica Bovina/análisis , Dodecil Sulfato de Sodio/análisisRESUMEN
To understand the bioaccessibility of the flavonoid quercetin we studied its interaction with bile salt micelles. The environmental sensitivity of quercetin's UV-visible absorption spectrum gave information about quercetin partitioning. Two quercetin absorption peaks gave complementary information: Peak A (240-280nm) on the intermicellar phase and Peak B (340-440nm) on the micellar phase. Thus, by altering pH, we showed that only non-ionised quercetin partitions into micelles. We validated our interpretation by studying quercetin's interaction with SDS micelles. Pyrene fluorescence and the quercetin UV-visible spectra show that the adsorption site for pyrene and quercetin in bile salt micelles is more hydrophobic than that for SDS micelles. Also, both quercetin and pyrene reported a higher critical micelle concentration for bile salts than for SDS. Our method of using a flavonoid as an intrinsic probe, is generally applicable to other lipophilic bioactives, whenever they have observable environmental dependent properties.
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Ácidos y Sales Biliares/química , Quercetina/química , Dodecil Sulfato de Sodio/química , Ácidos y Sales Biliares/análisis , Interacciones Hidrofóbicas e Hidrofílicas , Micelas , Quercetina/análisis , Dodecil Sulfato de Sodio/análisis , SolubilidadRESUMEN
The CMC is one of the fundamental characteristics of surfactants and its determination is crucial for detail understanding of micelles formation. In this study the CMC of SDS in presence of ACN was determined by two independent experimental techniques, capillary electrophoresis and fluorescence correlation spectroscopy (FCS). Yet, studies of SDS micellization in solutions containing ACN as organic modifier are sparse and inconsistent in literature. The measurements were performed for various ACN contents in the range of 0-50% v/v. At ACN contents of up to 10% v/v the CMC is lower when compared to the aqueous solution, while increasing ACN content causes a significant increase of the CMC. Formation of micelles was observed up to ACN concentrations of 35% v/v, which is in contrast to most of the reports in literature. Based on the results of the FCS experiments, we were able to confirm that presence of ACN causes a gradual increase of the size of the micelles with increasing concentration of SDS. Simultaneously, we proved that the classical conductivity approach for the determination of the CMC does not yield reliable results in the presence of higher content of an organic modifier such as ACN.
Asunto(s)
Acetonitrilos/química , Micelas , Dodecil Sulfato de Sodio/análisis , Solventes/química , Electroforesis Capilar/métodos , Espectrometría de Fluorescencia/métodos , AguaRESUMEN
Vibrational sum-frequency generation (SFG) spectroscopy has become an established technique for in situ surface analysis. While spectral recording procedures and hardware have been optimized, unique data analysis routines have yet to be established. The SFG intensity is related to probing geometries and properties of the system under investigation such as the absolute square of the second-order susceptibility χ((2)) (2). A conventional SFG intensity measurement does not grant access to the complex parts of χ((2)) unless further assumptions have been made. It is therefore difficult, sometimes impossible, to establish a unique fitting solution for SFG intensity spectra. Recently, interferometric phase-sensitive SFG or heterodyne detection methods have been introduced to measure real and imaginary parts of χ((2)) experimentally. Here, we demonstrate that iterative phase-matching between complex spectra retrieved from maximum entropy method analysis and fitting of intensity SFG spectra (iMEMfit) leads to a unique solution for the complex parts of χ((2)) and enables quantitative analysis of SFG intensity spectra. A comparison between complex parts retrieved by iMEMfit applied to intensity spectra and phase sensitive experimental data shows excellent agreement between the two methods.