Alginate microparticles as carriers for the UV filter 2-ethylhexyl 4-methoxycinnamate: Influence on photostability.

OBJECTIVE: The organic ultraviolet UVB filter 2-ethylhexyl 4-methoxycinnamate (EHMC) was encapsulated in microparticles (MPs) of sodium alginate and co-loaded with vitamin E (Vit.E) by an extrusion process using an aerodynamically assisted jetting (AAJ) methodology. The aim was to assess the effect of encapsulation concerning UVB filter release from the MPs and its photochemical stability. METHODS: The EHMC photostability was analysed by exposing the samples (both MPs in aqueous dispersion and incorporated in a cream preparation) during 1 h to simulated solar light. For the MPs (empty-MP, EHMC-MP and EHMC + Vit.E-MP), the morphology and size were characterized; while in the case of the encapsulated samples, the amount of EHMC-loading was determined. The release of EHMC was evaluated by adding EHMC-MP or EHMC + Vit.E-MP to 65% ethanol in water under mechanical stirring at 32°C. RESULTS: All MPs showed a homogeneous size distribution with a median of 90.5 ± 2.5 µm for EHMC-MP and 70.4 ± 1.14 µm for EHMC + Vit.E-MP. The encapsulation efficiency was 92.9% and 99.4% for EHMC-MP and EHMC + Vit.E-MP, respectively. The observed release from the MPs was lower than the dissolution of the pure UV filter. EHMC-MP and EHMC + Vit.E-MP were successfully incorporated into a cream formulation, with no evidence of phase separation or colour modification. Upon simulated light exposure, the photoisomerization/phototransformation of EHMC encapsulated in MPs and Vit.E-MP decreased as compared to free EHMC, both in aqueous dispersion and as a cream. The conformational ratio of the isomers (Z-/E-EHMC) was found to be the lowest in the presence of Vit.E. CONCLUSION: This work demonstrates that use of these alginate microparticulate carriers could enhance the effectiveness of sunscreen preparations containing this UVB filter.

OBJECTIF: Le filtre ultraviolet organique D'UVB 2-éthylhexyl 4-methoxycinnamate (EHMC) a été encapsulé dans des microparticules (MPs) d'alginate de sodium et co-chargé avec la vitamine E (Vit.E) par un processus d'extrusion utilisant une méthode de jet assisté aérodynamique (AAJ). L'objectif était d'évaluer l'effet de l'encapsulation concernant la libération du filtre UVB par les MPs et sa stabilité photochimique. MÉTHODES: La photostabilité EHMC a été analysée en exposant les échantillons (les deux MPs en dispersion aqueuse et incorporés dans une préparation de type crème) pendant 1 h à la lumière solaire simulée. Pour les MPs (MP-vide, EHMC-MP et EHMC - Vit.E-MP), la morphologie et la taille ont été caractérisées; tandis que dans le cas des échantillons encapsulés, la quantité de charge EHMC a été déterminée. Le relargage de l'EHMC a été évalué en ajoutant EHMC-MP ou EHMC - Vit.E-MP à 65% d'éthanol dans l'eau sous agitation mécanique à 32°C. RÉSULTATS: Tous les MP ont montré une distribution homogène de taille avec une médiane de 90,5 +- 2,5 µm pour EHMC-MP et de 70,4 +- 1,14 µm pour l'EHMC et Vit.E-MP. L'efficacité de l'encapsulation était de 92,9 % et 99,4 % pour EHMC-MP et EHMC - Vit.E-MP, respectivement. Le relargage observé des MPs était inférieur à la dissolution du filtre UV pur. EHMC-MP et EHMC - Vit.E-MP ont été incorporés avec succès dans une formulation de crème, sans aucune preuve de séparation de phase ou de modification des couleurs. Après exposition à la lumière simulée, la photoiomérisation/phototransformation de l'EHMC encapsulée dans les MPs et Vit.E-MP a diminué par rapport à l'EHMC libre, à la fois dans la dispersion aqueuse et la crème. Le rapport conformationnel des isomères (Z-/E-EHMC) s'est avéré le plus bas en présence de Vit.E.

Enhancement of trans-resveratrol photostability by encapsulation in lipid microparticles: in vitro and in vivo studies.

Lipid microparticles (LMs) loaded with the antioxidant polyphenol, trans-resveratrol were developed in order to enhance its photostability in topical formulations. The LMs were prepared by the melt emulsification technique, using tristearin as the lipidic material and hydrogenated phosphatidylcholine as the surfactant. The obtained microparticles were characterized by optical microscopy and release studies. The trans-resveratrol loading was 10.8% (w/w). Free or microencapsulated trans-resveratrol was introduced in model topical formulations (cream and hydrogel) and irradiated with a solar simulator. The light-induced degradation of trans-resveratrol was significantly reduced by incorporation into the LMs both in the cream (the trans-resveratrol loss decreased from 34.3% to 19.9%) and in the hydrogel (the trans-resveratrol decomposition decreased from 15.4% to 9.4%) vehicles. Moreover, the in vitro (i.e., antioxidant action) and in vivo (i.e., anti-inflammatory action) biological activities of trans-resveratrol in the cream preparation were not altered by the encapsulation process.

Comparative evaluation of the effect of permeation enhancers, lipid nanoparticles and colloidal silica on in vivo human skin penetration of quercetin.

BACKGROUND/AIM: The aim of this study was to evaluate emulsions containing a penetration enhancer, lipid nanoparticles (LNs) or colloidal silica as systems to improve the topical delivery of the flavonoid quercetin. METHODS: The skin penetration of quercetin was investigated in vivo on human volunteers by tape stripping. Quercetin-loaded LNs were prepared using hot high-pressure homogenization and characterized by means of dynamic light scattering and release studies. The location of the silica nanoparticles in the skin was determined by inductively coupled plasma mass spectrometry assay of silicon in the stratum corneum strips. RESULTS AND CONCLUSIONS: The penetration enhancer diethylene glycol monoethyl ether did not produce any significant increase in the fraction of the applied quercetin dose permeated in vivo into human stratum corneum (17.1 ± 3.2%) compared to the control emulsion (18.1 ± 2.3%). A greater but statistically nonsignificant accumulation of the flavonoid in the human horny layer (21.2 ± 2.9% of the applied dose) was measured following topical application of quercetin-loaded LNs (mean particle size: 527 nm). On the other hand, the addition of colloidal silica (average particle diameter: 486 nm) to the emulsion (2%, w/w) significantly increased the in vivo uptake of quercetin by the human stratum corneum to 26.7 ± 4.1% of the applied dose, the enhancing effect on permeation being more marked in the deepest horny layer strips. The measured in vivo skin penetration profile of colloidal silica showed that silica particles diffused down to the intermediate region of the human horny layer and hence could act as carrier for quercetin.

Encapsulation of the UV filters ethylhexyl methoxycinnamate and butyl methoxydibenzoylmethane in lipid microparticles: effect on in vivo human skin permeation.

BACKGROUND: Lipid microparticles loaded with the UVB filter ethylhexyl methoxycinnamate (EHMC) and the UVA filter butyl methoxydibenzoylmethane (BMDBM) were evaluated for their effect on the sunscreen agent's percutaneous penetration. METHODS: Microparticles loaded with EHMC or BMDBM were prepared by the melt emulsification technique using stearic acid or glyceryl behenate as lipidic material, respectively, and hydrogenate phosphatidylcholine as the surfactant. Nonencapsulated BMDBM and EHMC in conjunction with blank microparticles or equivalent amounts of the 2 UV filters loaded in the lipid microparticles were introduced into oil-in-water emulsions and applied to human volunteers. Skin penetration was investigated in vivo by the tape-stripping technique. RESULTS: For the cream with the nonencapsulated sunscreen agents, the percentages of the applied dose diffused into the stratum corneum were 32.4 ± 4.1% and 30.3 ± 3.3% for EHMC and BMDBM, respectively. A statistically significant reduction in the in vivo skin penetration to 25.3 ± 5.5% for EHMC and 22.7 ± 5.4% for BMDBM was achieved by the cream containing the microencapsulated UV filters. The inhibiting effect on permeation attained by the lipid microparticles was more marked (45-56.3% reduction) in the deeper stratum corneum layers. CONCLUSIONS: The reduced percutaneous penetration of BMDBM and EHMC achieved by the lipid microparticles should preserve the UV filter efficacy and limit potential toxicological risks.

Comparative evaluation of different substrates for the in vitro determination of sunscreen photostability: spectrophotometric and HPLC analyses.

Polymethylmethacrylate (PMMA) plates and Transpore(TM) tapes were compared as substrates for the in vitro evaluation of photostability of commercial sunscreen products. The sun care preparations were applied respectively on Transpore(TM) tapes and PMMA plates and their sun protection factors (SPF) and UVA protection parameters [UVA/UVB ratio, critical wavelength, UVA protection factor (UVA-PF)] were measured by transmission spectroscopy, before and after irradiation with simulated sunlight. No significant differences were observed in the UV protection parameters measured on Transpore(TM) tapes or PMMA plates, before exposure to the solar simulator. Conversely, after irradiation, the SPF values of the sun care products exhibited marked variations between the two substrates, the decrease in SPF being greater on PMMA plates (31.3-63.1%) than on Transpore(TM) tapes (10.4-23.8%). Differences between the two substrates were detected also for the UVA protection parameters, although they were significant only for the UVA-PF. The tested samples were assayed also by high-performance liquid chromatography (HPLC) to assess the extent of photodegradation of the UV filters present in the examined formulations. The results showed that for the PMMA plates, the light-induced decrease in SPF, as determined by spectrophotometry, fitted well with the percentage loss of ethyl hexyl methoxycinnamate (the only photounstable UVB filter present) measured by HPLC. Moreover, for the PMMA substrate, the UVA-PF percentage reduction was consistent with the percentage degradation of butyl methoxydibenzoylmethane (the only photounstable UVA filter present) determined by HPLC. On the other hand, poor correlation between spectrophotometric and HPLC analyses was observed on Transpore(TM) tapes. Therefore, PMMA plates are more reliable than Transpore(TM) tapes as substrates for in vitro photodegradation tests of sunscreen products by transmission spectroscopy.

Solid lipid microparticles for the stability enhancement of the polar drug N6-cyclopentyladenosine.

The objective of this study was to prepare solid lipid microparticles (SLMs) loaded with the polar adenosine A1 receptor agonist N6-cyclopentyladenosine (CPA). The microparticles were produced by the conventional hot emulsion technique, using different lipidic carriers (tristearin, glyceryl behenate and stearic acid) and hydrogenated phosphatidylcholine as the surfactant. The controlled release of CPA was achieved only with stearic acid microparticles. This phenomenon has been attributed to direct acid-base interactions between the basic nitrogen atoms of CPA and stearic acid. These SLMs were characterized by release studies, scanning electron microscopy and powder X-ray diffraction analyses. The obtained particles showed proper features in terms of morphology and size distribution (3.2-10.3microm), with a drug loading of 0.15+/-0.04%. The influence of the SLMs carrier system on CPA stability was investigated in vitro using human whole blood. The degradation kinetic of microparticle-entrapped CPA was significantly lower from that measured for the free CPA. The overall results indicate that it was possible to achieve the encapsulation and controlled release of a polar drug, such as CPA, within a lipid matrix without resorting to the complex methods generally used for the preparation of these systems.

In vivo and in vitro skin permeation of butyl methoxydibenzoylmethane from lipospheres.

Lipid microparticles (lipospheres) loaded with butyl methoxydibenzoylmethane (BMDBM), a widely used UV-A sunscreen agent, were prepared by melt technique and evaluated for skin permeation both in vivo, by tape stripping method, and in vitro, by a flow-through diffusion chamber. Following in vivo human skin application of an O/W emulsion containing 2% of BMDBM loaded in lipospheres, 15% of the applied sunscreen accumulated in the uppermost layers of the stratum corneum without remarkably modifying the skin permeation of the unencapsulated sunscreen. These results were found to be predicted by an in vitro methodology involving the diffusion of BMDBM through a lipophilized synthetic membrane into a hydrophilic receptor phase, simulating the viable epidermis better than an ethanolic receptor phase.

Poly(D,L-lactide) nanoencapsulation to reduce photoinactivation of a sunscreen agent.

The use of sunscreens is the 'gold standard' for protecting the skin from ultraviolet light. Octyl methoxycinnamate (OMC) is one of the most widely used UVB filter but it can act as a sensitizer or photoallergen. When exposed to sunlight, OMC can change from the primary trans-form to cis-form and the isomerization, not reversible, conducts to a reduction of the UVB filtering efficiency because the trans-form has a higher extinction coefficient. Photostability is the most important characteristic of effective sunscreens and it can be influenced by formulation ingredients and by applying technological strategies. In this work, photostability experiments, performed on emulsion-gels containing different percentages of OMC free or loaded in poly(D,L-lactide) nanoparticles, were carried out. The presence of a polymeric envelop may act to protect the active ingredient. In this study, the influence of poly(D,L-lactide) matrices on the photochemical stability of the sunscreen agent was investigated. As highlighted in this study, free OMC in different formulations has different photoisomerization degree. Moreover, a dissimilar behaviour was observed by studying different sunscreen concentrations in the same cosmetic formulation. Photostability results show a significant reduction in photoisomerization degree for formulations containing sunscreen loaded in nanoparticles, highlighting that the encapsulation is a suitable strategy to improve OMC photostability. Moreover, sun protection factor (SPF) results show that the UVB filter protective power is also maintained after encapsulation.

Influence of complexation with cyclodextrins on photo-induced free radical production by the common sunscreen agents octyl-dimethylaminobenzoate and octyl-methoxycinnamate.

The influence of complexation with hydroxypropyl-beta-cyclodextrin (HP-beta-CD) or beta-cyclodextrin (beta-CD) on the photo-induced production of free radicals by the sunscreen agents octyl-dimethylaminobenzoate (ODAB), oxybenzone (OB) and octyl-methoxycinnamate (OMC) was investigated. The formation of radical species during irradiation was detected by spin-trapping electron paramagnetic resonance (EPR) spectroscopy. 2,2,6,6-tetramethylpiperidine-1-oxyl, nitroxide radical (TEMPO) and 5,5-dimethyl-1-pyrroline-N-oxide (DMPO) were used a spin-traps. Following the 4-h illumination with simulated sunlight, OB did not generate radicals. On the other hand, photoexcitation of solutions containing ODAB or OMC produced a marked decrease (>40%) of the TEMPO signal intensity, demonstrating the formation of carbon-centred radicals. In addition, the results obtained on irradiation of ODAB solutions containing DMPO as spin-trap indicated the generation of oxygen-centred radicals. Complexation of ODAB with HP-beta-CD and OMC with beta-CD markedly inhibited (>64%) the formation of free radicals generated by the sunscreens on exposure to simulated sunlight. Therefore, inclusion of ODAB and OMC into the cyclodextrin cavities minimizes their photosensitising potential.

Hypo-excitability of cortical areas in patients affected by Friedreich ataxia: a TMS study.

The aim of the study was to explore excitability of a motor and a non-motor (visual) area in patients affected by Friedreich ataxia and to correlate neurophysiological data with clinical parameters. Seven patients (3M/4F) and ten healthy controls (5M/5F) participated in the study. The hot-spot for activation of right abductor pollicis brevis was checked by means of a figure-of-eight coil and the motor threshold (MT) on this point was recorded. The phosphene threshold (PT) was measured by means of a focal coil over the occipital cortex as the lower intensity of magnetic stimulation able to induce the perception of phosphenes. The patients showed a significantly higher mean PT (p<.03) and MT values (p<.001) than controls. In all but one patient unable to perceive phosphenes (42% vs. 50% of controls), TMS at 100% intensity did not elicit motor response at rest. The difference in percentage of patients (57.1%) and controls (100%) with motor responses was nearly significant. The size of GAA1 expansion showed significant correlations with PT and MT values. The results of our study showed that FA patients had reduced cortical activation, involving both the motor and the visual cortex. The cortical involvement in these patients seems to be mainly genetically determined. The study provides the first evidence of cortical dysfunction in patients with genetically defined Friedreich ataxia.

Reversed-phase HPLC study on the in vitro enzymic degradation of dermorphin.

A high-performance liquid chromatographic (HPLC) method for the separation of the opioid heptapeptide dermorphin and related fragments has been developed. The chromatographic system was applied in the study of the kinetics of degradation of dermorphin (Der) in various tissues. Der was found to be extremely resistant to human and rat plasma (T 1/2 greater than 180 min). Upon incubation with homogenates of rat brains and kidneys, Der was cleaved with a half-life of 20.8 +/- 2.2 min and 2.4 +/- 0.3 min respectively. The catabolite formed was identified, in both tissues, as the N-terminal tetrapeptide H-Tyr-D-Ala-Phe-Gly-OH. The stability to rat kidney and brain of the N-terminal hexa- and pentapeptides and of the [4 psi 5, NHCO] Der analogue was also investigated. The nature of the enzyme systems involved in the in vitro degradations is discussed.

In vitro degradation study of polyester microspheres by a new HPLC method for monomer release determination.

Biodegradable polyesters have increasing importance as materials used for the preparation of microspheres. The knowledge of their degradation process is important to prepare microparticulate delivery systems with suitable drug release rates. In this work an in vitro degradation study of empty and drug loaded microspheres is described. Three different polyesters were used: two poly-d, l-lactides of different molecular weight and a poly-d, l-lactide-co-glycolide (50:50). Diazepam has been chosen as the model drug. Solvent evaporation and spray-drying were used as preparation methods. To study the polymer degradation process, a new HPLC method is proposed for the direct and (in the case of the copolymer) simultaneous determination of the monomer(s): lactic acid (LA) and glycolic acid (GA). SEM and particle size analysis highlight the different characteristics of the particles, depending on their preparation method: spray-dried spheres result to be always smaller with respect to particles obtained by solvent evaporation. The results obtained indicate in particular that: the preparation methods play an important role in determining the degradation behaviour of microspheres, as unloaded spray-dried particles are characterized by a higher monomer release rate with respect to microspheres obtained by solvent evaporation; PLGA spheres degrade faster than PDLLA microparticles, according to the higher hydrophilicity of the copolymer; the two monomers are released at a different rate in the case of PLGA (faster for GA, slower for LA); the presence of diazepam increases the polymer degradation rate, with respect to empty particles.

HPLC assay of conjugated bile acids in human fluids using on-line sample pretreatment on a standard isocratic chromatograph.

A rapid and simple on-line purification procedure was developed for determining the conjugated bile acids in human fluids by high-performance liquid chromatography (HPLC). Biological samples were diluted and directly injected without further treatment onto a pre-column dry-packed with 40-microns octadecylsilica and installed at the injector loop position. After washing the pre-column with 40% methanol in acetate buffer and then with water, the retained compounds were back-flushed by the mobile phase onto the analytical column during the normal course of chromatography. The method was found to be accurate and reproducible. The levels of conjugated bile acids in serum, duodenal bile and gastric juice from patients with hepatobiliary and gastric diseases were determined by on-line pre-column clean-up and reversed-phase HPLC.

Phacovitrectomy without prone posture for full thickness macular holes.

AIMS: To investigate the role of phacovitrectomy surgery without prone posture for stage 2 and 3 macular holes. METHODS: A pilot study was performed on 20 patients (20 eyes) having phacoemulsification lens removal and vitrectomy surgery with 20% C(2)F(6) tamponade. Patients were advised to avoid lying on their backs for 10 days following surgery but no other posturing instructions were given. Closure rates and improvement in visual acuity were compared with a group of historical controls in whom phacovitrectomy with gas tamponade and face down posturing was performed. RESULTS: Anatomical hole closure was noted in 18 of the 20 eyes (90%). 19 eyes (95%) showed an improvement of at least 0.3 logMAR units. This compares favourably with the postured group in which anatomical hole closure was noted in 11 of 13 eyes (85%) and nine of 13 eyes (69%) showed an improvement of at least 0.3 logMAR units. CONCLUSION: Combined surgery facilitates the use of a large gas bubble. Sufficient tamponade of the hole occurs for closure without prone posturing. Combined surgery prevents patients posturing and returning for cataract surgery.

Determination of sunscreen agents in cosmetic products by supercritical fluid extraction and high-performance liquid chromatography.

A rapid supercritical fluid extraction (SFE) procedure for the isolation of five of the most common sunscreen agents (2-ethylhexyl-p-dimethylaminobenzoate, 2-hydroxy-4-methoxybenzophenone, 2-ethylhexyl-p-methoxycinnamate, 4-methylbenzylidene camphor and 4-tert.-butyl-4'-methoxydibenzoylmethane) from cosmetic products is described. Investigation of the factors affecting the extraction efficiency in SFE indicated that sunscreen recoveries were affected mainly by the supercritical CO2 pressure and by the trapping method. The sunscreens were analyzed by reversed-phase high-performance liquid chromatography after a 10-min extraction of the cosmetic product with CO2 at 250 bar and 40 degrees C, using sequential glass surface and C18 sorbent as collection system. A quantitative comparison of SFE with a liquid extraction procedure was performed on commercial cosmetics. The SFE method yielded recoveries higher than 94.8% compared with conventional liquid extraction and exhibited a precision better than 5.3% relative standard deviation. Moreover, SFE minimized sample handling, reduced the consumption of harmful solvents and afforded a more effective purification of the cosmetic matrices.

Determination of glycolic acid in cosmetic products by solid-phase extraction and reversed-phase ion-pair high-performance liquid chromatography.

A procedure has been developed for the assay of glycolic acid in cosmetic products by reversed-phase high-performance liquid chromatography using an ion-pairing method. After dissolution in tetrahydrofuran-water (90:10, v/v), samples were purified by solid-phase extraction using silica-based strong anion-exchange cartridges and analysed directly on an Ultrasphere ODS column with UV detection at 210 nm and methanol-phosphate buffer (2:98, v/v), containing tetrabutylammonium iodide, as the mobile phase. Recovery of glycolic acid from different cosmetic matrices was between 92.4 and 96.2% and the precision of the method was better than 5.4% relative standard deviation. The procedure is rapid, simple, selective and it is suitable for routine analyses of commercial cosmetics.

Determination of free bile acids in pharmaceutical preparations by packed column supercritical fluid chromatography.

A method was developed for the baseline separation of common free bile acids by supercritical fluid chromatography. A phenylbonded silica column, with UV detection at 210 nm, and carbon dioxide modified with methanol as the mobile phase were used. The influence of the stationary phase, modifier concentration, temperature, column pressure, and modifier identity on retention was studied. The separation obtained is at least eight times faster than those achieved for similar mixtures by the existing chromatographic techniques. This new procedure is applicable to the assay of ursodeoxycholic acid and chenodeoxycholic acid in capsule and tablet formulations. Individual dosage forms were disintegrated in methanol and an aliquot of the resulting suspension was filtered for the supercritical fluid chromatographic analysis. The method is rapid, accurate, and reproducible.

Determination of vitamin A, vitamin E, and their esters in tablet preparations using supercritical fluid extraction and HPLC.

A rapid and precise method for the isolation of vitamins A and E and their acetate or palmitate esters from table matrices was developed using supercritical fluid extraction (SFE). The vitamins were analyzed by nonaqueous reversed-phase high-performance liquid chromatography after a 15-min extraction of the dosage form with supercritical carbon dioxide at 40 degrees C and at a pressure of 250 atm. A quantitative comparison of SFE with an established liquid extraction procedure was performed on commercial tablet formulations. Vitamin recoveries of over 95.6% compared with conventional liquid extraction were achieved by the SFE technique with a much shorter sample preparation time (15 min vs 2h). Moreover, the automated SFE process minimized the number of sample handling operations, drastically reduced the consumption of harmful solvents, and provided mild extraction conditions for the analysis of the labile vitamins. The described SFE method is suitable for quality control analyses of pharmaceutical tablets.

Gabexate mesylate inhibition of serine proteases: thermodynamic and computer-graphics analysis.

The inhibitory effect of gabexate mesylate, which is used therapeutically in the treatment of pancreatitis and disseminated intravascular coagulation, and as a regional anticoagulant agent for hemodialysis, has been measured on bovine factor Xa, bovine alpha-thrombin, human Lys77-plasmin, human urinary kallikrein, human urokinase, porcine pancreatic beta-kallikrein-B, and bovine beta-trypsin catalyzed hydrolysis of p-nitrophenyl esters of N-alpha-carbobenzoxy-L-arginine and N-alpha-carbobenzoxy-L-lysine. On the basis of enzyme:gabexate mesylate affinities, the serine proteases can be arranged as follows: human urinary kallikrein approximately porcine pancreatic beta-kallikrein-B much less than bovine beta-trypsin approximately bovine factor Xa approximately human Lys77-plasmin approximately human urokinase approximately bovine alpha-thrombin. The mode of binding of gabexate mesylate to the serine proteases conforms to the active-reactive site geometries observed in their complexes with natural and synthetic inhibitors. Differences in gabexate mesylate affinities for these proteases reflect structural differences at their primary specificity subsite, which have been investigated by comparative analysis of amino acid sequences and by computer-graphics techniques.

Wild Amaranthus caudatus seed oil, a nutraceutical resource from Ecuadorian flora.

Seed oil of wild Amaranthus caudatus from Ecuador was analyzed for determining the tocopherol, fatty acid, and sterol contents. The data obtained were compared with the analogous chemical profile of seed oil of Italian A. caudatus with the objective of evaluating the nutraceutical and alimentary potential of the Ecuadorian matrix. Supercritical fluid and ultrasound-enhanced extractions were performed on both matrices. Qualitative and quantitative determinations of tocopherols were performed by HPLC, whereas GC and GC-MS were used to determine the fatty acid composition and sterols, respectively. Supercritical fluid extraction at 400 atm was the most efficient extraction method in terms of both total yield extract and tocopherol yield. Seeds of Ecuadorian of A. caudatus contained higher levels of tocopherols than Italian samples, whereas the fatty acid composition and sterol content were similar. From the obtained results it can be suggested that seed oil of wild Ecuadorian A. caudatus can prove to be an effective nutraceutical and alimentary resource and a valid alternative to the European varieties.