RESUMEN
Due to their ability to recognize carbohydrate structures, lectins emerged as potential receptors for bacterial lipopolysaccharides (LPS). Despite growing interest in investigating the association between host receptor lectins and exogenous glycan ligands, the molecular mechanisms underlying bacterial recognition by human lectins are still not fully understood. We contributed to fill this gap by unveiling the molecular basis of the interaction between the lipooligosaccharide of Escherichia coli and the dendritic cell-specific intracellular adhesion molecules (ICAM)-3 grabbing non-integrin (DC-SIGN). Specifically, a combination of different techniques, including fluorescence microscopy, surface plasmon resonance, NMR spectroscopy, and computational studies, demonstrated that DC-SIGN binds to the purified deacylated R1 lipooligosaccharide mainly through the recognition of its outer core pentasaccharide, which acts as a crosslinker between two different tetrameric units of DC-SIGN. Our results contribute to a better understanding of DC-SIGN-LPS interaction and may support the development of pharmacological and immunostimulatory strategies for bacterial infections, prevention, and therapy.
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The evaluation of Bacteroides vulgatus mpk (BVMPK) lipopolysaccharide (LPS) recognition by DC-SIGN, a key lectin in mediating immune homeostasis, has been here performed. A fine chemical dissection of BVMPK LPS components, attained by synthetic chemistry combined to spectroscopic, biophysical, and computational techniques, allowed to finely map the LPS epitopes recognized by DC-SIGN. Our findings reveal BVMPK's role in immune modulation via DC-SIGN, targeting both the LPS O-antigen and the core oligosaccharide. Furthermore, when framed within medical chemistry or drug design, our results could lead to the development of tailored molecules to benefit the hosts dealing with inflammatory diseases.
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The aroma characterization of 58 unroasted cocoa beans from 22 different geographical origins was performed by head space solid phase micro-extraction (HS-SPME) combined with gas chromatography-mass spectrometry (GC-MS). Sampling is representative of the average world production (America, Africa, and Southeast Asia). Analysis of cocoa beans before and after roasting were performed to follow the aroma modification with the aim to achieve a cocoa volatile fingerprint and a discrimination model based on beans origin. A total of 57 volatiles was identified in unroasted cocoa beans, while 71 volatiles were identified in roasted cocoa beans. The compounds belong to several chemical groups including esters, alcohols, organic acids, aldehydes, ketones and pyrazines. Datasets were submitted to multivariate statistical analysis (Principal Component Analysis, PCA). Results allowed to discriminate unroasted cocoa beans based on their geographical origin: samples coming from African countries were separated from samples of American regions, whereas samples from Southeast Asia lie between the other two continents suggesting that Asian samples have intermediate characteristics between African and South American cocoa beans. PCA, applied on the corresponding roasted samples, showed that although the same roasting treatment has been applied to all the samples, the differences among the unroasted samples were also maintained in the aromatic profile after roasting. The discrimination model based on volatile fingerprint combined with chemometric tools, showed interesting potential for origin authentication of both unroasted and roasted cocoa beans.
Asunto(s)
Cacao/química , Semillas/química , Compuestos Orgánicos Volátiles/análisis , África , Alcoholes/análisis , Aldehídos/análisis , Américas , Asia Sudoriental , Chocolate/análisis , Ésteres/análisis , Fermentación , Manipulación de Alimentos , Cromatografía de Gases y Espectrometría de Masas , Cetonas/análisis , Odorantes/análisis , Olfato , Microextracción en Fase SólidaRESUMEN
Galdieria phlegrea is a polyextremophilic red alga belonging to Cyanidiophyceae. Galdieria phlegrea C-phycocyanin (GpPC), an abundant light-harvesting pigment with an important role in energy capture and transfer to photosystems, is the C-phycocyanin (C-PC) with the highest thermal stability described so far. GpPC also presents interesting antioxidant and anticancer activities. The X-ray structure of the protein was here solved. GpPC is a [(αß)3]2 hexamer, with the phycocyanobilin chromophore attached to Cys84α, Cys82ß and Cys153ß. Details of geometry and interaction with solvent of the chromophores are reported. Comparison with the structure of a C-PC in the entire Porphyridium purpureum phycobilisome system reveals that linker polypeptides have a significant effect on the local structure of the chromophores environment. Comparative analyses with the structures of other purified C-PCs, which were carried out including re-refined models of G. sulphuraria C-PC, reveal that GpPC presents a significantly higher number of inter-trimer salt bridges. Notably, the higher number of salt bridges at the (αß)3/(αß)3 interface is not due to an increased number of charged residues in this region, but to subtle conformational variations of their side chains, which are the result of mutations of close polar and non-polar residues.
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Ficocianina/química , Rhodophyta/enzimología , Temperatura , Cristalografía por Rayos X , Estabilidad de Enzimas , Metilación , Modelos Moleculares , Ficocianina/metabolismo , Conformación ProteicaRESUMEN
C-phycoyanins are abundant light-harvesting pigments which have an important role in the energy transfer cascade of photosystems in prokaryotic cyanobacteria and eukaryotic red algae. These proteins have important biotechnological applications, since they can be used in food, cosmetics, nutraceutical, pharmaceutical industries and in biomedical research. Here, C-phycocyanin from the extremophilic red alga Galdieria phlegrea (GpPC) has been purified and characterized from a biophysical point of view by SDS-PAGE, mass spectrometry, UV-Vis absorption spectroscopy, circular dichroism and intrinsic fluorescence. Stability against pH variations, addition of the oxidizing agent hydrogen peroxide and the effects of temperature have been also investigated, together with its in cell antioxidant potential and antitumor activity. GpPC is stable under different pHs and unfolds at a temperature higher than 80 °C within the pH range 5.0-7.0. Its fluorescence spectra present a maximum at 650 nm, when excited at 589 nm. The protein exerts interesting in cell antioxidant properties even after high temperature treatments, like the pasteurization process, and is cytotoxic for A431 and SVT2 cancer cells, whereas it is not toxic for non-malignant cells. Our results assist in the development of C-phycocyanin as a multitasking protein, to be used in the food industry, as antioxidant and anticancer agent.
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Fenómenos Químicos , Ficocianina/química , Secuencia de Aminoácidos , Antineoplásicos/química , Antineoplásicos/aislamiento & purificación , Antineoplásicos/farmacología , Antioxidantes/química , Antioxidantes/aislamiento & purificación , Antioxidantes/farmacología , Cromatografía Liquida , Concentración de Iones de Hidrógeno , Ficocianina/aislamiento & purificación , Ficocianina/farmacología , Estabilidad Proteica , Especies Reactivas de Oxígeno/metabolismo , Rhodophyta/química , Espectrometría de Masas en Tándem , TemperaturaRESUMEN
Black soldier fly (BSF, Hermetia illucens) represents a valuable source of biomolecules and it also constitutes an economic way to valorise residual biomasses. BSF prepupae contain high amounts of lipids (37% DM basis). The present investigation aimed at studying the composition of BSF lipids and the effect of killing/storage on their quality. The main fatty acid was lauric acid, sterols were represented primarily by beta-sitosterol and campesterol. Global fatty acid and sterol profiles, determined by GC-MS, were only slightly affected by the killing procedure, while lipid classes distribution, determined by 1H NMR, strongly changed. Prepupae killed by freezing showed a drastic reduction of acylglycerols during storage and a relevant release of free fatty acids, likely due to activation of lipases. On the contrary, prepupae killed by blanching have a stable lipid fraction constituted mainly by triacylglycerols. Therefore, killing procedure strongly influences BSF oil composition and the potential applications.
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Manipulación de Alimentos/métodos , Homicidio , Lípidos/análisis , Simuliidae/química , Animales , Composición Corporal , Dípteros , Ácidos Grasos/análisis , Congelación , Glicéridos/análisis , Larva/química , Esteroles/análisisRESUMEN
In this study we investigated the oligopeptide pattern in fermented cocoa beans and derived products after simulated gastrointestinal digestion. Peptides in digested cocoa samples were identified based on the mass fragmentation and on the software analysis of vicilin and 21 KDa cocoa seed protein sequences, the most abundant cocoa proteins. Quantification was carried out by liquid chromatography/electrospray ionisation mass spectrometry (LC/ESI-MS) using an internal standard. Sixty five peptides were identified in the digested samples, including three pyroglutamyl derivatives. The in vitro angiotensin-converting enzyme (ACE)-inhibitory activity of cocoa digests were tested, demonstrating a high inhibition activity, especially for digestates of cocoa beans. The peptides identified were screened for their potential ACE inhibitory activity through an in silico approach, and about 20 di-, three- and tetra-peptides actually present in our samples were predicted as active. Two of the potentially active peptides were chemically synthesized and then assessed for their inhibitory activity by using the ACE in vitro assay. These peptides demonstrated an ACE inhibitory activity, however, that was too weak to explain alone the high activity of cocoa digestates, suggesting a synergic effect of all cocoa peptides. As a whole, results showed that an average chocolate portion (30 g) ensures an amount of peptides after digestion that, assuming complete absorption, could reach almost a complete inhibition of ACE.
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Inhibidores de la Enzima Convertidora de Angiotensina/metabolismo , Cacao/química , Digestión , Oligopéptidos/aislamiento & purificación , Cromatografía Líquida de Alta Presión , Humanos , Simulación del Acoplamiento Molecular , Espectrometría de Masa por Ionización de ElectrosprayRESUMEN
Cyclopropane fatty acids (CPFAs) are unusual fatty acids of microbial origin, recently detected in milk and dairy products. CPFAs have been demonstrated to be interesting molecular markers for authentication of dairy products obtained without ensiled feeds. Moreover, they can also be recognized as a new secondary component of human diet. Information is lacking on the presence of cyclic fatty acids in other food sources. Cyclopropane fatty acids have been detected by GC-MS analysis in cheese and other animal fats in concentration ranging from 200 to 1000 mg/kg fat, but in some cases, the complex fatty acid profile and the possible presence of interfering peaks make the separation not straightforward and the quantification uneasy. Therefore, a new reliable 1H NMR method was developed to detect and measure CPFA content in different foods of animal origin, based on the detection of the characteristic signals of cyclopropane ring. The 1H NMR (600 MHz) method showed detection limits comparable with those of full scan GC-MS, and it allowed the identification and quantitation of the cyclopropane fatty acids in different foods.
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Black soldier fly (BSF, Hermetia illucens) constitutes an economic way to convert residual biomasses into a valuable source of biomolecules, such as proteins, lipids and chitin. The present investigation was undertaken to evaluate the feasibility of applying different extraction protocols, either chemical extractions or enzymatic assisted extraction, to recover pure fat, protein and chitin fractions. First, exact proximate composition, total amino acids, fatty acids profile, and N-acetylglucosamine content of the prepupae samples were determined. BSF prepupae biomass contained, expressed on dry weight, 32% proteins, 37% lipids, 19% minerals, 9% chitin. The lipid fraction was easily recovered by organic solvents, while the most challenging issue was the separation of protein from chitin. The best separation was obtained by alkali extraction of proteins (96% of protein recovered) albeit with loss in their integrity as indicated by the measurement of the degree of hydrolysis with the o-phthaldialdehyde method. To avoid protein damage in alkali media, a stepwise protein extraction adopting milder conditions was also explored based on Osborne fractionation method, allowing the recovery of >85% of BSF high purity and high quality proteins, and the obtainment of chitin-enriched fraction as well. The possibility of using an enzymatic assisted extraction of proteins was also explored, obtaining a maximum nitrogen solubilisation in the best case (with Bacillus licheniformis protease) of about 60%. In this latter case, the chitin fraction obtained also had a significant residual protein content.
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Quitina/aislamiento & purificación , Manipulación de Alimentos/métodos , Proteínas de Insectos/aislamiento & purificación , Lípidos/aislamiento & purificación , Pupa/metabolismo , Simuliidae/metabolismo , Animales , Composición Corporal , Fraccionamiento Químico , Hidrolasas/metabolismo , Hidrólisis , Valor Nutritivo , Péptido Hidrolasas/metabolismo , Proteolisis , Pupa/crecimiento & desarrollo , Simuliidae/crecimiento & desarrolloRESUMEN
Lactic acid bacteria are commonly used in dairy industries to acidify milk and to enhance the flavour of the end products thanks to their metabolisms. The formation routes of aroma compounds mainly rely on the specific ability of different species and strains to convert precursors derived from carbohydrate and amino acids catabolism. It is well known that the strains largely involved in the aroma formation of the very appreciated Italian long ripened cheeses belong to the Lactobacillus casei group and origin from raw milk. In this study, a spontaneous fermentation of Parmigiano Reggiano raw milks was carried out to isolate new strains potentially usable as adjunctive aromatic starter. For this reason, specific selection criteria were chosen to isolate strains belonging to L. casei, and L. paracasei species. An integrated approach, by mean of impedance microbiology and SPME GC-MS analysis, was applied to investigate the acidifying performance and the production of volatile compounds of seven strains in UHT whole milk. One of these strains, L. paracasei 4341, appear to be the most interesting one from the technological point of view both for its acidifying and aromatic features. This approach could be employed for selection of the aromatic strains to be potentially used as adjunct starter in dairy sector.
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Queso/microbiología , Manipulación de Alimentos/métodos , Lacticaseibacillus casei/metabolismo , Microbiología de Alimentos , Cromatografía de Gases y Espectrometría de Masas , Microextracción en Fase Sólida , Temperatura , Compuestos Orgánicos Volátiles/análisis , Compuestos Orgánicos Volátiles/química , Compuestos Orgánicos Volátiles/metabolismoRESUMEN
The lipid fraction of Asiago Protected Designation of Origin (PDO) cheese was analyzed to identify specific biomarkers of its main production systems through a canonical discriminant analysis. The three main production systems of the cheese were considered. Two were located in the upland (UL): pasture-based (P-UL) vs hay-based total mixed rations (H-UL). The third was located in the lowland (LL) and processed milk from cows fed maize silage-based rations (maize silage lowland: MS-LL). The discriminant analysis selected nine fatty acids and vitamin A as lipid biomarkers useful to separate the three production systems. High contents of conjugated linoleic acids, anteiso-C15:0, and vitamin A were discriminant factors for P-UL cheese. The separation between H-UL and MS-LL cheese was less marked with the former having the higher content of conjugated linoleic acids and some polyunsaturated n-6 fatty acids and with the latter being identified by cyclopropane fatty acid and C9:0.
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Bovinos/metabolismo , Queso/análisis , Ácidos Grasos/metabolismo , Alimentación Animal/análisis , Animales , Biomarcadores/análisis , Análisis Discriminante , Femenino , Leche/química , Leche/metabolismo , Ensilaje/análisis , Vitamina A/análisis , Vitamina A/metabolismo , Zea mays/química , Zea mays/metabolismoRESUMEN
Cyclopropane fatty acids (CPFA), as lactobacillic acid and dihydrosterculic acid, are components of bacterial membranes and have been recently detected in milk and in dairy products from cows fed with corn silage. In this paper, a specific quantitative gas chromatography-mass spectrometry (GC-MS) method for the detection of CPFA in cheeses was developed, and the quality parameters of the method (limit of detection, limit of quantitation, and intralaboratory precision) were assessed. Limit of detection and quantitation of CPFA were, respectively, 60 and 200 mg/kg of cheese fat, and the intralaboratory precision, determined on three concentration levels, satisfied the Horwitz equation. This method was applied to 304 samples of PDO cheeses of certified origin, including Parmigiano Reggiano (Italy), Grana Padano (Italy), Fontina (Italy), Comté (France), and Gruyère (Switzerland). Results showed that CPFA were absent in all of the cheeses whose Production Specification Rules expressly forbid the use of silages (Parmigiano Reggiano, Fontina, Comté, and Gruyère). CPFA were instead present in variable concentrations (300-830 mg/kg of fat) in all of the samples of Grana Padano cheese (silages admitted). A mix of grated Parmigiano Reggiano and Grana Padano was also prepared, showing that the method is able to detect the counterfeiting of Parmigiano Reggiano with other cheeses up to 10-20% Grana Padano content. These results support the hypothesis that CPFA can be used as a marker of silage feedings for cheeses, and the data reported can be considered a first attempt to create a database for CPFA presence in PDO cheeses.
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Queso/análisis , Ciclopropanos/análisis , Ácidos Grasos/análisis , Contaminación de Alimentos/análisis , Cromatografía de Gases y Espectrometría de Masas/métodos , Animales , Bovinos , Europa (Continente) , Ensilaje/análisisRESUMEN
Peptides and amino acids generated during cocoa bean fermentation are the most important precursors for the development of cocoa aroma, however cocoa oligopeptide fraction is under-investigated. In this study, we perform a deep investigation of the presence of oligopeptides in unfermented, under fermented, and well-fermented cocoa beans from all of the main producing countries, with the aim to obtain a better definition of cocoa quality and a deeper comprehension of biochemical changes occurring during fermentation. Oligopeptides were determined by UPLC/ESI-MS and 35 low-molecular weight peptides were identified and subjected to semi-quantitative analysis. Results showed that fermented cocoas can be differentiated from unfermented cocoas by their possession of a higher total amount of oligopeptides and a lower ratio of vicilin to 21kDa cocoa seed albumin peptides. A variability in the peptide pattern was observed also among well-fermented cocoa samples of different geographical origin, suggesting diversified proteolytic activities.
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Cacao/química , Fermentación , Oligopéptidos/análisis , Albúminas/química , Aminoácidos/química , Geografía , Peso Molecular , Proteínas de Almacenamiento de Semillas/químicaRESUMEN
Atmospheric double-barrier discharge (DBD) plasma technology is a promising tool in the food industry as an alternative to traditional food preservation methods. However, the effect of the reactive species generated during the treatment on the content of bioactive compounds in food is still little studied, and there are no data concerning potential deleterious effects of DBD-treated foods on human cells. Some functional properties of DBD-treated minimally processed Pink Lady apples were evaluated in comparison with untreated samples through different in vitro and ex vivo tests. Plasma treatment caused only a slight reduction of antioxidant content and antioxidant capacity (up to 10%), mainly limited to the amphiphilic fraction. Noteworthy, treated apple polyphenol extracts did not reduce cell viability and did not suppress the beneficial physiological cell response to oxidative stress in terms of reactive oxygen species production and phase II enzyme activation in human cultured colonocytes.
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A survey was carried out to determine the presence of cyclopropane fatty acids (CPFA) in various dairy products. CPFA such as lactobacillic acid and dihydrosterculic acid are components of bacterial membranes and have been recently detected in milk from cows fed with maize silage. In this paper about 200 dairy samples comprising cow, sheep, and goat milk, cheese, yogurt/fermented milk, and butter were analyzed. Results showed that cow milks were generally positive to CPFA (0.014-0.105% of total fatty acids), while goat, yak, and sheep milks were negative. Experimental yogurt and fermented milks showed the same CPFA content of the starting milk. Positive to CPFA were also the majority of samples of commercial butter and cheeses, except some PDO cheeses as Parmigiano-Reggiano and Fontina, cheeses from mountain regions, and goat and sheep cheeses. These data suggest that the presence of CPFA in dairy products could be used as a marker of silage feeding.
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Ciclopropanos/análisis , Productos Lácteos/análisis , Ácidos Grasos/análisis , Leche/química , Alimentación Animal , Animales , Bacterias/química , Bacterias/ultraestructura , Mantequilla/análisis , Bovinos , Pared Celular/química , Queso/análisis , Fermentación , Cromatografía de Gases y Espectrometría de Masas , Cabras , Lactobacillus/metabolismo , Lactobacillus/ultraestructura , OvinosRESUMEN
This study reports for the first time the use of (1)H NMR technique combined with chemometrics to study the metabolic profile of cocoa (Theobroma cacao L.) beans of different varieties, origin and fermentation levels. Results of PCA applied to cocoa bean (1)H NMR dataset showed that the main factor influencing the cocoa bean metabolic profile is the fermentation level. In fact well fermented brown beans form a group clearly separated from unfermented, slaty, and underfermented, violet, beans, independently of the variety or geographical origin. Considering only well fermented beans, the metabolic profile obtained by (1)H NMR permitted to discriminate between some classes of samples. The National cocoa of Ecuador, known as Arriba, showed the most peculiar characteristics, while the samples coming from the African region showed some similar traits. The dataset obtained, representative of all the classes of soluble compounds of cocoa, was therefore useful to characterise fermented cocoa beans as a function of their origin and fermentation level.
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Cacao/química , Imagen por Resonancia Magnética/métodos , Fermentación , Espectroscopía de Protones por Resonancia MagnéticaRESUMEN
Different grades of genuine and counterfeit Fraxinus excelsior exudates, marketed as natural sweeteners or mild laxatives, were evaluated for their proximate composition and for saccharidic, organic acids, lipidic and phenolic profile by means of GC-MS and (1)H NMR. Genuine samples contained mannitol (39-48 g/100 g, according to the grade), fructose (9-16 g/100 g), glucose (2-3.7 g/100 g), sorbitol (0,5-0,6 g/100 g), galactose (0.02-0.74 g/100 g), oligosaccharides as mannotriose (13-22 g/100 g) and stachyose (1-11 g/100 g), and traces of myo-inositol, mannose, sucrose. On the contrary, counterfeit samples contained mostly mannitol and sorbitol, with traces of fructose, glucose and mannose. Differences in ash, total polyphenolic content and fatty acid composition allowed a quick identification of counterfeit products, confirmed by a distinct mono-, oligosaccharidic and phenolic pattern. Elenolic acid (63-1628 mg/kg), tyrosol (15-774 mg/kg), homovanillic acid (2,39-52.8 mg/Kg), dopaol (0.8-63 mg/kg), pinoresinol (4.2-18.5 mg/kg) and fraxetin (0.25-11.64 mg/kg), albeit showing a wide concentration range, were the most abundant substances detected in the phenolic fraction of Fraxinus manna, while esculetin, p-hydroxybenzoic acid, 4-hydroxyphenacetic acid, 3,4 hydroxybenzoic acid, hydroxy-pinoresinol, medioresinol and siringaresinol were present in low amounts. The polyphenolic profile may be used as a marker for authentication and should be considered in the evaluation of nutritional and health properties ascribed to Fraxinus manna.
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Fraxinus/química , Hexosas/análisis , Oligosacáridos/análisis , Extractos Vegetales/química , Exudados de Plantas/química , Polifenoles/análisis , Alcoholes del Azúcar/análisis , Cumarinas/análisis , Ácidos Grasos/análisis , Furanos/análisis , Ácido Homovanílico/análisis , Lignanos/análisis , Alcohol Feniletílico/análogos & derivados , Alcohol Feniletílico/análisis , Extractos Vegetales/normas , Piranos/análisisRESUMEN
Content of cyclopropyl and ω-cyclohexyl fatty acids of microbial origin, respectively, from silage and rumen, was determined by GC-MS and confirmed by (1)H NMR in more than 200 milk samples from dairy cows fed with different forages. Cyclopropyl fatty acids (about 0.1% of milk fat) were detected for the first time in milk and they were present only in milk samples from cows fed with forages containing maize silage, which is not allowed to produce milk for some PDO cheeses as Parmigiano-Reggiano. Their determination can be proposed as a quality parameter of milk or feed and it can be useful especially to distinguish cheeses sold as Parmigiano Reggiano from others cheeses. The content of ω-cyclohexyl tridecanoic acid varied from 0.0% to 0.15% of milk fat, and it was higher in milk samples from cows fed with diets richer in cereal meals.
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Alimentación Animal/análisis , Queso/análisis , Ácidos Grasos/química , Leche/química , Fenómenos Fisiológicos Nutricionales de los Animales , Animales , Bovinos , Digestión , Estructura Molecular , Control de CalidadRESUMEN
The occurrence of triterpene pentacyclic acids in plants is extensive, but little is known about their availability in commercial extracts. A simple GC-MS method for the simultaneous determination of betulinic, corosolic, maslinic, oleanolic and ursolic acids was developed and applied to 38 different commercial plant extracts sold as ingredients for dietary supplements. A suitable protocol was set up to perform routine control of a diverse array of samples with different botanical, chemical and physical characteristics. Remarkable quantities of corosolic acid were found in dried extracts from aerial parts of Lagerstroemia speciosa and Ortosiphon stamineus (14233 and 1132 mg/kg, respectively), while oleanolic acid was abundant in O. stamineus and Crataegus monogyna flowers (2774 and 2339 mg/kg); ursolic was identified in O. stamineus, C. monogyna, L. speciosa and Arctostaphylos uva-ursi leaves (7773, 4165, 2108 and 1034 mg/kg). Only L. speciosa was rich in maslinic acid (4958 mg/kg), while minor amounts of betulinic acid (257 and 80 mg/kg) were detected in L. speciosa and C. monogyna extracts. Lower quantities of triterpenic acids were identified in dried extracts of Harpagophyton procumbens root, propolis, Punica granatum root, Styrax benzoin, Vaccinium myrtillus fruits and Vitis vinifera seeds. Decoctions and fluid extracts lacked or contained very low amounts of triterpenic acids. Results are discussed in terms of quality and safety of these ingredients.