RESUMEN
Fourier-Transform mid-infrared (FTIR) spectroscopy offers a strong candidate screening tool for rapid, non-destructive and early detection of unauthorized virgin olive oil blends with other edible oils. Potential applications to the official anti-fraud control are supported by dozens of research articles with a "proof-of-concept" study approach through different chemometric workflows for comprehensive spectral analysis. It may also assist non-targeted authenticity testing, an emerging goal for modern food fraud inspection systems. Hence, FTIR-based methods need to be standardized and validated to be accepted by the olive industry and official regulators. Thus far, several literature reviews evaluated the competence of FTIR standalone or compared with other vibrational techniques only in view of the chemometric methodology, regardless of the inherent characteristics of the product spectra or the application scope. Regarding authenticity testing, every step of the methodology workflow, and not only the post-acquisition steps, need thorough validation. In this context, the present review investigates the progress in the research methodology on FTIR-based detection of virgin olive oil adulteration over a period of more than 25 years with the aim to capture the trends, identify gaps or misuses in the existing literature and highlight intriguing topics for future studies. An extensive search in Scopus, Web of Science and Google Scholar, combined with bibliometric analysis, helped to extract qualitative and quantitative information from publication sources. Our findings verified that intercomparison of literature results is often impossible; sampling design, FTIR spectral acquisition and performance evaluation are critical methodological issues that need more specific guidance and criteria for application to product authenticity testing.
Asunto(s)
Olea , Proyectos de Investigación , Aceite de Oliva/análisis , Espectroscopía Infrarroja por Transformada de Fourier/métodos , Quimiometría , Aceites de Plantas/química , Contaminación de Alimentos/análisisRESUMEN
The last years, non-targeted fingerprinting by Fourier-transform infrared (FT-IR) spectroscopy has gained popularity as an alternative to classical gas chromatography (GC)-based methods because it may allow fast, green, non-destructive and cost-effective assessment of quality of essential oils (EOs) from single plant species. As the relevant studies for Laurus nobilis L. (bay laurel) EO are limited, the present one aimed at exploring the diagnostic potential of FT-IR fingerprinting for the identification of its botanical integrity. A reference spectroscopic dataset of 97 bay laurel EOs containing meaningful information about the intra-species variation was developed via principal component analysis (PCA). This dataset was used to train a one-class model via soft independent modelling class analogy (SIMCA). The model was challenged against commercial bay laurel and non-bay laurel EOs of non-traceable production history. Overall, the diagnostic importance of spectral bands at 3060, 1380-1360, 1150 and 1138 cm-1 was assessed using GC-FID-MS data. The findings support the introduction of FT-IR as a green analytical technique in the quality control of these often mislabeled and/or adulterated precious products. Continuous evaluation of the model performance against newly acquired authentic EOs from all producing regions is needed to ensure validity over time.
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Laurus/química , Aceites Volátiles/química , Espectroscopía Infrarroja por Transformada de Fourier/métodos , Conjuntos de Datos como Asunto , Cromatografía de Gases y Espectrometría de Masas , Aceites Volátiles/normas , Análisis de Componente Principal/clasificación , Control de CalidadRESUMEN
In previous works on authentic samples of saffron of known history (harvest and processing year, storage conditions, and length of time) some biomarkers were proposed using both FT-IR and NMR metabolomics regarding the shelf life of the product. This work addresses the difficulties to trace back the "age" of commercial saffron samples of unknown history, sets a limit value above which these products can be considered substandard, and offers a useful tool to combat saffron mislabeling and fraud with low-quality saffron material. Investigations of authentic and commercial saffron samples of different origin and harvest year, which had been stored under controlled conditions for different lengths of time, allowed a clear-cut clustering of samples in two groups according to the storage period irrespectively of the provenience. In this respect, the four-year cut off point proposed in our previous work assisted to trace back the "age" of unknown samples and to check for possible mislabeling practices.
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Crocus/química , Metabolómica/métodos , Espectroscopía de Protones por Resonancia Magnética , Control de Calidad , Espectroscopía Infrarroja por Transformada de FourierRESUMEN
Saffron is a spice comprised of the dried stigmas and styles of Crocus sativus L. flowers and, since it is very expensive, it is frequently adulterated. So far, proteomic tools have never been applied to characterize the proteome of saffron or identify possible cases of fraud. In this study, 1D-Gel Electrophoresis was carried out to characterize the protein profile of (i) fresh stigmas and styles of the plant; (ii) dried stigmas and styles from different geographical origins (Spanish, Italian, Greek and Iranian) that had been stored for various periods of time after their processing; and (iii) two common plant adulterants, dried petals of Carthamus tinctorius L. and dried fruits of Gardenia jasminoides Ellis. A selective protein extraction protocol was applied to avoid interference from colored saffron metabolites, such as crocins, during electrophoretic analyses of saffron. We succeeded in separating and assigning the molecular weights to more than 20 proteins. In spite of the unavailability of the genome of saffron, we were able to identify five proteins by Peptide Mass Fingerprinting: phosphoenolpyruvate carboxylase 3, heat shock cognate 70 KDa protein, crocetin glucosyltransferase 2, α-1,4-glucan-protein synthase and glyceraldehydes-3-phosphate dehydrogenase-2. Our findings indicate that (i) few bands are present in all saffron samples independently of origin and storage time, with amounts that significantly vary among samples and (ii) aging during saffron storage is associated with a reduction in the number of detectable bands, suggesting that proteases are still active. The protein pattern of saffron was quite distinct from those of two common adulterants, such as the dried petals of Carthamus tinctorius and the dried fruits of Gardenia jasminoides indicating that proteomic analyses could be exploited for detecting possible frauds.
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Carthamus tinctorius/metabolismo , Crocus/química , Gardenia/metabolismo , Proteínas de Plantas/aislamiento & purificación , Crocus/metabolismo , Electroforesis en Gel de Poliacrilamida , Flores/metabolismo , Mapeo Peptídico , Proteínas de Plantas/metabolismo , Proteómica , Especificidad de la EspecieRESUMEN
The present study aims to examine whether and to what extent the bioaccessibility of the major saffron apocarotenoids, namely crocetin sugar esters (CRTSEs), is affected by the presence of strong water-soluble antioxidants, ingredients of the herbs found in commercial tea blends with saffron. An in vitro digestion model was applied to infusions from these products to investigate the possible changes. All of the studied infusions were rich in total phenols (9.9-22.5 mg caffeic acid equivalents/100 mg dry infusion) and presented strong DPPH radical scavenging activity regardless of the composition of the corresponding herbal blends. RP-HPLC-DAD and LC-MS analysis enabled the grouping of the infusions into hydroxycinnamic acid-rich and in flavan-3-ol-rich ones. CRTSEs in herbal tea infusions were found to be significantly more bioaccessible (66.3%-88.6%) than those in the reference saffron infusion (60.9%). The positive role of strong phenolic antioxidants (caffeic acid, rosmarinic acid) on the stability of CRTSEs was also evidenced in model binary mixtures. On the contrary, cinnamic acid, exerting no antioxidant activity, did not have such an effect. Our findings suggest that strong radical scavengers may protect the crocetin sugar esters from oxidation during digestion when present in excess.
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Antioxidantes/química , Carbohidratos/química , Carotenoides/química , Crocus/química , Extractos Vegetales/química , Cromatografía Líquida de Alta Presión , Ésteres , Fenoles/química , Vitamina A/análogos & derivadosRESUMEN
In recent years, there has been a growing demand for minimally processed foods that offer health benefits and premium sensory characteristics. This trend has led to increased consumption of virgin (VOs) and cold-pressed (CPOs) oils, which are rich sources of bioactive substances. To meet consumer needs for new oil products conferring multi-functional properties over a longer storage period, the scientific community has been revisiting traditional enrichment practices while exploring novel fortification technologies. In the last four years, the interest has been growing faster; an ascending number of annually published studies are about the addition of different plant materials, agri-food by-products, or wastes (intact or extracts) to VOs and CPOs using traditional or innovative fortification processes. Considering this trend, the present review aims to provide an overview and summarize the key findings from relevant papers that were retrieved from extensively searched databases. Our meta-analysis focuses on exposing the most recent trends regarding the exploitation of VOs and CPOs as substrates, the fortification agents and their form of use, as well as the fortification technologies employed. The review critically discusses possible health claim and labeling issues and highlights some chemical and microbial safety concerns along with authenticity issues and gaps in quality specifications that manufacturers have yet to address. All these aspects are examined from the perspective of developing new oil products with well-balanced techno-, senso- and bio-functional characteristics.
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Aceites de Plantas , Verduras , Aceites de Plantas/química , Manipulación de Alimentos , Oxidación-Reducción , Estrés OxidativoRESUMEN
Essential oils (EOs) find application as flavoring agents in the food industry and are also desirable ingredients as they possess preservative properties. The Mediterranean diet involves the use of a lot of herbs and spices and their products (infusions, EOs) as condiments and for the preservation of foods. Application of EOs has the advantage of homogeneous dispersion in comparison with dry leaf use in small pieces or powder. Among them, Laurus nobilis (bay laurel) L. EO is an interesting source of volatiles, such as 1,8-cineole and eugenol, which are known for their preservative properties. Its flavor suits cooked red meat, poultry, and fish, as well as vegetarian dishes, according to Mediterranean recipes. The review is focused on its chemistry, quality control aspects, and recent trends in methods of analysis and activity assessment with a focus on potential antioxidant activity and applications to olive industry products. Findings indicate that this EO is not extensively studied in comparison with those from other Mediterranean plants, such as oregano EO. More work is needed to establish authenticity and activity methods, whereas the interest for using it for the preparation of flavored olive oil or for the aromatization and preservation of table oils must be further encouraged.
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Wheat is rich in non-starch polysaccharides (NSP) and their degradation in poultry diets is promoted by exogenous carbohydrases. The objective here was to evaluate the effect of adding an intrinsically thermostable xylanase on wheat-based diets for laying hens in yolk color, carotenoid and fatty acid profiles of eggs. A total of 128 laying hens were used for 12 weeks. They were randomly allocated to four dietary treatments with different levels of xylanase: T1: control (no xylanase), T2: 30,000 U/g, T3: 45,000 U/g and T4: 90,000 U/g, with 32 birds, 16 replicates per treatment (2 birds/replicate). At the end of the experimental period, egg yolk color index, redness (a*) and yellowness (b*) of egg yolks were found significantly higher in all the enzyme supplemented diet groups (T2, T3, T4) compared with the control (T1). Canthaxanthin levels were significantly higher in T3 than T1 (p < 0.05). Total n-3, n-6 and total polyunsaturated fatty acids (FAs) were significantly higher in T4 compared with the control (p < 0.01), while the reverse trend was evidenced for monounsaturated FAs. Additionally, total n-3 FAs were higher in the T2 than T1 (p < 0.005). Overall, the results showed that exogenous xylanase enzyme supplementation in wheat-based diets for laying hens contribute to maintaining egg yolk color. Overall, exogenous xylanase enzyme supplemented at all levels in wheat-based laying hens' diets improved egg yolk color compared to the control diet. The enzyme supplemented at the higher level (90,000 U/g) improved polyunsaturated and reduced monounsaturated egg yolk fatty acid content.
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The pan-European distributed Research Infrastructure for Promoting Metrology in Food and Nutrition (METROFOOD-RI) has evolved in the frame of the European Strategy Forum on Research Infrastructures (ESFRI) to promote high-quality metrology services across the food chain. The METROFOOD-RI comprises physical facilities and electronic facilities. The former includes Reference Material plants and analytical laboratories (the 'Metro' side) and also experimental fields/farms, processing/storage plants and kitchen-labs (the 'Food' side). The RI is currently prepared to apply for receiving the European Research Infrastructure Consortium (ERIC) legal status and is organised to fulfil the requirements for operation at the national, European Union (EU) and international level. In this view, the METROFOOD-RI partners have recently reviewed the scientific plan and elaborated strategic priorities on key thematic areas of research in the food and nutrition domain to which they have expertise to contribute to meet global societal challenges and face unexpected emergencies. The present review summarises the methodology and main outcomes of the research study that helped to identify the key thematic areas from a metrological standpoint, to articulate critical and emerging issues and demands and to structure how the integrated facilities of the RI can operate in the first five years of operation as ERIC.
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In this study, it was investigated whether integration of microwave-heating into the pretreatment step of persimmon juice processing allows the concomitant production of both functional juice and added-value solid residue from the Diospyros Kaki "Jiro" cultivar. In this direction, persimmon pulp was treated under three different microwave-heating conditions (0.7, 4.2, and 8.4 kJ/g) prior to enzymatic maceration and compared to the non-heated material. Irrespective of microwave energy employed, the proposed hybrid treatment was highly efficient in terms of juice yield (70% w/w). The mildest heating conditions resulted in juice and residue that were both of inferior quality. Intensification of the microwave energy reduced the microbial load of the juice up to 2-log without compromising the content in total soluble solids, sugars, and L-ascorbic acid. Under the most drastic conditions, the juice was enriched in gallic acid, polyphenols, and potent DPPHâ scavengers, but its orange color faded and was more acidic. In parallel, the solid juice residue retained pro-vitamin A carotenoids (~278 µg retinol activity equivalents) and low-methoxy pectin (9 g/100 g DW). Overall, our findings can assist the efforts of the local juice processing industry to utilize persimmon fruits through energy-efficient technologies in a sustainable approach.
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Kinetics of individual crocetin ester degradation in aqueous extracts of saffron upon thermal treatment in the dark has been studied. Special attention has been paid to the comparison between saffron extracts and aqueous solutions of a crocetin ester rich fraction, with a lower stability of the latter observed. The degradation reaction was the same for all crocetin esters whether they were in saffron extracts or whether they were purified, although it was affected by external factors that modified their kinetic and thermodynamic parameters, making some of them less stable than others.
Asunto(s)
Carotenoides/química , Crocus/química , Manipulación de Alimentos/métodos , Calor , Oscuridad , Ésteres/análisis , Cinética , Termodinámica , Vitamina A/análogos & derivadosRESUMEN
The stepwise approach takes advantage of simple, versatile, low-cost screening tools that can be applied to several posts of the saffron trade chain to specifically detect adulteration with carminic acid (CA). This natural dye is of insect origin and should not be present in Kosher and Halal foods such as saffron. For gross adulteration levels (>25.0%, w/w) reaction with diphenylamine-sulfuric acid was found adequate to indicate the presence of extraneous matter but not its identity. FT-IR analysis of the dry material combined with chemometrics served to rapidly sort out samples containing >10.0% CA without any sample pretreatment except grinding. Aqueous extracts prepared according to ISO 3632-2 were then examined by tristimulus colorimetry and derivative UV-Vis spectrometry to detect adulteration down to the level of 2.0% (w/w). Determination of CA down to 0.2%, w/w was achieved by RP-HPLC-DAD using aqueous acetonitrile elution solvent (pH=2.8).
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Carmín/análisis , Crocus/química , Contaminación de Alimentos/análisis , Cromatografía Líquida de Alta Presión/métodos , Colorantes/análisis , Análisis de los Alimentos/métodos , Espectrofotometría Ultravioleta , Espectroscopía Infrarroja por Transformada de Fourier , Flujo de TrabajoRESUMEN
Examination of the crocin bleaching assay performance and in-house validation were focused on probe and test compound characteristics, conditions for peroxyl radical generation, reaction monitoring, and expression of results. HPLC and spectrometric examination showed that any authentic commercial saffron (origin, grade) can be used for probe preparation given that (a) interferences, such as tocopherols, are removed, (b) working solution concentration is adequately adjusted, and (c) stock probe solution changes during storage are not neglected. As suggested by log P values, calculated for a great number of radical scavengers (AHs), any AH more polar than Trolox (common reference compound) can be tested in the aqueous environment of the assay. AH activities order obeyed principles of structure-activity relationships. The assay was robust toward preheating of the azo-initiator (2,2'-azobis(2-aminopropane) dihydrochloride). Reaction monitoring through periodic UV-vis spectra recording was very informative. An alternative expression of results as "percent inhibition of crocin bleaching value", % Inh = [(DeltaA(0) - DeltaA)/DeltaA(0))] x 100, is proposed for [AH]/[crocin] = 1, instead of the so far used k(rel) values. The above findings also lead to analysis cost and time reduction.
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Carotenoides/química , Depuradores de Radicales Libres/análisis , Fenómenos Químicos , Química Física , Cromanos , Cromatografía Líquida de Alta Presión , Crocus/química , Cinética , Peróxidos/química , Reproducibilidad de los Resultados , EspectrofotometríaRESUMEN
The applicability of the crocin bleaching assay (CBA) to structure-activity relationship (SAR) studies of a great number (n = 39) of selected phenolic compounds was thoroughly investigated. The focus was on the activity of hydroxybenzoic, hydroxyphenylacetic, hydroxyphenylpropanoic, and hydroxycinnamic acids. Other assays [oxygen radical absorbance capacity (ORAC), lipid oxidation] were applied when necessary. Hydroxybenzoic acids were less active than the respective simple phenols. The position of the -COOH group relative to hydroxyl substituents was critical. The number and position of the -OH groups governed the order and size of activity within the subgroup of these acids. Gallic acid was the most active, being 1.6- and 3.4-fold superior to protocatechuic and syringic acids, respectively. The effect of proximity of the -COOH group to the phenyl ring was more distinct for 3,4-guaiacol acids (ferulic >> dihydroferulic congruent with homovanillic > vanillic) than for 3,4-catechol ones (caffeic >> protocatechuic > or = dihydrocaffeic congruent with homoprotocatechuic). Compounds such as vanillin, tyrosol, ferulic acid derivatives, rosmarinic acid, and quercetin were examined to reinforce discussion on the basis of physical organic chemistry principles. Taking into account the acidity of most compounds, the CBA-derived order of activity was meaningful.
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Carotenoides/química , Depuradores de Radicales Libres/química , Fenoles/química , Fenómenos Químicos , Química Física , Ácidos Cumáricos/química , Ácido Gálico/química , Hidroxibenzoatos/química , Hidroxilación , Fenilacetatos/química , Relación Estructura-ActividadRESUMEN
The scavenging behavior of a series of catechol and guaiacol acid derivatives toward DPPH(*) was examined having as a starting point the order of activity derived on the basis of theoretically calculated BDE values. The studied compounds were protocatechuic, homoprotocatechuic, dihydrocaffeic, and caffeic acids and also vanillic, homovanillic, dihydroferulic, and ferulic acids. Catechol and guaiacol were used as reference compounds. Observations from the parallel study were made with regard to structural features (number and position of hydroxyl groups and the side-carbon chain characteristics) that regulated the behavior of the compounds experimentally. The exceptional DPPH(*) scavenging behavior observed for homoprotocatechuic acid in ethanol and for caffeic acid in acetonitrile could not be supported by the respective BDE values. Ferulic was the most active among guaiacolic acids, whereas dihydroferulic exhibited the highest stoichiometry. Ionizable carboxylic groups seem to affect considerably the relative order of activity as was also evidenced using the ORAC assay. Questions raised about the validity of widely accepted views on criteria for SARs are discussed with regard to literature findings.
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Catecoles/química , Depuradores de Radicales Libres/química , Guayacol/química , Picratos , Ácido 3,4-Dihidroxifenilacético/química , Compuestos de Bifenilo , Ácidos Cafeicos/química , Ácidos Cumáricos/química , Relación Estructura-Actividad , TermodinámicaRESUMEN
Near-infrared reflectance spectroscopy has been applied for the first time to saffron spice to determine the chemical composition and geographical origin of 111 samples from the there main producers' countries: Iran, Greece, and Spain. The validation procedures with the results obtained by UV-vis and HPLC-DAD measurements demonstrated that this technique is appropriate to determine the following parameters: moisture and volatile content, coloring strength, (250 nm), and (330 nm), established on the ISO 3632 Technical Specification Normative and used to certify saffron quality in the international market. Also, it can be used to estimate the content of the five main crocetin glycosides, the compounds responsible for saffron color, the best correlations being for trans-crocetin di-(beta-D-gentibiosyl) ester (R2= 0.93), trans-crocetin (beta-D-glucosyl)-(beta-D-gentibiosyl) (R2= 0.94), and picrocrocin (R2= 0.92), the compound accepted as responsible for saffron bitterness. Finally, a discriminant analysis among the three geographical origins reveals that Iranian samples are the most different, whereas Greek and Spanish samples are more similar. All of these results reveal that NIRS spectroscopy has an enormous potential for its application to saffron quality control as the results are obtained in 2 min and without any sample manipulation.
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Crocus/química , Espectroscopía Infrarroja Corta , Carotenoides/análisis , Cromatografía Líquida de Alta Presión , Glicósidos/análisis , Grecia , Irán , Control de Calidad , España , Vitamina A/análogos & derivadosRESUMEN
The Crocin Bleaching Assay (CBA) appears in literature as an in vitro method for measuring antioxidant and prooxidant capacity of model dietary antioxidants, food formulations, pharmaceuticals, and biological samples. The assay is based on simple competitive reactions between a colored probe, crocin, and the test compounds/constituents for scavenging peroxyl radicals generated after thermolysis of a water-soluble azo-initiator. So far, several researchers in the fields of food chemistry, nutrition and clinical biochemistry have sporadically addressed critical views about advantages, limitations and potential field of CBA application. This chapter presents step-by-step critical aspects of CBA in order to assist standardization of its performance. Detailed procedures for calculation of two attributes of peroxyl radical scavenging reactions, the relative rate constant and "total antioxidant capacity", are also presented.
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Antioxidantes/análisis , Bioensayo/métodos , Blanqueadores/química , Carotenoides/química , Oxidantes/metabolismo , Cromatografía Líquida de Alta Presión , Crocus/química , Concentración 50 Inhibidora , Cinética , Modelos Lineales , Peróxidos/química , Extractos Vegetales/química , Polvos , Estándares de Referencia , Suero/metabolismo , Soluciones , Espectrofotometría UltravioletaRESUMEN
The present study aimed to extend application of the FT-MIR technique to the quality control of traded saffron that suffers various types of fraud or mislabelling. Spectroscopic data were obtained for samples stored for different periods in the dark. Samples with the highest quality according to ISO 3632 specifications produced a typical spectrum profile (reference set). Principal Component Analysis (PCA) of spectroscopic data for this set along with HPLC-DAD analysis of major apocarotenoids assisted identification of FT-IR bands that carry information about desirable sensory properties that weaken during storage. The band at 1028cm(-1), associated with the presence of glucose moieties, along with intensities in the region 1175-1157cm(-1), linked with breakage of glycosidic bonds, were the most useful for diagnostic monitoring of storage effects on the evaluation and test set samples. FT-IR was found to be a promising, sensitive and rapid tool in the fight against saffron fraud.
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Crocus/química , Flores/química , Contaminación de Alimentos/análisis , Espectroscopía Infrarroja por Transformada de Fourier/métodos , Almacenamiento de Alimentos , Análisis de Componente Principal , Control de CalidadRESUMEN
An on-line HPLC (high performance liquid chromatography) method for the rapid screening of individual antioxidants in mixtures was developed using crocin as a substrate (i.e. oxidation probe) and 2,2'-azobis(2-amidinopropane dihydrochloride (AAPH)) in phosphate buffer (pH 7.5) as a radical generator. The polyene structure of crocin and AAPH-derived peroxyl radicals resemble the lipidic substrates and radicals found in true food more closely than the popular, albeit artificial, DPPH (1,1-diphenyl-2-picrylhydrazyl) and ABTS(+) (2,2-azinobis-(3-ethylbenzothiazoline-6-sulfonate)) do. After separation by a C18 (octadecyl silica) column and UV (ultraviolet) detection, antioxidative analytes react with peroxyl radicals at 90°C and the inhibition of crocin oxidation (i.e. bleaching) is detected as a positive peak by an absorbance detector at 440 nm. The method is simple, uses standard instruments and inexpensive reagents. It can be applied for isocratic HPLC runs using mobile phases containing 10-90% organic solvent in water, weak acids or buffers (pH 3.5-8.5). With baseline correction, gradient runs are also feasible. The radical scavenging activity of several natural antioxidants and a green tea extract was studied. After optimisation of conditions such as reagent concentrations and flows, the limit of detection varied from 0.79 to 7.4 ng, depending on the antioxidant.
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Antioxidantes/análisis , Carotenoides/química , Cromatografía Líquida de Alta Presión/métodos , Cinética , Límite de Detección , Espectrofotometría UltravioletaRESUMEN
Data for antiradical properties of saffron extract and its bioactive constituents (crocins, crocetin) are limited and poorly discussed in comparison with those of extracts containing potent scavengers. Further examination was sought using the Folin-Ciocalteu (F-C) reagent and various free radical species produced in cell-free or cell model systems. Oregano and turmeric methanol extracts, rich in well established scavengers, and also crocetin, rosmarinic acid, and curcumin, representing the major types of constituents in the three studied extracts, were used as "reference". On the same weight basis, saffron extract activity was found to be rather negligible in all cell-free systems with regard to that found for reference ones. On the contrary, in the human monocyte system, saffron extracts or free crocetin were found to reduce ROS production as effectively as the phenolic antioxidants. Our findings point out that saffron extracts exhibit a remarkable intracellular antioxidant activity that cannot be revealed using assays repeatedly applied to the evaluation of phenolic-type antioxidants.