Technological, sensory, nutritional and bioactive potential of pan breads produced with refined and whole grain buckwheat flours.

The nutritional quality and bioactive potential of breads made with partial replacement of refined wheat flour (RWF) with 30% or 45% refined buckwheat flour (RBF) or whole buckwheat flour (WGBF) was assessed through mineral bioaccessibility, starch digestibility, dietary fiber content and bioactive potential by determining rutin and quercetin levels during processing. Moreover, technological quality and sensory acceptance were also evaluated. Breads made with 30% or 45% WGBF showed higher mineral and fiber contents compared to the control, while the formulations with RBF showed higher bioaccessibility. No changes were observed in the rutin levels of the dough before and after fermentation, but after baking, rutin and quercetin levels increased. The highest starch hydrolysis was found in the formulation containing 45% RBF. The formulations made with 30% RBF or 30% WGBF were well accepted by consumers. Our study shows interesting results, as few studies report the effect of processing on bioactive compounds.

Multivariate optimization results in an edible extract from Ilex paraguariensis unexplored residues with a high amount of phenolic compounds.

Ilex paraguariensis A. St. Hil. plants are used for the preparation of food and drinks which are widely consumed worldwide. During the harvest season of these plants, 2-5 ton hec-1 of agricultural residue is generated, which remains underutilized. Therefore, this study aimed to obtain an edible extract with high content of bioactive compounds and antimicrobial properties from the agricultural residue of I. paraguariensis for industrial use in food applications. The extraction conditions were optimized through a multivariate experimental design using ethanol:water. The extracted compounds were characterized by HPLC-ESY-QTOF-MS. In the optimal extraction conditions, 55 compounds were extracted, including 8 compounds that were not previously reported in I. paraguariensis. The method proved to be simple, fast, economical and environmentally friendly, with the use of green solvents. This optimization allowed for the extraction of 15.07 g of phenolic compounds per 100 g of residue. The extract showed high antioxidant activity and the capacity to inhibit Staphylococcus aureus. Results indicate that it is possible to obtain an edible extract with a high content of bioactive compounds, particularly phenolic compounds, from the I. paraguariensis residue, which has high prospects for the valorization of unexplored natural resources.

Herbs and herbal infusions: Determination of natural contaminants (mycotoxins and trace elements) and evaluation of their exposure.

Herbal infusions are amongst the world's most popular and widely enjoyed beverages, due to both large variety and convenience. However, natural contaminants, such as mycotoxins and trace elements can accumulate in aromatic herbs, which may have serious food safety and public health implications. In this study, the presence of mycotoxins, as well as the content of trace elements was evaluated in herbs and herbal infusions commercialized in Brazil. For the determination of fourteen mycotoxins, including the emerging mycotoxins enniatins (EN), beauvericin (BEA), and sterigmatocystin (STE), a liquid-chromatography tandem mass spectrometry (LC-MS/MS) method was validated. Overall, 42 out of 58 herb samples (72%) were contaminated, being BEA the most usual mycotoxin, present in 43% of the samples, followed by STE and HT-2 toxin, present in 37% and 24% of the samples, respectively. In herbal infusions, the occurrence of mycotoxins was 88% lesser than those verified in raw products. Despite these low levels, the hazard quotient (HQ) calculated revealed a potential health concern for HT-2 in infusions. The margin of exposure values for aflatoxins (AF), and ochratoxin A (OTA) from six herbal infusions were below 10,000, indicating also potential health risks. The twenty-one trace elements comprising toxic elements such as arsenic (As), cadmium (Cd), and lead (Pb) were determined in herb raw materials by inductively coupled plasma-mass spectrometry (ICP-MS). The levels of trace elements in herbs were very varied, with aluminum (Al) presenting the highest amount. The levels of legislated elements (As, Cd, Pb) analyzed in herbs were lower than 3.03 µg g-1 (Pb), thus not exceeding the legal limits defined for herbal medicinal by both European and Brazilian pharmacopeia.

In vitro digestion of spermidine and amino acids in fresh and processed Agaricus bisporus mushroom.

Mushrooms are significant sources of amino acids and bioactive amines; however, their bioaccessibility can be affected by processing and during in vitro digestion. Fresh Agaricus bisporus mushroom was submitted to cooking and canning and samples were submitted to in vitro gastric and gastric-intestinal digestions. An UHPLC method was used for the simultaneous determination of 18 free amino acids, 10 biogenic amines and ammonia in the samples. Fresh mushroom contained 14 free amino acids, with predominance of alanine and glutamic acid; spermidine was the only amine detected; and ammonia was also detected. Spermidine levels were not affected by cooking, but there was a significant loss (14%) after canning. Spermidine levels were not affected by the in vitro gastric and intestinal digestion, suggesting full bioaccessibility. There was a significant decrease on total amino acids levels after cooking and canning, with higher losses for aspartic and glutamic acids in cooked and for aspartic acid and valine in canned mushrooms. After the in vitro gastric and intestinal digestions, the total levels of amino acids increased and two additional amino acids (arginine and methionine) were detected. During in vitro digestion many of the amino acids were released mainly in the intestinal phase. After in vitro digestion, amino acids per gram of protein of mushrooms are adequate for most FAO amino acid pattern for adults. Multivariate analysis confirmed that protein hydrolysis in processed mushrooms is higher in intestinal phase. Bioaccessibility data for spermidine in A. bisporus is a novelty and increase the value of this food.

Yellow sweet potato flour: use in sweet bread processing to increase carotene content and improve quality

ABSTRACT Yellow sweet potato is mostly produced by small farmers, and may be a source of energy and carotenoids in the human diet, but it is a highly perishable crop. To increase its industrial application, yellow sweet potato flour has been produced for use in bakery products. This study aimed to evaluate the technological quality and the carotenoids content in sweet breads produced with the replacement of wheat flour by 0, 3, 6, and 9% yellow sweet potato flour. Breads were characterized by technological parameters and β-carotene levels during nine days of storage. Tukey's test (p<0.05) was used for comparison between means. The increase in yellow sweet potato flour concentrations in bread led to a decrease of specific volume and firmness, and an increase in water activity, moisture, orange coloring, and carotenoids. During storage, the most significant changes were observed after the fifth day, with a decrease in intensity of the orange color. The β-carotene content was 0.1656 to 0.4715 µg/g in breads with yellow sweet potato flour. This work showed a novel use of yellow sweet potato in breads, which brings benefits to consumers' health and for the agricultural business.

Yellow sweet potato flour: use in sweet bread processing to increase ß-carotene content and improve quality.

Yellow sweet potato is mostly produced by small farmers, and may be a source of energy and carotenoids in the human diet, but it is a highly perishable crop. To increase its industrial application, yellow sweet potato flour has been produced for use in bakery products. This study aimed to evaluate the technological quality and the carotenoids content in sweet breads produced with the replacement of wheat flour by 0, 3, 6, and 9% yellow sweet potato flour. Breads were characterized by technological parameters and ß-carotene levels during nine days of storage. Tukey's test (p<0.05) was used for comparison between means. The increase in yellow sweet potato flour concentrations in bread led to a decrease of specific volume and firmness, and an increase in water activity, moisture, orange coloring, and carotenoids. During storage, the most significant changes were observed after the fifth day, with a decrease in intensity of the orange color. The ß-carotene content was 0.1656 to 0.4715 µg/g in breads with yellow sweet potato flour. This work showed a novel use of yellow sweet potato in breads, which brings benefits to consumers' health and for the agricultural business.

Brazilian Capsicum peppers: capsaicinoid content and antioxidant activity.

BACKGROUND: Capsicum peppers are known as a source of capsaicinoids, phenolic compounds and antioxidants. Brazilian Capsicum peppers are important spices used in foods worldwide. However, little information is available on the chemical composition and antioxidant activity of these peppers. RESULTS: Capsaicin, dihydrocapsaicin, total phenolic compounds and antioxidant activity were investigated in extracts of three Brazilian peppers: Capsicum frutescens, C. chinense and C. baccatum var. pendulum, in two different harvest years and at two ripening stages. The bioactive compound content was dependent on harvest year, and changes in the concentration profiles were found for capsaicin. Mature fruits of C. chinense harvested in the first year had the highest capsaicin concentration (2.04 mg g-1 fresh pepper), and mature fruits of C. frutescens harvested in the same first year had the highest dihydrocapsaicin content (0.95 mg g-1 fresh pepper). Mature fruits of C. frutescens harvested in the first year showed the major total phenolic compound content (2.46 mg g-1 fresh pepper). The total phenolic compound content was directly related to antioxidant activity. CONCLUSION: Our results suggest that phenolic compounds significantly contribute to the antioxidant activity of the investigated peppers. Also, these data add valued novel information that enhances current knowledge of Brazilian pepper fruits. © 2017 Society of Chemical Industry.

Multiclass pesticide analysis in fruit-based baby food: A comparative study of sample preparation techniques previous to gas chromatography-mass spectrometry.

With the aim to develop a new gas chromatography-mass spectrometry method to analyze 24 pesticide residues in baby foods at the level imposed by established regulation two simple, rapid and environmental-friendly sample preparation techniques based on QuEChERS (quick, easy, cheap, effective, robust and safe) were compared - QuEChERS with dispersive liquid-liquid microextraction (DLLME) and QuEChERS with dispersive solid-phase extraction (d-SPE). Both sample preparation techniques achieved suitable performance criteria, including selectivity, linearity, acceptable recovery (70-120%) and precision (⩽20%). A higher enrichment factor was observed for DLLME and consequently better limits of detection and quantification were obtained. Nevertheless, d-SPE provided a more effective removal of matrix co-extractives from extracts than DLLME, which contributed to lower matrix effects. Twenty-two commercial fruit-based baby food samples were analyzed by the developed method, being procymidone detected in one sample at a level above the legal limit established by EU.

Neurotoxicity of Dietary Supplements from Annonaceae Species.

Dietary supplements containing plant materials of Annonaceae species (Annona muricata L., A. squamosa L., A. mucosa JACQ., A. squamosa × cherimola Mabb.) were extracted by hot, pressurized ethyl acetate and analyzed for their effect in vitro on Lund human mesencephalic neurons. Cell viability was measured by the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay, and cell death was determined by lactate dehydrogenase levels. Three supplements strongly decreased the cell viability at extract concentrations of 1 µg/mL, of which 1 decreased cell viability at 0.1 µg/µL. Also, strong neuronal toxicities of these supplements were found. Cell death was observed at concentrations of 10 µg/mL. The degree of toxicity was comparable to the ones found in Annonaceous fruit extracts. Two fruit pulps of Annonaceae (A. muricata and A. squamosa) showed a reduction in cell viability at lower concentrations. The fruit pulp extract of A. muricata revealed the strongest neurotoxic effect, with 67% cell death at a concentration of 1 µg/mL. A high reduction in cell viability coupled with pronounced cell death was found at 0.1 µg/mL for an Annonaceous seed extract. These results demonstrate that the intake of dietary supplements containing plant material from Annonaceae may be hazardous to health in terms of neurotoxicity.

Use of multivariate statistical techniques to optimize the separation of 17 capsinoids by ultra performance liquid chromatography using different columns.

In this work a multivariate statistical tool (Derringer and Suich optimization) was proposed for the separation of seventeen capsinoids (natural and synthetic) using the UHPLC-DAD chromatography. Capsinoids were analyzed at 280 nm. The variables optimized were the mobile phase (water (0.1% acetic acid as solvent A) and acetonitrile (0.1% as solvent B)), gradient time and flow rate. Two columns with different length (50 and 100 mm) were used for the chromatographic separation. The two columns used properly separated the seventeen capsinoids, however the 100 mm column length showed a better chromatographic separation with a shorter run time and smaller peak widths. These results provided better values of limit of detection and quantification for the 100 mm column length. The better conditions of separation with the 100 mm column length were established with: initial mobile phase with 41.8% of solvent B; 3.96 min of linear gradient time to reach 100% of solvent B; flow rate of 0.679 mL min(-1). A validation of the method has been done with excellent values of repeatability (RSD<1.92) and intermediate precision (RSD<3.92). The developed method has been applied to real samples. Capsiate has been identified and quantified in some varieties of peppers.

Comprehensive characterization of lipids from Amazonian vegetable oils by mass spectrometry techniques.

An integrative approach in mass spectrometry (MS) comprising gas chromatography coupled to MS (GC-MS), ultra-efficiency liquid chromatography coupled to MS (UPLC-MS) and easy ambient sonic-spray ionization MS (EASI-MS) is proposed for the comprehensive characterization of Amazonian oils. Coconut, andiroba and castor seed oils, which are vastly sold in markets of the Amazonian region of Brazil, were selected as a representative test set. These oils were found to contain several lipids such as triacylglycerides (TAGs), fatty acids (FAs), phytosterols and limonoids. In the analyzed samples 30 different TAGs, 11 FAs, 6 phytosterols and 7 limonoids were identified. The antioxidant capacity (AOC) of the oils, as measured by their oxygen radical absorbance capacity (ORAC), was also used to evaluate their potential biological properties as well as their possible consumption as food. Edible virgin coconut oil was the most active (0.720±0.001 Trolox eq./mmol), whereas considerable lower activity was observed for andiroba and castor seed oils. The antimicrobial activities of the oils were also recorded against a panel of pathogenic bacteria and fungi in which andiroba oil was the only one that was active, solely against Enterococcus aeruginosa.

Application of Kohonen neural network for evaluation of the contamination of Brazilian breast milk with polychlorinated biphenyls.

Due to the tendency of polychlorinated biphenyls (PCB) to accumulate in matrixes with high lipid content, the contamination of the breast milk with these compounds is a serious issue, mainly to the newborn. In this study, milk samples were collected from breastfeeding mothers belonging to 4 Brazilian regions (south, southeast, northeast and north). Twelve PCB were analyzed by HS-SPME-GC-ECD and the corresponding peak areas were correlated to the answers to a questionnaire of general habits, breastfeeding and characteristics of the living places. To realize this exploratory analyze, self-organizing maps generated applying Kohonen neural network were applied. It was possible to verify the occurrence of different PCB congeners in the breast milk relating to the region of the Brazil that the breastfeeding lives, the proximity to an industry, the proximity to a contaminated river or sea, the type of milk (colostrum, foremilk and hindmilk) and the number of past pregnancies.

Chemometrics optimization of carbohydrate separations in six food matrices by micellar electrokinetic chromatography with anionic surfactant.

Multivariate statistical design modeling and the Derringer-Suich desirability function analysis were applied to micellar electrokinetic chromatography (MEKC) results with anionic surfactant to separate carbohydrates (CHOs) in different food matrices. This strategy has been studied with success to analyze compounds of difficult separation, but has not been explored for carbohydrates. Six procedures for the analysis of different sets of CHOs present in six food matrices were developed. The effects of pH, electrolyte and surfactant concentrations on the separation of the compounds were investigated using a central composite design requiring 17 experiments. The simultaneous optimization of the responses for separation of six sets of CHOs was performed employing empirical models for prediction of optimal resolution conditions in six matrices, condensed milk, orange juices, rice bran, red wine, roasted and ground coffee and breakfast cereal samples. The results indicate good separation for the samples, with appropriate detectability and selectivity, short analysis time, low reagent cost and little waste generation, demonstrating that the proposed technique is a viable alternative for carbohydrate analysis in foods.

Nutritional status of patients with biliary atresia and autoimmune hepatitis related to serum levels of vitamins A, D and E.

CONTEXT: Chronic liver disease may induce to malabsorption of lipids and fat-soluble vitamins, leading to injury of nutritional status. OBJECTIVES: To evaluate the nutritional status of pediatric-age patients with autoimmune hepatitis and biliary atresia related to serum levels of vitamins A, D and E and the disease severity. METHODS: This controlled transverse study, evaluated the patients with autoimmune hepatitis and biliary atresia and a reference group paired by sex and age. The patients underwent anthropometric evaluation, alimentary inquiry and determination of serum levels of vitamins A, D and E by high performance liquid chromatography. The Mann-Whitney test, Spearman correlation coefficients and variance analysis (ANOVA) were utilized for data treatment, regarding significant difference if P<0.05. RESULTS: The highest nutritional deficit was observed in patients with biliary atresia, mainly with cholestasis. The serum levels of vitamins A and E for the reference group changed as a function of age. The serum levels of vitamins A, D and E were higher in reference group than in patients with biliary atresia and autoimmune hepatitis together or separately. There were not difference in the serum levels of vitamins A, D and E between biliary atresia groups with cholestasis and without cholestasis. It was verified correlation between weight/age, triceps skinfold thickness, subscapular skinfold thickness, midarm circumference, midarm fat area values and vitamin A serum levels, as well as between all anthropometric indicators and vitamin E in patients with autoimmune hepatitis and biliary atresia. CONCLUSION: The patients with biliary atresia and cholestasis presented the highest nutritional injury. The patients with biliary atresia and autoimmune hepatitis presented lower serum levels of vitamins A, D and E that in control group. There is a directly proportional correlation between vitamin serum levels, mainly vitamin E, and all anthropometric variables of biliary atresia and autoimmune hepatitis groups.

Nutritional status of patients with biliary atresia and autoimmune hepatitis related to serum levels of vitamins A, D and E / Estado nutricional de pacientes com atresia biliar e hepatite autoimune e relação com os níveis séricos de vitaminas A, D e E

CONTEXT: Chronic liver disease may induce to malabsorption of lipids and fat-soluble vitamins, leading to injury of nutritional status. OBJECTIVES: To evaluate the nutritional status of pediatric-age patients with autoimmune hepatitis and biliary atresia related to serum levels of vitamins A, D and E and the disease severity. METHODS: This controlled transverse study, evaluated the patients with autoimmune hepatitis and biliary atresia and a reference group paired by sex and age. The patients underwent anthropometric evaluation, alimentary inquiry and determination of serum levels of vitamins A, D and E by high performance liquid chromatography. The Mann-Whitney test, Spearman correlation coefficients and variance analysis (ANOVA) were utilized for data treatment, regarding significant difference if P<0.05. RESULTS: The highest nutritional deficit was observed in patients with biliary atresia, mainly with cholestasis. The serum levels of vitamins A and E for the reference group changed as a function of age. The serum levels of vitamins A, D and E were higher in reference group than in patients with biliary atresia and autoimmune hepatitis together or separately. There were not difference in the serum levels of vitamins A, D and E between biliary atresia groups with cholestasis and without cholestasis. It was verified correlation between weight/age, triceps skinfold thickness, subscapular skinfold thickness, midarm circumference, midarm fat area values and vitamin A serum levels, as well as between all anthropometric indicators and vitamin E in patients with autoimmune hepatitis and biliary atresia. CONCLUSION: The patients with biliary atresia and cholestasis presented the highest nutritional injury. The patients with biliary atresia and autoimmune hepatitis presented lower serum levels of vitamins A, D and E that in control group. There is a directly proportional correlation between vitamin serum levels, mainly vitamin E, and all anthropometric...

CONTEXTO: As doenças hepáticas crônicas podem induzir à má absorção de lipídios e vitaminas lipossolúveis e levar ao comprometimento do estado nutricional. OBJETIVOS: Avaliar o estado nutricional e relacionar com os níveis séricos de vitaminas (A, D e E) e a gravidade da doença em pacientes com atresia biliar e hepatite autoimune na faixa etária pediátrica. MÉTODOS: O estudo foi transversal controlado e foram avaliados os pacientes com hepatite autoimune e atresia biliar e um grupo controle pareado por sexo e idade. Foi realizada avaliação antropométrica, aplicação do inquérito alimentar e determinação dos níveis séricos das vitaminas A, D e E pela técnica de cromatografia líquida de alta eficiência. Foram empregados os testes de Mann-Whitney, o coeficiente de correlação de Spearman e análise de variância (ANOVA), sendo considerada diferença significativa se P<0,05. RESULTADOS: O déficit nutricional mais grave foi observado nos pacientes com atresia biliar, principalmente com colestase. Em relação às vitaminas, no grupo controle, constatou-se que os níveis séricos das vitaminas A e E variaram com a idade. Os níveis séricos das vitaminas A, D e E foram maiores no grupo controle em relação aos pacientes com atresia biliar e hepatite autoimune em conjunto ou separadamente. Verificou-se a correlação do peso/idade, prega cutânea tricipital, prega cutânea subescapular, circunferência braquial, área adiposa braquial com a vitamina A e de todos os indicadores antropométricos com a vitamina E nos pacientes com hepatite autoimune e atresia biliar em conjunto. CONCLUSÕES: Os pacientes com atresia biliar e colestase apresentaram o maior comprometimento nutricional. Os pacientes com atresia biliar e hepatite autoimune possuíram menores níveis séricos das vitaminas A, D e E do que o grupo controle. Existe uma correlação diretamente proporcional, principalmente da vitamina E com todos as variáveis antropométricas do grupo de AB e HAI em conjunto.

Validation of methodology for simultaneous determination of synthetic dyes in alcoholic beverages by capillary electrophoresis.

In this work a method of analysis for synthetic dyes was developed using capillary electrophoresis in alcoholic beverages. The analyses were carried out with fused silica capillary, with 73 cm effective length, at 35 degrees C, buffer phosphate solution of 10 mmol/L with sodium dodecyl sulphate 10 mmol/L, pH 11, and +25 kV of voltage. For dye analyses, three wavelengths in the visible region were used for the qualitative and quantitative determination of the 11 synthetic dyes allowed in Brazil: 450, 525 and 625 nm for the yellow, red and blue dyes, respectively. The detection limits varied from 0.4 to 2.5 microg/mL and the quantification limits varied from 1.3 to 7.1 microg/mL. The average recovery was 92.6 and 104.0% at two levels of concentration. Repeatability for standards and spiked sample showed that the calculated values were greater than the observed values, demonstrating the precision of the method. The proposed and validated method was used to analyze some alcoholic beverage samples, consisting of 12 red wines, 9 coolers, 6 aromatized spirits, 7 bitters, 3 cocktails and 8 liquors from different Brazilian manufacturers. The results showed the coolers, bitters and red wines did not have synthetic dyes, but dyes were found in six of the eight analyzed liquor samples. In all the samples of cocktails and spirits, the presences of dyes were observed. No analyzed sample exceeded the limit established by Brazilian legislation (maximum 30 mg/100 mL).

Characterization of must and wine of six varieties of grapes by direct infusion electrospray ionization mass spectrometry.

Samples of must derived from six different varieties of grapes taken during the fermentation process, as well as the respective wine samples directly after the end of the malolactic fermentation, were analyzed by direct infusion negative ion mode electrospray ionization mass spectrometry (ESI-MS). Diagnostic ions for must were different from those of wine samples, although small variations for each of the grape varieties were also detected. The addition of unfermented must or sugar to wine could also be clearly detected. The spectra were acquired in a few minutes per sample, indicating that ESI-MS can be used for high-throughput analysis of samples and should prove useful for quality control during and after the fermentation process.

Validação de metodologia por cromatografia líquida de alta eficiência (CLAE)para análise de ácido fólico adicionado em farinhas de trigo e milho / Determination of folic acid in enriched wheat and corn flour by means of high performance liquid chromatography

O enriquecimento de alimentos com ácido fólico tem se tornado uma prática comum em todo o mundo. Recentemente, no Brasil, a ANVISA publicou uma resolução (Resolução nº 344) que obriga o enriquecimento de farinha de trigo e milho com essa vitamina. Dessa forma, metodologias analíticas capazes de avaliar ácido fólico nesses alimentos, tornam-se necessárias. O objetivo do trabalho foi a validação de uma metodologia para análise de ácido fólico em farinhas de trigo e milho enriquecidas, utilizando-se a técnica de cromatografia líquida de alta eficiência. O ácido fólico foi extraído das farinhas com solução de hidróxido de potássio e acetonitrila. Seguiu-se a etapa de purificação com ácido tricloroacético. Para a etapa cromatográfica utilizou-se como fase móvel um sistema gradiente composto por solução acidificada/acetonitrila e coluna de C18 para separação e eluição da vitamina. A detecção foi feita a 290nm e a quantificação por padronização externa. A metodologia mostrou-se eficiente com taxas de recuperação entre 92 e 98%, boa repetibilidade (CV 0,60 a 0,98%) e limites de detecção e quantificação de 1,3ng/g e 2,6ng/g, respectivamente

Validação de metodologia para a determinação de ácido fólico em margarina enriquecida e avaliação da estabilidade da vitamina durante estocagem / Validation of methodology for determination of folic acid enriched margarine and evaluation of the stability of the vitamin during stockage

Com a finalidade de amenizar os problemas relacionados à carência de ácido fólico, muitos produtos alimentícios estão sendo submetidos ao processo de enriquecimento. Portanto, tornam-se necessárias metodologias analíticas capazes de avaliar o comportamento dessa vitamina, quando adicionada a esses produtos. O objetivo deste trabalho foi a validação de uma metologia para a determinação de ácido fólico adicionada à margarina, utilizando-se a técnica de cromatografia líquida de alta eficiência. A vitamina foi extraída da matriz com solução KOH, seguida por etapa de limpeza com ácido tricloacético. O ácido fólico foi separado utilizando-se coluna C18 e solução acidificada e acetonitrila. A detecção foi feita a 290 nm e a quantificação por padronização externa. A metodologia mostrou-se eficiente com taxas de recuperação entre 94 e 97%, boa repetibilidade (CV de 0,5 a 1,4) e limites de detecção e quantificação de 1,3ng/g e 2,6ng/g, respectivamente. No estudo de vida-de-prateleira realizado, durante 4 meses de estocagem, resultou em perda de grande parte da vitamina, aproximadamente 55%