Validation of a confirmatory analytical method for the determination of aflatoxins B₁, B₂, G₁ and G₂ in foods and feed materials by HPLC with on-line photochemical derivatization and fluorescence detection

A sensitive and selective analytical method for the simultaneous separation and quantitative determination of aflatoxins B1, B2, G1 and G2 in foodstuffs and materials for feed has been validated. The method is based on high performance liquid chromatography with on-line post-column photochemical derivatization and fluorescence detection. The chromatographic separation of aflatoxins was accomplished using a C18 column eluted with an isocratic mobile phase consisting of water, methanol and acetonitrile. The sample preparation required a simple extraction of aflatoxins with MeOH/H2O (80:20, v/v) and a purification step by immunoaffinity column cleanup. The total analysis time, including sample preparation and chromatographic separation, did not exceed 40 min with a run time of 10 min. The on-line photochemical derivatization ensures better results in terms of simplicity, sensitivity and reproducibility with respect to chemical derivatization techniques, and provides an increase of the peak resolution and an extent of automation in comparison with the electrochemical ones. The procedure for the determination of aflatoxins in food samples and cereals for animal consumption was extensively validated following Regulation (EC) No. 882/2004. Detection limits in wheat bran samples of 0.08 μg kg1 for AFB1, 0.02 μg kg1 for AFB2, 0.16 μg kg1 for AFG1 and 0.04 μg kg1 for AFG2 were attained. The method allows high recovery with mean values ranging from 72 to 94% and it satisfies the necessary requirements for sensitivity, linearity, selectivity, precision and ruggedness, demonstrating the conformity of the method with provisions of Regulation (EC) No. 401/2006.     

Assessing the reactivity of free chlorine constituents Cl₂, Cl₂O, and HOCl toward aromatic ethers

Cl(2) and Cl(2)O are highly reactive electrophiles capable of influencing rates of disinfection byproduct (DBP) precursor chlorination in solutions of free available chlorine (FAC). The current work examines how organic compound structure influences susceptibility toward chlorination by Cl(2) and Cl(2)O relative to the more abundant (but less reactive) electrophile HOCl. Chlorination rates and products were determined for three aromatic ethers, whose reactivities with FAC increased in the order: 3-methylanisole <1,3-dimethoxybenzene <1,3,5-trimethoxybenzene. Varying solution conditions (pH, [FAC], [Cl(-)]) permitted quantification of regiospecific second-order rate constants for formation of each product by Cl(2), Cl(2)O, and HOCl. Our results indicate that as the reactivity of methoxybenzenes decreases, the importance of Cl(2) and Cl(2)O (relative to HOCl) increases. Accordingly, Cl(2) and Cl(2)O are likely to play important roles in generating DBPs that originate from natural organic matter (NOM) constituents of somewhat moderate reactivity. As [Cl(2)] is proportional to [Cl(-)] and [Cl(2)O] is proportional to [HOCl](2), ramifications for DBP control measures may differ significantly for these precursors compared to more reactive NOM moieties likely to react predominantly with HOCl. In particular, the role of chloride as a chlorination catalyst challenges its traditional classification as an "inert" electrolyte in water treatment processes.     

Comparison of methods and optimisation of the analysis of fumonisins B₁ and B₂ in masa flour, an alkaline cooked corn product

A comparison study of different extraction and clean-up procedures for the liquid chromatographic analysis of fumonisins B(1) (FB(1)) and B(2) (FB(2)) in corn masa flour was performed. The procedures included extraction (heat or room temperature) with acidic conditions or EDTA-containing solvents, and clean-up by immunoaffinity or C18 solid-phase extraction columns. Thereafter an analytical method was optimised using extraction with an acidic mixture of methanol-acetonitrile-citrate/phosphate buffer, clean-up through the immunoaffinity column and determination of fumonisins by liquid chromatography with automated pre-column derivatisation with o-phthaldialdehyde reagent. Recovery experiments performed on yellow, white and blue masa flours at spiking levels of 400, 800 and 1200 μg kg(-1) FB(1) and of 100, 200 and 300 μg kg(-1) FB(2) gave overall mean recoveries of 99% (±6%) for FB(1) and 88% (±6%) for FB(2). Good recoveries (higher than 90% for both FB(1) and FB(2)) were also obtained with corn tortilla chips. The limits of quantification of the method (signal-to-noise ratio of 10) were 25 μg kg(-1) for FB(1) and 17 μg kg(-1) for FB(2). The method was tested on different commercial corn masa flours as well as on white and yellow corn tortilla chips, showing fumonisin contamination levels (FB(1) + FB(2)) up to 1800 μg kg(-1) (FB(1) + FB(2)) in masa flour and 960 μg kg(-1) in tortilla chips. Over 30% of masa flours originating from Mexico exceeded the European Union maximum permitted level.     

Cheers,proost, saúde: Cultural,contextual and psychological factors of wine and beer consumption in Portugal and in the Netherlands

Wine and beer consumption are an integral part of European culture: Southern Europe is associated with wine and Northern Europe is associated with beer. When consumed in moderation, these alcoholic beverages can be part of a balanced and healthy diet. In the 1990s, non-alcoholic beer (NAB), which has no cultural roots, became available in the market. This review identifies determinants for consumption of wine, beer, and NAB, using data on consumption patterns from Portugal and the Netherlands. Since the 1960s the image of Portugal as a wine country declined, whereas the image of the Netherlands as a beer country remained stable. In each country beer is now the most consumed alcoholic beverage and is mainly a men's beverage, whereas wine is the second most consumed and is consumed by both genders. Cultural differences define Portuguese as “outdoors, everyday drinkers”, within a meal context, and Dutch as “at home, weekend drinkers.” Wine is perceived as the healthiest beverage, followed by NAB, and regular beer. Motivation for consumption is related to context: wine for special occasions, beer for informal occasions, and NAB for occasions when alcohol is not convenient. Moderate wine and beer consumption seems to be surrounded by positive emotions.

This review is relevant for public health, for industry market strategies, and identifies opportunities of future research on drinking behaviour.     

Guaraná, a supplement rich in caffeine and catechin,modulates cytokines: evidence from human in vitro and in vivo protocols

Guaraná powder is an antiobesogenic supplement; however, its effect on inflammatory biomarkers has not yet been determined. Therefore, this study analysed whether guaraná supplementation can differentially modulate the levels of proinflammatory cytokines [interleukin 6 (IL-6), tumour necrosis factor-alpha, interleukin 1 beta (IL-1β), interferon-gamma (Ig-γ)] and anti-inflammatory interleukin 10 (IL-10) from in vitro and in vivo protocols. In the pilot in vitro protocol, human peripheral blood mononuclear cells were exposed to guaraná, as well as to resveratrol, quercetin and ascorbic acid as positive controls. The effect of guaraná on cytokine levels was also evaluated in culture medium supplemented with glucose and insulin. A randomised, placebo-controlled in vivo assay was also performed to evaluate the potential influence of guaraná on the blood cytokine levels of 14 healthy volunteers supplemented for 14 days. The effect of guaraná was similar to that of resveratrol, a known anti-inflammatory molecule, decreasing IL-1β, IL-10 and Ig-γ levels and increasing IL-10 levels compared to those of the control group. The in vitro insulin supplementation potentiated the effect of guaraná on some cytokines. A decreasing effect on the blood inflammatory cytokine levels, along with an increase in IL-10 levels, was also observed in volunteers supplemented with guaraná. In conclusion, guaraná positively modulates cytokines associated with inflammatory metabolism.     

Deoxynivalenol in wheat,maize, wheat flour and pasta: surveys in Hungary in 2008–2015

Among Fusarium mycotoxins, deoxynivalenol (DON) is the most common contaminant in case of cereals and cereal-based foods in Hungary. In this study, Hungarian wheat (n = 305), maize (n = 108), wheat flour (n = 179) and pasta (n = 226) samples were analysed (N = 818). The samples were collected during 2008–2015 in Hungary. Applied methods of analysis were enzyme-linked immunosorbent assay and liquid-chromatography coupled with a mass spectrometer. Results were compared and evaluated with Hungarian weather data. Among cereal samples, in 2011, wheat was contaminated with DON (overall average ± standard deviation was 2159 ± 2818 µg kg^?1), which was above the maximum limit (ML). In case of wheat flour and pasta, no average values above ML were found during 2008–2015, but higher DON contamination could be observed in 2011 as well (wheat flour: 537 ± 573 µg kg^?1; pasta: 511 ± 175 µg kg^?1).     

化学合成维生素B1,B2,B6,BC和B13研究进展

综述了化学合成盐酸硫胺、核黄素、吡哆醇、叶酸和乳清酸研究进展。     

Determination of Free Arginine, Glutamine, and β-alanine in Nutritional Products and Dietary Supplements

A method for the determination of free arginine, glutamine, and β-alanine in nutritional products and dietary supplements is described. The amino acids are derivatized with the fluorescent tag 9-fluorenylmethoxycarbonyl (FMOC), and the derivatives are then determined by reversed phase HPLC. Method suitability was defined by experimental assessments of linearity (R ² > 0.999), precision (day-to-day RSD ≤ 1.0%), accuracy (spike recoveries=98.7%–101.8%, n = 18), and selectivity (baseline resolution from the other common amino acids). The method provides for an accurate and precise quantification of the three amino acids, when present at concentrations >0.2% (w/w) in nutritional products and dietary supplements. Assessments of free l-glutamine stability in three different reconstituted powder products, as performed by the method, found recoveries >97% through 24 h at room temperature.     

Associations between immune competence,stress responsiveness,and production in Holstein-Friesian and Holstein-Friesian × Jersey heifers reared in a pasture-based production system in Australia

The objectives of this study were to assess antibody and cellular immune responses in first-lactation dairy heifers reared under a pasture-based production system and to investigate associations between immune competence and stress responsiveness, health, and productivity. A commercial vaccine was used to induce antibody and cellular immune responses and, based on measured responses, animals were classified as above average, average, and below average for each trait independently and in combination (overall immune competence). Overall immune competence phenotypic rankings were generated by combining standardized residual values for antibody and cellular responses measured in individual heifers. Cortisol responses to handling and yarding were used to assess stress responsiveness, worm egg counts were used to assess internal parasite burden, somatic cell counts to assess udder health and estimates of total daily milk volume, and milk fat and milk protein contents to assess productivity. A total of 393 Holstein-Friesian and Holstein-Friesian × Jersey crossbred heifers from 2 herds practicing seasonal calving in a pasture-fed production system were enrolled in the study. The immune competence phenotypes of individual heifers were assessed before their first mating and their subsequent performance during their first lactation was monitored. Animals with below-average antibody and cellular immune competence had higher serum cortisol concentrations compared with their counterparts following handling, suggesting they had a reduced ability to cope with management-induced stress. Furthermore, a significant negative (favorable) correlation was observed between antibody responses and stress responsiveness. Similarly, correlations between antibody responses and internal parasite burden were significant and negative (favorable). No correlations were observed between overall immune competence and milk yield, milk fat, and milk protein content.     

Effects of ascorbic acid, rosemary, and Origanox™ in preventing bone marrow discoloration in beef lumbar vertebrae in aerobic and anaerobic packaging systems

Treatments of 1.25% or 2.5% ascorbic acid (AA); 0.1% or 0.2% rosemary; or a combination of 0.15% Origanox™ + 0.3% AA on beef lumbar vertebrae (n = 72) were evaluated for their effectiveness in preventing bone discoloration in retail packages. Vertebrae were held either 6 or 14 d postmortem; cut into 2.54-cm sections; and packaged into either polyvinyl chloride film overwrap (PVC), high-oxygen (80% O₂/20% CO₂) modified-atmosphere packages (MAP), or ultra-low-oxygen (70% N₂/30% CO₂) MAP. Controls and vertebrae treated with 0.1% or 0.2% rosemary discolored significantly, whereas 0.15% Origanox™ + 0.3% AA was effective through d 2 of display in PVC and high-oxygen MAP. In both high- and ultra-low-oxygen MAP, 1.25% AA was as effective as 2.5% AA in preventing bone discoloration. Vertebrae treated with AA or packaged in ultra-low-oxygen MAP had the largest a* values over display time. Bone discoloration was effectively prevented with AA, especially at the 2.5% concentration.     

Short communication: Relationships among plasma and milk vitamin B12, plasma free fatty acids,and blood β-hydroxybutyrate concentrations in early lactation dairy cows

This study was undertaken to evaluate the relationship between plasma and milk concentrations of vitamin B₁₂ as well as the relationship between plasma or milk concentrations of vitamin B₁₂ and plasma concentration of free fatty acids (FFA) or blood concentration of β-hydroxybutyrate (BHB) of early lactating Ayrshire (AY) and Holstein (HO) cows. A total of 44 dairy herds (7 AY and 37 HO herds) and 62 AY (21 in first, 19 in second, and 22 in third and more lactations) and 228 HO (51 in first, 74 in second, and 103 in third and more lactation) cows between 3 and 40 d in milk were involved in the study. Hand-stripped milk samples and blood samples were taken 6 h after the morning milking. Milk and plasma samples were analyzed for vitamin B₁₂ concentration and plasma samples were analyzed for FFA concentration. A handheld device was used for blood BHB concentration determination. Thresholds for elevated plasma FFA concentration and hyperketonemia were set at ≥0.70 and ≥1.2 mmol/L, respectively. Vitamin B₁₂ concentration in milk of AY primiparous cows [2,557 (1,995–3,276) pg/mL] was lower than in milk from HO primiparous cows [3,876 (3,356–4,478) pg/mL], whereas no difference was observed among other parities and breeds. Regardless of breeds, plasma concentration of vitamin B₁₂ of first and second parities was lower than plasma concentration of third and more lactation cows. Milk vitamin B₁₂ concentration was positively correlated with plasma vitamin B₁₂ concentration, but the relationship was stronger for AY (ρ averaging 0.63) than for HO cows (ρ averaging 0.36). For AY and HO breeds, a significant relationship between milk or plasma vitamin B₁₂ concentrations and plasma FFA concentration (ρ between 0.29 and 0.59) was observed. Moreover, cows with elevated plasma FFA concentration had greater milk and plasma vitamin B₁₂ concentrations than cows with normal plasma FFA concentration. No relationship between vitamin B₁₂ concentration in milk or plasma and blood BHB concentration and hyperketonemia was noted. In summary, milk is not well correlated with plasma vitamin B₁₂ concentration for HO. It could be hypothesized that elevated plasma concentration of FFA could have a negative effect on the use of vitamin B₁₂ by cow cells, which increases the concentration of the vitamin in plasma and its secretion in milk.     

Production, reproduction, health, and growth traits in backcross Holstein × Jersey cows and their Holstein contemporaries

A total of 648 purebred Holstein and 319 backcross Holstein × Jersey dairy cattle were compared for production, reproduction, health, linear type, and growth traits. Animals were born between 2003 and 2009 and were housed in the University of Wisconsin–Madison Integrated Dairy Facility. All animals had Holstein dams; lactating dams were mated to unproven Holstein sires to produce purebred (control) Holsteins or to unproven F₁ Jersey × Holstein crossbred sires to produce backcross animals, whereas nulliparous dams were mated to proven Holstein sires to produce purebred (other) Holsteins. Traits were analyzed using mixed linear models with effects of season of birth, age of dam, sire, birth year of sire, days in milk, lactation, and linear type score evaluator. Control Holsteins had greater 305-d milk yield (12,645 vs. 11,456 kg), 305-d mature equivalent milk yield (13,420 vs. 12,180 kg), peak daily milk yield (49.5 vs. 46.4 kg), total lactation milk yield (11,556 vs. 10,796 kg), and daily fat-corrected milk yield (43 vs. 40 kg) compared with backcrosses. Days open and services per conception as a heifer or cow did not differ between control Holsteins, other Holsteins, or backcrosses. The proportion of first-parity births that required assistance was less in control Holsteins than in backcross cows (3.7 vs. 11.2%). The incidence of scours or respiratory problems in calves did not differ between control Holsteins, other Holsteins, and backcrosses, nor did the incidence of mastitis, injury, or feet problems. Control Holstein heifers were heavier (629 vs. 557 kg), with greater hip height (145 vs. 139 cm), body length (167 vs. 163 cm), heart girth (205 vs. 198 cm), and hip width (54 vs. 53 cm) at 22 mo of age. On a 50-point scale for linear type traits, Holsteins were larger in stature compared with backcrosses (41 vs. 28), had wider rumps (37 vs. 33), and wider rear udders (34 vs. 32). Results of this study suggest that backcross Holstein × Jersey cattle have decreased production but fail to demonstrate an advantage in health and reproduction compared with purebred Holsteins.     

Oxidative stress,NF-κB signaling,NLRP3 inflammasome,and caspase apoptotic pathways are activated in mammary gland of ketotic Holstein cows

Ketosis is a serious metabolic disorder characterized by systemic and hepatic oxidative stress, inflammation, and apoptosis, as well as reduced milk yield. Because of the paucity of data on mammary responses during ketosis, the aim of this study was to evaluate alterations in oxidative stress, NF-κB signaling, NLRP3 inflammasome, and caspase apoptotic pathways in mammary gland of dairy cows with ketosis. Blood, mammary gland tissue, and milk samples were collected from healthy cows [Control, blood concentration of β-hydroxybutyrate (BHB) <0.6 mM, n = 10] and cows with subclinical ketosis (SCK, blood concentration of BHB >1.2 mM and <3 mM, n = 10) or clinical ketosis (CK, blood concentration of BHB >3 mM, n = 10) at median 8 d in milk (range = 6–12). Compared with Control, serum concentration of glucose was lower (3.91 vs. 2.86 or 2.12 mM) in cows with SCK or CK, whereas concentrations of fatty acids (0.25 vs. 0.57 or 1.09 mM) and BHB (0.42 vs. 1.81 or 3.85 mM) were greater. Compared with Control, the percentage of milk fat was greater in cows with SCK or CK. In contrast, the percentage of milk protein was lower in cows with SCK or CK. We detected no differences in milk lactose content across groups. Compared with Control, activities of glutathione peroxidase, superoxide dismutase, and catalase were lower in mammary gland tissue of cows with SCK or CK. In contrast, concentrations of hydrogen peroxide and malondialdehyde were greater in cows with SCK or CK. Compared with Control, mRNA abundances of TNFA, IL6, and IL1B were greater in mammary tissues of cows with SCK or CK. In addition, activity of IKKβ and the ratio of phosphorylated inhibitor of κBα to IκBα, and of phosphorylated NF-κB p65 to NF-κB p65, were also greater in mammary tissues of cows with SCK or CK. Subclinical or clinical ketosis also led to greater activity of caspase 1 and protein abundance of caspase 1, NLRP3, Bax, caspase 3, and caspase 9. In contrast, abundance of the antiapoptotic protein was lower in SCK or CK cows. The data indicate that the mammary gland of SKC or CK cows undergoes severe oxidative stress, inflammation, and cell death.     

Method development and survey of Sudan I–IV in palm oil and chilli spices in the Washington,DC, area

Sudan I, II, III and IV dyes are banned for use as food colorants in the United States and European Union because they are toxic and carcinogenic. These dyes have been illegally used as food additives in products such as chilli spices and palm oil to enhance their red colour. From 2003 to 2005, the European Union made a series of decisions requiring chilli spices and palm oil imported to the European Union to contain analytical reports declaring them free of Sudan I–IV. In order for the USFDA to investigate the adulteration of palm oil and chilli spices with unapproved colour additives in the United States, a method was developed for the extraction and analysis of Sudan dyes in palm oil, and previous methods were validated for Sudan dyes in chilli spices. Both LC-DAD and LC-MS/MS methods were examined for their limitations and effectiveness in identifying adulterated samples. Method validation was performed for both chilli spices and palm oil by spiking samples known to be free of Sudan dyes at concentrations close to the limit of detection. Reproducibility, matrix effects, and selectivity of the method were also investigated. Additionally, for the first time a survey of palm oil and chilli spices was performed in the United States, specifically in the Washington, DC, area. Illegal dyes, primarily Sudan IV, were detected in palm oil at concentrations from 150 to 24 000 ng ml^–1. Low concentrations (< 21 µg kg^–1) of Sudan dyes were found in 11 out of 57 spices and are most likely a result of cross-contamination during preparation and storage and not intentional adulteration.     

Aflatoxin B1 in eggs and chicken livers by dispersive liquid–liquid microextraction and HPLC

A rapid, low-cost and simple technique has been developed for the determination of aflatoxin B₁ (AFB₁) in eggs and livers using high-performance liquid chromatography (HPLC) with UV detection. In this study, the presence of AFB₁ was investigated in 150 eggs and 50 chicken livers from the local market of Tabriz, Iran. AFB₁ was extracted with a mixture of acetonitrile:water (80:20) and cleaned up by dispersive liquid–liquid microextraction which is a very economical, fast and sensitive method. AFB₁ was quantified by HPLC-UV without need for any complex derivatisation in samples to enhance the detection. The results showed that 72% of the liver and 58% of the egg samples were contaminated with AFB₁ ranging from 0.30 to 16.36 µg kg ^?1. limit of detection and limit of quantification for AFB₁ were 0.08 and 0.28 µg kg ^{? 1}, respectively. The proposed method is suitable for fast analysing of AFB₁ in egg and liver samples.