Levels of synthetic antioxidants (ethoxyquin, butylated hydroxytoluene and butylated hydroxyanisole) in fish feed and commercially farmed fish

Several synthetic antioxidants are authorized for use as feed additives in the European Union. Ethoxyquin (EQ) and butylated hydroxytoluene (BHT) are generally added to fish meal and fish oil, respectively, to limit lipid oxidation. The study was conducted to examine the concentrations of EQ, BHT and butylated hydroxyanisole (BHA) in several commercially important species of farmed fish, namely Atlantic salmon, halibut and cod and rainbow trout, as well as concentrations in fish feed. The highest levels of BHT, EQ and BHA were found in farmed Atlantic salmon fillets, and were 7.60, 0.17 and 0.07 mg kg(-1), respectively. The lowest concentrations of the synthetic antioxidants found were in cod. The concentration of the oxidation product ethoxyquin dimer (EQDM) was more than ten-fold higher than the concentration of parent EQ in Atlantic salmon halibut and rainbow trout, whereas this dimer was not detected in cod fillets. The theoretical consumer exposure to the synthetic antioxidants EQ, BHA and BHT from the consumption of farmed fish was calculated. The contribution of EQ from a single portion (300 g) of skinned fillets of the different species of farmed fish would contribute at most 15% of the acceptable daily intake (ADI) for a 60 kg adult. The consumption of farmed fish would not contribute measurably to the intake of BHA; however, a 300 g portion of farmed Atlantic salmon would contribute up to 75% of the ADI for BHT.     

Nitrate in leafy vegetables,culinary herbs,and cucumber grown under cover in Estonia: content and intake

The content of nitrate in leafy vegetables, culinary herbs, and cucumber was determined during the years 2006–2008. All samples of Estonian origin, except white cabbage, were grown under cover. Seasonal differences in nitrate concentrations were observed in lettuce and spinach. Nitrate concentrations in lettuce were 22% and those in spinach were 24% higher in winter crops compared with samples collected in summer. The mean nitrate level was 3023 mg kg^–1 for fresh lettuce and 2337 mg kg^–1 for spinach. On average, 11.6% of fresh lettuce and spinach samples nitrate concentration exceeded the maximum level specified in European Commission Regulation No. 1881/2006. The mean levels were 999 mg kg^–1 for imported iceberg lettuce and 1287 mg kg^–1 for frozen spinach, which are below the maximum European Commission limits. Parsley, dill, basil, thyme, and rucola contained high concentrations of nitrate from mean levels of 2134 mg kg^–1 for parsley up to 8150 mg kg^–1 for rucola. Mean nitrate concentrations ranged from 382 to 1115 mg kg^–1 for white cabbage and Chinese cabbage, respectively. The per capita mean daily intake of nitrates related to the consumption of leafy vegetables, culinary herbs, and cucumber for the whole Estonian population was 31.3 mg day^–1, which comprised 14.2% of the acceptable daily intake (ADI).     

Effects of oral exposure of pigs to deoxynivalenol (DON) sulfonate (DONS) as the non-toxic derivative of DON on tissue residues of DON and de-epoxy-DON and on DONS blood levels

The Fusarium toxin deoxynivalenol (DON) is of outstanding importance in pig nutrition because of its frequent occurrence in cereal grains at levels high enough to cause adverse effects such as a decrease in feed intake and impairment of the immune system. Thus, simple decontamination procedures would be useful. The present study aimed to examine the effects of wet preservation of triticale contaminated with DON and zearalenone (ZON) with sodium metabisulphite (SBS) on the treatment-related non-toxic derivative of DON (DON-sulfonate, DONS), and on ZON and its metabolites in blood and various physiological specimens of piglets. The uncontaminated control triticale (CON) and the DON-contaminated triticale (FUS) were included in the diets either untreated or SBS treated (CON-SBS, FUS-SBS) and fed to piglets for 28 days starting from weaning. The diet concentrations for DON were 0.156, 0.084, 2.312 and 0.275 mg kg^?1, for DONS were <0.05, <0.05, <0.05 and 1.841 mg kg^?1, and for ZON were <0.001, 0.006, 0.017, and 0.016 mg kg^?1 for each of CON, CON-SBS, FUS and FUS-SBS, respectively. DONS was present in the blood of piglets fed the FUS-SBS at a median concentration of 15.5 ng ml^?1 (3–67 ng ml^?1), while the median DON concentration amounted to 2 ng ml^?1 (0–5 ng ml^?1) at the same time. The median DON concentration in the blood of piglets fed the FUS diet reached a median concentration of 10.5 ng ml^?1 (5–17 ng ml^?1). Moreover, the relative differences between the DON concentrations in other physiological specimens (muscle, liver, kidney, bile and urine) in piglets fed the FUS-SBS and the FUS diet were comparable with the blood DON concentration differences. Although these differences can be taken as an indication for DONS stability after absorption and distribution further studies examining DONS in these other physiological specimens directly are necessary to substantiate this conclusion. Moreover, ZON and α-zearalenol could only be detected in bile and urine where their levels were not influenced by the SBS treatment.     

Determination of polycyclic aromatic hydrocarbons (PAHs) in commonly consumed Nigerian smoked/grilled fish and meat

Smoking and/or grilling, when carried out with traditional methods involving direct contact with wood combustion fumes, is responsible for high contamination levels with carcinogenic polycyclic aromatic hydrocarbons (PAHs). The aim of this work was to investigate the PAH content of different smoked or grilled meat and fish products commonly consumed in Nigeria. A rapid method involving microwave-assisted saponification and simultaneous extraction followed by solid-phase extraction (SPE), high-performance liquid chromatography (HPLC) separation and spectrofluorometric detection was employed. Samples that were smoked or grilled using traditional systems, which use a wood fire, were heavily contaminated with benzo[a]pyrene (BaP) at levels ranging from 2.4 to 31.2 µg kg^?1 wet weight. Considerably lower contamination levels were found in samples smoked or grilled in the laboratory using a charcoal fire (BaP from 0.7 to 2.8 µg kg^?1 wet weight). The health risk associated with a daily consumption of 100 g of these products was also evaluated using the margin of exposure (MOE) approach. MOE values lower than 10,000 were obtained for all smoked/grilled commercial samples, indicating a potential concern for consumer health.     

Residues of antibacterial drugs in honey from the Italian market

Antibacterial drugs are used worldwide for the control of American and, less often, European foulbrood. Their administration is mostly uncontrolled and applied without approved protocols and instructions for use as well as precautionary recommendations. Consequently, this practice is responsible for the contamination of beehive products and contributes to the problem of food safety. According to this situation, 4672 analyses were carried out on 5303 honeys collected from 2001 to 2007. These samples were investigated for antibacterial residues of tetracyclines, sulphonamides, streptomycin, chloramphenicol and tylosin. Honeys were classified according to their origin: imported honey and honey from the Italian market. In the last group (only for samples collected from 2001 to 2004), another type of honey was distinguished: that of local honey. A total of 6.3% of all samples were positive for the antibacterial drugs analysed; in particular, 6.8% of imported honeys and 6.1% of honeys on the Italian market. Only 1.7% of local honey had antibacterial residues. These results are indicative of a rather frequent presence of antibacterial drug residues in both Italian and imported honeys. Furthermore, the data showed that among the active substances analysed, sulphonamides are the most used antibacterial substance followed by tetracyclines, streptomycin, tylosin, and chloramphenicol. Finally, a continuous monitoring programme is needed, accompanied by an education programme to beekeepers on proper hive management.     

Estimated daily intakes of butylated hydroxyanisole (BHA), butylated hydroxytoluene (BHT) and tert-butyl hydroquinone (TBHQ) antioxidants in Korea.

The study was conducted to establish the estimated daily intake (EDI) of antioxidants such as butylated hydroxyanisole (BHA), butylated hydroxytoluene (BHT) and tert-butyl hydroquinone (TBHQ) in Korea. The EDIs were obtained from two sources. One of the estimations was based on the analytical determination of BHA, BHT and TBHQ in 12 food categories (ten food categories for TBHQ) and on individual dietary intake data obtained from the National Health and Nutrition Survey in 1998 (n=11 525, age > 1 year). The other EDIs of BHA, BHT and TBHQ were based on the maximum permitted levels specified in national food standards in Korea and on individual dietary intake data obtained from the National Health and Nutrition Survey in 1998 (n=11 525, age > 1 year). To establish the EDIs based on the analytical determination and on individual dietary intake data, 133 food samples in 12 food categories were selected from the foods considered to be representative sources of BHA, BHT and TBHQ in the Korean diet. Selected samples were analysed by GC with FID. BHA was not detected in any of the samples analysed. BHT and TBHQ were detected in the samples, but the levels were significantly lower than their maximum limits. The EDIs1 of BHT, and TBHQ for average consumers were 0.0156(-3), and 0.0012(-3) mg kg(-1) body weight bw day(-1) and as a proportion of the ADI were 0.0052 and 0.0002%, respectively. For 95th percentile consumers, the EDIs of BHT and TBHQ were 0.0080 and 0.0006 mg kg(-1) bw day(-1), and as a proportion of the ADI were 2.67 and 0.09%, respectively. EDIs for BHA, BHT and TBHQ based on the maximum permitted levels and on individual dietary intake data were 0.04, 0.04 and 0.04 mg kg(-1) bw day(-1), respectively. The EDIs of BHA, BHT and TBHQ for average consumers ranged from 6.00 to 14.42% of the ADI of each antioxidant. According to these results, the EDIs of BHA, BHT and TBHQ in Korea were significantly lower than ADI of these antioxidants established by the JECFA.     

Comparison of extraction solvents and conditions for herbicide residues in milled rice with liquid chromatography-diode array detection analysis (LC-DAD)

Different extraction procedures and clean-up methods were compared in order to develop a sample preparation procedure for the multi-residue analysis of six post-emergence herbicides (metsulfuron methyl, bensulfuron methyl, pyrazosulfuron ethyl, bentazone, bispyribac sodium and cyhalofop butyl) in rice grains followed by liquid chromatography-diode array detection (LC-DAD). Optimum results were obtained dispersing milled rice grain in water, followed by the addition of 1% acetic acid in acetonitrile, MgSO₄ and sodium acetate as a modification of the quick, easy, cheap, effective, rugged and safe (QuEChERS) method but no primary and secondary amine (PSA) sorbent was added due to the acidic nature of the herbicides. The method was further expanded to other post-emergence herbicides (quinclorac, clomazone and propanil). Except for quinclorac, which cannot be analysed with this method, the recoveries of the other eight herbicides were in the range 73–111%, with relative standard deviations lower than 12%. Limits of detection (LODs) ranged from 0.03 to 0.08 mg kg^?1. A single analyst can extract twelve samples in 4 h. The method presented here allows the simultaneous residue determination of the most common post-emergence herbicides employed in cultivating rice. It is simple, rapid, sensitive, and can be applied routinely to polished rice grain herbicide residue analysis.     

Monitoring programme on nitrates in vegetables and vegetable-based baby foods marketed in the Region of Valencia,Spain: levels and estimated daily intake

This study was carried out to determine the current levels of nitrates in vegetables and vegetable-based baby foods (a total of 1150 samples) marketed in the Region of Valencia, Spain, over the period 2000–2008, and to estimate the toxicological risk associated with their intake. Average (median) levels of nitrate in lettuce, iceberg-type lettuce and spinach (1156, 798 and 1410 mg kg^?1 w/w, respectively) were lower than the maximum limits established by European Union legislation. Thirteen fresh spinach samples exceeded the regulatory limits. Median nitrate values in other vegetables for which a maximum limit has not been fixed by the European Commission were 196, 203, 1597, 96, 4474 and 2572 mg kg^?1 w/w (for potato, carrot, chard, artichoke, rucola and lamb's lettuce, respectively). The estimated nitrate daily intakes through vegetables consumption for adult, extreme consumers and children were found to be about 29%, 79.8% and 15.1%, respectively, of the acceptable daily intake (3.7 mg kg^?1). The levels (median = 60.4 mg kg^?1 w/w) found in vegetable-based baby foods were, in all cases, lower the maximum level proposed by European Union legislation. The estimated nitrate daily intake through baby foods for infants between 0–1 and 1–2 years of age were 13% and 18%, respectively, of the acceptable daily intake.     

Survey of aflatoxins in retail samples of whole and ground black and white peppercorns

A total of 126 local and imported samples of commercial white and black pepper in Malaysia were analysed for aflatoxins B₁, B₂, G₁ and G₂ (AFB1, AFB2, AFG1, AFG2) content using high-performance liquid chromatography (HPLC) with a fluorescence detector (FD). An acetonitrile–methanol–water (17 : 29 : 54; v/v) mixture was used as a mobile phase and clean-up was using an immunoaffinity column (IAC). Seventy out of 126 (55.5%) samples were contaminated with total aflatoxins, although only low levels of aflatoxins were found ranging from 0.1 to 4.9 ng g^?1. Aflatoxin B₁ showed the highest incidence of contamination and was found in all contaminated samples. There was a significant difference between type of samples and different brands (p < 0.05). The results showed black peppers were more contaminated than white peppers.     

Monitoring of ractopamine concentration in the mixture of this feed additive with vitamin mineral complex and with swine feed by HPLC

Ractopamine (RAC) analysis at all stages in the feed chain until its final mixing into swine feed is necessary to ensure the safety of all meat consumers and to decrease waste and the cost of supplementation of feed. Two suitable HPLC methods were developed and validated for RAC determination in vitamin mineral complex (VMC) and in swine feed. Both methods employed reverse-phase (C18 column at 40°C) and isocratic elution, but with some modifications to the methods. Validation parameters, such as selectivity, linearity, precision, trueness and robustness, were shown to be within the acceptable range. Therefore, the developed methods can be successfully applied for the monitoring of RAC concentrations in samples of VMC and swine feed ensuring economy to producers and security to consumers of swine meat.     

Assessment of the dietary habits and polycyclic aromatic hydrocarbon exposure in primary school children

Thirty Italian children, 7–9 year aged, living in Naples were investigated on their dietary habits and on polycyclic aromatic hydrocarbon (PAH) exposure by a food diary-questionnaire and one week duplicate diet sample analyses. Daily total food consumption mean value was 632 ± 215 g day^?1, median value 613 g day^?1. The daily energy intake and the diet composition meanly agreed with the official guidelines for the Italian children. Sixteen PAHs were simultaneously detected and, according to the European Food Safety Authority (EFSA) approach, benzo[a]pyrene; benzo[a]pyrene + chrysene (PAH2); PAH2 + benz[a]anthracene + benzo[b]fluoranthene (PAH4); PAH4 + benzo[k]fluoranthene + benzo[ghi]perylene + dibenz[a, h]anthracene + indeno[1,2,3-cd]pyrene (PAH8) were considered in evaluating the children's dietary exposure to PAHs. The benzo[a]pyrene (BaP) median concentrations in foods varied from 0.06 to 0.33 µg kg^?1. Only three samples of cooked foods (one fish and two meat samples) exceeded legal limits fixed by the European Union for BaP. Daily median intakes of benzo[a]pyrene, PAH2, PAH4, and PAH8 were 153; 318; 990; 1776 ng day^?1; their median exposure values were 5; 10; 28; 54 ng kg^?1 bw day^?1. The Margins of Exposure (MOEs) in median consumers agreed with the EFSA safety values except for PAH8.     

Formation of fumonisins and other secondary metabolites by Fusarium oxysporum and F. proliferatum: a comparative study

The principal aim of this study was to estimate the formation of fumonisins (FB₁ and FB₂), moniliformin (MON), and ergosterol (ERG) by Fusarium oxysporum and Fusarium proliferatum, while the formation of beauvericin (BEA) was estimated by the latter Fusarium species only. Moreover, the effect of temperature on the biosynthesis of mycotoxins was also evaluated. Fumonisins were formed by F. proliferatum, with the highest yield at 18°C (720.0–1976.6 µg g^?1 for FB₁, 74.2–670.8 µg g^?1 for FB₂) and only by three of four F. oxysporum strains at a very low level (0.02–4.77 µg g^?1 for FB₁, 0.02–2.15 µg g^?1 for FB₂). The amount of MON formed by F. proliferatum was the highest (p < 0.001) at 32°C (3056.87 µg g^?1), while MON biosynthesis by F. oxysporum was lower 227.54 µg g^?1 (p < 0.001). BEA was produced by F. proliferatum with the highest level at 25°C (p < 0.001). ERG–recognized as an indicator of fungal biomass development and as a consequence of mycotoxin formation–was found at the highest concentration at a biosynthesis temperature of 25°C for F. proliferatum and F. oxysporum (p < 0.001).     

12‐Lipoxygenase activity in the muscle tissue of Atlantic mackerel (Scomber scombrus) and its prevention by antioxidants

Lipoxygenase was prepared from Atlantic mackerel muscle using differential centrifugation, ammonium sulphate precipitation and gel permeation (phenyl Sephadex G‐50) column chromatography. The crude lipoxygenase enzyme preparation was characterised by sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS‐PAGE), which showed two prominent bands with molecular weights of 119 and 125 kDa. Fractions collected from the chromatography column were tested for enzyme activity by reacting with arachidonic acid and determining the production of hydroxyeicosatetraenoic acid (12‐HETE) using reverse phase HPLC and GC–MS. The 12‐HETE peak was absent from the fresh arachidonic acid control sample and from arachidonic acid treated with heat‐inactivated lipoxygenase. Esculetin, a known inhibitor of lipoxygenase, inhibited the production of 12‐HETE from the reaction of lipoxygenase with arachidonic acid, thus confirming that the enzyme was lipoxygenase. The HETE peak was partially reduced in the presence of antioxidants, namely synthetic butylated hydroxytoluene (BHT) and natural antioxidants vitamins C and E. The presence of lipoxygenase in Atlantic mackerel muscle indicates the possibility that the lipid oxidation mechanism is initiated enzymatically in chilled and frozen stored fillets of mackerel and that this oxidative deterioration could be inhibited by antioxidants (BHT, vitamins C and E) which are used widely in the food industry. © 2001 Society of Chemical Industry     

高效液相色谱法测定牛奶中林可霉素的残留量

研究建立牛奶中林可霉素残留量的高效液相色谱测定方法。样品用水相提取,离心分离,经C18固相萃取柱净化,在210nm波长下用二极管矩阵检测器检测。色谱柱为Agilent ZORBOX SBC18柱(4.6×250mm,5μm),以0.05mol/L硼砂溶液(用磷酸调节pH6.0)-甲醇(55:45,V/V)为流动相,流速1.0ml/min。林可霉素浓度在1.0～10.0μg/g范围内与峰面积呈良好的线性关系,平均回收率为82.9%～93.1%,变异系数为1.45%～4.58%,最低检测限为0.5μg/ml。本方法简便、快速、准确。     

Usage pattern and exposure assessment of food colours in different age groups of consumers in the State of Uttar Pradesh,India

The present study aims to investigate the nature and levels of colours in food items and to undertake risk assessment vis-à-vis intake among different age groups of consumers in the State of Uttar Pradesh, India. A total of 478 edible foodstuffs were analysed, and of six permitted colours, Sunset Yellow FCF (SSYFCF) and Tartrazine were most popular, and two non-permitted colours, namely Metanil Yellow and Rhodamine B, were encountered. The study showed a marked improvement in the trend of use of non-permitted colours over previous surveys, with 90% foods now resorting to approved food colours. However, 59% of foods employing permitted colours exceeded the maximum allowable limit, with average quantities crossing the threshold of 100 mg kg^?1 in most food commodities. The intake of SSYFCF exceeded the acceptable daily intake (ADI) for children and adolescents by 88% and 39%, respectively, and was statistically significant when analysed by error bars and distribution curves. In adults, SSYFCF saturated 59% of the ADI. For Carmoisine, Tartrazine and Ponceau 4R, saturation of ADI ranged from 27.4% to 90.3% in children and adolescents and from 10.8% to 47.6% in adult subjects. These results indicate that children and adolescents are more vulnerable to higher intakes of food colours compared with the adult population. Allowing a uniform level of all colours in foods under Indian rules, notwithstanding wide variations of 250-fold in their allocated ADIs, could be one reason for the higher intake and hence only technological need-based levels of individual colours are desired to be prescribed.     

Characterization,antigenicity and detection of fish gelatine and isinglass used as processing aids in wines

Fish gelatine (FG) and isinglass (IG) are widely used in the pharmaceutical industry and as ingredients or processing aids in food production. Both products are the focus of interest since several countries, particularly the member states of the EU and Japan, USA, Australia and New Zealand, have introduced special labelling regulations for allergenic foodstuffs, such as fish and products thereof. Thus, there is a demand for a reliable and sensitive method for the detection of FG and IG in foodstuffs. In this study, the characterization of various FGs and IGs, polyclonal antibodies raised against fish collagen and the development of a sensitive indirect ELISA for the detection of FG and IG is described. The ELISA method detected ≤0.11 µg ml^?1 from all FGs and IGs tested. The indirect ELISA was applied to various experimental wines where FG and IG had been used as processing aids and to several commercial wines. No residues of FG and IG were detected in the experimental wines additionally treated with bentonite, a strong adsorbent for proteins, and successive filtration. Additionally, no residues were found in the commercial wines. However, amounts of up to 0.33 µg ml^?1 were found in some experimental wines not treated with bentonite and successive filtration. Therefore, wines may contain traces of FG and IG that were used as processing aids during wine production. However, treatment with bentonite in combination with additional filtration had a clear impact on these residues.     

高效液相色谱法测定鸡蛋、牛奶和猪肉中环丙氨嗪及三聚氰胺的实验研究

本实验研究建立用NH2柱和97%乙腈水溶液做流动相洗脱测定鸡蛋、猪肉和牛奶中环丙氨嗪和三聚氰胺残留量的高效液相色谱法,样品均质后经NaOH和20%氨水乙腈溶液重复提取2次、浓缩、过C18固相萃取小柱净化等处理后上机检测,环丙氨嗪和三聚氰胺得到了很好的分离,各自的保留时间分别是8min和12min。标准曲线在0.01～1.0μg/ml浓度范围内呈线性相关,环丙氨嗪、三聚氰胺标准曲线的相关系数(r)分别为0.9999、0.9994。检测限为0.02mg/kg,定量限低于0.05mg/kg。     

Occurrence of ochratoxin A and citrinin in Czech cereals and comparison of two HPLC methods for ochratoxin A detection

The aims of the study were to obtain information about the occurrence of ochratoxin A (OTA) and citrinin (CIT) in cereals harvested in the Czech Republic and to compare two analytical procedures for detecting OTA. A total of 34 cereal samples, including two matrix reference materials (R-Biopharm, Germany), were analysed. The results were compared with the limit for raw cereal grains used as a foodstuff according to Commission Regulation No. 1881/2006, which allows a maximum OTA level of 5 µg kg^?1. Compared were two methods based on the high-performance liquid chromatography principle, one using the immunoaffinity columns OchraTest? (VICAM) and the second based on solvent partition (PART), both followed by fluorescence detection. The highest OTA contents were found in two barley samples. According to the method employed, the results for the first sample (malting barley) were VICAM = 31.43 µg kg^?1 and PART = 44.74 µg kg^?1. For the second sample (feeding barley) they were VICAM = 48.63 µg kg^?1 and PART = 34.40 µg kg^?1. Two samples of bread wheat had an OTA content approaching the legal limit (VICAM = 4.71 µg kg^?1 and PART = 6.03 µg kg^?1; VICAM = 4.12 µg kg^?1 and PART = 3.95 µg kg^?1). CIT was analysed using the PART method only, and its highest content (93.64 µg kg^?1) was found for the malting barley sample with high OTA content (44.74 µg kg^?1 as analysed using PART).     

An investigation into the occurrence of the toxic amino acid gizzerosine in fish meal

A method is presented whereby the toxic amino acid gizzerosine can be determined in fish meals to a lower limit of 10 mg kg^?1. Fish meal was hydrolysed with hydrochloric acid and, after concentration by freeze drying, derivatised with dansyl chloride solution without further clean-up. A fraction containing the tridansyl derivative of gizzerosine was separated and collected by HPLC and from this the didansyl derivative was separated after treatment with formic acid on the same column as previously using the same mobile phase. Seventeen fish meals, some causing severe gizzard erosion in chickens, were analysed but in none of them could gizzerosine at or above this level be found.