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.     

Trimethylamine oxide,dimethylamine, trimethylamine and formaldehyde levels in main traded fish species in Hong Kong

Levels of trimethylamine oxide (TMAO), dimethylamine (DMA), trimethylamine (TMA) and formaldehyde (FA) were studied in 266 different fishes, including fresh/frozen raw whole fishes of 89 different species that traded in Hong Kong, China. Determination of TMAO can confirm the source of DMA and FA if present in the sample. These samples were purchased from different commercial outlets between April and August 2007. All samples of raw whole fish were identified for their species by the Agriculture, Fisheries and Conservation Department. The content of TMAO was determined by high-performance liquid chromatography (HPLC) coupled with a chemiluminescent nitrogen detector. The possible decomposition products of TMAO, DMA and TMA were analysed by headspace solid-phase micro-extraction gas chromatography-mass spectrometry (HS-SPME-GC-MS), while FA was conducted by steam distillation then quantified by a HPLC. The range for TMAO of all samples was <5–3800 mg kg^?1 with median of 970 mg kg^?1, while the endogenous enzymatic cleavage products DMA, TMA and FA were in the range of <2–320, <1–190 and <1–160 mg kg^?1, respectively. These cleavage products were mainly found in three fish species, Harpadon nehereus, Saurida elongata and Saurida tumbil, that belong to the family Synodontidae (Lizardfishes) and subfamily Harpadontinae. Besides, freshwater fish species, namely, Micropterus salmoides, Oreochromis niloticus niloticus and Siniperca chuatsi, were found to contain TMAO in the range of 510–760, 85–720 and 400–640 mg kg^?1, respectively.     

高效液相色谱法同时测定食品中的12种抗氧化剂

目的：建立一种快速、准确测定食品中12 种抗氧化剂的高效液相色谱法。方法：样品用正己烷溶解,用含抗坏血酸棕榈酸盐(AP)的饱和乙腈萃取,以反相C18 柱为分离柱,以甲醇- 乙腈- 乙酸- 水体系为流动相进行梯度洗脱,采用高效液相色谱- 紫外检测器于280nm 定量检测。结果：12 种抗氧化剂在36min 内完全分离,线性范围为0.2～200mg/L(r=0.9981～0.9999),定量限为0.2～1.0mg/kg,回收率为81.13%～107.57%,相对标准偏差为0.26%～4.52%(n=7)。结论：本方法准确、可靠、简便、检出限低,适合分析大批量样品。     

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.     

芹菜提取物中芹菜素的HPLC与HPTLC定量分析研究

通过采用高效液相色谱(HPLC)、紫外光谱和黄酮颜色特征反应对芹菜提取物主要成分芹菜素进行定性鉴定,同时建立定量测量芹菜素的高效薄层色谱(HPTLC)方法.结果表明:芹菜提取物的主要成分是黄酮醇.芹菜素,与芹菜素标准品具有相同的光谱和色谱性质;采用G60高效硅胶板,以氯仿-甲醇-水(18:2.3:0.35)为展开剂,芹菜素标样与试样有相同的Rf值为0.65;用最小二乘法作线性回归,芹菜素含量与斑点峰面积的标准曲线回归方程:y=15138x 7594.8,R2=0.993,线性范围:0.332～0.996μg/斑点.精密度实验RSD(n=5)=1.33%,平均回收率97.82%.     

Trace elements in farmed fish (Cyprinus carpio,Ctenopharyngodon idella and Oncorhynchus mykiss) from Beijing: implication from feed

Concentrations of 30 trace elements, Li, V, Cr, Mn, Fe, Ni, Cu, Mo, Zn, Se, Sr, Co, Al, Ti, As, Cs, Sc, Te, Ba, Ga, Pb, Sn, Cd, Sb, Ag, Tm, TI, Be, Hg and U in major cultured freshwater fish species (common carp-Cyprinus carpio, grass carp-Ctenopharyngodon idella and rainbow trout-Oncorhynchus mykiss) with the corresponding feed from 23 fish farms in Beijing, China, were investigated. The results revealed that Fe, Zn, Cu, Mn, Sr, Se were the major accumulated essential elements and Al, Ti were the major accumulated non-essential elements, while Mo, Co, Ga, Sn, Cd, Sb, Ag, Tm, U, TI, Be, Te, Pb and Hg were hardly detectable. Contents of investigated trace elements were close to or much lower than those in fish from other areas in China. Correlation analysis suggested that the elemental concentrations in those fish species were relatively constant and did not vary much with the fish feed. In comparison with the limits for aquafeeds and fish established by Chinese legislation, Cd in 37.5% of rainbow trout feeds and As in 20% of rainbow trout samples exceeded the maximum limit, assuming that inorganic As accounts for 10% of total As. Further health risk assessment showed that fish consumption would not pose risks to consumers as far as non-essential element contaminants are concerned. However, the carcinogenic risk of As in rainbow trout for the inhabitants in Beijing exceeded the acceptable level of 10^?4, to which more attention should be paid.     

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.     

超高效液相色谱-串联质谱法测定鱼油软胶囊中的合成抗氧化剂

目的建立超高效液相色谱-串联质谱法(ultra performance liquid chromatography-tandem mass spectrometry,UPLC-MS/MS)测定鱼油软胶囊中叔丁基对苯二酚(tert-butylhydroquinone,TBHQ)、2,6-二叔丁基-4-甲基苯酚(butylated hydroxytoluene,BHT)、丁羟基茴香醚(butyl hydroxyanisole,BHA)和没食子酸丙酯(propyl gallate,PG)4种合成抗氧化剂的含量。方法采用乙腈为溶剂和涡旋方式对152批次鱼油软胶囊进行提取;UPLC以乙腈-水为流动相进行梯度洗脱,质谱采用电喷雾离子源负离子模式(electrospray ionization,ESI-)及多反应监测模式(multiple reaction monitoring,MRM)进行测定。结果 PG、TBHQ、BHA和BHT在0.50~20μg/mL浓度范围内具有良好的线性关系(r0.997),在2、8和16 mg/kg3个不同浓度加标水平下测得回收率为88.8%~94.2%,RSD为1.6%~4.2%(n=6),检出限为0.2μg/kg。结论该方法灵敏度高、抗干扰性强且准确性好,可适用于鱼油软胶囊中PG、TBHQ、BHA和BHT合成抗氧剂的检测。     

Formaldehyde in cultivated mushrooms: a negligible risk for the consumer

Following the detection of formaldehyde in cultivated mushrooms, an evaluation was carried out to assess whether its presence in food poses a risk to public health. Formaldehyde, a carcinogenic chemical, has a broad range of industrial applications and, hence, exposure to formaldehyde is ubiquitous through diverse consumer goods, food, the air, etc. The observed levels of formaldehyde in mushrooms are lower than the levels reported for vegetables, fruit, meat, fish and dairy products. On the basis of available data, a rough estimate of the dietary exposure to formaldehyde was performed. The exposure through the consumption of cultivated mushrooms (approximately 0.19 µg kg^?1 body weight day^?1 on average, consumers only) appeared to be small compared with the total dietary intake of formaldehyde (approximately 99.0 µg kg^?1 body weight day^?1, total population). Based on comparison with toxicological safety limits for chronic exposure and given that formaldehyde is carcinogenic only through inhalation and not by ingestion, it can be concluded that the dietary exposure to formaldehyde is not a cause for concern.     

Effect of some habitual cooking processes on the domoic acid concentration in the cockle (Cerastoderma edule) and Manila clam (Ruditapes philippinarum)

The effect of cooking on the concentration and burden of domoic acid in two bivalve molluscs was studied. The Manila clam (Ruditapes philippinarum) and cockle (Cerastoderma edule) were subjected to steaming and boiling, respectively. In both cases, factorial plans were used to evaluate the effects of common cooking methods and the variations likely to take place during the cooking procedure (cooking time and salt concentration in both species, in addition to ethanol percentage in Manila clam). The domoic acid concentration and toxin content were affected by cooking in very different ways in the two species studied. The cockle lost a significant part of its domoic acid content, while the clam did not. Since the weight of the soft tissues in cooked bivalves was lower than in the raw samples in both species, the toxin concentration decreased less than the toxin burden in the cockle, while it increased in the clam, where the toxin burden did not change significantly. Among the cooking variables tested, only cooking time had a noticeable effect on the domoic acid content in the clam and cockle, with the bivalves that were cooked for a longer time having smaller amounts of toxin. It is clear that cooking affects the toxin concentration in bivalves in a way that is species specific. This characteristic must be taken into account when evaluating epidemiological information, establishing allowable toxin levels and in cases where pre-processing treatments such as cooking or similar methods are used in monitoring systems.     

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 patulin occurrence in apple juice and apple products in Catalonia,Spain, and an estimate of dietary intake

This study was conducted to assess patulin exposure in the Catalonian population. Patulin levels were determined in 161 apple juice samples, 77 solid apple-based food samples and 146 apple-based baby food samples obtained from six hypermarkets and supermarkets from twelve main cities of Catalonia, Spain. Patulin was analysed by a well-established validated method involving ethyl acetate extraction and direct analysis by high-performance liquid chromatography (HPLC) with ultraviolet light detection. Mean patulin levels for positive samples in apple juice, solid apple-based food and apple-based baby food were 8.05, 13.54 and 7.12 µg kg^?1, respectively. No samples exceeded the maximum permitted levels established by European Union regulation. Dietary intake was separately assessed for babies, infants and adults through a Food Frequency Questionnaire developed from 1056 individuals from Catalonia. Babies were the main group exposed to patulin, however no risk was detected at these levels of contamination. Adults and infants consumers were far from risk levels. Another approach to determine estimated exposure was conducted through Monte Carlo simulation that distinguishes variability in exposures from uncertainty of distributional parameter estimates.     

Estimate of intake of benzoic acid in the Belgian adult population

An exposure assessment was performed to estimate average daily benzoic acid intake for Belgian adults. Food consumption data were retrieved from the national food-consumption survey. As a first step, individual food-consumption data were multiplied with the maximum permitted use levels for benzoic acid per food group (Tier 2). As a second step, a label survey to identify the foods where benzoic acid is effectively used as an additive and a literature review of the possible occurrence of benzoic acid as a natural substance were performed. With this information, a refined list of foods was drafted for the quantification of benzoic acid, which was performed by a high-performance liquid chromatography (HPLC) method, optimized and validated for this purpose. Individual food-consumption data were then multiplied with the actual average concentrations of benzoic acid per food group (Tier 3). Usual intakes were calculated using the Nusser method. The mean benzoic acid intake was 1.58 mg kg^?1 body weight day^?1 (Tier 2) and 1.25 mg kg^?1 body weight day^?1 (Tier 3). In Tier 2, men exceeded the acceptable daily intake (ADI) of 5 mg kg^?1 body weight day^?1 at the 99th percentile. The greatest contributors to the benzoic acid intake were soft drinks. Benzoic acid as a natural substance represents only a small percentage of the total intake. The results show that actual benzoic acid intake is very likely to be below the ADI. However, there is a need to collect national food-consumption data for children as they might be more vulnerable to an excessive intake.     

Occurrence of patulin and its dietary intake through pear,peach, and apricot juices in Italy

Pb, Cd and Hg in muscles of flathead mullet (Mugil cephalus), collected from Bocca di Magra, La Spezia (Ligurian Sea, Mediterranean Sea, Italy), were determined using graphite furnace atomic absorption spectrometry after microwave digestion for Pb and Cd and direct mercury analyser for Hg. Average Pb concentrations varied in the range 0.20–0.24 mg/kg, whereas Cd and Hg levels were negligible. None of the tested 200 samples exceeded the European regulatory limits as set by EC 1881/2006 and 420/2011. Metal concentrations in fish muscles were assessed for human consumption according to provisional tolerable weekly intake. The estimated values of Pb, Cd and Hg in M. cephalus’s edible parts in this study were below the values established by the Joint WHO/FAO Expert Committee on Food Additives. Therefore, it can be concluded that there is no health problem in human consumption.     

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.     

Effect of the inclusion of adsorbents on aflatoxin B1 quantification in animal feedstuffs

The extraction efficiency of aflatoxin B₁ (AFB₁) in cattle feed containing nine adsorbents (ADSs) was investigated using two organic/aqueous solvents composed of methanol/water (80/20 v/v; MeOH) and acetone/water (85/15 v/v; AC). Samples were obtained including a highly AFB₁-contaminated (HC) and a low-level AFB₁-contaminated (LC) feedstuff (15.33 and 7.57 µg kg^–1, respectively), nine ADSs (four clay minerals; one yeast cell wall-based product; one activated carbon and three commercial ADS products) at two different levels of inclusion (10 and 20 g kg^?1). After solvent extraction and immunoaffinity column clean-up, all samples were analysed for AFB₁ by high-performance liquid chromatography (HPLC) with fluorescence detection. For each contamination level (HC and LC), the data obtained were analysed using a factorial arrangement in a completely randomized design. Means were compared with the correspondent controls using the Dunnett's test. No statistical difference was found in AFB₁ levels of feedstuffs not containing ADSs when extracted with AC or MeOH, even if numerically higher values were obtained with AC. A dose-dependent effect (p < 0.01) of ADSs inclusion was observed on AFB₁ recoveries that were lower when the higher ADS level (20 g kg^?1) was included in the HC and LC feedstuffs. Higher AFB₁ recoveries were obtained using AC compared with MeOH, both in HC (75.0% versus 12.0%, respectively) and in LC (84.0% versus 22.8%, respectively) ADSs containing feedstuffs. However, when the activated carbon and the sodium bentonite were included in feeds, lower AFB₁ concentrations with respect to control values (p < 0.001 and <0.05, respectively) were obtained also using AC. The data obtained in this study indicate that routine use of the MeOH solvent for AFB₁ analysis of unknown feedstuffs, can produce misleading results if they contain an ADS.