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.     

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.     

Estimates of the theoretical maximum daily intake of phenolic antioxidants BHA, BHT and TBHQ in Brazil

The theoretical maximum daily intakes (TMDI) of the phenolic antioxidants butylated hydroxyanisole (BHA), butylated hydroxytoluene (BHT) and tertbutyl hydroquinone (TBHQ) in Brazil were estimated using food consumption data derived from a household economic survey and a packaged goods market survey. The estimates were based on maximum levels of use of the food additives specified in national food standards. The calculated intakes of the three additives for the mean consumer were below the ADIs. Estimates of TMDI for BHA, BHT and TBHQ ranged from 0.09 to 0.15, 0.05 to 0.10 and 0.07 to 0.12mg/kg of body weight, respectively. To check if the additives are actually used at their maximum authorized levels, analytical determinations of these compounds in selected food categories were carried out using HPLC with UV detection. BHT and TBHQ concentrations in foodstuffs considered to be representive sources of these antioxidants in the diet were below the respective maximum permitted levels. BHA was not detected in any of the analysed samples. Based on the maximal approach and on the analytical data, it is unlikely that the current ADI of BHA (0.5mg/kg body weight), BHT (0.3mg/kg body weight) and TBHQ (0.7mg/kg body weight) will be exceeded in practice by the average Brazilian consumer.     

Migration analysis,antioxidant, and mechanical characterization of polypropylene-based active food packaging films loaded with BHA,BHT, and TBHQ

Polypropylene (PP) based active composite films were prepared by adding butylated hydroxy anisole (BHA), butylated hydroxytoluene (BHT), and tertiary butylated hydroquinone (TBHQ) antioxidants using the extrusion molding process. All concentrations of BHT, 2% to 3% BHA, and 3% TBHQ significantly increased the tensile strength (TS) of the composite films compared with control films. Increasing antioxidant concentration decreased TS values for BHT films, whereas an opposite trend was observed for BHA and TBHQ films. BHA at < 2%, BHT at > 2%, and TBHQ at all added concentrations significantly reduced elongation at break (E_b) of the composite films compared to control films. Water vapor permeability (WVP) of 1% BHT film was not significantly different from control. However, other antioxidants especially at increased concentrations significantly increased WVP values. TBHQ films with 300% to 662% increase had the highest WVP and BHT films with 5% to 81% increase had the lowest WVP among composite films. All three antioxidants had a negative effect on the transparency of the films; however the effect of BHA at higher concentrations was greater. The antioxidants did not change the color attributes of the films. Films containing all antioxidants showed 2,2-diphenyl-1-picrylhydrazyl radical-scavenging activity, which increased with increase in their concentration, especially for those containing 3 wt.% BHT and TBHQ. Overall, incorporating BHA and BHT into a PP matrix improved mechanical, barrier, antioxidant properties, and film appearance and consequently were proposed for the development of antioxidant active PP films. TBHQ film is not recommended for food packaging because of its weak mechanical properties (lower E_b and TS values, higher WVP, and greater migration).     

Kinetics of Growth and Inhibition of Listeria monocytogenes in the Presence of Antioxidant Food Additives

Listeria monocytogenes strain Scott A in Tryptose Broth was treated with 100-300 ppm butylated hydroxyanisole (BHA), 300-700 ppm butylated hydroxytoluene (BHT) and 10-30 ppm tertiary butylhydroquinone (TBHQ). Resulting growth curves were fitted using the logistic model, and growth parameters [lag period (LP), generation time (GT), and maximum growth (MG)] were calculated. BHA and BHT inhibited Listeria monocytogenes by increasing LP and GT and decreasing MG. Extent of inhibition was concentration-dependent for cultures with BHA, but not with BHT. TBHQ at 10-30 ppm increased LP but did not affect other parameters. LP increased exponentially with increased BHA or TBHQ in Listeria culture. Concentrations of additive required to increase LP by one order of magnitude were 240 ppm for BHA and 26 ppm for TBHQ.     

气相色谱--质谱法测定XO酱中BHA、BHT和TBHQ

采用气相色谱—质谱(全扫描方式)测定XO酱中丁基羟基茴香醚(BHA)、二丁基羟基甲苯(BHT)和叔丁基对苯二酚(TBHQ)。样品用甲醇振荡萃取，以DB—5MS为分析柱。3种组分的回收率在85％～99％之间，相对标准偏差小于8．4％；样品中BHA、BHT和TBHQ的检测限分别为0．05、0．05和0．10(mg／kg)。该法简单、快速、准确，可用于XO酱等基质复杂的富油食品中BHA、BHT和TBHQ的检测和确证。     

Assessment of estimated daily intakes of benzoates for average and high consumers in Korea.

A study was performed to evaluate the estimated daily intakes (EDI) of benzoates for the average and high (90th percentile) consumers by age and sex categories in Korea. The estimation of daily intakes of benzoates was based on individual dietary intake data from the National Health and Nutrition Survey in 1998 and on the determination of benzoates in eight food categories. The EDI of benzoates for average consumers of different age groups ranged from 0.009 to 0.025 mg kg(-1) bw day(-1). For high consumers, the range of EDI of benzoates was 0.195-1.878 mg kg(-1) bw day(-1). The intakes represented 0.18-0.50% of the acceptable daily intake (ADI) of benzoates for average consumers and 3.9-37.6% of the ADI for high consumers. Foods that contributed most to the daily intakes of benzoates were mixed beverages and soy sauce in Korea.     

Gas Chromatographic Determination of Butylated Hydroxyanisole, Butylated Hydroxytoluene and Tertiarybutyl Hydroquinone in Soybean Oil

Oils containing different levels of the most common phenolic antioxidants, butylated hydroxyanisole (BHA), butylated hydroxytoluene (BHT), and tertiarybutyl hydroquinone (TBHQ) from 0–200 ppm were analyzed by a new gas chromatographic method. The antioxidants in oils were isolated by nitrogen gas purging, collected on Tenax GC coated with polymetaphenoxylene, and then separated and quantitated by gas chromatography. This method requires neither extraction of antioxidants from oil nor derivatization of antioxidants. The correlation coefficients (r) between gas chromatographic peak heights or peak area and their concentrations in oils were 0.99 for BHA, BHT, and TBHQ. This simple gas chromatographic method can determine as little as 10 ppm pf BHA, BHT, or TBHQ in oils in an hour.     

SYNERGISTIC ACTIVITY OF CAPELIN PROTEIN HYDROLYSATES WITH SYNTHETIC ANTIOXIDANTS IN A MODEL SYSTEM

Capelin protein hydrolysates (CPHs) were examined in a β-carotene-linoleate model system together with synthetic antioxidants, namely butylated hydroxyanisole (BHA), butylated hydroxytoluene (BHT), and tert-butyl hydroquinone (TBHQ). Addition of CPHs was at 0.25 and 1 mg per 5 mL in the above emulsion. BHA and BHT were added at levels of 2.5 and 5 μg and TBHQ at 5 and 10 μg. Absorbance of model emulsion system at 470 nm was recorded every 30 min for 2 h. CHPs and synthetic antioxidants inhibited oxidation of linoleic acid effectively. In the experiment with BHA, the influence of CPHs was significant (p<0.01) for the samples incubated for 60, 90, and 120 min; and in experiments with BHT and TBHQ during the entire incubation period. The influence of added BHA and BHT on inhibition of bleaching of β-carotene in the emulsion system was significant (p <0.01) during the incubation period and after 60, 90 and 120 min when TBHQ was used. A synergistic effect was observed only for CPHs and TBHQ when incubation time was 60, 90, and 120 min.     

Effect of BHA, BHT, TBHQ and PG on Growth and Toxigenesis of Selected Aspergilli

The antifungal effect of butylated hydroxyanisole (BHA), butylated hydroxytoluene (BHT), tertiary butylhydroquinone (TBHQ) and propyl gallate (PC) alone or in combination on three toxigenic strains of aspergilli (NRRL 2999, NRRL 4123, NRRL 5835) and three nontoxigenic strains of aspergilli (NRRL 5521, NRRL 5917, NRRL 5918) was examined in a solid medium and in salami. BHT and PG (0.001,0.005,0.01,0.02g per plate) did not inhibit growth, sporulation, and toxigenesis of all six cultures. Aflatoxin production by toxigenic aspergilli (B₁, B₂, G₁ and G₂) in the presence of BHA, TBHQ, and a combination of BHA and TBHQ was reduced significantly (P < 0.05). In salami BHA, TBHQ alone or in combination at 100 ppm significantly (P < 0.05) decreased the aflatoxin production by aspergilli when compared to control samples. A combination of BHA and TBHQ showed synergistic inhibition in both studies (solid medium and salami studies).     

Rapid micropreparation procedure for the gas chromatographic-mass spectrometric determination of BHT, BHA and TBHQ in edible oils

A rapid, accurate and organic solvent saving procedure has been developed for the GC/MS determination of three phenolic antioxidants butylated hydroxytoluene (BHT), butylated hydroxyanisole (BHA) and butyl hydroquinone (TBHQ) in vegetable oils. The method involves two-step microextraction and a centrifugal procedure in a 2 mL autosampler vial, consuming only 50 mg sample and total 1 mL acetonitrile. Recoveries of the phenolic antioxidants when spiked to soya bean oil, peanut oil and cereal cooking oil at 50, 200 and 250 mg/kg, respectively were in the ranges 95.6-104.3% for BHT, 99.7-107.5% for BHA and 93.6-103.8% for TBHQ with the relative standard deviation (RSD) were less than 3% for their independent measurements. The developed method was repeatable and could be applied to determine trace amounts of phenolic antioxidants in vegetable oils.     

Assessment of Korean consumer exposure to sodium saccharin,aspartame and stevioside

The dietary intakes of sodium saccharin, aspartame and stevioside were estimated on the basis of food consumption data of the Korean consumer and the concentration of sweeteners in processed foods. Results were compared with the acceptable daily intake (ADI) of sweeteners. Among the 28 food categories for which the application of sodium saccharin, aspartame and stevioside is permitted in Korea, they were detected in 5, 12 and 13 categories, respectively. The estimated daily intake (EDI) of sodium saccharin and aspartame were high in infants and children, whereas the EDI of stevioside was high in adolescents and adults. The most highly consumed sweetener was aspartame, and the highest EDI/ADI ratio was found for sodium saccharin. The main food categories contributing to sweetener consumption were beverages, including alcoholic beverages. For most Korean consumers, the EDIs were no greater than 20% of their corresponding ADI; however, the EDI of sodium saccharin for conservative consumers aged 1–2 years reached 60% of their ADI.     

GC-MS法同时检测葡萄酒中10种防腐剂和抗氧化剂

利用离子阱气相色谱-质谱联用仪,优化了色谱、质谱仪器参数,利用质谱全扫描技术建立了同时检测葡萄酒中山梨酸、苯甲酸、脱氢乙酸、叔丁基对苯二酚（TBHQ）、叔丁基羟基茴香醚（BHA）、2,6-二叔丁基对甲酚（BHT）、对羟基苯甲酸甲酯、对羟基苯甲酸乙酯、对羟基苯甲酸丙酯、对羟基苯甲酸丁酯等10种添加剂的方法。10种添加剂的检测限分别为0．11mg/L、0．072mg／L、0．046mg／L、0．035mg／L、0．032mg／L、0．020mg／L、0．035mg／L、0．042mg／L、0．035mg,／L、0．035mg／L,方法的精密度为2．9％～4．6％、回收率为82％～101％,应用于葡萄酒中食品添加剂的检测,效果良好,检测效率大大提高。     

Synergistic antioxidant and antibacterial activity of rosemary plus butylated derivatives

Antioxidant and antibacterial activity of a methanol rosemary extract (RE) containing 30% carnosic acid (CA), 16% carnosol (COH) and 5% rosmarinic acid (RA) was studied in vitro alone and in combination with the antioxidant food additives butylated hydroxytoluene (BHT) and butylated hydroxyanisole (BHA). The antioxidant efficiency of the extract, CA, and RA, was determined by a kinetic analysis of the 2,2-diphenyl-2-picrylhydrazyl hydrate radical (DPPH·) scavenging activity. RE showed two different rate slopes in the reduction of DPPH· vs. time curve, which correlated with the distinct behaviours of RA and CA; pure RA reached the plateau more rapidly than CA. A synergistic antioxidant effect between RE and BHT was demonstrated by isobolographic analysis and a synergistic interaction of RE with BHA to inhibit Escherichia coli and Staphylococcus aureus growth was observed. Therefore, rosemary not only enhances the antioxidant efficiency of BHA and BHT, but also the antibacterial effect of BHA; allowing a decrease from 4.4 to17 folds in the amounts of the synthetic compounds used.     

Two novel synthetic antioxidants for deep frying oils

Lauryl tert-butylated hydroquinone (LTBHQ) and its oxidized compound, lauryl tert-butylated quinone (LTBQ) were synthesized from tert-butylated hydroquinone (TBHQ) and lauryl alcohol. Their antioxidant activities were investigated. At temperatures higher than 140 °C, the antioxidant activity of LTBHQ and LTBQ was higher than TBHQ. In emulsions, these two compounds had stronger antioxidant activity than TBHQ, butylated hydroxytoluene (BHT) and butylated hydroxyanisole (BHA).     

Daily intake assessment of saccharin, stevioside, D-sorbitol and aspartame from various processed foods in Korea

This study was carried out to estimate the daily intakes (EDIs) of artificial sweeteners such as saccharin, stevioside, D-sorbitol and aspartame in order to evaluate the safety of the artificial sweeteners in Korea. A total of 274 food samples were selected from the foods considered to be representative sources of artificial sweeteners in the Korean diet and analysed by using HPLC with evaporative light scattering and ultraviolet detectors. In case of aspartame, the reference values were used without instrumental analysis. The EDIs of saccharin, stevioside, D-sorbitol and aspartame for average consumers were 0.028, 0.008, 4.9 and 0.14 mg kg^-1 body weight day^-1, respectively, and as a proportion of the acceptable daily intake (ADI) were not higher than 1% of ADI of the Joint FAO/WHO Expert Committee on Food Additives (JECFA). For 90th percentile consumers, the EDIs of saccharin, stevioside, D-sorbitol and aspartame were 2.0, 0.20, 141 and 4.6 mg kg^-1 body weight day^-1, respectively, and as a proportion of the ADI, the EDIs of saccharin and aspartame were 40.7% and 11.4% of the ADI set by the JECFA, respectively. Because JECFA did not assign ADIs for stevioside and D-sorbitol, the values for these sweeteners were not compared. According to these results, the EDIs of artificial sweeteners such as saccharin and aspartame in Korea are significantly lower than ADI set by the JECFA.     

THERMAL STABILITY OF SOME COMMERCIAL NATURAL AND SYNTHETIC ANTIOXIDANTS AND THEIR MIXTURES

The thermal stability of some commercially available antioxidants alone or in binary or ternary mixtures was investigated in order to find the optimal combination of antioxidants for food processing. The synthetic antioxidants used were butylated hydroxyanisole (BHA), butylated hydroxytoluene (BHT), propyl gallate (PG) and tertiary-butyhydroquinone (TBHQ). Other antioxidants such as ascorbyl palmitate (AP), mixed tocopherols (TOCO) and monoacylglycerol citrate (MGC) were also employed. The effectiveness of these antioxidants and their combinations was assessed using the Rancimat test by measuring the induction period for the oxidation of sunflower oil after heating at 180C for 1 h and comparing it to the oxidation kinetics of the oil without added antioxidants. All antioxidants and their binary or ternary mixtures showed different degrees of thermal instability. TBHQ individually, among all the examined antioxidants, showed the highest thermal stability. On the other hand, AP as an antioxidant in sunflower oil exhibited low stability during heating. The thermal stability of AP could be enhanced by the addtion of BHA or BHT in binary mixtures at a ratio of 1:3 (w/w). In addition, the ternary mixture of AP, TOCO and MGC (65:25:10) in sunflower oil also showed a higher stability to thermal inactivation. However, the ternary mixture containing 0.13% AP, 0.05% TBHQ and 0.02% MGC provided the optimal protection during thermal treatment. Although a combination of 0.13% BHT, 0.05% AP and 0.02% MG was very effective synergistically at room temperature, it showed a higher susceptibility to thermal inactivation. It was     

油脂及其制品中BHA、BHT、PG和TBHQ快速测定方法的研究

研究了采用高效液相色谱仪同时测定油脂及食品中BHA、BHT、PG、TBHQ的检测方法。样品经甲醇提取、过滤后即可直接测定。检测波长 ,2 80nm ;流动相 ,V(甲醇 )∶V(质量分数 1%乙酸 ) =40∶60 ;梯度 ,40 %～ 10 0 %甲醇 /7 5min ,10 0 %甲醇保持 4 5min。 4种组分的回收率在 85 8%～ 10 1 5 %之间 ,相对标准偏差 (RSD) <5 5 4%。该法简单、快速、准确 ,可用于油脂及其制品中BHA、BHT、PG和TBHQ的日常检测。     

Determination and confirmation of five phenolic antioxidants in foods by LC/MS and GC/MS

Identification and determination of butylated hydroxyanisole (BHA), butylated hydroxytoluene (BHT), nordihydroguaiaretic acid (NDGA), propyl gallate (PG) and tert-butylhydroquinone (TBHQ) by means of LC/MS and GC/MS were examined. These five phenolic antioxidants were detected as their pseudo-molecular ions [M-H]- by LC/MS using a Shim-pack FC-ODS column with drying gas. Moreover, BHA, BHT and TBHQ were detected based on their mass fragment ions by GC/MS. Decomposition of TBHQ, NDGA and PG during analysis could be prevented by the addition of L-ascorbic acid (AsA) to the extraction solvent. All five antioxidants were extracted from nikuman, olive oils, peanut butter, pasta sauce and chewing gum with a mixture of acetonitrile-2-propanol-ethanol (2:1:1) containing 0.1% AsA (AsA mixture), which had been cooled in a freezer and filtered. One part filtrate and 5 parts water were mixed and placed on a Mega-Bond Elut C18 cartridge, except in the case of chewing gum. Lipids in foods were removed on a C18 cartridge by washing with 5 mL of 5% acetic acid, and antioxidants were eluted with 5 mL of AsA mixture. The antioxidants spiked into nikuman, olive oil, peanut butter, pasta sauce and chewing gum were successfully identified and their concentrations determined by LC/MS, and GC/MS with good recoveries.     

Quantitative determination of the phenolic antioxidants using voltammetric techniques

Synthetic antioxidants, such as butylated hydroxyanisole (BHA), butylated hydroxytoluene (BHT) and tert-butylhydroquinone (TBHQ), show sensitivity to voltammetric waves. The waves of these antioxidants, however, are seriously overlapped and it is difficult to determine them simultaneously. The influence of different parameters (working electrode, supporting electrolyte, pH, voltammetric technique) was evaluated in a quantitative simultaneous determination of three antioxidants in alcoholic mixtures and real sample foods. Glassy carbon (GC) and platinum (Pt) working electrodes were investigated as mediators of oxireduction reactions. Two supporting electrolytes were investigated: Britton-Robinson 0.1 mol l⁻¹ buffer (pH 2.0) and HCl 0.1 mol l⁻¹ (pH 2.0) both with 2 g l⁻¹ (p/v) of methanol. In this paper, voltammetric conditions for the analysis of up to three-component mixtures of antioxidant present at levels: 2.0-100.0 mg l⁻¹ for BHA, 4.0-100.0 mg l⁻¹ for TBHQ and 2.0-20.0 mg l⁻¹ for BHT at GC in HCl 0.1 mol l⁻¹ and 8.0-120.0, 10.0-130.0 and 4.0-30.0 mg l⁻¹ for BHA, TBHQ and BHT, respectively, at Pt in the same supporting electrolyte. The results show that for real food samples, the parameters investigated were satisfactory for quantitative determination using square wave voltammetry (SWV) without chemometric approaches and without suffering overlapping problems.