Antioxidant and pro-oxidant activity of green tea extracts in marine oils

The activity of green tea extracts (GTE) on the oxidation of refined, bleached and deodorized (RBD) seal blubber oil (SBO) and menhaden oil (MHO) was examined under Schaal oven conditions at 65 °C. Progression of oxidation was monitored using weight gain, peroxide value (PV) and 2-thiobarbituric acid-reactive substances (TBARS) data. GTE exhibited a pro-oxidant effect in both oils examined, perhaps due to the catalytic effect of their chlorophyll constituents. Therefore, in follow-up experiments, a column chromatographic technique was employed to remove chlorophyll from GTE. The resultant dechlorophyllized green tea extract (DGTE) was applied to both SBO and MHO at 100, 200, 500 and 1000 ppm levels. The antioxidant activity of DGTE was compared with the effects of the commonly-used antioxidants such as butylated hydroxyanisole (BHA), butylated hydroxytoluene (BHT) and tert-butylhydroquinone (TBHQ) at 200 ppm and α-tocopherol at 500 ppm. DGTE at ≥200 ppm exhibited excellent antioxidant activity in both oils and its efficacy was higher than that of BHA, BHT and α-tocopherol, but less than that of 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。     

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.     

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.     

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.     

食用油中5种合成抗氧化剂同时检测方法研究

建立了基于高效液相色谱的一种简单、快速、准确定量分析25份食用油中没食子酸丙酯(PG)、没食子酸十二酯(DG)、叔丁基对苯二酚(TBHQ)、丁基化羟基茴香醚(BHA)和二丁基羟基甲苯(BHT) 5种合成抗氧化剂的方法。结果显示,5种合成抗氧化剂(PG、DG、TBHQ、BHA和BHT)的质量浓度在10~70μg/m L范围内与色谱峰面积呈良好线性关系,相关系数均大于0.999,检出限为0.27~0.34μg/kg,加标回收率为84.56%~101.12%,相对标准差在1.27%~4.82%之间。该方法具有良好的精确度和准确度,且在13 min内可完成5种合成抗氧化剂的分离,能够适用于食用油中合成抗氧化剂的检测分析。     

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).     

Control of Lipid Oxidation in Cooked Ground Pork with Antioxidants and Dinitrosyl Ferrohemochrome

A number of antioxidants were added to ground meat prior to cooking to achieve oxidative stability. The 2-thiobarbituric acid (TBA) numbers of cooked-meat samples after storage for up to 35 days were determined. Of the antioxidants tested, butylated hydroxyanisole (BHA), butylated hydroxytoluene (BHT), propyl gallate (PG), t-butylhydro-quinone (TBHQ), trihydroxybutyrophenone (THBP), nordihydro-guaiaretic acid (NDGA), catechol, and ethoxyquin were effective at a level of 200 ppm giving TBA values of less than one. BHA, TBHQ, and ethoxyquin were equally effective at a level of 30 ppm. Ascorbyl acetal and ascorbyl palmitate retarded lipid oxidation effectively (TBA values of slightly over one after five weeks of storage). The cooked cured-meat pigment showed some antioxidant effect which increased with increasing concentration, and at 18–24 ppm was better than that of 200 ppm of α-tocopherol.     

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.     

Ascorbyl Palmitate Efficacy in Enhancing the Accelerated Storage Stability of Canola Oil

The effect of various levels of ascorbyl palmitate (AP) and of butylated hydroxyanisole/toluene (BHA/BHT) on the accelerated storage stability of canola oils was determined by sensory, gas liquid chromatographic (GLC) and chemical evaluations. In Schaal oven tests (65°C, 0–16 days), chemical, GLC and trained sensory panel data indicated that 200 ppm AP retarded autoxidation in stored canola oils. Monoglyceride citrate (MGC) addition to oils containing 200 ppm AP did not enhance oil stability. Fluorescent light tests (7500 lux, 24°C, 0–24 hr) showed that 200 ppm AP, with or without MGC, had a limited effect in protecting canola oil from photooxidation. BHA/ BHT, at 100 ppm each, with MGC, did not improve canola oil stability.     

Qualitative and Quantitative Effects of Antioxidants on the Flavor Stability of Oil

Qualitative and quantitative effects of the common antioxidants, BHA, BHT, PG and TBHQ on the rate of dissolved free oxygen disappearance in soybean oil during storage were studied. The order of effectiveness was BHA, BHT, PG and TBHQ with TBHQ as the most effective. Statistical analyses of the results showed that the effects of BHA and BHT were not significantly different from each other at the 5% level, but BHA or BHT was different from PG or TBHQ, and PG was different from TBHQ. The antioxidant effectiveness of levels of 0, 50, 100, 150 and 200 ppm BHA, BHT, PG or TBHQ was significantly different from one another at the 5% level. The higher the amounts of antioxidants added, the slower the rate of dissolved oxygen disappearance in the oil.     

ENHANCEMENT OF THE SHELF-LIFE OF FLAVORED BUTTER OILS BY SYNTHETIC ANTIOXIDANTS

The shelf-life of flavored butter oils was enhanced by adding permitted antioxidants butylated hydroxyanisole (BHA) and butylated hydroxytoluene (BHT) at 0.02% level. Such samples competed well with conveitional ghee. Tertbuthydroquinone (TBHQ) (permitted antioxidant for vegetable fats) showed the greatest antioxidant potency and could be recommended even at a very low level (0.005%) for imparting a longer shelf-life to flavored butter oils.     

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.     

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.     

电分析法同时快速测定植物油中TBHQ、BHA和BHT的研究

研究抗氧化剂TBHQ、BHA和BHT在玻碳电极上的伏安性质,三者的循环伏安图表明:3个峰形良好的氧化峰分别对应:TBHQ的氧化峰电位为590 mV,BHA的氧化峰电位为710 mV,BHT的氧化峰电位为1 050 mV,只有一个还原峰,电位为-90 mV。通过实验优化了电分析法测定植物油样抗氧化剂的分析技术参数。借助最小偏二乘法(PLS),利用电化学分析法预测了人工模拟试样中的3种抗氧化剂的含量,预测结果及加标回收率结果较好。利用所开发的方法测定了实际油样中的抗氧化剂的含量,其回收率在94%~104.1%,结果表明该法同时测定植物油中2种及以上的抗氧化剂是可行的,而且无需提取、分离及清洗等预处理程序,应用潜力巨大。     

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).     

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     

Analysis of synthetic phenolic antioxidants in edible oils by micellar electrokinetic capillary chromatography

Synthetic antioxidants most used in oil-based food to avoid oxidation processes, butylated hydroxyanisole (BHA), butylated hydroxytolueno (BHT), and dodecyl gallate (DG), were analyzed in edible oils using micellar electrokinetic capillary chromatography (MECK) with bis-(2-ethylhexyl) sodium sulfosuccinate as the pseudostationary phase.

Studies involving solid-phase and liquid–liquid extraction were performed to find the best sample treatment before injection into the electrophoretic system. The best methodology for the isolation of antioxidants was extraction with acetonitrile from edible oil diluted with hexane. A method that allows the determination of the antioxidants present in these samples was proposed. With this method BHA, BHT and DG were evaluated at levels permitted in the European Union.     

Cloud-point extraction and reversed-phase high-performance liquid chromatography for the determination of synthetic phenolic antioxidants in edible oils

A cloud-point extraction (CPE) method using Triton X-114 (TX-114) nonionic surfactant was developed for the extraction and preconcentration of propyl gallate (PG), tertiary butyl hydroquinone (TBHQ), butylated hydroxyanisole (BHA), and butylated hydroxytoluene (BHT) from edible oils. The optimum conditions of CPE were 2.5% (v/v) TX-114, 0.5% (w/v) NaCl and 40 min equilibration time at 50 °C. The surfactant-rich phase was then analyzed by reversed-phase high-performance liquid chromatography with ultraviolet detection at 280 nm, using a gradient mobile phase consisting of methanol and 1.5% (v/v) acetic acid. Under the studied conditions, 4 synthetic phenolic antioxidants (SPAs) were successfully separated within 24 min. The limits of detection (LOD) were 1.9 ng mL(-1) for PG, 11 ng mL(-1) for TBHQ, 2.3 ng mL(-1) for BHA, and 5.9 ng mL(-1) for BHT. Recoveries of the SPAs spiked into edible oil were in the range 81% to 88%. The CPE method was shown to be potentially useful for the preconcentration of the target analytes, with a preconcentration factor of 14. Moreover, the method is simple, has high sensitivity, consumes much less solvent than traditional methods, and is environment-friendly. Practical Application: The method established in this article uses less organic solvent to extract SPAs from edible oils; it is simple, highly sensitive and results in no pollution to the environment.     

QuEChERS-高效液相色谱法快速检测食用油中4种抗氧化剂

目的 建立QuEChERS-高效液相色谱法(high performance liquid chromatography, HPLC)快速测定食用油中没食子酸丙酯(propyl gallate, PG), 特丁基对苯二酚(tertiary butylhydroquinone, TBHQ), 叔丁基对羟基茴香醚(butyl hydroxyanisd, BHA), 2,6-二叔丁基对甲酚(butylated hydroxytoluene, BHT)4种抗氧化剂的分析方法。方法 样品用乙腈提取, 经C18-无水MgSO4粉末净化, 色谱柱分离, 以0.5%甲酸水溶液和甲醇为流动相进行梯度洗脱, 二极管阵列检测器(diode array detector, DAD)检测, 检测波长280 nm。结合保留时间和紫外光谱进行定性分析, 峰面积进行定量分析。结果 4种抗氧化剂质量浓度在2.0~100 mg/L范围内线性良好, 相关系数r2＞0.9994; 在不同种类食用油中3个添加水平的平均回收率为78.8%~94.5%, 相对标准偏差为1.1%~6.8%。结论 该方法简便、快捷、节约溶剂, 适用于食用油中快速检测4种抗氧化剂。