• Korean Journal of Food Science and Technology
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• v.19 no.1
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• pp.54-60
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• 1987

Butylated hydroxyanisole (BHA) and butylated hydroxytoluene (BHT) were tested for their effectiveness in inhibiting the growth of microorganisms. Among the test microorganisms, the growth of three yeasts (Saccharomyces cerevisiae, Kloeckera apiculata, Cryptococcus hungaricus), three bacteria (Bacillus subtilu, Lactobacillus cases, Escherichia colt) and two molds (Aspergillus oryzae. Penicillium sp.) was progressively decreased as concentrations of BHA were increased. A. oyzae was completely inhibited with 100ppm of BHA and a majority of the test microorganisms (S. cerevisiae, K. apiculata. C. hungaricus, B. subtilis, A. oryzae) were completely inhibited by 150 ppm of BHA. The growths of L. casei, E. coli and Penicillium sp. were not affected as much as those of other microorganisms by BHA. Final cell yiedls were becoming lower as the concentration of BHA increased. The growth of C. hungaricus and L. casci was slightly inhibited by BHT. Other microorganisms were not effected by the test concentrations of BHT.

• Molecular & Cellular Toxicology
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• v.3 no.3
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• pp.172-176
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• 2007

Butylated hydroxytoluene (BHT) is widely used antioxidant food additives. It has been extensively studied for potential toxicities. BHT appears adverse effects in liver and thyroid. In this study, we evaluated the genetic toxicity of BHT with more advanced methods, in vitro mouse lymphoma assay $tk^{+/-}$ gene assay (MLA) and in vivo mouse supravital micronucleus (MN) assay. BHT did not appear the significantly results in the absence and presence of metabolic activation system with MLA. Also, in vivo testing of BHT yielded negative results with supravital MN assay. These results suggest that BHT itself was not generally considered genotoxic.

• Toxicological Research
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• v.5 no.1
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• pp.9-15
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• 1989

The effect of butylated hydroxytoluene (BHT) and its major metabolite, 3, 5-di-tert-butyl-4-hydroxybenzoic acid (BHT-acid) on the uptake of taurocholate into hepatocytes was studied using the primary culture of rat hepatocytes. Hepatocyte were isolated by an in situ collagenase perfusion technique and maintained as a monolayer in serum-free meadia for 24 hours before use. The uptake of taurocholate was saturable with an apparent Km of 12.8+2.8 MuM and Vmax of 0.18+0.01 nmol/mg/min. Both BHT and BHT-acid inhibited the hepatocellular uptake of taurocholate when they were added to the culture.

• Asian-Australasian Journal of Animal Sciences
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• v.23 no.10
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• pp.1276-1281
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• 2010

The present study was conducted to investigate the effects of butylated hydroxytoluene (BHT) supplementation on diluted, cooled and frozen-thawed ram spermatozoa. After primary evaluation of collected ejaculates, only semen samples with motility of more than 70% and sperm concentration higher than $3{\times}10^3$ sperm/ml were used for cryopreservation. The selected semen samples were then pooled and diluted 1:4 with Tris Citrate Fructose Yolk (TCFY) extender supplemented with different concentrations of BHT (0.5, 10, 2.0 and 3.0 mM). As the control, semen was diluted and frozen in the diluent without BHT. Motility, progressive motility, viability, membranes and acrosome integrity were evaluated after dilution (part 1), cooling (part 2) and freezing and thawing (part 3). The results of the first part of the experiment showed that there were no significant difference between treatments in the motility, progressive motility, viability, membranes and acrosome integrity of spermatozoa, but the results with 2.0 mM BHT were slightly better than obtained with other levels of BHT and control extender. Significantly better results (p<0.05) were observed in the second part of the experiment for cooled spermatozoa characteristics, when extender was supplemented with 2.0 and 3.0 mM BHT. Furthermore, the results obtained in the third part of the experiment indicated that, after freezing and thawing, all evaluated semen characteristics were improved significantly (p<0.05) by increasing BHT levels, with the best results obtained for extender containing 2 mM BHT. Comparison of these results with those of control diluent, the effects of supplementation were significantly (p<0.01) better. However, the higher concentration of BHT (3.0 mM) reduced the motility, acrosomal integrity, viability and hypo-osmotic swelling response of spermatozoa compared to extender containing 2.0 mM BHT. In conclusion, the results obtained in this study showed that the semen quality of rams was improved when BHT was added to extender used before the freezing process.

• Journal of Nutrition and Health
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• v.23 no.1
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• pp.11-18
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• 1990

Sprague-Dawley male rats were fed the diet of p/s 4.0(soybean oil : I), p/s 0.08(Beef tallow : II) at the level of 15% fat until 8 weeks after weaning. I & II groups were divided into 4 sub-groups by diets with or without 0.3% butylated hydroxytoluene(BHT). 2-AAF was injected at the age of $5_{1/2}$, 6, $5_{1/2}$, 7 weeks. MFO system enzyme(cytochrome p-450, cytochrome p-450 reductase, cytochrome b5) activities and lipid peroxide were determined from isolated liver microsome. 2-AAF injected young rats had growth retardatiion. Lipid peroxide values were not influenced greatly by dietary fat, 2-AAF and BHT. Cytochrome p-450 contents were increased in I-BHT-AAF & II-AAF groups by 2-AAF and its contents were not affected by BHT. But cytochrome p-450 and cytochrome p-450 reductase were not increased in soybean oil diet ybean oil groups. Cytochrome b5 was not influenced by dietary fat, 2-AAF and BHT. Cytochrome p-450 and lipid peroxide, cytochrome p-450 reductase and cytochrome b5, which transfer to MFO system, appeared to have positive correlations(r=0.2474, r=0.2475, p<0.05) each other. This result suggests that MFO system metabolizing 2-AAF was influenced by dietary fats and BHT. 2-AAF induced growth retardation in young rats.

• Archives of Pharmacal Research
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• v.10 no.4
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• pp.223-227
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• 1987

The effects of butylated hydroxyanisole and butylated hydroxytoluene on the immune status in normal male were evaluated. They exhibited significant decrease in the circulating leukocyte counts. Relative spleen and thymus weights were slightly decreased, but not stratistically significant. These were, however, significant liver hypertrophies in theier exposed mice. Splenic IgM PFCs per one million cells in 1/20 LD50 BHA and BHT exposed mice were significantly reduced IgM PFCs per spleen were similar tothose of control, except in 1/20 LD50 BHA exposed mice, where they were significantly suppressed. The precise nature of the inhibition is not clear. Direct cytotoxicity is not responsible for the depressed antibody response, even following relatively high doses of them, because the changes in spleen cellularity are not significant. Both substances, however, did not show any effects on the arthus reaction and delayed hypersensitivity reaction induced by heat aggreagted bovine serum albumin, and in vivo phagocytosis of colloidal carbon. In the light of the present results, in vivo antibody response as well as in vitro, may be sensitive to BHA of the present results, in vivo antibody response as well as in vitro, amy be sensitie to BHA and BHT. Further elucidation of the precise nature of antibody suppression in their exposed mice, is warranted.

• Toxicological Research
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• v.11 no.2
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• pp.215-221
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• 1995

This paper examines the effects of dietary polyunsaturated fatty acid/saturated fatty acid (p/s) ratios and butylated hydroxytoluene (BHT) on the hepatic microsomaI mixed-function oxidase sy. stem in 2~acetylaminofiuorene (2-AAF) treated rats. Sprague-Dawley male rats were fed the diet of beef tallow (p/s 0.08), beef tallow plus soybean oil (p/s 1.0), and soybean oil (p/s 4.0) at the level of 15%fat and with or without 0.3% BHT. After 2-AAF was injected twice at the ages of 23 and 27 weeks, cholesterol/phospholipid molar ratio, thiobarbituric acid reactive substances (TBARS) level, cytochrome P450, cytochrome $b_5$, NADPH-cytochrome $b_5$, and NADPH-cytochrome c reductase activity were measured from isolated hepatic microsomal fractions. In the beef tallow (p/s 0.08) and beef tallow plus soybean oil (p/s 1.0) groups, cholesterol/phospholipid molar ratio showed decreasing tendency by 2-AAF and BHT. Cytochrome P-450 content was decreased in the group of soybean oil (p/s 4.0) and NADPH-cytochrome c reductase activity was increased by 2-AAF and BHT in all the dietary groups. While TBARS levels were increased by 2-AAF in all the dietary groups, they were reduced by BHT in the soybean oil (p/s 4.0) group. These results suggest that long term intake of soybean oil (p/s 4.0) diet induced changes in the nature of microsomal membrane and induced less cytochrome P-450, low level feeding of BHT increased cytochrome c reductase activity and lowered microsomal lipid peroxidation levels, which were increased by 2-AAF treatment.

• Applied Microscopy
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• v.27 no.2
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• pp.197-215
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• 1997

A morphologic study on the effect of butylated hydroxytoluene (BHT) on experimental hepatocarcinogenesis induced by 3'-methyl-4-dimethyl aminoazobenzene (3'-MeDAB) was investigated. A total of 110 Sprague-Dowley male rats weighting about 200 g each were used for the experiment, and divided into 4 groups; the 3'-MeDAB, BHT, 3'-MeDAB/BHT treated group, and the control group. Four to eight rats of each group were sacrified on the 4th, 8th, 14th and 16th experimental weeks, with continuous pelletized feeding containing 0.09% 3'-MeDAB and 0.5% BHT. The liver was examined by transmission and scanning electron microscopy. The results were as follows; Electron microscopically, the fine structure of the hepatocytes remained consistently abnormal up to 16 weeks after the 3'-MeDAB treatment. There was no significant difference in the groups observed earlier than in the ones observed later. Many subcellular changes were observed : nuclear change, decreased glycogen, mitochondrial abnormalities, disaggregated rough endoplasmic reticulum, marked proliferation of smooth endoplasmic reticulum, dilatation and distortion of bile canaliculi, increased lysosomes, apoptotic bodies, migration of bile ductule cell. In the BHT treated group, the ultrastructural changes of hepatocytes were not significant, except for the lipid droplets and proliferated smooth endoplasmic reticulum among hepatocytes depending on the experimental duration. The various subcellular changes of 3'-MeDAB/BHT treated groups were simillar to those of the 3'-MeDAB treated group, but the degree of changes in the 3'-MeDAB/ BHT treated group decreased compared with those of the 3'-MeDAB treated group. These results suggest that dietary butylated hydroxytoluene has a protective/inhibitory effect on the hepatocarcinogenesis induced by 3'-methyl-4-dimethyl -aminoazobenzene.

• Journal of the Society of Cosmetic Scientists of Korea
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• v.25 no.3
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• pp.67-86
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• 1999

Ascorbic acid which has various physiological benefits as the functional substance is easily oxidized and destroyed by the structural instability. Liposome encapsulated pure ascorbic acid was prepared for the sake of the constraint of oxidation. The influence of cholestrol or $\beta$-sitosterol on the stabilization of liposome was investigated. Butylated hydroxytoluene(BHT), tertiary butylhydroquinone(TBHQ), $\alpha$ -glycosyl rutin and natural concentrated tocopherol were used for constraint of oxidation of ascorbic acid. The presence of cholesterol or $\beta$-sitosterol decreased oxidation of ascorbic acid. That results were thought that cholesterol or $\beta$-sitosterol so increased rigidity of bilayer that the leakage of vitamin C decreased. As a result the oxidation and degradation of vitamin C were constrained. At 0.3w/w% cholesterol content the most stable liposome was formulated. The whole antioxidant that used at the research constrained oxidation of ascorbic acid. The antioxidation for ascorbic acid increased in order of tertiary butylhydroquinone, $\alpha$-glycosyl rutin, butylated hydroxytoluene and natural concentrated tocopherol. But u -glycosyl rutin is preferable to tertiary butylhydroquinone which was the most effective in antioxidation as the antioxidant of ascorbic acid which was utilized in cosmetics and pharmacy.

• Korean Journal of Food Science and Technology
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• v.14 no.3
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• pp.283-288
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• 1982

식품에 항 산화제로 첨가시키는 BHT와 BHA의 독성에 대해 고찰하였다. BHT와 BHA는 실험동물에서 간, 폐, 신장, 순환계, 생식계, 등에 여러 영향을 주며 이러한 영향은 동물의 종류와 성에 따라 상당한 차이를 나타내고 있다. 대부분 실험에 사용된 BHT와 BHA의 양이 인체가 매일 식품으로부터 흡수할 수 있는 양보다는 훨씬 많은 양을 사용하였기 때문에 동물실험의 결과로 인체에 유독성을 직접 판단하기는 어렵고 이들의 안전성에 대해서는 보다 체계적인 연구가 앞으로 요구된다. 미량의 BHT와 BHA를 장기간 복용하였을 때 생기는 부작용에 대한 연구가 필요하며 또한 이들 항 산화제는 약물대사에 관여하는 효소의 합성내지 활성도를 증가시키는 것으로 알려져 있기 때문에 체내에서 다른 약물의 대사 및 독성에 어떤 상호 작용을 가져오는 지에 대한 면밀한 연구가 요구되고 있다. 더불어 본 논문은 현시점에서 이들 항산화제의 식품에 첨가여부를 논쟁하고자 하는 의도가 전혀 없음을 명백히 하는 바이다.