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In the present study, we synthesized a diglyceride (CLA-DG) from glycerol and conjugated linoleic acid (CLA). CLA-DG contained about 66% CLA as fatty acids. The safety of CLA-DG was assessed in acute and subchronic studies and in genotoxicity assays. In a single-dose acute study of CLA-DG, male and female rats were administered 5000 mg/kg body weight. CLA-DG caused no adverse effects and body weight gain and food consumption were within the normal range, thus the LD50 of CLA-DG was determined to be greater than 5000 mg/kg. In a 90-day subchronic study (500, 1000, and 2000 mg/kg b.w./day, oral via gavage) in male and female rats, no significant adverse effects in food consumption, body weight, mortality, clinical chemistry, hematology, gross pathology, and histopathology were observed. CLA-DG did not show any potential to induce gene mutations in reverse mutation tests using Salmonella typhimurium TA98, TA100, TA1535, TA1537, and Escherichia coli WP2uvrA strains. CLA-DG did not induce chromosomal aberrations in cultured Chinese hamster lung cells. The results from these studies support the safety of CLA-DG.

To develop a new functional food material, edible crude saponin was isolated from soybean cake using HP-20 resin and its anticancer effect and immune-activity were investigated. The saponin significantly inhibited the growth of cancer cells such as A549, MCF-7 and SW480 at a concentration of 1,000 g/mL. Morphological changes was observed in the AS49 cells surface treated with the saponin of 1,000 g/mL concentration. The proliferation of mouse spleen cells treated with saponin was increased in a dose-dependent manner compared with untreated control cells until the concentration of 1 g/mL but decreased at higher concentrations than that. The NO production in marcrophage cell lines (RAW 264.7) treated with saponin was increased in a dosedependent manner compared with untreated control cells.

The aims of this study were to investigate the antioxidative and antitumor effects of crude polysaccharide fraction from Pleurtus eryngii (CPPE). CPPE inhibited autoxidation of linoleic acid at 1,000 g/mL concentration, and the inhibitory rates of lipid peroxidation in rat liver microsome were 5.28, 9.40, and 32.5% at 10, 100, and 1,000 g/mL concentrations, respectively. After treatment with CPPE for 72 hours, the inhibitory rates against MCF-7, A549 and AGS cell lines showed 42.3, 33.4 and 26.7% at concentration of 1,000 g/mL, respectively. Results of CPPE treatment at 100 and 300 mg/kg/day for 7 days in sarcoma-180 bearing-mice showed survival rates of 70 and 90%, respectively. Body weights of mice treated with CPPE were significantly decreased when compared with the control.

In an ongoing study, rat and human urine have been examined for the presence of malondialdehyde (MDA) derivatives as indicators of the nature of lipid peroxidative damage caused by this compoundin vivo. MDA in urine was found to be present mainly in the form of two lysine adducts, one acetylated and the other unacetylated, reflectingin vivo reactions with tissue proteins. Two minor metabolites were identified as adducts with the phospholipid bases serine and ethanolamine and a third one as an adduct with the nucleic acid base guanine. The identification of an MDA adduct with deoxyguanosine (dG-MDA) among the products of hydrolysis of rat liver DNA suggested the possible occurrence of this compound in urine. In the present study dG-MDA was identified in rat and in human urine, and a high-performance liquid chromatographic method utilizing fluorescence detection was developed for its estimation. The method is sensitive to 1 pmol of dG-MDA and requires a minimum of 1 mL of rat urine or 5 mL of human urine. Its rate of excretion by five-week-old rats (28.54±2.28 nmol/kg/24 h) (mean±SEM) was higher than that for nine-week-old rats (6.29±1.02) and much higher than that for adult humans (0.40±0.05). The results indicate that, as reported for 8-hydroxy-deoxyguanosine, dG-MDA excretion is related to metabolic rate. Excretion of dG-MDA by the rat, like the excretion of total MDA, declines during growth on a body weight basis at a rate similar to the decrease in resting energy metabolism. In contrast to other MDA derivatives excreted in rat urine, vitamin E deficiency had no effect on the excretion of dG-MDA. Together with evidence that the dG-MDA content of rat liver DNA likewise is unaffected by vitamin E depletion or by administration of catalysts ofin vivo lipid peroxidation, these findings indicate that DNA is protected from lipid peroxidative damage, possibly through conservation of the vitamin E associated with the lipids of the nuclear membrane.

This study was carried out to develop a cost-, labor- and efficiency-effective elimination method of pesticide residues in ginseng extract. A two-phase partition method between soybean oil and distilled water or aqueous ginseng extract was employed for the elimination of pesticide residues. Content of the pesticides was determined by gas chromatography with electron capture or nitrogen phosphorus detector. A total of 15 pesticides representing four categories (organochlorine, organophosphorus, carbamate, pyrethroid) were spiked (ca. 2 ppm) to 2 ml soybean oil in a test tube and the oil was mixed with 6 ml distilled water or 10% aqueous ginseng extract. The test tubes were then vortexed (2 min) and centrifuged at 3000 rpm for 15 min to separate the oil and aqueous layers. Each layer was harvested and subjected to quantitative analysis of pesticides. The average distribution ratio of the pesticides to the oil layer was 94.4 +/- 6.7% in the mixture of the oil and distilled water, and 105.5 +/- 6.6% in the mixture of the oil and ginseng extract. No significant qualitative and quantitative change of ginsenosides, the active ingredients of Panax ginseng, was observed in the ginseng extract before and after the oil treatment. These results suggest that two-phase partition chromatography between soybean oil and the aqueous phase is a cost-, labor- and efficiency-effective reliable method for the elimination of pesticide residues in ginseng extract.

The effect of age and peroxidative stress on the concentration of a deoxyguanosine malondialdehyde adduct (dG-MDA) in rat tissues was investigated. Vitamin E deficiency had not effect on the dG-MDA content of liver DNA in rats fed a diet containing 10% corn oil. When 2% cod liver oil was added to this diet, the dG-MDA content of liver DNA doubled in the positive controls fed a high level of vitamin E (100 ppm dl-α-tocopherol), and there was a further increase when vitamin E was deleted. Neither iron nitrilotriacetate administration nor choline deficiency had any effect on the dG-MDA content of liver DNA. Carbon tetrachloride had a lowering effect. The failure of iron or carbon tetrachloride administration and of vitamin E deficiency to increase liver dG-MDA is consistent with their failure in previous experiments to affect the urinary excretion of dG-MDA. In contrast, these forms of peroxidative stress produce large increments in the urinary excretion of MDA adducts with lysine, reflecting increased formation and degradation of MDA-modified proteins. DNA appears to be protected from modification by MDA produced at extranuclear sites. The frequency of dG-MDA in different tissues of 4-month-old rats varied markedly: brain ≫ liver > kidneys and testes. Higher concentrations of dG-MDA were found in the liver and kidneys, but not the testes, of 25-month-old rats. The determinants of the concentration of dG-MDA in DNA merit further investigation.