Chaniphun Butryee

Effect of processing on the flavonoid content and antioxidant capacity of Citrus hystrix leaf

The objective of the present study was to compare fresh (F) use and the effects of boiling (B) and deep-fat frying (DF) on the leaf of Citrus hystrix on total phenolic content, the types and amounts of flavonoids and their total antioxidant capacities (TAC), as measured by three different assays: oxygen radical absorption capacity, ferric reducing/antioxidant power, and scavenging effect on the 2,2-diphenyl-1-picrylhydrazyl free radical. Boiling decreased TAC values on the three assays. The amount of total flavonoids calculated as aglycone equivalents of eight identified flavonoids (cyanidin, myricetin, peonidin, quercetin, luteolin, hesperetin, apigenin and isorhamnetin) determined by high-performance liquid chromatography was 1,129 (DF), 1,104 (F) and 549 (B) mg/100 g freeze-dried weight (dry matter exclude fat). Hesperetin was the predominant flavonoid. The total phenolic content expressed as grams of gallic acid equivalents/100 grams fresh weight (excluding fat) was 2.0, 1.9 and 1.8 in F, DF and B samples, respectively. These results suggest that method of processing can significantly affect the content of flavonoids and their TAC values.

Anticlastogenic effect of Eryngium foetidum L. assessed by erythrocyte micronucleus assay.

The aim of this study was to investigate the anticlastogenicity as well as the clastogenicity of Eryngium foetidum leaf (EF) using the in vivo mouse peripheral blood erythrocyte micronucleus assay. Eighty ICR male mice were fed AIN-76 diet supplemented with ground freeze-dried EF at 0.0%, 0.8%, 1.6% and 3.2% for 2 weeks prior to the administration of both direct-acting, mitomycin C (MMC), and indirect-acting, 7, 12-dimethylbenz(a) anthracene (DMBA) clastogens. Peripheral blood samples were collected from mice just before administration of clastogen and at 24 and 48 h thereafter for MMC. Blood samples were collected at the same times and after 72 h for DMBA. Then, reticulocytes in blood samples were counted using fluorescent microscopy. The results indicated that EF had no clastogenic effect in mice. All doses of diets supplemented with EF decreased the number of micronucleated peripheral reticulocytes in all the MMC-treated groups in a dose dependent manner, but significant reduction was found only at 1.6% and 3.2% EF in the DMBA-treated groups. It can be concluded that EF has no clastogenicity, but possesses anticlastogenic potential against both direct- and indirect-acting types of clastogen in mice.

A 24-Weeks Toxicity Study of Eryngium foetidum Linn. Leaves in Mice.

Eryngium foetidum Linn. leaves (EF) are widely used in Thailand and many countries throughout Asia as a culinary seasoning and a traditional medicine. However, adverse effect of high dose consumption in long duration has not been evaluated. The aim of this study was to investigate chronic toxicity of EF in mice. Thirty-two ICR male mice were divided into 4 groups of 8 mice each. The mice were fed AIN-76 rodent diet, or AIN-76 rodent diet supplemented with ground freeze-dried EF at 0.8%, 1.6% and 3.2% that is equivalent to approximately 35, 73 and 155 times that of human consumption, respectively, at 97.5 percentile for a period of 24 weeks. At the end of experiment, the mice were euthanized and blood samples were collected for hematological and biochemical evaluations. Necropsy was performed while visceral organs such as lung, liver, kidneys, spleen etc. were collected, weighed and histopathologically examined. Blood urea nitrogen (BUN) results of mice in 1.6% and 3.2% EF diet groups were significantly higher than the BUN of control group. No significant difference was noted in other biochemical and hematological properties between the treatment groups and control; all results were within normal range. Histopathology of almost all visceral organs showed no significant changes. However, tubulonephrosis and chronic interstitial nephritis were observed in the groups treated with 1.6% and 3.2% EF diet. Body weight was reduced significantly at week 12 to week 20 when compared to the control group while relative kidney weights were significantly increased. In conclusion, the consumption of EF in diet at high doses illustrated the adverse effect on some biochemical parameters and histopathology in mice. Our findings suggested that EF daily consumption for 24 weeks, at higher doses than the 0.8% EF diet (35 times of human consumption), might cause adverse effect on kidney function in mice.

Chemopreventive Effects of Eryngium foetidum L. Leaves on COX-2 Reduction in Mice Induced Colorectal Carcinogenesis

ABSTRACT To investigate the potential effects of Eryngium foetidum Linn. leaves (EF) in colitis-induced colorectal carcinogenesis in mice by azoxymethane (AOM) and dextran sulfate sodium (DSS), 39 ICR male mice were studied and divided into 6 groups. The mice were received a modified AIN-76 diet in Group 1, whereas Group 2 was given an AOM, DSS, and AIN-76 diet. Groups 3 and 4 were fed with 0.8% and 3.2% freeze-dried EF with AIN-76 diets, for 5 wk. Groups 5 and 6 were fed with 0.8% and 3.2% EF diets for 5 wk during AOM/DSS administration. The mice were necropsied at Week 20 and their colons were collected. The results indicated that the incidences of tumors in Groups 2, 5, and 6 was 100%, 75%, and 88%, with multiplicities (mean ±SE) of 3.75 ±0.92, 2.38 ± 0.96 and 4.25 ± 0.79, respectively. Interestingly, there was a significant difference in COX-2 expression in mice received 3.2% EF in their diet, but the proliferative cell nuclear antigen index and iNOS protein expression were not significantly different. We concluded that EF at a dose level of 3.2% in their diet had a preventive effect on colorectal carcinogenesis via the proinflammatory cytokine, COX-2.

Antimutagen Formation and Nitrite Scavenging by Unprocessed or Processed Fruit and Vegetable Fibers

Salmonella typhimurium strains TA 98 and TA 100 were used in carrying out the Ames test for the study of antimutagen formation. A variety of fruit and vegetable fibers were tested upon the mutagen-generating reaction between aminopyrene and sodium nitrite in a gastric simulation. The nitrite-scavenging capability (NSC) of such fibers was also determined. Thai style cooking or the maturity of fruits and vegetables (raw and ripe) had no significant effect on the formation of antimutagen, detected with aminopyrene plus nitrite, or on the NSC. The scavenging activity of fibers was found to range from 0.1 to 0.25 fig nitrite per mg fiber. The incorporation of selected fibers into western-style baked products, and deep-fat frying of starchy foods, resulted in a reduction of the NSC.