Squalene was identified by gas chromatography-mass spectrometry and high-performance liquid chromatography (HPLC) spiking analyses in the supercritical CO(2) extracts of freeze-dried abscisic leaves of Terminalia catappa L. When the freeze-dried abscisic, senescent, mature, and immature leaves and seeds were subjected to supercritical CO(2) extraction at 40 degrees C and 3000 psi and HPLC quantitation, squalene contents were 12.29, 2.42, 1.75, 0.9, and 0% in the extracts and corresponding to 1499, 451, 210, 65, and 0 microg/g in the freeze-dried sample, respectively. When the extracts were applied for antioxidative characterization by supplementation in an iron/ascorbate system with linoleic acid and in a pork fat storage system for inhibition of conjugated diene hydroperoxide (CDHP) formation or in a free radical scavenging system with 1,1-diphenyl-2-picryl-hydrazyl (DPPH), the extracts of leaves exhibited potent antioxidative and DPPH scavenging activities and increased with an increase of leaf maturity. However, the seed extracts only exhibited potent inhibition of CDHP formation and very low DPPH scavenging activity.
Natural antimutagens may prevent cancer and are therefore of great interest to oncologists and the public at large. Phytochemicals are potent antimutagen candidates. When the Ames test was applied to examine the antimutagenic potency of supercritical carbon dioxide (SC-CO(2)) extracts of Terminalia catappa leaves at a dose of 0.5 mg/plate, toxicity and mutagenicity were not detected. The antimutagenic activity of SC-CO(2) extracts increased with decreases of temperature (60, 50, and 40 degrees C) and pressure (4000, 3000, and 2000 psi) used for extraction. The most potent antimutagenicity was observed in extracts obtained at 40 degrees C and 2000 psi. At a dose of 0.5 mg of extract/plate, approximately 80% of the mutagenicity of benzo[a]pyrene (B[a]P, with S-9) and 46% of the mutagenicity of N-methyl-N '-nitroguanidine (MNNG, without S-9) were inhibited. Media supplemented with SC-CO(2) extracts at a range of 0-500 microg/mL were used to cultivate human hepatoma (Huh 7) and normal liver (Chang liver) cells. The viability of the cells was assayed by measuring cellular acid phosphatase activity. A dose-dependent growth inhibition of both types of cells was observed. The SC-CO(2) extracts were more cytotoxic to Huh 7 cells than to Chang liver cells. The observation that SC-CO(2) extracts of T. catappa leaves did not induce mutagenicity at the doses tested while exhibiting potent antimutagenicity and were more cytotoxic to human hepatoma cells than to normal liver cells is of merit and warrants further investigation.
Status of equol excretion is likely a reflection of incidence risk of sex-hormone related diseases and deserves research attention. In this study, urine samples collected from 182 volunteers after soymilk ingestion were subjected to equol quantification using GC-MS. As categorized into male, female, and both genders and each category further classified into 6 age (year) categories, namely, <20, 21 to 30, 31 to 40, 41 to 50, >51 and overall, a trend indicating that the younger age the higher equol producer ratio was observed. For the >51 subjects, the corresponding producer ratios of males and females were 35.3% and 50.0%, respectively. Among the volunteers, 20 nonproducers were further recruited to ingest 1000 mL soymilk weekly for 16 wk and urines were analyzed bi-weekly. As resulted, 8 of 20 nonproducers were induced to become equol producers. The observed change of nonproducers to equol producers induced by repeating ingestions of soymilk is of merit from the viewpoint of healthcare.
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