The in Vivo Antioxidant Effects of (-)-Epigallocatechin-3-Gallate Consumption in Healthy Postmenopausal Women Measured by Urinary Excretion of Secondary Lipid Peroxidation Products.

The present study was carried out to determine whether the consumption of epigallocatechin (EGCG), the major bioactive green tea catechin, exerts a positive effect on lowering in vivo lipid peroxidation, a measure of oxidative stress, in healthy postmenopausal women. Urinary excretion of secondary lipid peroxidation products, a measure of in vivo lipid peroxidation, was determined in 40 participants randomly assigned to consume a green tea catechin extract (843.0 ± 44.0 mg EGCG/d) or placebo capsules for 12 months. Urine samples were analyzed for individual polar and nonpolar lipophilic aldehydes and related carbonyl compounds by high-performance liquid chromatography (HPLC) at the beginning and at the end of the 12-month intervention period. Results show that two nonpolar aldehydes, nonanal and decatrienal, were both 48% lower (p < 0.005) following consumption of EGCG. These results indicate that a modest degree of in vivo antioxidant activity exists with long-term EGCG consumption, which could slightly limit oxidative damage associated with lipid peroxidation and the onset and progression of chronic diseases.

Kinetics of Forming Aldehydes in Frying Oils and Their Distribution in French Fries Revealed by LC-MS-Based Chemometrics.

In this study, the kinetics of aldehyde formation in heated frying oils was characterized by 2-hydrazinoquinoline derivatization, liquid chromatography-mass spectrometry (LC-MS) analysis, principal component analysis (PCA), and hierarchical cluster analysis (HCA). The aldehydes contributing to time-dependent separation of heated soybean oil (HSO) in a PCA model were grouped by the HCA into three clusters (A1, A2, and B) on the basis of their kinetics and fatty acid precursors. The increases of 4-hydroxynonenal (4-HNE) and the A2-to-B ratio in HSO were well-correlated with the duration of thermal stress. Chemometric and quantitative analysis of three frying oils (soybean, corn, and canola oils) and French fry extracts further supported the associations between aldehyde profiles and fatty acid precursors and also revealed that the concentrations of pentanal, hexanal, acrolein, and the A2-to-B ratio in French fry extracts were more comparable to their values in the frying oils than other unsaturated aldehydes. All of these results suggest the roles of specific aldehydes or aldehyde clusters as novel markers of the lipid oxidation status for frying oils or fried foods.

Whole grain consumption has a modest effect on the development of diabetes in the Goto-Kakisaki rat.

Epidemiological evidence suggests that whole grain intake is associated with reduced risk of type 2 diabetes. However, studies of individual whole grains on the prevention of type 2 diabetes are lacking. The objective of the present study was to examine the effect of different whole grains on type 2 diabetes in an animal model of type 2 diabetes, the Goto-Kakisaki (GK) rat. GK rats were fed either a basal diet or a whole grain-containing diet for 5 months. Whole grain diets contained 65 % whole grain flours of wheat, barley, oats or maize. After 2 months of feeding, fasting plasma glucose concentrations were lower in the wheat, barley and oats groups, compared with the basal group, whereas glycated Hb was significantly greater in the wheat group compared with other groups. Feeding of whole barley and maize increased plasma C-peptide concentrations compared with whole wheat at 2 months. There was a trend in the improvement of insulin resistance with a consumption of barley and oats diets at 2 months (P = 0·06) compared with the basal diet. Oxidative stress markers, urinary thiobarbituric acid-reactive substances and 8-isoprostane, did not improve with whole grain intake at 2 months. At 5 months, whole grain diets did not differ from the basal diet in glycaemic control, insulin secretion, oxidative stress and preservation of pancreatic ß-cell mass. These results suggest that the consumption of whole grains may offer modest benefit early in the development of type 2 diabetes, but this benefit is lost with further development of the disease.

Quantitative analysis of the oxidative DNA lesion, 2,2-diamino-4-(2-deoxy-beta-D-erythro-pentofuranosyl)amino]-5(2H)-oxazolone (oxazolone), in vitro and in vivo by isotope dilution-capillary HPLC-ESI-MS/MS.

A major DNA oxidation product, 2,2-diamino-4-[(2-deoxy-beta-D-erythro-pentofuranosyl)amino]-5(2H)-oxazolone (oxazolone), can be generated either directly by oxidation of dG or as a secondary oxidation product with an intermediate of 8-oxo-7,8-dihydro-2'-deoxyguanosine (8-oxo-dG). Site-specific mutagenesis studies indicate that oxazolone is a strongly mispairing lesion, inducing approximately 10-fold more mutations than 8-oxo-dG. While 8-oxo-dG undergoes facile further oxidation, oxazolone appears to be a stable final product of guanine oxidation, and, if formed in vivo, can potentially serve as a biomarker of DNA damage induced by oxidative stress. In this study, capillary liquid chromatography-electrospray ionization tandem mass spectrometry (HPLC-ESI-MS/MS) methods were developed to enable quantitative analysis of both 8-oxo-dG and oxazolone in DNA from biological sources. Sensitive and specific detection of 8-oxo-dG and oxazolone in enzymatic DNA hydrolysates was achieved by isotope dilution with the corresponding 15N-labeled internal standards. Both nucleobase adducts were formed in a dose-dependent manner in calf thymus DNA subjected to photooxidation in the presence of riboflavin. While the amounts of oxazolone continued to increase with the duration of irradiation, those of 8-oxo-dG reached a maximum at 20 min, suggesting that 8-oxo-dG is converted to secondary oxidation products. Both lesions were found in rat liver DNA isolated under carefully monitored conditions to minimize artifactual oxidation. Liver DNA of diabetic and control rats maintained on a diet high in animal fat contained 2-6 molecules of oxazolone per 10(7) guanines, while 8-oxo-dG amounts in the same samples were between 3 and 8 adducts per 10(6) guanines. The formation of oxazolone lesions in rat liver DNA, their relative stability in the presence of oxidants and their potent mispairing characteristics suggest that oxazolone may play a role in oxidative stress-mediated mutagenesis.

Conjugated linoleic acid, cis-9,trans-11, is a substrate for pulmonary 15-lipoxygenase-1 in rat.

The objective of this study was to determine whether two of the major conjugated linoleic acid (CLA) isomers, cis-9,trans-11 (c9,t11) and trans-10,cis-12 (t10,c12), are possible substrates for pulmonary 15-lipoxygenase-1 (15-LOX-1) and, therefore, they are also involved in the production of 13(S)-hydroxyoctadecadienoic acid [13(S)-HODE] in biological systems. 13(S)-HODE, a major bioactive metabolite of linoleic acid, is an important intracellular signal agent and is involved in cell proliferation and differentiation in various biological systems. Nordihydroguaiaretic acid (NDGA), a known LOX inhibitor, was used as a control for measuring 15-LOX-1 enzyme activity. It was found that c9,t11-CLA was 25% as active as linoleic acid as a substrate for 15-LOX-1; however, t10,c12-CLA was not a substrate for 15-LOX-1 as measured by 13(S)-HODE production. The authenticity of the production of 13(S)-HODE from c9,t11 as a substrate was established by isolation and cochromatography with pure standard on HPLC using non-radioactive and [14C]-c9,t11-CLA.

Effect of high stearic acid containing fat on markers for in vivo lipid peroxidation.

The objective of this study was to determine the effect of high stearic acid (SA) diets versus high polyunsaturated fatty acid (PUFA) diets on several measures of lipid peroxidation in vivo. Sprague-Dawley rats were fed diets that differed only in the fat source (8% by weight) for 19 weeks. High SA fats were beef tallow (BT) and cocoa butter (CB), high PUFA fats were soybean oil (SO) and menhaden oil (MO). Urine was analyzed for lipophilic aldehydes, the secondary products of lipid peroxidation, by HPLC. Decreases (P<0.05) were found for 4 nonpolar lipophilic aldehydes and related carbonyl compounds (NPC) and 4 polar lipophilic aldehydes and related carbonyl compounds (PC) when the BT-fed group was compared to the SO-fed group. Decreases were also found to be significant for total NPC (P<0.01) and total PC (P<0.05) between BT and SO-fed groups. Serum increase in resistance to oxidation (P<0.01) was found in the BT group when compared to the SO group. The differences in urine and serum measurements in the present experiment indicate lower level of lipid peroxidation in vivo due to the consumption of high SA containing BT diet compared to high PUFA containing SO diet without raising serum triglycerides and cholesterol levels significantly for the BT-fed groups.

Nonradiometric HPLC measurement of 13(S)-hydroxyoctadecadienoic acid from rat tissues.

A major bioactive metabolite of linoleic acid formed by the action of 15-lipoxygenase-1 is 13(S)-hydroxy-cis-9, trans-11-octadecadienoic acid (13(S)-HODE). 13(S)-HODE is an important intracellular signal agent and is involved in cell proliferation and differentiation in various biological systems. Separation and quantification of 13(S)-HODE from biological materials has previously been achieved only by using radiolabeled linoleic acid as the substrate and two serially connected or two separate HPLC columns to achieve separation of 13(S)-HODE. In the current method, separation and quantification of 13(S)-HODE was achieved by use of a normal-phase HPLC and a solvent system containing hexane/isopropanol/acetonitrile/acetic acid (800/8/30/1, v/v) using isocratic elution with detection at 235 nm. With the currently described method, good separation from unreacted interfering compounds and quantification for 13(S)-HODE were achieved within 35 min with a minimum detection limit of 0.5 ng per injection.

Protection of superoxide-induced cholesterol oxidation by antioxidants in protic conditions.

It was found in previous experiments that superoxide in the presence of added hydrogen peroxide in protic conditions produces oxysterols. The oxysterols formed under these conditions were 7beta-ketocholesterol, 7alpha-hydroxycholesterol and 7beta-hydroxycholesterol. In the present experiments, the inhibitory effects of three antioxidants, alpha-tocopherol (alpha-T), butylated hydroxyanisole (BHA) and butylated hydroxytoluene (BHT), on the oxidation of cholesterol in the presence of superoxide anion, water and hydrogen peroxide were investigated. It was found that BHA had the highest antioxidant activity on cholesterol oxidation, followed by alpha-T and BHT. The presence of antioxidants markedly retarded the formation of 7-ketocholesterol. The formation of 7beta-hydroxycholesterol or 7alpha-hydroxycholesterol was also reduced, but to a lesser degree.