Recently, research on natural antioxidants has become increasingly active in various fields. Accordingly, numerous articles on natural antioxidants, including polyphenols, flavonoids, vitamins, and volatile chemicals, have been published. Assays developed to evaluate the antioxidant activity of plants and food constituents vary. Therefore, to investigate the antioxidant activity of chemical(s), choosing an adequate assay based on the chemical(s) of interest is critical. There are two general types of assays widely used for different antioxidant studies. One is an assay associated with lipid peroxidations, including the thiobarbituric acid assay (TBA), malonaldehyde/high-performance liquid chromatography (MA/HPLC) assay, malonaldehyde/gas chromatography (MA/GC) assay, beta-carotene bleaching assay, and conjugated diene assay. Other assays are associated with electron or radical scavenging, including the 2,2-diphenyl-1-picrylhydrazyl (DPPH) assay, 2,2'-azinobis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) assay, ferric reducing/antioxidant power (FRAP) assay, ferrous oxidation-xylenol orange (FOX) assay, ferric thiocyanate (FTC) assay, and aldehyde/carboxylic acid (ACA) assay. In this review, assays used recently were selected for extended discussion, including discussion of the mechanisms underlying each assay and its application to various plants and foods.

Total chlorogenic acids of nine isomers from seven commercial green and roasted coffee beans ranged from 34.43 +/- 1.50 to 41.64 +/- 3.28 mg/g and from 2.05 +/- 0.07 to 7.07 +/- 0.16 mg/g, respectively. Methanol/water (7:3) extracts from four commercial green coffee beans roasted at different conditions (230 degrees C, 12 min; 24 degrees C, 14 min; 250 degrees C, 17 min; and 250 degrees C, 21 min) were also analyzed for chlorogenic acids. The total chlorogenic acid found in green coffee beans ranged from 86.42 +/- 2.04 to 61.15 +/- 1.40 mg/g. Total chlorogenic acids present were reduced in accordance with the intensity of roasting conditions. When green beans were roasted at 230 degrees C for 12 min and at 250 degrees C for 21 min, total chlorogenic acid content was reduced to nearly 50% and to almost trace levels, respectively. The results indicate that roasting conditions play an important role in chlorogenic acid content in roasted coffee beans. A general correlation between total caffeoylquinic acids and pH was observed.

The volatile chemicals in dichloromethane extracts from green coffee beans, roasted at 230 degrees C for 12 min (light), at 240 degrees C for 14 min (medium), at 250 degrees C for 17 min (city), or at 250 degrees C for 21 min (French), were analyzed by gas chromatography and gas chromatography-mass spectrometry. Among the 52 volatile compounds identified, the major compounds were 5-hydroxymethylfurfural, furfuryl alcohol, and 6-methyl-3,5-dihydroxy-4H-pyran-4-one in light-roasted beans; furfuryl alcohol, 5-hydroxymethylfurfural, and gamma-butyrolactone in medium-roasted beans; furfuryl alcohol, gamma-butyrolactone, and 2-acetylpyrrole in city-raosted beans; and gamma-butyrolactone, furfuryl alcohol, and catechol in French-roasted beans. Furfural derivatives and furanones were yielded in relatively high concentrations under mild roasting conditions and then reduced at higher roasting intensities. More pyridines and pyrroles were formed by high roasting intensities than by mild roasting intensities. Chlorogenic acid degradation products, phenols, and a lactone were produced more by high roasting intensities than by low roasting intensities. The results of the present study suggest that controlling the roasting conditions according to the formation of particular chemicals can prepare a roasted coffee with preferable flavor.

We cloned a salicylic acid/benzoic acid carboxyl methyltransferase gene, OsBSMT1, from Oryza sativa. A recombinant OsBSMT1 protein obtained by expressing the gene in Escherichia coli exhibited carboxyl methyltransferase activity in reactions with salicylic acid (SA), benzoic acid (BA), and de-S-methyl benzo(1,2,3)thiadiazole-7-carbothioic acid (dSM-BTH), producing methyl salicylate (MeSA), methyl benzoate (MeBA), and methyl dSM-BTH (MeBTH), respectively. Compared to wild-type plants, transgenic Arabidopsis overexpressing OsBSMT1 accumulated considerably higher levels of MeSA and MeBA, some of which were vaporized into the environment. Upon infection with the bacterial pathogen Pseudomonas syringae or the fungal pathogen Golovinomyces orontii, transgenic plants failed to accumulate SA and its glucoside (SAG), becoming more susceptible to disease than wild-type plants. OsBSMT1-overexpressing Arabidopsis showed little induction of PR-1 when treated with SA or G. orontii. Notably, incubation with the transgenic plant was sufficient to trigger PR-1 induction in neighboring wild-type plants. Together, our results indicate that in the absence of SA, MeSA alone cannot induce a defense response, yet it serves as an airborne signal for plant-to-plant communication. We also found that jasmonic acid (JA) induced AtBSMT1, which may contribute to an antagonistic effect on SA signaling pathways by depleting the SA pool in plants.

4(5)-Methylimidazole has received the attention of federal and state regulatory agencies because of its carcinogenicity and common presence in foods and beverages. In the present study, the formation of 4(5)-methylimidazole in Maillard reaction model systems consisting of D-glucose/NH(3), L-rhamnose/NH(3), methylglyoxal/NH(3), and methylglyoxal/formaldehyde/NH(3) was investigated. 4(5)-Methylimidazole was formed at levels ranging from 0.49 to 0.71 mg/mL in the d-glucose/NH(3) model system. The formation of 4(5)-methylimidazole was slightly higher in the L-rhamnose/NH(3) system (0.91 mg/mL) than in the d-glucose/NH(3) system (0.71 mg/mL) under the conditions used in the present study. A methylglyoxal/NH(3) system produced significantly higher levels of 4(5)-methylimidazole (5.70 mg/mL), suggesting that methylglyoxal is an important precursor of 4(5)-methylimidazole. Ammonolysis of methylglyoxal, which is one of the glucose degradation products, was proposed to form formamide, which subsequently reacted with 2-aminopropanal (α-aminocarbonyl intermediate) formed from methylglyoxal to give 4- or 5-methylimidazole. The levels of 4(5)-methylimidazole found in commercial cola soft drinks range from 0.30 μg/mL (brand 3) to 0.36 μg/mL (brands 1 and 5).