Studies on the constituents of Chenopodium quinoa seeds: isolation and characterization of new triterpene saponins

Six triterpenoid saponins were isolated from the edible grain quinoa, which is seeds of Chenopodium quinoa (Chenopodiaceae). Following are their structures: phytolaccagenic acid 3-O-[alpha-L-arabinopyranosyl-(1' '-->3')-beta-D-glucuronopyranosyl]-28-O-beta-D-glucopyranoside (1); phytolaccagenic acid 3-O-[beta-D-glucopyranosyl-(1' '-->3')-alpha-L-arabinopyranosyl]-28-O-beta-D-glucopyranoside (2); phytolaccagenic acid 3-O-[beta-D-glucopyranosyl-(1' "-->3' ')-beta-D-xylopyranosyl-(1' '-->2')-beta-D-glucopyranosyl]-28-O-beta-D-glucopyranoside (3); phytolaccagenic acid 3-O-[beta-D-glucopyranosyl-(1' "-->2' ')-beta-D-glucopyranosyl-(1' '-->3')-alpha-L-arabinopyranosyl]-28-O-beta-D-glucopyranoside (4); oleanolic acid 3-O-[alpha-L-arabinopyranosyl-(1' '-->3')-beta-D-glucuronopyranosyl]-28-O-beta-D-glucopyranoside (5); and oleanolic acid 3-O-[beta-D-glucopyranosyl-(1' '-->3')-alpha-L-arabinopyranosyl]-28-O-beta-D-glucopyranoside (6). The oleanane-type saponins (5, 6) were isolated for the first time in this plant, two of the phytolaccagenane (1, 3) were new compounds and two (2, 4) were previously found in quinoa. The structures were characterized on the basis of hydrolysis and spectral evidence, including 1D- and 2-D NMR (HMQC and HMBC) and ESI-MS analyses.     

凤凰单枞茶树吸收土壤铬和锰的规律研究

[目的]探究凤凰单枞茶树对土壤中铬和锰的吸收规律,为综合防治茶园土壤Cr和Mn污染提供科学依据。[方法]对粤东揭阳市揭东县宝山茶园凤凰单枞茶树和土壤进行采样,运用原子吸收分光光度计法测定茶叶、茶茎和茶根中Cr和Mn的含量,并采用乙酸提取法和原子吸收分光光度计法测定土壤样品中酸可提取态Cr和Mn的含量。[结果]茶树各器官Cr含量由高到低分布情况依次为:茶根>茶茎>茶叶;而Mn含量分布情况依次为:茶叶>茶茎>茶根。茶叶中Cr和Mn含量与土壤中酸可提取态Cr和Mn呈显著正相关性,茶茎中Cr含量与土壤中酸可提取态Cr也呈显著正相关性;茶叶、茶茎和茶根中的Cr含量均与土壤中有机质含量呈显著正相关性,茶根中的Mn含量与土壤pH值呈显著负相关性,茶叶中的Mn与土壤有机质呈显著正相关性。[结论]通过调节土壤酸可溶解态Cr和Mn,土壤pH值和有机质含量,从而控制茶树吸收Cr和Mn的量,可以达到防治茶叶重金属污染,提高茶叶质量的目的。     

New oleanane saponins in Chenopodium quinoa

Six triterpenoid saponins were isolated from the seeds of Chenopodium quinoa (Chenopodiaceae). Their structures were as follows: phytolaccagenic acid 3-O-[alpha-L-arabinopyranosyl-(1' '-->3')-beta-D-glucuronopyranosyl]-28-O-beta-D-glucopyranoside (1); spergulagenic acid 3-O-[beta-D-glucopyranosyl-(1-->2)-beta-D-glucopyranosyl-(1-->3)-alpha-L-arabinopyranosyl-28-O-beta-D-glucopyranoside (2); hederagenin 3-O-[beta-D-glucopyranosyl-(1-->3)-alpha-L-arabinopyranosyl]-28-O-beta-D-glucopyranoside (3); phytolaccagenic acid 3-O-[beta-D-glucopyranosyl-(1-->4)-beta-D-glucopyranosyl-(1-->4)-beta-D-glucopyranosyl]-28-O-beta-D-glucopyranoside (4); hederagenin 3-O-[beta-D-glucopyranosyl-(1-->4)-beta-D-glucopyranosyl-(1-->4)-beta-D-glucopyranosyl]-28-O-beta-D-glucopyranoside (5); and spergulagenic acid 3-O-[alpha-L-arabinopyranosyl-(1' '-->3')-beta-D-glucuronopyranosyl]-28-O-beta-D-glucopyranoside (6). Saponins 5 and 6 are new. The structures were characterized on the basis of hydrolysis and spectral evidence, including IR, UV, optical rotations, 1D- and 2D-NMR (HMQC and HMBC), ESIMS, and FABMS analyses.     

HPLC analysis of naturally occurring methylated catechins, 3' '- and 4' '-methyl-epigallocatechin gallate, in various fresh tea leaves and commercial teas and their potent inhibitory effects on inducible nitric oxide synthase in macrophages

(-)-Epigallocatechin-3-gallate (EGCG), a major polyphenol of green tea, undergoes substantial biotransformation to species that includes the methylated compounds. Recent studies have demonstrated that the methylated EGCG has many biological activities. In this study, we have investigated the composition of the three O-methylated EGCG derivatives, (-)-epigallocatechin-3-O-(3-O-methyl)gallate (3' '-Me-EGCG), (-)-epigallocatechin-3-O-(4-O-methyl)gallate (4' '-Me-EGCG) and (-)-4'-methyl epigallocatechin-3-O-(4-O-methyl)gallate (4',4' '-di-Me-EGCG) in tea leaves which were picked from various species and at various seasons, ages of leaves, locations, and fermentation levels. Higher levels of 3' '-Me-EGCG and 4' '-Me-EGCG were detected in Chinshin-Kanzai (a species of Camellia sinensis) cultivated in the mountain area of Sansia, Taipei County, Taiwan. Also, these O-methylated EGCG levels were found to be higher in autumn and winter than in spring and summer. The young leaves were found to be richer in the O-methylated compounds than old leaves and the amount of O-methylated EGCG was higher in unfermented longjin green tea than in semifermented oolong tea. However, the fermented black tea and puerh tea did not contain these compounds. 4',4' '-diMe-EGCG could not be detected in either fresh tea leaves or commercial tea leaves. We also found that 3' '-Me-EGCG has a higher inhibitory effect on the nitric oxide generation and inducible nitric oxide synthase (iNOS) expression as compared with EGCG, while 4' '-Me-EGCG and 4',4' '-di-Me-EGCG were less effective.     

Inhibitory effects of tea catechins and O-methylated derivatives of (-)-epigallocatechin-3-O-gallate on mouse type IV allergy

The inhibitory effects of tea catechins, the O-methylated derivatives of (-)-epigallocatechin-3-O-gallate (EGCG), and the polyphenol extracts from tea leaves (Camellia sinensis L.) on oxazolone-induced type IV allergy in male ICR mice were investigated. Four major tea catechins and two O-methylated derivatives, (-)-epigallocatechin-3-O-(3-O-methyl)gallate (EGCG3' 'Me) and (-)-epigallocatechin-3-O-(4-O-methyl)gallate (EGCG4' 'Me), showed significant inhibitory effects on mouse type IV allergy after a percutaneous administration at a dose of 0.13 mg/ear. Among tea catechins, the compounds including galloyl moieties, such as EGCG and (-)-epicatechin-3-O-gallate (ECG), showed the strongest inhibitory activities on mouse type IV allergy. The inhibitory activities of EGCG3' 'Me and EGCG4' 'Me were higher than that of EGCG at a dose of 0.05 mg/ear. Polyphenol extract from tea leaves of Benihomare cultivar, which includes EGCG3' 'Me, strongly inhibited mouse type IV allergy after percutaneous administration in comparison with that from Yabukita cultivar, which does not include EGCG3' 'Me, at doses of 0.05 and 0.13 mg/ear. EGCG3' 'Me is thought to contribute, at least in part, to the inhibitory ability of Benihomare tea leaves on mouse type IV allergy. EGCG and the polyphenol extracts from Benihomare and Yabukita tea leaves also inhibited mouse type IV allergy by oral administration at 1 h before the sensitization and at 1 h before the challenge with oxazolone. Therefore, daily intake of tea drinks could have potential to prevent type IV allergy.     

Changes of isoflavones during the growth cycle of Lupinus albus

The objective of this study was to monitor the changes in isoflavone content in different plant organs (leaves, stems, roots) during the crop growth stage of three cultivars of Lupinus albus (white lupin) under field conditions, taking into account sowing time effects (autumn and early spring) and cultivar effects. Three sampling dates (from late vegetative to late grain growth stages) were evaluated. Seven isoflavones and four flavonoids were identified by LC-ESI-MS analysis. The isoflavone content was higher in leaves than in stems, and it was highest before flowering, whereas it decreased during maturity. Autumn-sown plants showed higher isoflavone content than early spring-sown plants, especially in late vegetative and early reproductive stages. Genistein 7- O-glucoside was the main isoflavone of leaves and stems in the late vegetative stages of early spring sowing, whereas genistein was the main isoflavone under autumn sowing. Variation among cultivars affected only marginally the total isoflavone content. No isoflavones were detected in seeds.     

Seasonal,spatial, and interspecific variation in quercetin in Apocynum venetum and Poacynum hendersonii,Chinese traditional herbal teas

Quercetin is of particular importance as it has been found to have functions of suppressing tumors, reducing blood pressure, and scavenging free radicals. It is one of the major flavonoids in Apocynum venetum and Poacynum hendersonii, whose leaves have long been used as traditional herbal teas in China and Japan. Both species are also cultivated as fiber plants because of their outstanding quality of phloem fiber in stems. To obtain high output of both quercetin and fiber, it is necessary to optimize harvesting time for their leaves. Thus, understanding the developmental patterns of quercetin in leaves and fiber in stems is crucial to achieving this goal. In the present study, temporal and interspecific variations in quercetin in the leaves between A. venetum and P. hendersonii and spatial variation among P. hendersonii populations were studied by HPLC during the period from April to October in 1999. The results show that the content of quercetin in both species reached its highest level in summer and its lowest in autumn. The quercetin content in the leaves of P. hendersonii was generally higher than that of A. venetum no matter when their leaves were harvested. There was significant difference in quercetin content among three geographical populations of P. hendersonii, which might be the result of climatic difference-cooler climate might favor accumulation of quercetin in the leaves of P. hendersonii. Furthermore, the developmental patterns of total phenolics in the leaves of the two species were the same as that of the quercetin, that is, summer is an optimal harvesting season for both quercetin and other phenolics. The results obtained here suggest that P. herdersonii is a better material for herbal tea or pharmaceutical purposes, and that the best harvest time of its leaves should be summer.     

Enhancement of Rooibos (Aspalathus linearis) aqueous extract and antioxidant yield with fungal enzymes

The leaves and stems of the Rooibos plant ( Aspalathus linearis) are used for the production of an herbal tea known for its health promoting properties, which have been linked to its flavonoid content but which is substantially reduced by the traditional processing method employed. Selected food-grade fungi were screened for their potential to improve the yield of soluble matter extracted from rooibos plant material. Fungal cocktails of hydrolyzing enzymes enhanced either the yield of soluble solids ( Lentinula edodes and Rhizopus oryzae cultured in yeast peptone-wheat straw medium) or the yield in antioxidants from fermented rooibos ( R. oryzae cultured in potato dextrose or yeast peptone-wheat straw medium). When applied to green rooibos, L. edodes (cultured in yeast peptone-wheat straw medium) enhanced the release of soluble solids as well as color formation, leading to semifermented rooibos with a relatively high aspalathin content, compared to fermented rooibos.     

Phenolic metabolites from honeybush tea (Cyclopia subternata)

Cyclopia subternata is one of the 24 Cyclopia species that are used to brew honeybush tea, a unique South African herbal beverage with a pleasant taste and flavor. It contains various antioxidants, very low tannin content, and no caffeine. Many health properties are associated with regular consumption of the tea. Honeybush infusions have been noted as a tonic for colds and influenza, catarrh, and pulmonic tuberculosis and is becoming well-known for its effectiveness in alleviating menopausal symptoms in women. "Unfermented" leaves of C. subternata contain pinitol, shikimic acid, p-coumaric acid, 4-glucosyltyrosol, epigallocatechin gallate, the isoflavone orobol, the flavanones hesperedin, narirutin, and eriocitrin, a glycosylated flavan, the flavones luteolin, 5-deoxyluteolin, and scolymoside, the xanthone mangiferin, and the flavonol C-6-glucosylkaempferol. The structures were elucidated by spectroscopic and spectrometric analysis.     

Puerins A and B, two new 8-C substituted flavan-3-ols from Pu-er tea

Pu-er tea is a special treated fermented tea produced from crude green tea, which is prepared from the leaves of Camellia sinensis var. assamica. It is a traditional beverage having been used in China, particularly the southern areas, for a long time. Chemical investigation led to the identification of two new 8-C substituted flavan-3-ols, puerins A (1) and B (2), and two known cinchonain-type phenols, epicatechin-[7,8-bc]-4alpha-(4-hydroxyphenyl)-dihydro-2(3H)-pyranone (3) and cinchonain Ib (4), together with other seven known phenolic compounds, 2,2',6,6'-tetrahydroxydiphenyl (5), (-)-epicatechin-8-C-beta-d-glucopyranoside (6), (-)-epicatechin (EC) (7), (-)-epigallocatechin (EGC) (8), (+/-)-gallocatechin (GC) (9), gallic acid (10), and myricetin (11), in addition to caffeine (12). Their structures were determined by spectroscopic methods. The compounds 1-5, which might be formed in the postfermentative process of Pu-er tea, were isolated from tea and theaceous plants for the first time.     

Correlations of oil and protein with isoflavone concentration in soybean [Glycine max (L.) Merr.

Twelve isoflavones were detected by high-performance liquid chromatography in seeds of 17 soybean [Glycine max (L.) Merrill] cultivars grown at three locations. 6' '-O-Malonyldaidzin and 6' '-O-malonylgenistin together constituted 71-81% of total isoflavones, which ranged in concentration from 2038 to 9514 microg/g and averaged 5644 microg/g across locations and cultivars. The total as well as several individual isoflavones had a moderate negative correlation with oil across locations and cultivars. Six cultivars had a moderate or strong negative correlation of total isoflavones with oil. Five cultivars had a moderate or strong positive correlation of total isoflavones with protein. These results suggest that judicious selection of germplasm for soybean breeding may facilitate development of soybean lines with desirable isoflavone concentrations.     

Natural fungicides from Ruta graveolens L. leaves,including a new quinolone alkaloid

Bioassay-directed isolation of antifungal compounds from an ethyl acetate extract of Ruta graveolens leaves yielded two furanocoumarins, one quinoline alkaloid, and four quinolone alkaloids, including a novel compound, 1-methyl-2-[6'-(3' ',4' '-methylenedioxyphenyl)hexyl]-4-quinolone. The (1)H and (13)C NMR assignments of the new compound are reported. Antifungal activities of the isolated compounds, together with 7-hydroxycoumarin, 4-hydroxycoumarin, and 7-methoxycoumarin, which are known to occur in Rutaceae species, were evaluated by bioautography and microbioassay. Four of the alkaloids had moderate activity against Colletotrichum species, including a benomyl-resistant C. acutatum. These compounds and the furanocoumarins 5- and 8-methoxypsoralen had moderate activity against Fusarium oxysporum. The novel quinolone alkaloid was highly active against Botrytis cinerea. Phomopsis species were much more sensitive to most of the compounds, with P. viticola being highly sensitive to all of the compounds.     

茶园施肥模式对茶叶黄酮类及糖苷类代谢物含量的影响

  【目的】  研究施肥模式对茶叶生化成分含量和黄酮类及糖苷类代谢物累积的影响,为实现茶叶优质高产提供科学依据。  【方法】  田间试验设在四川茶树种植典型区域—雅安,试验连续进行了3年,设置不施肥 (CK)、常量化肥 (N 585 kg/hm2,TF)、有机肥替代25%化肥氮 (OF)、减施25%氮肥 (SF) 4个处理。分析了不同处理茶叶主要品质成分含量,并基于非靶向代谢组学检测结果分析了黄酮类及糖苷类代谢物累积差异。  【结果】  与CK处理相比,TF、OF、SF处理茶氨酸、游离氨基酸含量分别提高了7.22%～13.40%、23.15%～25.50%,可溶性糖和茶多酚含量分别降低了8.17%～13.86%、6.08%～11.49%,表儿茶素、表没食子儿茶素、芸香苷等10种代谢物累积水平显著下降。与TF处理相比,SF处理茶氨酸、咖啡碱和水浸出物含量分别降低了5.45%、5.97%、8.91%,茶多酚、可溶性糖、游离氨基酸含量和酚氨比无显著差异;芍药素-3-O-葡萄糖苷、肉桂萜醇C1-19-葡萄糖苷、查耳酮樱花素等7种代谢物累积水平显著降低,川陈皮素、牡荆素-4'-O-α-L-吡喃鼠李糖苷、(S)-橙花醇-3-O-[a-L-鼠李糖吡喃醇-(1->2)-β-D-吡喃葡萄糖苷]和Ranupenin3-rutinoside等4种代谢物累积水平显著增加。与TF处理相比,OF处理茶叶咖啡碱含量降低了5.73%;茶氨酸、游离氨基酸、可溶性糖、茶多酚、水浸出物含量差异不显著;10种黄酮苷、2种黄酮、6种萜苷、5种甾体皂甙和2种脂酰苷共25种代谢物累积水平显著增加,以川陈皮素、飞燕草素-3-(6-对香豆酰基半乳糖苷)增幅较大,分别为2280.20%、1355.11%。4个处理相比,OF处理下有15种黄酮类及糖苷类代谢物累积水平显著高于CK、TF和SF处理。  【结论】  有机肥替代25%化肥氮降低了茶叶咖啡碱含量,提高了川陈皮素、飞燕草素、槲皮素、芹黄素、山奈酚、牡荆素等多种黄酮类代谢物和去氢大豆皂甙I、珠子参苷R2、孕甾烷甙等多种糖苷组分累积水平。减施25%氮肥降低了茶氨酸、咖啡碱、水浸出物含量和糖苷类代谢物 (如杨梅酮-3-O-洋槐糖苷、芍药素-3-O-葡萄糖苷、肉桂萜醇C1-19-葡萄糖苷等)累积水平,对茶多酚、可溶性糖、游离氨基酸含量影响较小。有机肥替代部分化肥氮有利于茶叶黄酮类及糖苷类物质累积,单纯减少氮肥投入会降低茶叶品质。     

Polyphenolic content in different plant parts of soy cultivars grown under natural conditions

Roots, cotyledons, leaves, stems, pods, and seeds of three soy cultivars were analyzed for their content of isoflavones, flavonols, coumarins, and phenolic acid derivatives with three samplings during a three-month period. The extracts were analyzed by HPLC/DAD and HPLC/MS, allowing us to confirm the presence of daidzein and genistein derivatives as the major isoflavones and to characterize coumarins, most flavonols and phenolic acid derivatives. Seeds exhibited the highest content of isoflavones: 12.61 g/kg of dry weight (DW) in cv. Emiliana; 8.97 g/kg of DW in cv. Elvir; 4.49 g/kg of DW in cv. Kure, and roots are the only part with coumarins, ranging from 4.08 g/kg of DW (cv. Emiliana) to 1.29 g/kg of DW (cv. Elvir) for the longest sampling period. Leaves, pods, and stems have flavonols, and in particular leaves showed: 7.28 g/kg of DW in cv. Emiliana; 6.57 g/kg of DW in cv. Elvir; 7.08 g/kg of DW in cv. Kure. The high content of isoflavones found in the seeds could be ascribed to the natural conditions under which the soy plants were grown.     

不同施氮水平下不同氮利用效率小黑麦植株氮素积累分配特性

采用土培盆栽试验,以小黑麦氮高效利用品种‘Clxt82’、‘PI429186’和氮低效利用品种‘Clxt74’为材料,研究0(不施氮)、0.033 g(N)·kg-1(低氮)和0.066 g(N)·kg-1(正常氮)3个不同施氮水平下,各生长时期氮素在器官间和器官内不同功能性氮素分配的特性。结果表明:氮高效利用品种在氮素不足的条件下优势更明显,抽穗期高效利用品种和低效利用品种间生物量的差异随施氮量的增加而减小,在不施氮、低氮和正常供氮时‘Clxt82’、‘PI429186’地上部生物量分别为‘Clxt74’的1.55倍、1.19倍、1.06倍和1.79倍、1.35倍、1.30倍。不同生育时期,小黑麦氮积累量均随施氮量的增加而显著增加,低氮和正常供氮处理,在分蘖期、拔节期氮高效利用品种氮积累量均显著高于低效利用品种,而在抽穗期差异则不大。随施氮量的增加,氮素在叶片和穗部的分配比例减小,在茎的分配比例增大;分蘖期和拔节期,氮高效利用品种茎中氮素分配比例小于低效利用品种,叶片氮素分配比例则大于低效利用品种。抽穗期氮高效利用品种穗部氮素分配比例大于低效品种,而叶部则相反。各生育时期各器官不同形态氮素含量总体上随施氮量的增加而增加。不施氮和低氮处理,拔节期氮高效品种‘Clxt82’、‘PI429186’叶片营养性氮含量是低效品种‘Clxt74’的1.31倍、1.76倍和1.12倍、1.35倍,而结构性氮含量则是低效品种的86.12%、64.01%和80.82%、71.51%;抽穗期氮高效品种‘Clxt82’、‘PI429186’叶片营养性氮含量是低效品种‘Clxt74’的1.01倍、1.11倍和1.04倍、1.13倍,结构性氮含量为低效品种‘Clxt74’的74.99%、63.08%和75.78%、62.84%;各时期品种间功能性氮素含量差异不大。低氮条件下氮高效利用品种通过降低结构性氮素含量、增加营养性氮素含量来满足氮素的利用和体内循环。     

Epimerization of tea catechins and O-methylated derivatives of (-)-epigallocatechin-3-O-gallate: relationship between epimerization and chemical structure

Epimerization at C-2 of O-methylated catechin derivatives and four major tea catechins were investigated. The epimeric isomers of (-)-epicatechin (I), (-)-epicatechin-3-O-gallate (II), (-)-epigallocatechin (III), (-)-epigallocatechin-3-O-gallate (IV), and (-)-epigallocatechin-3-O-(3-O-methyl)gallate (V) in green tea extracts increased time-dependently at 90 degrees C. The epimerization rates of authentic tea catechins in distilled water are much lower than those in tea infusion or in pH 6.0 buffer solution. The addition of tea infusion to the authentic catechin solution accelerated the epimerization, and the addition of ethylenediaminetetraacetic acid, disodium salt (Na(2)EDTA) decreased the epimerization in the pH 6.0 buffer solution. Therefore, the metal ions in tea infusion may affect the rate of epimerization. The proportions of the epimers to authentic tea catechins [III, IV, V, and (-)-epigallocatechin-3-O-(4-O-methyl)gallate (VI)] in pH 6.0 buffer solution after heating at 90 degrees C for 30 min were 42.4%, 37.0%, 41.7%, and 30.4%, respectively. These values were higher than those of I and II (23.5% and 23.6%, respectively). The O-methylated derivatives at the 4'-position on the B ring of IV and VI were hardly epimerized. These results suggest that the hydroxyl moiety on the B ring of catechins plays an important role in the epimerization in the order 3',4',5'-triol type > 3',4'-diol type > 3',5'-diol type.     

Effects of the environment, cultivar, maturity, and preservation method on red clover isoflavone concentration

Red clover (Trifolium pratense L.) contains isoflavones that are of interest because of their benefits for human health as well as their adverse effects on the fertility of farm animals. A series of field experiments was conducted in Sainte-Anne-de-Bellevue, QC, Canada, to determine the effects of the environment, cultivar, plant maturity, plant part, and preservation method on the concentration of the two predominant isoflavones in red clover, formononetin and biochanin A. In a multi-year, multisite trial, the total isoflavone concentration in 10 cultivars ranged between 8,923 and 12,753 microg g(-1) of DM averaged across sites, harvests, and years. Despite strong environmental effects, the cultivar "Start" consistently had the lowest isoflavone concentrations, with few differences observed among other cultivars. Across stages of maturity, leaves were found to have the highest isoflavone concentration followed by stems and inflorescences (11,970, 4,896, and 3,297 microg g(-1) of DM, respectively). Changes in isoflavone concentrations with increasing maturity varied depending on the plant part. Overall, highest isoflavone concentrations were found in leaves and stems during the vegetative stages, with the formononetin concentration declining until plants initiated flowering, especially in stems, with concentrations then stabilized in both parts. Upon initiation, inflorescences contained similar isoflavone concentrations than leaves, but concentrations decreased rapidly during flower development to fall even below those observed in stems. Inflorescences then had isoflavone concentrations that were as much as 11 times lower than leaves. Fresh herbage contained higher formononetin and total isoflavone concentrations than did silage and hay (14,464, 12,200, and 11,604 microg g(-1) of DM, respectively). The isoflavone concentration in field-grown red clover is thus high but can be affected by a range of agronomic factors.     

Effect of temperature, elevated carbon dioxide, and drought during seed development on the isoflavone content of dwarf soybean [Glycine max (L.) Merrill] grown in controlled environments

The effects of elevated temperature, carbon dioxide, and water stress on the isoflavone content of seed from a dwarf soybean line [Glycine max (L.) Merrill] were determined, using controlled environment chambers. Increasing the temperature from 18 degrees C during seed development to 23 degrees C decreased total isoflavone content by about 65%. A further 5 degrees C increase to 28 degrees C decreased the total isoflavone content by about 90%. Combining treatments at elevated temperature with elevated CO(2) (700 ppm) and water stress to determine the possible consequences of global climate change on soybean seed isoflavone content indicated that elevated CO(2) at elevated temperatures could partially reverse the effects of temperature on soybean seed isoflavone content. The addition of drought stress to plants grown at 23 degrees C and elevated CO(2) returned the total isoflavone levels to the control values obtained at 18 degrees C and 400 ppm CO(2). The promotive effects of drought and elevated CO(2) at 23 degrees C on the 6' '-O-malonygenistin and genistin levels were additive. The individual isoflavones often had different responses to the various growth conditions during seed maturation, modifying the proportions of the principal isoflavones. Therefore, subtle changes in certain environmental factors may change the isoflavone content of commercially grown soybean, altering the nutritional values of soy products.     

Flavone C-glycoside,phenolic acid,and nitrogen contents in leaves of barley subject to organic fertilization treatments

From the leaves of barley, Hordeum vulgare, one new flavone C-glucoside and three known flavone glucosides were isolated and characterized by (1)H and (13)C NMR and MALDI-TOF-MS. The novel flavone C-glucoside was isovitexin 7-O-beta-[6' "-O-(E)-p-coumaroyl]glucoside (6' "-coumaroylsaponarin), and the known compounds were isovitexin 7-O-beta-[6' "-O-(E)-feruloyl]glucoside, isoorientin 7-O-beta-[6' "-O-(E)-feruloyl]glucoside, and tricin 7-O-beta-glucoside. The sum of all the flavone glycosides and soluble phenolic acids in the leaves decreased with increased rate of plant nutrients given in animal manure and with increased crop yield. All of the major phenylpropanoids showed the same general response to nutrient level. The concentration of nitrogen in the leaves was not directly related to nutrient application or to contents of phenylpropanoids.     

Bilberry adulteration using the food dye amaranth

Vaccinium myrtillus or bilberry fruit is a commonly used herbal product. The usual method of determining the anthocyanin content is a single-wavelength spectrophotometric assay. Using this method, anthocyanin levels of two extracts were found to be 25% as claimed by the manufacturers. When high-performance liquid chromatography (HPLC) was used, however, one extract was found to contain 9% anthocyanins probably not derived from V. myrtillus but from an adulterant. This adulterant was subsequently identified, using HPLC, mass spectroscopy, and nuclear magnetic resonance, as amaranth, that is, 3-hydroxy-4-[(4-sulfo-1-naphthalenyl)azo]-2,7-naphthalenedisulfonic acid trisodium salt-a synthetic dark red sulfonic acid based naphthylazo dye. As described in this study, if deliberate adulteration occurs in an extract, a single-wavelength spectrophotometric assay is inadequate to accurately determine the levels of compounds such as anthocyanins. Detection of deliberate adulteration in commercial samples thus requires the use of alternative, more sophisticated, methods of analysis such as HPLC with photodiode array detection as a minimum.