Determination of anthocyanins from camu-camu (Myrciaria dubia) by HPLC-PDA, HPLC-MS, and NMR

Camu-camu [Myrciaria dubia (HBK) McVaugh] is a small fruit native to the Amazonian rain forest. Its anthocyanin profile has now been investigated for the first time. Fruits from two different regions of the S?o Paulo state, Brazil, were analyzed. The major anthocyanins were isolated by high-speed countercurrent chromatography. HPLC-PDA, HPLC-MS/MS, and 1H NMR were used to confirm the identity of the main anthocyanins of camu-camu. Cyanidin-3-glucoside was identified as the major pigment in the fruits from both regions, representing 89.5% in the fruits produced in Iguape and 88.0% in those from Mirandópolis, followed by the delphinidin-3-glucoside, ranging between 4.2 and 5.1%, respectively. Higher total anthocyanin contents were detected in the fruits from Iguape (54.0 +/- 25.9 mg/100 g) compared to those from Mirandópolis (30.3 +/- 6.8 mg/100 g), most likely because of the lower temperatures in the Iguape region.     

Chemical and functional characterization of Italian propolis obtained by different harvesting methods

The composition and antioxidant activity of Italian poplar propolis obtained using three harvesting methods and extracted with different solvents were evaluated. Waxes, balsams, and resins contents were determined. Flavones and flavonols, flavanones and dihydroflavonols, and total phenolics were also analyzed. To characterize the phenolic composition, the presence of 15 compounds was verified through HPLC-MS/MS. The antioxidant activity was evaluated through 1,1-diphenyl-2-picrylhydrazyl radical and reducing power assays. The ability of propolis to inhibit lipid oxidation was monitored by analyzing hydroperoxide and TBARS formation in lipids incorporated into an oil-in-water (O/W) emulsion. Acetone shows the highest extraction capacity. Wedge propolis has the highest concentration of active phenolic compounds (TP = 359.1 ± 16.3 GAEs/g; TFF = 5.83 ± 0.42%; TFD = 7.34 ± 1.8%) and seems to be the most promising for obtaining high-value propolis more suitable to prepare high-quality dietary supplements (TBARS = 0.012 ± 0.009 mmol std/g; RP = 0.77 ± 0.07 TEs/g).     

Cuban and Brazilian red propolis: botanical origin and comparative analysis by high-performance liquid chromatography-photodiode array detection/electrospray ionization tandem mass spectrometry

Chemical composition of propolis depends on the specificity of the local flora at the site of collection and thus on the geographic and climatic characteristics of this place. This paper describes a comparative analysis of Cuban red propolis (CRP), Brazilian red propolis (BRP), and Dalbergia ecastophyllum exudates (DEE) by high-performance liquid chromatography with diode-array detection and tandem mass spectrometry. The aim of this study was to investigate the overall chemical profile and the botanical origin of red propolis and to suggest similarities and differences between samples collected in different tropical regions. Isoliquiritigenin (1), liquiritigenin and naringenin (2 and 17), isoflavones (3-4 and 16), isoflavans (5-7 and 18), and pterocarpans (8-13) were detected in CRP, BRP, and DEE, whereas polyisoprenylated benzophenones (PPBs) guttiferone E/xanthochymol (14a,b) and oblongifolin A (15) were detected only in BRP. Pigments responsible for the red color of DEE and red propolis were also identified as two C30 isoflavans, the new retusapurpurin B (19) and retusapurpurin A (20). PPBs and pigments were isolated and unambiguously characterized by 1D and 2D NMR analysis. These results show that red propolis samples from different tropical zones have a similar chemical composition. DEE is the main red propolis source, but the presence of PPBs in BRP suggests the contribution of different botanical sources for Brazilian samples. This chemical information is important for quality control of red propolis and its commercial products and for biological study.     

添加生物质炭对黄棕壤和红壤上油菜生长的影响

通过盆栽试验,探究施用不同比例的生物质炭对黄棕壤和红壤理化性质和油菜生长发育的影响。结果表明:黄棕壤和红壤在添加1%生物质炭后,两者的pH值、有效磷、速效钾、有机质含量均比对照显著增加。油菜根、茎、叶、角果、籽粒等干物重均增加,其中黄棕壤上油菜籽粒产量比对照提高114.8%,达到显著水平。添加生物质炭后,两种土壤上油菜各部位氮含量均有所下降,磷含量也呈降低趋势,钾含量则有所升高,其中红壤上油菜的变化幅度较黄棕壤显著。由于油菜干物重增加,两种土壤上油菜氮磷钾积累量在加入生物质炭后也有明显地提高。因此,施用1%生物质炭可以促进油菜生长,并有助于油菜对磷、钾等养分的吸收。     

Betalains in red and yellow varieties of the Andean tuber crop ulluco (Ullucus tuberosus)

The betalain pigments in ulluco (Ullucus tuberosus), a tuberous crop native to the Andes, have been investigated for the first time using LC-DAD-ESI-MS-MS(2) analyses. Five red, yellow, and red-spotted accessions introduced into New Zealand as a new food crop plus two red tetraploid lines were investigated. Thirty-two different betalains were identified. Both the yellow and red tubers were rich in yellow betaxanthins, and the most prominent among the 20 identified were histidine-betaxanthin, arginine-betaxanthin and glutamine-betaxanthin. Arginine-betaxanthin has been reported to occur naturally only once before and was found in yellow ulluco but not in the red tubers. Twelve betacyanins were found in red tubers, with roughly 50% of this content being betanin/isobetanin. Betacyanin levels were up to 70 microg/g fresh weight in red tubers, but were below quantifiable levels in yellow tubers. Betaxanthin levels were up to 50 microg/g fresh weight in yellow tubers. Interference by betacyanins in measuring levels of betaxanthins by visible spectrophotometry is discussed. Low concentrations of betalains were detected in leaves, whereas stems contained total levels similar to the tubers, with dopamine-betaxanthin and betanin being the major pigments. This is the first report describing both the betacyanin and betaxanthin patterns in a plant from the Basellaceae family.     

生物炭对我国南方红壤和黄棕壤理化性质的影响

为了探讨生物炭对不同土壤的改良效果,采用盆栽试验,研究了施用生物炭对我国南方两种代表性土壤(红壤和黄棕壤)理化性质的影响及其动态变化差异。结果表明:强酸性红壤施用生物炭能明显提高pH而降低其酸度,同时增加土壤的有机质、速效磷、速效钾和碱解氮含量,且随着生物炭施用量(生物炭量/土壤量:0、0.5%、1.0%、2.0%)的增加,改良效果不断加强;弱酸性黄棕壤施用生物炭也提高了土壤pH、有机质、速效磷、速效钾含量,但对该土壤中的碱解氮含量无明显影响。不同生物炭用量的效应存在较大差异,在2.0%时对两种土壤各理化性质影响均表现为最明显,红壤pH平均增加0.61,有机质、速效磷、速效钾、碱解氮分别平均提高203.4%、369.3%、368.0%、30.4%,而黄棕壤pH、有机质、速效磷、速效钾分别平均增加0.55、124.2%、57.5%、50.3%。因而,相同用量的生物炭对红壤的改良效应好于黄棕壤,且施用生物炭对两种土壤速效钾含量影响最大,其次是有机质、pH、速效磷、碱解氮。     

Isolation and structural characterization of new anthocyanin-derived yellow pigments in aged red wines

Two newly formed yellow pigments that revealed unique spectral features were detected and isolated from an aged Port red wine by TSK Toyopearl HW-40(s) gel chromatography and characterized by UV-visible spectrophotometry, 1H NMR and 13C NMR, and mass spectrometry (LC-ESI/MS). The UV-vis spectra of these pigments showed maximum absorption at 478 nm that is significantly hypsochromically shifted from those of original grape anthocyanins and other pyranoanthocyanins, exhibiting a more yellow hue in acidic solution. The structures of these pigments correspond to methyl-linked pyranomalvidin 3-glucoside and its respective coumaroyl glucoside derivative. They were shown to arise from the reaction between acetoacetic acid and genuine grape anthocyanins. Isolation and NMR identification using 1D and 2D NMR techniques are reported for the first time for this new family of anthocyanin-derived yellow pigments occurring in red wines.     

Chemical properties and pedogenesis of yellowish/yellow brown forest soils in the submontane and mountain zones of Kyushu district,southwestern Japan

Abstract

This study aimed to clarify pedogenetic processes and classification of yellowish Brown Forest Soils according to the Classification of Forest Soils in Japan and the Yellow Brown Forest soils according to the Unified Soil Classification System of Japan in the warm and cool temperate forest of Kyushu district, Japan. In addition, the study aimed to clarify a problem with the Unified Soil Classification System of Japan. Thirty-six soil profiles of Brown Forest Soils, including 13 yellowish Brown Forest Soils and 15 Yellow Brown Forest soils, were compared with regard to their chemical properties and the relationship with climatic conditions was assessed. The yellowish Brown Forest Soils had thin A horizons, low pH and low levels of free oxides in the B horizons, and a low amount of silica and a high aluminum and iron to silica ratio. These features were related to the paleo reddish weathering. The immaturely developed A horizon of the yellowish Brown Forest Soils was caused by these weathered, low-activity substances. The Yellow Brown Forest soils had low levels of active iron oxides and a low activity ratio of free iron oxides compared with the Haplic Brown Forest soils in the same thermal climatic conditions. The activity ratio of free iron oxides was correlated to mean annual air temperature with the carbon stocks and with many other chemical properties. Accordingly, classification of Brown Forest Soils was clearer according to thermal climatic conditions. The activity ratio of free iron oxides can become an effective index that distinguishes Yellow Brown Forest soils under warm temperate lucidophyllous forest and Haplic Brown Forest soils under cool temperate broad-leaved deciduous forest with considerable vertical soil zonality.     

Structure determination of partially deuterated carotenoids from intrinsically labeled vegetables by HPLC-MS and 1H NMR

The structures of biosynthetic deuterated carotenoids in labeled vegetables were investigated: (all-E)-lutein and (all-E)-beta-carotene from spinach, and (all-E)-beta-carotene and (all-E)-alpha-carotene from carrots. The vegetables were grown hydroponically using a nutrient solution enriched with deuterium oxide (D(2)O) and were extracted using matrix solid-phase dispersion (MSPD). Deuterium enrichment in the carotenoid molecules was determined by liquid chromatography-mass spectrometry (LC-MS). (all-E)-Lutein and (all-E)-beta-carotene in spinach showed partial deuteration from (2)H(1) to (2)H(12), with the abundance maximum at (2)H(5). (all-E)-beta-Carotene and (all-E)-alpha-carotene from carrots showed partial deuteration from (2)H(1) to (2)H(17), with the abundance maximum at (2)H(11). The (1)H NMR spectra of the four deuterated carotenoids showed additional signals for all methyl groups and decreased signal intensity for the olefinic protons and the methylene protons in the ring. These differences are due to isotopic effects and are based on the substitution of protons by deuterium atoms. The deuteration was distributed randomly throughout the carotenoid molecules.     

Variation in the selection of Rhizobium trifolii by varieties of red and white clover

The selection of Rhizobium trifolii by six varieties of red and white clover was examined under laboratory conditions. The varieties differed in their selection of strains and there was considerable variation within any one variety. The implications of such variation are discussed with particular reference to clover seed inoculation.     

Separation, characterization, and quantitation of phenolic acids in a little-known blueberry (Vaccinium arctostaphylos L.) fruit by HPLC-MS

The aim of this study was the qualitative and quantitative determination of free, ester, glycoside, and ester-bound phenolic acids in the blueberry (Vaccinium arctostaphylos L.) fruit. A method for the determination of the profile of phenolic acids of four different phenolic fractions in the fruit was developed using high-performance liquid chromatography-mass spectrometry (HPLC-MS). Thirteen compounds (gallic, protocatechuic, p-hydroxybenzoic, m-hydroxybenzoic, gentisic, chlorogenic, p-coumaric, caffeic, ferulic, syringic, sinapic, salicylic, and trans-cinnamic acids) were identified and quantified in the berry. These experimental results showed that the predominant phenolic acid in the fruit of V. arctostaphylos is caffeic acid in free and insoluble ester-bound forms and p-coumaric acid in soluble ester and glycoside forms. Seven phenolic acids were identified as hydroxybenzoic acid derivatives (HBAs) and four as hydroxycinnamic acid derivatives (HCAs). Total content of HBAs and HCAs in the four phenolic fractions constituted 30.1 and 69.9% of the free, 27.9 and 72.1% of the ester, 24.7 and 75.3% of the glycoside, and 51.7 and 48.3% of the ester-bound forms, respectively. Total phenolics as the sum of individual phenolic acids identified is 698.5 ng/g of fresh weight (fw) for the free, 3399.2 ng/g of fw for the ester, 3522.1 ng/g of fw for the glycoside, and 3671.6 ng/g of fw for the ester-bound phenolic fractions. The present results were compared with reported levels of phenolic acids in the fruits of different Vaccinium species. These data suggest that the fruit can be considered as a potentially good dietary source of phenolic acids.     

Identification of anthocyanin-flavanol pigments in red wines by NMR and mass spectrometry

Three newly formed Port wine pigments were isolated by Toyopearl HW-40(s) gel chromatography and semipreparative HPLC. Furthermore, the pigments were identified by mass spectrometry (LC/MS) and NMR techniques (1D and 2D). These anthocyanin-derived pigments showed UV-visible spectra different from those of the original grape anthocyanins. These pigments correspond to malvidin 3-glucoside linked through a vinyl bond to either (+)-catechin, (-)-epicatechin, or procyanidin dimer B3 [(+)-catechin-(+)-catechin]. NMR data of these pigments are reported for the first time.     

Assessment of the protein quality of nine northern adapted yellow and brown seed coated soybean cultivars by amino acid analysis

Accurate and detailed amino acid determinations were carried out on nine northern adapted soybean cultivars to ascertain whether their amino acid profiles could be used as potentially useful indices for assessing their protein quality. The cultivars were Maple Amber, Maple Donovan, Maple Glen, Maple Isle, Maple Presto, Maple Ridge, and three brown seed coat near-isogenic lines, Maple Presto Brown, Maple Ridge Brown, and Maple Arrow Brown. Their total protein and amino acid composition were compared with those of an established cultivar, Maple Arrow. Mean protein values for the new cultivars ranged from 30.1 to 33.1% compared to Maple Arrow, which was 33.2%. The total nitrogen content was also variable among these cultivars, ranging from 5.0 to 5.4%. All nine Maple series soybean cultivars were higher in their essential amino acid (EAA) content, that is, EAA(9) = 45.2-46.5%, than the FAO/WHO reference protein pattern value of EAA(9) = 33.9%, for a 2-5-year-old child. Each of the nine new soybean cultivars was limited only in methionine and to a lesser extent in valine and isoleucine and had a protein digestibility corrected amino acid score of 91% for all cultivars, compared to the value of egg protein (97%). These results suggest that the most accurate evaluation of protein quality in soybeans, and possibly other legumes and cereals, is by the protein digestibility-corrected amino acid score.     

土地利用方式影响黄棕壤有机质周转和分配

土地利用方式影响土壤有机质平衡和周转。选取南京信息工程大学农业气象实验站长期定位试验的3种利用方式即水稻小麦(稻麦)轮作、大豆小麦(豆麦)轮作和自然植被(休闲)土壤作为研究对象。将土壤筛分团聚体和进行密度分组,测定全土有机质含量和13C丰度、各组分有机质含量,计算全土有机质周转率和半衰期,以揭示土地利用方式对土壤有机质含量及稳定性的影响。结果表明:与休闲处理相比,稻麦轮作对全土有机质含量无明显影响,而豆麦轮作降低了全土有机质含量。稻麦轮作促进了微团聚体黏结形成大团聚体,增加了土壤团聚体的平均重量直径。密度组分的结果表明,稻麦轮作中土壤游离态轻组和闭蓄态轻组有机质含量均未见明显变化,这是保持全土有机质含量无明显增加的主控因素。稻麦轮作处理土壤有机质的高周转率和较短半衰期有利于土壤有机质的更新和养分的释放。豆麦轮作破坏了大团聚体,且有机质源输入较少,土壤有机质的周转率相对较低,不利于有机质的周转和保持其含量稳定。相关分析的结果证实,粒径大于1 mm团聚体、游离态轻组和闭蓄态轻组中有机质含量都与全土有机质含量显著正相关。上述结果表明,在黄棕壤区,稻麦轮作不仅可以保持土壤有机质的含量,还会促进土壤有机质的更新周转,从而提高土壤肥力水平,是未来农业可持续发展需要的科学有效耕作方式。     

Determination of ETU in tomatoes and tomato products by HPLC-PDA: evaluation of cleanup procedures

An HPLC-PDA method for the determination of ethylenethiourea (ETU), the main degradation product of the organic fungicides ethylene bis(dithiocarbamate)s (EBDCs), in tomatoes and tomato products is reported. Solid-matrix liquid-liquid (l-l) partitioning and separatory funnel l-l partitioning for the cleanup were examined. The effect of salt addition, pH, and phase ratio on analyte recovery at the cleanup step was studied. It was found that solid-matrix l-l partitioning afforded higher precision and more selective separation of the analyte. According to the method proposed, the samples were extracted with methanol/water (3:1, v/v) and cleaned up on an Extrelut 20 column. ETU was eluted with dichloromethane and separated on a reversed phase HPLC column. For tomato products with degrees Brix > 20 further purification through silica cartridge was adopted. The method was validated over the following ranges of concentrations: 0.01-0.5 mg/kg for tomatoes, 0.01-0.1 mg/kg for tomato juice, and 0.05-0.25 mg/kg for tomato paste. The accuracy (recoveries > 70%) and the precision obtained (%RSD < 10%) were satisfactory.     

Separation, characterization and quantification of phenolic compounds in blueberries and red and black currants by HPLC-DAD-ESI-MSn

The phenolic profile of four blueberry varieties (Vaccinium corymbosum L., cv. Toro, Legacy, Duke and Bluecrop) and two varieties (Rosenthal and Rovada) of red currants (Ribes rubrum L.) and black currants (Ribes nigrum L.) cultivated in Macedonia have been analyzed using HPLC coupled to diode-array detection and tandem mass spectrometry with electrospray ionization. A complex profile of anthocyanins, flavonols, flavan-3-ols and hydroxycinnamic acid derivatives has been assayed in acetone-acetic acid (99:1, v/v) extracts. Anthocyanins comprised the highest content of total phenolic compounds in currants (>85%) and lower and variety dependent in blueberries (35-74%). Hydroxycinnamic acid derivatives comprised 23-56% of total phenolics in blueberries and 1-6% in currants. Chlorogenic acid was the major hydroxycinnamic acid in blueberries, only in the Legacy variety, two malonyl-caffeoylquinic acid isomers were major components. Flavonols, mainly quercetin and myricetin glycosides, were a minor group, but glucosides of laricitrin and syringetin were also detected in the blueberry varieties counting for 10-34% of total flavonols. From flavan-3-ols, catechin was detected in most samples; the dimer B2 was specific for blueberries whereas epigallocatechin was detected in currants.     

Isolation, characterization, and antioxidant activity of bromophenols of the marine red alga Rhodomela confervoides

A total of 19 naturally occurring bromophenols, with six new and 13 known structures, were isolated and identified from the methanolic extract of the marine red alga Rhodomela confervoides. The new compounds were identified by spectroscopic methods as 3,4-dibromo-5-((methylsulfonyl)methyl)benzene-1,2-diol (1), 3,4-dibromo-5-((2,3-dihydroxypropoxy)methyl)benzene-1,2-diol (2), 5-(aminomethyl)-3,4-dibromobenzene-1,2-diol (3), 2-(2,3-dibromo-4,5-dihydroxyphenyl)acetic acid (4), 2-methoxy-3-bromo-5-hydroxymethylphenol (5), and (E)-4-(2-bromo-4,5-dihydroxyphenyl)but-3-en-2-one (6). Each compound was evaluated for free radical scavenging activity against DPPH (α,α-diphenyl-β-dipicrylhydrazyl) and ABTS [2,2'-azinobis(3-ethylbenzothiazoline-6-sulfonic acid)diammonium salt] radicals. Most of them exhibited potent activities stronger than or comparable to the positive controls butylated hydroxytoluene (BHT) and ascorbic acid. The results from this study suggest that R. confervoides is an excellent source of natural antioxidants, and inclusion of these antioxidant-rich algal components would likely help prevent the oxidative deterioration of food.     

Chemical and sensory characterization of DOC red wines from Marche (Italy) related to vintage and grape cultivars

Monomeric phenols, color and copigmentation parameters, pigments with different chemical structure, tannin, glucose, fructose, glycerol, ethanol, and organic acids were determined in DOC red wines from Marche (Italy), obtained during three different vintages ranging from 1996 to 2000. The intensity of the bitter and astringent tastes of the wines was determined with panel tastings. Lacrima di Morro and Vernaccia di Serrapetrona (obtained from local cultivars) were different from Rosso Piceno, Rosso Piceno Superiore, and Rosso Conero (produced from different percentages of Sangiovese and Montepulciano). Vernaccia, a red, sweet, "spumante" wine, was an outlier. Lacrima showed a low tannin content, a high content of small pigments and phenols, and a high ratio of copigmented color, which persisted after 3 years of aging. The chemical determinations accounted for a high percentage of variability of measured panel astringency, copigmented color, and measured wine absorbance at 520 nm. It was not possible to create a predictive model for bitterness.     

High-resolution MAS NMR and chemometrics: characterization of the ripening of Parmigiano Reggiano cheese

We performed a preliminary NMR investigation on grated Parmigiano Reggiano cheese of different ripening ages. Principal component analysis (PCA) and discriminant analysis (DA) were able to successfully group the analyzed samples according to their respective ages.