Determination of free amino compounds in betalainic fruits and vegetables by gas chromatography with flame ionization and mass spectrometric detection

Amino acids and amines are the precursors of betalains. Therefore, the profiles of free amino compounds in juices obtained from cactus pears [Opuntia ficus-indica (L.) Mill. cv. Bianca, cv. Gialla, and cv. Rossa], pitaya fruits [Selenicereus megalanthus (K. Schumann ex Vaupel) Moran, Hylocereus polyrhizus (Weber) Britton & Rose, and Hylocereus undatus (Haworth) Britton & Rose], and in extracts from differently colored Swiss chard [Beta vulgaris L. ssp. cicla (L.) Alef. cv. Bright Lights] petioles and red and yellow beets (B. vulgaris L. ssp. vulgaris var. conditiva Alef. cv. Burpee's Golden) were investigated for the first time. Amino compounds were derivatized with propyl chloroformate. While gas chromatography (GC) with mass spectrometry was used for peak assignment, GC flame ionization detection was applied for quantification of individual compounds. Whereas proline was the major free amino compound of cactus pear and pitaya fruit juices, glutamine dominated in Swiss chard stems and beets, respectively. Interestingly, extremely high concentrations of dopamine were detected in Swiss chard stems and beets. Furthermore, the cleavage of betaxanthins caused by derivatization in alkaline reaction solutions is demonstrated for the first time. Amino acids and amines thus released might increase the actual free amino compound contents of the respective sample. To evaluate the contribution of betaxanthin cleavage to total amino acid and amine concentration, isolated betaxanthins were derivatized according to the "EZ:faast" method prior to quantification of the respective amino compounds released. On a molar basis, betaxanthin contribution to overall amino compound contents was always below 6.4%.     

Function of sodium and potassium in growth of sodium-loving Amaranthaceae species

We observed that the growth of three Amaranthaceae species was promoted by sodium (Na), in the order dwarf glasswort (Salicornia bigelovii Torr.) >> Swiss chard (Beta Burgaris L. spp. cicla cv. Seiyou Shirokuki) > table beet (Beta vulgaris L. spp. vulgaris cv. Detroit Dark Red). In the present study, these Na-loving plants were grown in solutions containing 4 mol m^?3 nitrate nitrogen (NO₃-N) and 100 mol m^?3 sodium chloride (NaCl) and potassium chloride (KCl) under six Na to potassium (K) ratios, 0:100, 20:80, 40:60, 60:40, 80:20 and 100:0, to elucidate the function of Na and K on specific characteristics of Na-loving plants. The growth of dwarf glasswort increased with increasing Na concentration of the shoot, and the shoot dry weight of plants grown in 100:0 Na:K was 214% that of plants grown at 0:100. In Swiss chard and table beet, growth was unchanged by the external ratio of Na to K. The water content was not changed in Swiss chard or table beet by the external Na to K ratio. These observations indicate that both Na and K have a function in osmotic regulation. However, dwarf glasswort could not maintain succulence at 0:100; therefore, Na has a specific function in dwarf glasswort for osmotic regulation to maintain a favorable water status, and the contribution of K to osmotic regulation is low. NO₃-N uptake was promoted by Na uptake in dwarf glasswort and Swiss chard. NO₃-N uptake and transport to shoots was optimal at 100:0 in dwarf glasswort and at 80:20 in Swiss chard. These functions are very important for the Na-loving mechanism, and the contribution of K was lower in dwarf glasswort than in Swiss chard.     

Identification and quantification of betalains from the fruits of 10 mexican prickly pear cultivars by high-performance liquid chromatography and electrospray ionization mass spectrometry

Qualitative and quantitative analyses of betalain pigments in 10 cultivars/lines of prickly pear (Opuntia spp.) fruit grown in Mexico were conducted with reverse phase high-performance liquid chromatography-diode array detection (HPLC-DAD) coupled with electrospray mass spectrometry (ESI-MS). Betacyanins and betaxanthins were identified by comparison with the UV/vis and mass spectrometric characteristics as well as the retention times of semisynthesized reference betaxanthins. Data revealed that the ratio and concentration of betalain pigments are responsible for the color in the different cultivars, showing the highest betalains content in the fruit of purple colored Camuesa (O. robusta Wendl.) (8.1 mg/g dry fruit), which is comparable to that found in red beet Beta vulgaris L. ssp. Var. Pablo) (8.6 mg/g dry tissue). Yellow betalains were absent in Reyna (O. alba-carpa) prickly pear cultivar. A total of 24 known/unknown betalains were present in the prickly pear fruit samples studied, including 18 betaxanthins and 6 betacyanins. Our results indicate that prickly pear cultivars can be considered as a potential source of yellow and red natural colorants.     

Effect of pH on the uptake of cadmium,copper, and zinc from soilless media containing sewage sludge

Abstract

Swiss chard (Beta vulgaris cicla ’Fordhook Giant'), bean (Phaseolus vulgaris ’Hawksbury Wonder'), and Agrostis capillaris ’Parys’ were grown in soilless media composed of Pinus radiata bark and 0–41% by volume of dried sewage sludge containing [in mg/kg] cadmium (Cd) [22], copper (Cu) [1,116], and zinc (Zn) [1,121]. Sublots of all mixtures were adjusted to pH 4.5, 5.5, or 6.5. Shoot weights were not reduced by sludge addition but those of Swiss chard and bean shoots declined with pH. Shoot contents of Cu, Cd, and Zn increased with decreasing pH. Shoot Cu concentrations were always sub‐toxic, and in Swiss chard toxicity symptoms were not observed until shoot Zn content reached 1,300 mg/kg. Cadmium in Swiss chard shoots reached 22 mg/kg. Metal concentrations in the shoots were well correlated with total and extractable metals in the media only if pH was included in the regression equations.     

Identification of flavonoids and hydroxycinnamic acids in pak choi varieties (Brassica campestris L. ssp. chinensis var. communis) by HPLC-ESI-MSn and NMR and their quantification by HPLC-DAD

Twenty-eight polyphenols (11 flavonoid derivatives and 17 hydroxycinnamic acid derivatives) were detected in different cultivars of the Chinese cabbage pak choi ( Brassica campestris L. ssp. chinensis var. communis) by HPLC-DAD-ESI-MS(n). Kaempferol was found to be the major flavonoid in pak choi, glycosylated and acylated with different compounds. Smaller amounts of isorhamnetin were also detected. A structural determination was carried out by (1)H and (13)C NMR spectroscopy for the main compound, kaempferol-3-O-hydroxyferuloylsophoroside-7-O-glucoside, for the first time. Hydroxycinnamic acid derivatives were identified as different esters of quinic acid, glycosides, and malic acid. The latter ones are described for the first time in cabbages. The content of polyphenols was determined in 11 cultivars of pak choi, with higher concentrations present in the leaf blade than in the leaf stem. Hydroxycinnamic acid esters, particularly malic acid derivatives, are present in both the leaf blade and leaf stem, whereas flavonoid levels were determined only in the leaf blade.     

红梗叶甜菜种子发芽特性的研究

研究了不同处理条件下红梗叶甜菜种子的发芽特性。结果表明，用一定浓度的双氧水、硼酸、磷酸二氢钾浸种处理红梗叶甜菜种子，可提高种子的发芽率、发芽势和发芽指数，其中以浓度10g／L H2O2、0．5g／L H3BO3和5g／L KH2PO4处理较为适宜；赤霉素处理种子的发芽效果不明显；吸湿一回干处理对红梗叶甜菜种子发芽有良好的促进作用；红梗叶甜菜种球色泽、粒径大小与发芽的关系密切，黄色或黄褐色种球的发芽力显著高于黑色或褐色种球的，大、中粒种子的发芽力显著高于小粒种子的；叶甜菜种子不耐贮藏，发芽力下降较快，使用年限仅为1a。     

Analysis of furanone, pyranone, and new heterocyclic colored compounds from sugar-glycine model Maillard systems

Aqueous sugar (xylose or glucose)-glycine model systems were refluxed for 2 h with the pH maintained at 5. Reverse-phase HPLC of the total reaction products gave two resolved peaks (one of which was colored) for the xylose system and five resolved peaks (two of which were colored) for the glucose system. The components responsible for these peaks were isolated from the ethyl acetate extracts by semipreparative HPLC. Using mainly NMR, the colored compound from the xylose system was identified as the new 2-acetyl-6-(hydroxymethyl)-5,6-dihydro-4H-pyridinone. The colored compounds from the glucose system were most likely to be two novel cis/trans ring isomers of the related new compound 2-acetyl-6-hydroxy-7-(hydroxymethyl)-1,5,6,7-tetrahydro-4H-azepinone+ ++. These compounds are the first one-ring structures isolated from sugar-amino acid model systems that are reported to be colored. Two of the colorless components of the glucose system were identified, mainly by NMR experiments, as the related compounds 4-hydroxy-2-(hydroxymethyl)-5-methyl-3(2H)-furanone and 2, 3-dihydro-3,5-dihydroxy-6-methyl-4H-pyranone. The remaining compound from the glucose system and the colorless compound from the xylose system were identified as 5-(hydroxymethyl)furfural and 4-hydroxy-5-methyl-3(2H)-furanone, respectively.     

Color, betalain pattern, and antioxidant properties of cactus pear (Opuntia spp.) clones

Total phenolics, ascorbic acid, and betalain contents of differently colored cactus pear clones (nine Opuntia ficus-indica [L.] Mill. clones and one O. robusta Wendl. clone) were investigated and related to their respective antioxidant potential assessed by Trolox-equivalent antioxidant capacity (TEAC) and oxygen radical absorbance capacity (ORAC) assays. TEAC and ORAC values were very highly correlated with each other and also with values for total phenolics, betalain contents, and ascorbic acid concentrations. Total phenolics had the greatest contribution to ORAC and TEAC values. High-performance liquid chromatography (HPLC)-diode array detector (DAD)-tandem mass spectrometry (MS/MS) measurements of cactus pear juices permitted the differentiation of the clones based on variations in pigment patterns and betalain concentrations. The red and yellow betalains were absent in lime green colored cactus fruits. The ratio and concentration of these pigments were responsible for the yellow, orange, red, and purple colors in the other clones. Progeny of purple and lime green colored parents were characterized by 12% and 88% of plants bearing lime green and purple fruit, respectively. This implies that the genes for betalain production were lacking in the lime green fruits but could be provided by a parent with a complete set of genes, that is, purple fruits. Besides known pigments typical of Cactaceae, two unexpected betalains were identified. Whereas gomphrenin I was found for the first time in tissues of cactus plants, methionine-betaxanthin has never been described before as a genuine betalain. In addition to their alleged health-promoting properties, various combinations of yellow betaxanthins and red-purple betacyanins may allow the development of new food products without using artificial colorants.     

Growth promotion by sodium in amaranthaceous plants

The amaranthaceous dwarf glasswort, Swiss chard, table beet, spinach and Mexican tea were grown in solutions containing 0, 20, 40, 80, 120, 160, 180 and 200 mol m^?3 sodium chloride (NaCl). Maximum growth and increase of biomass production compared to that at 0 mol m^?3 of dwarf glasswort was observed at 200 mol m^?3 (245%), and Swiss chard (146%), table beet (128%) and spinach (138%) at 80 mol m^?3. The growth of these species increased with increasing sodium (Na) concentration of shoot until it reached 4.18, 2.42, 1.60 and 1.58 mol kg^?1, respectively. These indicate that the order of Na-loving character is dwarf glasswort >> Swiss chard > table beet = spinach. The water contents increased with increasing Na concentration until growth reached maximum in dwarf glasswort and Swiss chard. In these highly Na-loving species, Na is activity transported to shoots and utilized for producing osmotic pressure to absorb water.     

Heavy metals in crops as affected by soil types and sewage sludge rates

Abstract

A municipal sewage sludge was applied at three application rates to three soils in field lysimeters to study the effects of soil and sludge application rate on cadmium (Cd), copper (Cu), nickel (Ni), and zinc (Zn) absorbed by ‘Larker’ barley (Hordeum vulgare L.) and by Swiss chard (Beta vulgaris L. ‘Cicla'). Sludge applied at 20, 40, and 100 Mg/ha oven‐dry equivalent were mixed into the top 0.15 m of soil in each lysimeter. In addition, a nil sludge rate (control) received 125 kg N/ha. Metal contents were relatively low in barley grain, higher in barley straw, and highest in Swiss chard. Metal contents in plants increased with increasing sludge loading. Most plants grown on soil amended with the higher sludge rates were too high in Cd (greater than 0.5 mg/kg of dry feed material) to be suitable for animal consumption. No plant materials tested exceeded the suggested maximum Zn, Cu, or Ni levels.     

Identification of betalains from yellow beet (Beta vulgaris L.) and cactus pear [Opuntia ficus-indica (L.) Mill.] by high-performance liquid chromatography-electrospray ionization mass spectrometry

Betaxanthins, the yellow-orange water-soluble pigments from yellow beet (Beta vulgaris ssp. vulgaris cv. Bejo Zaden) and cactus pear (Opuntia ficus-indica cv. Gialla) have been investigated using an HPLC system compatible with mass spectrometry. Five novel betaxanthins were found and characterized as the immonium adducts of betalamic acid with serine, gamma-aminobutyric acid, valine, isoleucine, and phenylalanine. To enable concentration of betalain samples, desalting was performed by solid-phase extraction. With this technique, betacyanins could be separated from the betaxanthins using the pH-dependent retention characteristics of red and yellow betalains. The betaxanthin fraction was taken for the preparation of betalamic acid as a precursor for semisynthetic standards. The HPLC method was applied to yellow beet and cactus pear, revealing a more complex betalain profile than described earlier, thus proving its suitability for screening of betaxanthin-containing plants as potential sources for natural food colors.     

Zinc accumulation characteristics of some leafy vegetables

Abstract

Twelve leafy vegetables were grown on field plots treated with 0, 10, 50, 100, 200, 400, and 800 pounds Zn per acre incorporated into the 0‐ to 10‐inch soil depth. At harvest maturity the edible parts of the plants were sampled and analyzed for Zn.

Of the 12 crops grown, only Swiss chard and spinach showed any tendency to be Zn accumulators. Even these crops would tolerate extremely high levels of available soil Zn before accumulating what might be considered hazardous Zn concentrations. Swiss chard and spinach were the only crops which showed visible growth depression when grown on plots treated with 800 lbs. Zn per acre.     

Crocetin esters, picrocrocin and its related compounds present in Crocus sativus stigmas and Gardenia jasminoides fruits. Tentative identification of seven new compounds by LC-ESI-MS

Crocetin esters present in saffron (Crocus sativus L.) stigmas and in Gardenia jasminoides Ellis fruit are the compounds responsible for their color. Of the fifteen crocetin esters identified in this study, five new compounds were tentatively identified: trans and cis isomers of crocetin (beta-D-triglucoside)-(beta-D-gentibiosyl) ester, trans and cis isomers of crocetin (beta-D-neapolitanose)-(beta-D-glucosyl) ester, and cis crocetin (beta-D-neapolitanose)-(beta-D-gentibiosyl) ester. The most relevant differences between both species were a low content of the trans crocetin (beta-D-glucosyl)-(beta-D-gentibiosyl) ester, the absence of trans crocetin di-(beta-D-glucosyl) ester in gardenia, and its higher content of trans crocetin (beta-D-gentibiosyl) ester and cis crocetin di-(beta-D-gentibiosyl) ester. With the same chromatographic method it was possible to identify, in a single run, ten glycosidic compounds in saffron extracts with a UV/vis pattern similar to that of picrocrocin; among them, 5-hydroxy-7,7-dimethyl-4,5,6,7-tetrahydro-3H-isobenzofuranone 5-O-beta-D-gentibioside and 4-hydroxymethyl-3,5,5-trimethyl-cyclohexen-2-one 4-O-beta-D-gentibioside were tentatively identified for the first time in saffron. Of these ten glycosides, only the O-beta-D-gentibiosyl ester of 2-methyl-6-oxo-2,4-hepta-2,4-dienoic acid was found in gardenia samples, but it was possible to identify the iridoid glycoside, geniposide.     

Chemical characterization of Allium ursinum L. depending on harvesting time

Sulfur-containing compounds of ramson (Allium ursinum L.) are responsible for its traditional use in terms of culinary and medicinal purposes. Leaves and bulbs were investigated for their contents of cysteine sulfoxides (volatile precursors) as well as volatile compounds released from minced plant material. Plants were analyzed during the whole vegetation period, focused on the months from March to June. Additionally, within the dormancy period bulbs were analyzed again and alliinase activity was determined. The pattern of volatile compounds was analyzed both by SPME/GC-MS and by SDE/GC-MS. Compared to each other, SDE exhibited a wider spectrum of detectable volatile compounds. The quality and quantity of volatiles significantly depended on the time of harvest. The highest amounts of volatile precursors can be gained in March and April, shortly before flowering time (up to 0.4% of total cysteine sulfoxides). The main cysteine sulfoxides were alliin and isoalliin. It has been found that alliinase of A. ursinum exhibited properties similar to those of alliinase of garlic (Allium sativum L.), but differing in terms of substrate specificity.     

HPLC-DAD-MS/MS-ESI screening of phenolic compounds in Pieris brassicae L. Reared on Brassica rapa var. rapa L

The phenolic profiles of cabbage white butterfly ( Pieris brassicae L.; Lepidoptera: Pieridae) at different development stages (larvae, exuviae, and butterfly), its excrements, and its host plant Brassica rapa var. rapa L. were determined by high performance liquid chromatography- diode-array detector-mass spectrometry/mass spectrometry-electrospray ionization (HPLC-DAD-MS/MS-ESI). Twenty-five acylated and nonacylated flavonoid glycosides and ferulic and sinapic acids were identified in host plant, from which only 12 compounds were found in the excrements. In addition, the excrements showed the presence of sulfate flavonoids and other flavonoid glycosides that were not detected in the leaves. In the larvae kept without food for 12 h, only 3 compounds common to the plant material and 2 others, also present in the excrements, were characterized. The results indicate that deacylation, deglycosylation, and sulfating steps are involved in the metabolic process of P. brassicae and that its excrements may constitute a promising source of bioactive compounds, which could be used to take profit of this common pest of Brassica cultures.     

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.     

Anti-inflammatory effects of caper (Capparis spinosa L.) fruit aqueous extract and the isolation of main phytochemicals

Caper (Capparis spinosa L.) fruits have been used as food as well as folk medicine in the treatment of inflammatory disorders, such as rheumatism. The present study was carried out to study the anti-inflammatory activities of C. spinosa L. fruit (CSF) aqueous extract and to isolate main phytochemicals from its bioactive fractions. The CSF aqueous extract were separated into three fractions (CSF1-CSF3) by macroporous adsorption resins. The fractions CSF2 and CSF3 effectively inhibited the carrageenan-induced paw edema in mice. Systematic fractionation and isolation from CSF2+3 led to the identification of 13 compounds (1-13). Their chemical structures were elucidated by spectroscopic analyses including nuclear magnetic resonance (NMR) and mass spectrometry (MS) and literature comparisons. Major compounds found in the bioactive fraction CSF2+3 are flavonoids, indoles, and phenolic acids. To our knowledge, 8 of these 13 compounds (1-4, 6-7, 10, and 13) were identified from caper fruits for the first time. The anti-inflammatory effects of these purified compounds are currently under investigation.     

Elemental Composition of Twelve Plant Species Grown with Irradiated Municipal Sludge

The objective of this research was to determine the availability to plants of elements in sewage sludge irradiated with high-energy electrons for disinfection. Irradiated raw sludge (irradiated with an electron dose of 400 krad) and non-irradiated raw sludge were obtained from the Deer Island Wastewater Treatment Plant of Boston, Massachusetts. The sludges were used to grow 12 plant species under controlled-environmental conditions. The species were: bush bean (Phaseolus vulgaris L.), sweet corn (Zea mays L.), dill (Anethum graveolens L.), lettuce (Lactuca sativa L.), mustard (Brassica juncea Coss.), parsley (Petroselinum crispum Nym.), pumpkin (Cucurbita pepo L.), radish (Raphanus sativus L.), spinach (Spinacia oleracea L.), Swiss chard (Beta vulgaris cicla L.), tomato (Lycopersicon esculentum Mill.), wheat (Triticum aestivum L. em. Thell.). Plants were also grown with inorganic fertilizer added at recommended rates or with tap water. At the end of the experiment (44 days after planting), dry weight and elemental composition of the plants and soil (fine sandy loam, Typic Ustifluvent) were determined. There was no measurable difference in nutrient composition of soil treated with irradiated and non-irradiated raw sludge. In general, plants treated with irradiated raw sludge grew as well as plants treated with non-irradiated raw sludge. Most plants treated with sludge grew as well as plants treated with inorganic fertilizer. Elemental composition of plants treated with irradiated sludge was similar to that of plants treated with non-irradiated sludge. Sludge-treated dill, parsley, spinach, and Swiss chard had high concentrations of one or more of the following heavy elements: Zn, Cu, Ni, Cr. The results showed that plants grown with sludge irradiated with high-energy electrons were similar to plants grown with non-irradiated sludge.     

Identification of flavonol and xanthone glycosides from mango (Mangifera indica L. Cv. "Tommy Atkins") peels by high-performance liquid chromatography-electrospray ionization mass spectrometry

Flavonol O- and xanthone C-glycosides were extracted from mango (Mangifera indica L. cv. "Tommy Atkins") peels and characterized by high-performance liquid chromatography-electrospray ionization mass spectrometry. Among the fourteen compounds analyzed, seven quercetin O-glycosides, one kaempferol O-glycoside, and four xanthone C-glycosides were found. On the basis of their fragmentation pattern, the latter were identified as mangiferin and isomangiferin and their respective galloyl derivatives. A flavonol hexoside with m/z 477 was tentatively identified as a rhamnetin glycoside, which to the best of our knowledge, has not yet been reported in mango peels. The results obtained in the present study confirm that peels originating from mango fruit processing are a promising source of phenolic compounds that might be recovered and used as natural antioxidants or functional food ingredients.