Identification and occurrence of tryptamine- and tryptophan-derived tetrahydro-beta-carbolines in commercial sausages

The identification and occurrence of tetrahydro-beta-carbolines were studied in different kinds of commercial sausages including cooked, fresh, dry-fermented, and ripened sausages, such as salamis and Spanish chorizo, salchichon, fuet, and morcilla, both smoked and unsmoked. Four compounds were identified in several sausages by high-performance liquid chromatography-mass spectrometry (HPLC-MS): 1,2,3,4-tetrahydro-beta-carboline-3-carboxylic acid (1), 1-methyl-1,2,3,4-tetrahydro-beta-carboline-3-carboxylic acid diastereoisomers (2a,b), 1,2,3,4-tetrahydro-beta-carboline (3), and 1-methyl-1,2,3,4-tetrahydro-beta-carboline (4). The latter two (3 and 4) are now reported for the first time in meat products. The presence and occurrence of tetrahydro-beta-carbolines were highly variable depending on each particular sample of sausage, and it did not follow a single specific pattern. The concentration range taken as a sum of the four carbolines varied from undetectable levels to 33 microg/g, with the highest content found in ripened, dry-fermented, and smoked sausages (salami, chorizo, and morcilla) and the lowest in cooked sausages (Frankfurt). Formation of tetrahydro-beta-carbolines might occur during elaboration and the ripening process from a chemical condensation between tryptophan or tryptamine and aldehydes (formaldehyde and acetaldehyde). Smoked samples had higher concentrations of formaldehyde-derived 1,2,3,4-tetrahydro-beta-carboline-3-carboxylic acid (1) and 1,2,3,4-tetrahydro-beta-carboline (tryptoline) (3) than those unsmoked. Also, 1 and 3 were more concentrated in the outer part of the sausage, likely to be in contact with smoke. It is concluded that some dry-fermented and/or smoked sausages may be significant dietary sources of tetrahydro-beta-carbolines.     

Identification and occurrence of the novel alkaloid pentahydroxypentyl-tetrahydro-beta-carboline-3-carboxylic acid as a tryptophan glycoconjugate in fruit juices and jams

The novel carbohydrate-derived beta-carboline, 1-pentahydroxypentyl-1,2,3,4-tetrahydro-beta-carboline-3-carboxylic acid, was identified in fruit- and vegetable-derived products such as juices, jams, and tomato sauces. This compound occurred as two diastereoisomers, a cis isomer (the major compound) and a trans isomer, ranging from undetectable amounts to 6.5 microg/g. Grape, tomato, pineapple, and tropical juices exhibited the highest amount of this alkaloid (up to 3.8 mg/L), whereas apple, banana, and peach juices showed very low or nondetectable levels. This tetrahydro-beta-carboline was also found in jams (up to 0.45 microg/g), and a relative high amount was present in tomato concentrate (6.5 microg/g) and sauce (up to 1.8 microg/g). This beta-carboline occurred in fruit-derived products as a glycoconjugate from a chemical condensation of D-glucose and L-tryptophan that is highly favored at low pH values and high temperature. Production, processing treatments, and storage of fruit juices and jams can then release this beta-carboline. Fruit-derived products and other foods containing this compound might be an exogenous dietary source of this glucose-derived tetrahydro-beta-carboline.     

Tetrahydro-beta-carbolines, potential neuroactive alkaloids, in chocolate and cocoa

Tetrahydro-beta-carbolines (THbetaCs), potential neuroactive alkaloids, were found in chocolate and cocoa. 6-Hydroxy-1-methyl-1,2, 3,4-tetrahydro-beta-carboline (6OHMTHbetaC), 1,2,3, 4-tetrahydro-beta-carboline-3-carboxylic acid (THCA), 1-methyl-1,2,3, 4-tetrahydro-beta-carboline-3-carboxylic acid (MTCA) in both diastereoisomers (1S,3S and 1R,3S), and 1-methyl-1,2,3, 4-tetrahydro-beta-carboline (MTHbetaC), besides serotonin and tryptamine biogenic amines, were identified and quantified in dark chocolate, milk chocolate, cocoa, and chocolate-containing cereals by RP-HPLC-fluorescence and HPLC-MS. For each THbetaC, the concentration ranges were determined: 6OHMTHbetaC (0.16-3.92 microg/g), THCA (0.01-0.85 microg/g), 1S,3S-MTCA (0.35-2 microg/g), 1R,3S-MTCA (0.14-0.88 microg/g), and MTHbetaC (nd-0.21 microg/g). The highest content was generally found in chocolates and cocoas, but cereals containing chocolate also showed an appreciable amount of THbetaCs. The possible biological implications of this novel group of alkaloids in chocolate are discussed.     

1-methyl-1,2,3,4-tetrahydro-beta-carboline-3-carboxylic acid and 1,2, 3,4-tetrahydro-beta-carboline-3-carboxylic acid in fruits

1,2,3,4-Tetrahydro-beta-carboline-3-carboxylic acid (THCA) and 1-methyl-1,2,3,4-tetrahydro-beta-carboline-3-carboxylic acid (MTCA), as two diastereoisomers (1S,3S and 1R,3S), occurred in commercial fruits. Citrus fruits exhibited the highest content; other fruits contained very low levels or none at all. The content of MTCA was as follows: orange, 0.35-2.47 microg/g; lemon, 0.15-2.05 microg/g; grapefruit, 1.12-8.37 microg/g; mandarin, 0.57-2.5 microg/g; banana, nd-0.74 microg/g; pear, nd-0.017 microg/g; grape, 0.01-0.22 microg/g, tomato, 0.05-0.25 microg/g; and apple, nd-0.012 microg/g). THCA, if present, usually occurred at <0.05 microg/g. Fruit ripening and softening during storage were accompanied with a significant increase of MTCA, in both pears and bananas. Those and previous results confirm that foods are an important source of tetrahydro-beta-carbolines in humans.     

L-tryptophan reacts with naturally occurring and food-occurring phenolic aldehydes to give phenolic tetrahydro-beta-carboline alkaloids: activity as antioxidants and free radical scavengers

The reaction between the essential amino acid l-tryptophan and flavoring or naturally occurring phenyl and phenolic aldehydes was studied, and the alkaloidal reaction products were characterized by NMR and HPLC-MS. Benzaldehyde, vanillin, syringaldehyde, salicylaldehyde, and anisaldehyde condensed with l-tryptophan in aqueous-acidic media affording the corresponding phenolic tetrahydro-beta-carboline-3-carboxylic acid as two diastereoisomers, 1S,3S-cis and 1R,3S-trans. With the exception of benzaldehyde, the rest of the aldehydes needed heating conditions (70 degrees C) to significantly form tetrahydro-beta-carbolines over time with the cyclization highly favored at low pH. This suggests a likely formation of these compounds under conditions that may occur in foods, food processing, or cooking. The new phenolic tetrahydro-beta-carboline alkaloids were assayed, for the first time, for their activity as free radical scavengers and antioxidants and showed good antioxidant properties with Trolox equivalent antioxidant capacity (TEAC) values much higher than those of ascorbic acid and the water soluble vitamin E analogue, Trolox, in the 2,2'-azinobis(3-ethylbenzothiazoline)-6-sulfonic acid (ABTS) assay.     

Identification of sulfur volatiles in canned orange juices lacking orange flavor

The purpose of this study was to understand why some canned orange juices are not perceived as orange juice. Sensory flavor profile data indicated that the primary odor (orthonasal) attributes were tropical fruit/grapefruit, cooked/caramel, musty, and medicine. By comparison fresh-squeezed juice lacked these odor attributes. GC-O analysis found 43 odor-active components in canned juices. Eight of these aroma volatiles were sulfur based. Four of the 12 most intense aroma peaks were sulfur compounds that included methanethiol, 1-p-menth-1-ene-8-thiol, 2-methyl-3-furanthiol, and dimethyl trisulfide. The other most intense odorants included 7-methyl-3-methylene-1,6-octadiene (myrcene), octanal, 2-methoxyphenol (guaiacol), 2-ethyl-4-hydroxy-5-methyl-3(2H)-furanone (homofuraneol), (E)-non-2-enal, (E,E)-deca-2,4-dienal, 4-hydroxy-3-methoxybenzaldehyde (vanillin), and alpha-sinensal. Odorants probably responsible for the undesirable sensory attributes included grapefruit (1-p-menth-1-ene-8-thiol), cooked [2-ethyl-4-hydroxy-5-methyl-3(2H)-furanone, 4-hydroxy-2,5-dimethyl-3(2H)-furanone (Furaneol), and 3-(methylthio)propanal (methional)], musty [7-methyl-3-methylene-1,6-octadiene and (E)-non-2-enal], and medicine (2-methoxyphenol). The canned juices also lacked several aldehydes and esters normally found in fresh orange juice.     

Polyphenol composition and antioxidant activity of Kei-apple (Dovyalis caffra) juice

The polyphenolic and ascorbate (ASC) components as well as the antioxidant capacity of Kei-apple (Dovyalis caffra) juice were analyzed and compared to three other fruit juices. The Kei-apple juice had significantly the highest total polyphenolic concentrations (1013 mg gallic acid equivalent/L), and solid phase (C(18)) fractionation identified the majority of these polyphenols to be phenolic acids. The Kei-apple juice also had significantly the highest ASC concentrations (658 mg/L), which showed exceptional heat stability with very little conversion to dehydroascorbate (DHA). Antioxidant capacities of both the unfractionated fruit juices and their solid phase-extracted fractions, as determined by oxygen radical absorbance capacity and ferric reducing antioxidant power analyses, correlated well to the polyphenol concentrations. Gas chromatography-mass spectrometry analyses showed caffeic acid as the most abundant polyphenol present (128.7 mg/L) in the Kei-apple juice; it contributed to 63% of the total antioxidant capacity (of all of the individual compounds identified). Other notable polyphenols identified in higher concentrations included p-coumaric acid, p-hydroxyphenylacetic acid, and protocatechuic acid. Our results therefore support the putative high antioxidant value linked to this fruit and better define this potential in terms of the major antioxidants that exist in the Kei-apple.     

New constituents from noni (Morinda citrifolia) fruit juice

Morinda citrifolia L. (Rubiaceae), known as noni, has a long history of traditional use in the Hawaiian and Tahitian islands. More recently, an array of commercial noni fruit juice products are gaining popularity as dietary supplements, with claims of anticancer and immunostimulant activities. The biologically active principles of noni are not fully known. In continuation of work on the isolation of markers from dietary supplements, this paper reports the isolation of three new markers, namely, 1-O-(3'-methylbut-3'-enyl)-beta-D-glucopyranose (1), 1-n-butyl-4-(5'-formyl-2'-furanyl)methyl succinate (2), and 4-epi-borreriagenin (3), together with the known iridoid glycosides asperulosidic acid (4) and deacetylasperulosidic acid (5) and a mixture of 1-n-butyl-4-methyl-2-hydroxysuccinate (6a) and 1-n-butyl-4-methyl-3-hydroxysuccinate (6b), as well as a mixture of alpha- and beta-glucopyranose from noni fruit juice obtained from Puerto Rico. The structures of compounds were based on 1H and 13C NMR, mainly 2D NMR COSY, HMQC, HMBC, and NOESY experiments, and HRMS. Furthermore, samples from fresh-squeezed noni fruit juice from Japan revealed the presence of scopoletin (7), in addition to compounds 1-6, indicating no significant differences in the marker constituents of noni collected from Atlantic and Pacific regions.     

Stability and enhancement of berry juice color

Attractive color is one of the main sensory characteristics of fruit and berry products. Unfortunately, the color of red juices is unstable and easily susceptible to degradation, leading to a dull and weak juice color. This study was designed to investigate the color stability and copigmentation of four different berry juices enhanced by phenolic acids and commercial color enhancers. Phenolic acid enrichment improved and stabilized the color of the berry juices during storage. The commercial color enhancers immediately produced an intensive color to the juices, which, however, was not very stable. The color enhancement was intensive in strawberry and raspberry juices and effective in lingonberry and cranberry juices. Sinapic acid induced the strongest color in strawberry juice. Ferulic and sinapic acids improved raspberry juice color equally. Rosmarinic acid enhanced the color of lingonberry and cranberry juices the most. The addition of the simple cinnamic acids produced novel peaks to the end of the high-performance liquid chromatography chromatogram, indicating a formation of new compounds. It can be assumed that sinapic and ferulic acids formed new intramolecular copigmentation compounds with berry anthocyanins whereas rosmarinic acid stabilized anthocyanins intermolecularly.     

Transgenic overexpression of expansin influences particle size distribution and improves viscosity of tomato juice and paste

Suppression of the expression of a ripening-related expansin gene, LeExp1, in tomato enhanced fruit firmness and overexpression of LeExp1 resulted in increased fruit softening. Because of the incompletely understood relationship between fresh fruit texture and the consistency of processed products, we examined the effects of LeExp1 overexpression on the processing characteristics of tomato fruit. As determined by Bostwick consistency and by controlled strain rheometry, juices and pastes prepared from transgenic tomatoes with suppressed LeExp1 expression had a higher viscosity than preparations from control fruits. However, the viscosity of juice and paste prepared from fruit overexpressing LeExp1 was significantly greater than products from controls or lines with reduced LeExp1. Bostwick consistency increased by 9% (juice) and 6% (paste) in lines with suppressed LeExp1 expression but increased by 27.5% (juice) and 19.5% (paste) in lines overexpressing LeExp1, relative to controls. Determined by laser diffraction, the particles in juice and paste prepared from transgenic fruits with reduced LeExp1 expression were smaller, and preparations from fruits overexpressing LeExp1 had a size distribution indicating more large particles. Analysis of cell wall polysaccharides size indicated that LeExp1 overexpression enhanced depolymerization of water soluble pectins as well as tightly bound matrix glycans. LeExp1 overexpression may allow increased cell wall hydration, resulting in expanded particle size and increased viscosity of products. Because either LeExp1 suppression or overexpression leads to improved consistency, the interactions that contribute to optimal product rheological properties are complex.     

Changes in antioxidant and metabolite profiles during production of tomato paste

Tomato products and especially concentrated tomato paste are important sources of antioxidants in the Mediterranean diet. Tomato fruit contain well-known antioxidants such as vitamin C, carotenoids, flavonoids, and hydroxycinnamic acids. The industrial processing of this fruit into tomato paste involves several treatments that potentially affect the final profile of antioxidants and other metabolites in the commercial product. Here we have used both biochemical and metabolomic techniques to assess the effect of each separate step in the industrial production chain starting from fresh fruit to the final tomato paste. Material was collected from five independent tomato paste production events spread over two successive years. Samples comprised the intact ripe fruits and semifinished products after fruit-breaking, separation of the pulp from skin and seeds, evaporation, and finally after canning and pasteurization. The effect of each processing step was determined by different types of analysis. First, the total antioxidant capacity and total phenolic content were determined by commonly used spectrophotometric methods. Second, individual antioxidants in the extracts were identified and compared using an HPLC with online antioxidant detection. Third, in each sample the levels of the major individual antioxidants present, i.e., vitamin C, phenolic compounds (such as rutin and chlorogenic acid), tocopherols, and carotenoids, were quantified. Fourth, an untargeted metabolomic approach using LC-QTOF-MS was used to identify those production steps that have the largest impact on the overall metabolic profile in the final paste as compared to the original fruits. This multifaceted approach has revealed that each processing step induces specific alterations in the metabolic profile, as determined by the different analysis procedures, and that in particular the fruit-breaking step and the removal of seed and skin significantly affect the levels of antioxidants and many other metabolites present in commercial tomato paste.     

Abscisic acid related compounds and lignans in prunes (Prunus domestica L.) and their oxygen radical absorbance capacity (ORAC)

Four new abscisic acid related compounds (1-4), together with (+)-abscisic acid (5), (+)-beta-D-glucopyranosyl abscisate (6), (6S,9R)-roseoside (7), and two lignan glucosides ((+)-pinoresinol mono-beta-D-glucopyranoside (8) and 3-(beta-D-glucopyranosyloxymethyl)-2- (4-hydroxy-3-methoxyphenyl)-5-(3-hydroxypropyl)-7-methoxy-(2R,3S)-dihydrobenzofuran (9)) were isolated from the antioxidative ethanol extract of prunes (Prunus domestica L.). The structures of 1-4 were elucidated on the basis of NMR and MS spectrometric data to be rel-5-(3S,8S-dihydroxy-1R,5S-dimethyl-7-oxa-6-oxobicyclo[3,2,1]oct-8-yl)-3-methyl-2Z,4E-pentadienoic acid (1), rel-5-(3S,8S-dihydroxy-1R,5S-dimethyl-7-oxa-6-oxobicyclo[3,2,1]oct-8-yl)-3-methyl-2Z,4E-pentadienoic acid 3'-O-beta-d-glucopyranoside (2), rel-5-(1R,5S-dimethyl-3R,4R,8S-trihydroxy-7-oxa-6-oxobicyclo[3,2,1]oct-8-yl)-3-methyl-2Z,4E-pentadienoic acid (3), and rel-5-(1R,5S-dimethyl-3R,4R,8S-trihydroxy-7-oxabicyclo[3,2,1]- oct-8-yl)-3-methyl-2Z,4E-pentadienoic acid (4). The antioxidant activities of these isolated compounds were evaluated on the basis of oxygen radical absorbance capacity (ORAC). The ORAC values of abscisic acid related compounds (1-7) were very low. Two lignans (8 and 9) were more effective antioxidants whose ORAC values were 1.09 and 2.33 micromol of Trolox equiv/micromol, respectively.     

Functional characterization of enone oxidoreductases from strawberry and tomato fruit

Fragaria x ananassa enone oxidoreductase (FaEO), earlier putatively assigned as quinone oxidoreductase, is a ripening-induced, negatively auxin-regulated enzyme that catalyzes the formation of 4-hydroxy-2,5-dimethyl-3(2H)-furanone (HDMF), the key flavor compound in strawberry fruit by the reduction of the alpha,beta-unsaturated bond of the highly reactive precursor 4-hydroxy-5-methyl-2-methylene-3(2H)-furanone (HMMF). Here we show that recombinant FaEO does not reduce the double bond of straight-chain 2-alkenals or 2-alkenones but rather hydrogenates previously unknown HMMF derivatives substituted at the methylene functional group. The furanones were prepared from 4-hydroxy-5-methyl-3(2H)-furanone with a number of aldehydes and a ketone. The kinetic data for the newly synthesized aroma-active substrates and products are similar to the values obtained for an enone oxidoreductase from Arabidopsis thaliana catalyzing the alpha,beta-hydrogenation of 2-alkenals. HMMF, the substrate of FaEO that is formed during strawberry fruit ripening, was also detected in tomato and pineapple fruit by HPLC-ESI-MSn and became 13C-labeled when d-[6-13C]-glucose was applied to the fruits, which suggested that a similar HDMF biosynthetic pathway occurs in the different plant species. With a database search (http://ted.bti.cornell.edu/ and http://genet.imb.uq.edu.au/Pineapple/), we identified a tomato and pineapple expressed sequence tag that shows significant homology to FaEO. Solanum lycopersicon EO (SlEO) was cloned from cDNA, and the protein was expressed in Escherichia coli and purified. Biochemical studies confirmed the involvement of SlEO in the biosynthesis of HDMF in tomato fruit.     

Red grape juice inhibits iron availability: application of an in vitro digestion/caco-2 cell model

Adequate bioavailable Fe intake is essential for optimal growth and intellectual development of infants and children. Fruit juices are nutritious and popular drinks for infants and children and are known to contain Fe uptake inhibitors (e.g., polyphenolic compounds) and a dominant promoter, ascorbic acid. Ascorbic acid is naturally present in fruit juices and is added during processing to almost all juices found in supermarkets. With these facts taken into account, an in vitro digestion/Caco-2 cell culture model was developed to compare the effects of apple, pear, white grape, red grape, prune, grapefruir, and orange juices on iron bioavailability. In two series of experiments, juices from a local supermarket were combined with FeCl(3) or commercial infant cereal fortified with elemental iron and subject to simulated gastric and intestinal digestion. Caco-2 cell ferritin formation in response to exposure to the digests served as the measure of Fe uptake. The pear, apple, grapefruit, orange, and white grape juice significantly increased Fe bioavailability from FeCl(3). For the infant cereal studies, the apple, orange, pear, and white grape juices increased the Fe bioavailability of the infant cereal. In contrast, the red grape juice and prune juice had profound inhibitory effects on iron bioavailability. These inhibitory effects were likely due to high levels of polyphenolic compounds that bind and thereby prevent absorption of soluble Fe. These inhibitory compounds appeared to counteract the promotional effects of ascorbic acid as they were in considerable molar excess relative to ascorbic acid and Fe in the digest. From a nutritional standpoint, the results suggest that individuals in need of optimal Fe absorption should avoid red grape and prune juice or at least vary the types of juices consumed. Alternatively, individuals seeking to limit Fe uptake (e.g., hemochromatitics and astronauts) may be able to utilize red grape or prune juice as effective inhibitors of Fe uptake. Consumers should be aware that the compounds that inhibit Fe availability are also linked to anticancer benefits; thus, a dietary balance of the above juices may be optimal.     

Evaluation of phenolic compounds in commercial fruit juices and fruit drinks

The total phenolic content of 13 commercially available fruit juices and juice drinks, selected to represent the most popular juice flavors in the United Kingdom, were analyzed using the Folin-Ciocalteu assay. Individual phenolic compounds were identified and quantified using HPLC-PDA-MS2. The catechin content and degree of polymerization of proanthocyanidins were also analyzed. Purple grape juice contained the largest number of individual phenolic compounds and also the highest concentration of total phenolics. The main components were flavan-3-ols, anthocyanins, and hydroxycinnamates, which accounted for 93% of the total phenolic content. In contrast, white grape juice, which contained principally hydroxycinnamates, had the lowest total phenolic content. Antioxidant activity was measured using the ORAC and FRAP assays, and the data obtained were in broad agreement with total phenol content. In view of the recent findings of the Kame project indicating that long-term fruit juice consumption can provide protection against Alzheimer's disease (Dai et al. Am. J. Med. 2006, 379, 464-475), it is suggested that the protective effects may be enhanced by consumption of a combination of juices rich in phenolics and containing a diverse variety of individual phenolic compounds, namely, juices derived from purple grapes, grapefruit, cranberries, and apples.     

Analysis of organic acids in fruit juices by liquid chromatography-mass spectrometry: an enhanced tool for authenticity testing

Organic acid analysis plays a fundamental role in the testing of authenticity of fruit juices. Analytical methods used routinely for organic acids suffer from poor reproducibility, often give false positives/negatives for tartaric acid, and do not offer the possibility of analyte confirmation. There are conflicting reports in the literature on the presence/absence of tartaric acid in pomegranate juice, a potential indicator of adulteration with grape juice. In this work, a method based on stable isotope dilution liquid chromatography-tandem mass spectrometry is described for citric, malic, quinic, and tartaric acid in fruit juices. Validation data including precision and recovery in six types of juice are presented. Tartaric and quinic acids were confirmed in pomegranate juice at concentrations of 1-5 and ～1 mg/L, respectively. These concentrations are much lower than those resulting from adulteration with grape juice and apple juice, respectively, at the 5% level. A separate method for isocitric acid in orange juice based on the single standard addition method is also described.     

Rubus fruit juices affect lipid peroxidation in a Drosophila melanogaster model in vivo

The antioxidant capacity of red cloudberry (Rubus chamaemorus) juice correlates well with its phenolic content. The red berries have a markedly higher content of anthocyanins, particularly cyanidin and pelargonidin derivatives, than that found in the more common yellow fruit. Conversely, the yellow juice has higher ellagitannin content. A feeding study was conducted to show the in vivo effects of the juices on lipid peroxidation in a sensitive Drosophila melanogaster stock. In young female flies there were significant (P < 0.01) effects of cloudberry juice on lipid peroxidation. In young male flies significant (P < 0.05) effects were found on primary products (hydroxyperoxides) with yellow juice and on secondary products (ketodienes) with red juice. With the red juice, a significant (P < 0.05) decrease in ketodienes was found in both young and old males. This study demonstrates that the effects of berry antioxidants on lipid peroxidation are easily and rapidly tested in vivo with the sensitive Drosophila model.     

Occurrence of flavonols in tomatoes and tomato-based products

The flavonol contents of 20 varieties of tomato fruit were investigated in relation to variety, size, season, and country of origin. Ten commonly consumed tomato-based food products were also assessed. Free and conjugated flavonols were identified and quantified using reversed-phase HPLC. Ninety-eight percent of flavonols detected in tomatoes were found to occur in the skin. Tomatoes contained, primarily as conjugates, quercetin and kaempferol. The main quercetin conjugate was identified as rutin (quercetin 3-rhamnosylglucoside) by LC-MS. The total flavonol content of the different varieties of tomato that were analyzed varied from 1.3 to 22.2 microgram/g of fresh weight (fw). Smaller cherry tomato fruits originating from warm sunny climates, such as Spain and Israel, were found to contain the highest concentration of flavonols. Among the tomato-based products investigated, tomato juice and tomato purée were rich in flavonols, containing 14-16 microgram/mL and 70 microgram/g fw, respectively. In contrast to fresh tomatoes, most tomato-based products contained significant amounts of free flavonols.     

Betaines in fruits of Citrus genus plants

Numerous compounds, many of them osmolytes, were quantified in natural juices and in frozen concentrate juices from fruits of plants of the Citrus genus. L-proline, N-methyl-L-proline (hygric acid), N,N-dimethyl-L-proline (stachydrine), 4-hydroxy-L-prolinebetaine (betonicine), 4-hydroxy-L-proline, γ-aminobutyric acid (Gaba), 3-carboxypropyltrimethylammonium (GabaBet), N-methylnicotinic acid (trigonelline), and choline in the fruit juices of yellow orange, blood orange, lemon, mandarin, bitter orange (Citrus aurantium), chinotto (Citrus myrtifolia), and grapefruit were analyzed by sensitive HPLC-ESI-tandem mass spectrometry procedure. It was found that the most represented osmolytes in the juices, that is, L-proline, stachydrine, and betonicine, can be quantified with minimal sample preparation and short analysis time (about 1 min) also by flow injection analysis (FIA) ESI-MS/MS with the same results as obtained by HPLC ESI-MS/MS. In all of the juices, discrete amounts of choline and trigonelline were present. Conversely, GabaBet was always below detection limits. Notably, N-methyl-L-proline and 4-hydroxy-L-prolinebetaine, which were discovered for the first time in the juice of bergamot (Citrus bergamia Risso et Poit), are also present in all of the citrus juices examined.