Differentiation of bee pollen samples according to the betaines and other quaternary ammonium related compounds content by using a canonical discriminant analysis.

In the last years, an increase has been observed in the adulteration of bee pollen. Consequently, different tools are required to authenticate the origin of this product, such as a study of the profile and composition of a specific family of compounds. The present study investigates the potential of betaines and related compounds as markers of the apiary of origin and harvest period of 71 bee pollen samples. These were collected from four apiaries (Pistacho, Tío Natalio, Monte and Fuentelahiguera), located in the same geographical area (Guadalajara, Spain) and sampled during three consecutive harvest periods in the same year (April-May, June, July-August). They were analyzed by means of a previously developed methodology, which involved solvent extraction, hydrophilic interaction liquid chromatography coupled to mass spectrometry, and a statistical analysis of the data (canonical discriminant analysis). Variable amounts of betaines and related compounds were found in the samples, with four of these being identified in all of them (betonicine, betaine, trigonelline and choline); betonicine was the predominant compound in a concentration range of 264 to 52384 mg/kg. It was possible to statistically assign over 50 % of the samples to the corresponding apiary of origin, the best results being obtained for the Tío Natalio apiary (75 %); this classification was even better in the case of the harvest period, as more than 75 % of the samples were correctly assigned, and in two periods (April-May and June) a 90 % rate was obtained.

Effect of Different Drying Methods on the Nutritional Value of Hibiscus sabdariffa Calyces as Revealed by NMR Metabolomics.

Red mature calyces of Hibiscus sabdariffa were collected from 16 different locations in Meghalaya, India. Samples were processed using shade drying (SD) and tray drying (TD). NMR spectroscopy was used to assess the metabolic composition of the calyces. In this study, 18 polar metabolites were assigned using 1D and 2D NMR spectra, and 10 of them were quantified. Proximate analysis showed that the TD method is more efficient at reducing moisture and maintaining the ash content of the Hibiscus biomass. NMR metabolomics indicates that the metabolite composition significantly differs between SD and TD samples and is more stable in TD plant processing. The differences in post-harvest drying has a greater impact on the metabolite composition of Hibiscus than the plant location.

1H NMR Metabolic Profile of Scyphomedusa Rhizostoma pulmo (Scyphozoa, Cnidaria) in Female Gonads and Somatic Tissues: Preliminary Results.

The Mediterranean basin is one of the regions heavily affected by jellyfish bloom phenomena, mainly due to the presence of scyphozoans, such as Rhizostoma pulmo. The jellyfish have few natural predators, and their bodies represent an organic-rich substrate that can support rapid bacterial growth with great impact on the structure of marine food webs. In Asiatic countries, jellyfish are widely studied for their health benefits, but their nutritional and nutraceutical values still remain poorly characterized. In this study, the differences in the 1H NMR spectroscopy metabolic profiles of R. pulmo female gonads and body fractions (including umbrella and oral arms), in different sampling periods, were studied. For each body compartment both lipid and aqueous extracts were characterized and their 1H NMR metabolic profiles subjected to multivariate analysis. From a statistical analysis of the extracts, a higher contents of ω-3 polyunsaturated fatty acids (PUFAs), amino acid and osmolytes (homarine, betaine, taurine) with important roles in marine invertebrates were observed in female gonads, whereas umbrella and oral arms showed similar metabolic profiles. These results support a sustainable exploitation of the jellyfish for the extraction of bioactive compounds useful in nutraceutical, nutricosmetics, and functional food fields.

Metabolomic analyses of plasma and liver of mice fed with immature Citrus tumida peel.

In this study, we investigated the effects of dietary supplementation of Citrus tumida hort. ex Tanaka on food intake, body and fat tissue weights, and metabolic profiles of plasma and liver in mice. Supplementation with 5% (w/w) of peels of immature C. tumida (PIC) for 4 weeks significantly suppressed body weight gain and decreased adipose tissue weight in epididymal, perirenal, and subcutaneous fats. Metabolome analyses showed that 2-hydroxyvaleric acid levels were reduced in the blood plasma of mice fed with PIC. PIC supplementation significantly elevated dipeptide (Thr-Asp, Ser-Glu, and Ala-Ala), glucuronic acid, and S-methylglutathione levels, and significantly reduced betaine aldehyde levels in the liver. In conclusion, PIC supplementation affects the metabolism of fatty acids, pectin, glutathione, and choline, showing potential beneficial effects for metabolic syndrome and obesity. PIC may be developed as a functional food and used in the treatment of these diseases.

Modulation of agronomic and nutritional response of Pleurotus eryngii strains by utilizing glycine betaine enriched cotton waste.

BACKGROUND: This study aimed to evaluate the possibility of cotton waste enrichment with glycine betaine (GB) for production of two strains (P9, P10) of king oyster (Pleurotus eryngii). Cotton waste was used as (100%) control (T0 = cotton waste) and augmented with various combinations of GB, (T1 = 2 mmol L-1 , T2 = 4 mmol L-1 , T3 = 6 mmol L-1 , T4 = 8 mmol L-1 and T5 = 10 mmol L-1 ). The response of king oyster to GB was evaluated by earliness, yield, biological efficiency (BE), minerals (nitrogen, phosphorus, potassium, zinc (Zn), copper (Cu), magnesium (Mg), manganese (Mn), iron (Fe), sodium (Na), calcium (Ca)), total sugars, total soluble solids, reducing sugars, non-reducing sugars, ascorbic acid, proximate (crude protein, carbohydrates, crude fibers, ash, fats) content of fruiting body and Fourier-transform infrared (FTIR) spectroscopy analysis compared with the control substrate (cotton waste). RESULTS: The earliness, yield, and BE were higher as compared to control substrate and increased with an augmentation in the concentration of GB within the cotton waste. Two strains showed (on dry weight basis) 33.9-54.9 mg g-1 nitrogen, 6.8-12.5 mg g-1 phosphorus, 16.9-25.1 mg g-1 potassium, 40.5-64.2 mg kg-1 Zn, 17.1-37.3 mg kg-1 Cu, 1174-1325 mg kg-1 Mg, 20.1-29.1 mg kg-1 Mn, 129-265 mg kg-1 Fe, 779-835 mg kg-1 Ca), 6.3%-11.3% total sugars, 7.3-14.9 °Brix total soluble solids, 2.1-7.3% reducing sugars, 10.4-18.1% crude protein, 3.6-4.4% crude fiber and 5.6-16.7 mg (100 g)-1 on various concentration of GB enrich cotton waste. Cotton waste enriched with GB significantly affected nutritional profile of king oyster mushroom. CONCLUSION: The results revealed that GB enriched cotton waste can be used as an innovative substrate to enhance the yield and quality of king oyster mushroom. © 2019 Society of Chemical Industry.

Chemometric analysis of Amaranthus retroflexus in relation to livestock toxicity in southern Australia.

Amaranthus retroflexus L., an introduced invasive weed in southern Australia, has been associated with acute renal failure and/or mortality in a number of livestock species. While its leaves, flowers and stems are generally reported to contain high levels of nitrogen, few studies have fully characterised the chemical composition of A. retroflexus foliage with respect to mammalian toxicity. We performed extensive metabolic profiling of stems, leaves, roots and inflorescence tissues of A. retroflexus collected from three spatially and/or temporally distinct toxicity outbreaks, and report on the 1) composition of primary and secondary metabolites in methanolic extracts of A. retroflexus tissues using HPLC and HPLC-MS QToF and 2) chemometric analysis of A. retroflexus extracts in relation to the associated toxin(s). All tissues of A. retroflexus possessed an abundance of N-containing metabolites, particularly quaternary ammonium compounds which were identified as betaines, two of which (valine betaine and isoleucine betaine) are rarely encountered in plants. Cytotoxicity to murine fibroblasts was highest in extracts of leaf tissue and was associated with a single, a small modified peptide with high similarity to N-acetyl-L-α-aspartyl-L-alanyl-L-α-aspartyl-L-α-glutamyl-O-(carboxymethyl)-L-tyrosyl-L-leucinamide, a synthetic phosphotyrosyl mimic involved in cell signaling processes. One possible mode of action leading to acute renal failure in grazing livestock by a modified peptide such as this is proposed.

[Fast determination of betaine in Lycii Fructus by ultra-performance convergence chromatography-tandem mass spectrometry].

Over the past 2500 years, Lycii Fructus has been widely used as a functional food and tonic in Chinese herbal medicine. It can nourish the liver and kidneys, moisten the lungs, and improve eyesight. In this study, a new rapid and sensitive method has been developed for the quantitative determination of betaine (index compound) in Lycii Fructus by using ultra-performance convergence chromatography-tandem mass spectrometry (UPC2-MS). The separation of betaine was successfully achieved on an ACQUITY UPC2 BEH 2-EP column (150 mm×2.1 mm, 1.7 µm), with isocratic elution by a CO2-methanol (80:20, v/v) solvent at a flow rate of 0.7 mL/min. The conditions used in the separation process were as follows:modifier, 0.1% (v/v) formic acid in methanol; column temperature, 40℃; backpressure, 1.31×107 Pa; injection volume, 1 µL; and retention time, 3 min. The MS system was equipped with an electrospray ionization (ESI) ion source and operated in the selected ion recording (SIR) and positive ion mode. Under the abovementioned conditions, the calibration curve was obtained. The linear range of detection was 0.5-50.0 µg/mL, with a correlation coefficient of 0.9992, and the limit of detection (LOD) was found to be 0.013 µg/mL. The validity of the method was tested by analyses of precision, repeatability, stability, and accuracy (average recovery:96.3%). Finally, the developed method was applied to analyze 11 batches of samples. The results indicated that this method was suitable for evaluating the quality of Lycii Fructus.

Toxicity of Natural Deep Eutectic Solvent Betaine:Glycerol in Rats.

Natural deep eutectic solvents (NaDES) are new natural solvents in green chemistry that in some cases have been shown to allow better extraction of plant bioactive molecules compared to conventional solvents and higher phenolic compound absorption in rodents. However, there is a serious lack of information regarding their in vivo safety. The purpose of this study was to verify the safety of a NaDES (betaine:glycerol (1:2 mole ratio) of water) extract from green coffee beans, rich in polyphenols. Twelve 6-week-old male Wistar rats were randomized into two groups of 6 animals each and twice daily gavaged for 14 days either with 3 mL of water or 3 mL of phenolic NaDES extract. Oral administration of phenolic NaDES extract induced mortality in two rats. In addition, it induced excessive water consumption, reduced dietary intake and weight loss, hepatomegaly, and plasma oxidative stress associated with high blood lipid levels. In conclusion, this work demonstrated the toxicity of oral administration of the selected NaDES under a short-term condition. This occurs despite the fact that this NaDES extract contains polyphenols, whose beneficial effects have been shown. Therefore, complementary work is needed to find the best dose and formulation of NaDES that are safe for the environment and animals and ultimately for humans.

New approaches for extraction and determination of betaine from Beta vulgaris samples by hydrophilic interaction liquid chromatography-tandem mass spectrometry.

Betaine is one of most studied biologically active compounds, due its role in the main biological processes. Although it may be found in several plants and roots, such as the Beta vulgaris family, present in typical diets, just a few analytical methods have been developed for its extraction from roots. A new, quick and effective procedure for the isolation and determination of betaine from two different varieties of B. vulgaris (red and gold) is presented. For betaine extraction, an accelerated solvent extraction (ASE) was coupled with solid-phase extraction. For betaine determination, a separation method based on hydrophilic interaction chromatography coupled with tandem mass spectrometry was optimized for a sensible detection of betaine by means of experimental design. Recoveries were about 93%, with RSD <5%, for both the matrices, without evidence of interfering species. The total content of betaine in extracts of various parts of plants (juice, peel, root) have been determined, obtaining concentrations in the range 3000-4000 mg/L for the juice and in the range 2-5 mg/g for the pulp and for the peel. The B. vulgaris gold species exhibited a higher concentration of betaine, compared to the red variety. Additionally, a micro extraction by packed sorbent technique and a modified quick, easy, cheap, rugged and safe (QuEChERS) procedure, were also tested and compared. Despite the lower recoveries of the latter, with respect to the ASE/SPE procedure (75-89%, RSD <1.5%), the ease of the method, which can be applied without the SPE purification procedure, can represent a positive improvement. Graphical abstract Determination of betaine from Beta vulgaris samples.

Polar Lipids Analysis of Cultured Phytoplankton Reveals Significant Inter-taxa Changes, Low Influence of Growth Stage, and Usefulness in Chemotaxonomy.

The high lipid diversity of microalgae has been used to taxonomically differentiate phytoplankton taxa at the class level. However, important lipids such as phospholipids (PL) and betaine lipids (BL) with potential chemotaxonomy application in phytoplankton ecology have been scarcely studied. The chemotaxonomy value of PL and BL depends on their intraspecific extent of variation as microalgae respond to external changing factors. To determine such effects, lipid class changes occurring at different growth stages in 15 microalgae from ten different classes were analyzed. BL occurred in 14 species and were the less affected lipids by growth stage with diacylglyceryl-hydroxymethyl-N,N,N-trimethyl-b-alanine (DGTA) showing the highest stability. PL were more influenced by growth stage with phosphatidylcholine (PC), phosphatidylglycerol (PG), and phosphatidyletanolamine (PE) declining towards older culture stages in some species. Glycolipids were the more common lipids, and no evident age-related variability pattern could be associated to taxonomic diversity. Selecting BL and PL as descriptor variables optimally distinguished microalgae taxonomic variability at all growth stages. Principal coordinate analysis arranged species through a main tendency from diacylglyceryl-hydroxymethyl-N,N,N-trimethyl-b-alanine (DGCC) containing species (mainly dinoflagellates and haptophytes) to DGTA or PC containing species (mainly cryptophytes). Two diatom classes with similar fatty acid profiles could be distinguished from their respective content in DGTA (Bacillariophyceae) or DGCC (Mediophyceae). In green lineage classes (Trebouxiophyceae, Porphyridophyceae, and Chlorodendrophyceae), PC was a better descriptor than BL. BL and PL explained a higher proportion of microalgae taxonomic variation than did fatty acids and played a complementary role as lipid markers.

Evaluation of potential cationic probes for the detection of proline and betaine.

Osmoregulants are the substances that help plants to tolerate environmental extremes such as salinity and drought. Proline and betaine are two of the most commonly studied osmoregulants. An indirect UV CE method has been developed for simultaneous determination of these osmoregulants. A variety of reported probes and compounds were examined as potential probes for the indirect detection of proline and betaine. Mobility and UV-absorption properties highlighted sulfanilamide as a potential probe for indirect analysis of proline and betaine. Using 5 mM sulfanilamide at pH 2.2 with UV detection at 254 nm, proline and betaine were separated in less than 15 min. The LODs for proline and betaine were 11.6 and 28.3 µM, respectively. The developed method was successfully applied to quantification of these two osmoregulants in spinach and beetroot samples.

Betalain and betaine composition of greenhouse- or field-produced beetroot ( Beta vulgaris L.) and inhibition of HepG2 cell proliferation.

The composition of betalain, red or yellow pigments, and betaine (trimethylglycine or glycinebetaine) of nine beetroot ( Beta vulgaris L.) cultivars produced in the greenhouse or field was studied. Inhibition of HepG2 cell proliferation by betanin and betaine was also tested. Four predominant betalains, two betacyanins (betanin and isobetanin) and two betaxanthins (vulgaxanthin I and miraxanthin V), were isolated and quantified. Betanin and vulgaxanthin I were the major compounds in red and yellow beetroot extracts, respectively, and they comprised >90% of the betalain content in the tested cultivars. The total betalain content of beetroots produced from the field was between 650 and 800 µg/g fresh weight, approximately 25% higher than those from the greenhouse. The betaine content of the beetroot grown in the field was between 3.0 and 4.8 mg/g fresh weight, approximately 20% higher than in plants from the greenhouse. There was great variation among the cultivars with respect to their contents of betalains and betaine. In vitro cancer cell cytotoxicity was evaluated using a 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide assay on HepG2 cells after exposure to betanin and betaine at concentrations ranging from 0 to 400 µg/mL and from 0 to 800 µg/mL for 48 h, respectively. Betanin resulted in a 49% inhibition of HepG2 cell proliferation at 200 µg/mL, and betaine yielded a 25% inhibition at 800 µg/mL, implying a higher cytotoxicity of betanin compared with betaine. The results indicated that the contents of health-beneficial compounds in beetroots, betalains and betaine, could be increased by modifying the growing conditions and that betanin and betaine extracted from beetroots had some anticancer effects against HepG2 cells.

Cereal foods are the major source of betaine in the Western diet--analysis of betaine and free choline in cereal foods and updated assessments of betaine intake.

Betaine and its precursor choline are important components of one-carbon metabolism, remethylating homocysteine into methionine and providing methyl groups for DNA methylation. Cereals are the main source of betaine in the diet, though there is little literature available on the content of betaine in cereal products, nor on betaine intake from cereals. Betaine and free-choline concentrations were measured by liquid-chromatography with tandem mass spectrometry in a wide range of commercially available cereal foods and cereal fractions. Whole grain wheat and related fractions were the best overall common source of betaine, while the pseudocereal quinoa had the highest amount of betaine measured (3900 µg/g). Based on estimates of dietary intake data cereal foods provide approximately 60-67% of betaine in Western diets, and 20-40% of betaine in South-East Asian diets. Average intake of betaine was 131 mg/d, well below those used in intervention studies using betaine to lower blood homocysteine.

Olive oil by capillary electrophoresis: characterization and genuineness.

Olive oil, obtained from Olea europaea L. (Oleaceae) fruits, is an important ingredient in the Mediterranean diet. The purpose of this paper is to review and evaluate olive oil analysis using capillary electrophoresis (CE). This review covers a selection of the literature published on this topic over the past decade. The current state of the art of the topic is evaluated, with special emphasis on separation conditions, analysis purpose, and analytes investigated. CE has been used to characterize or to carry out authenticity studies. Particular attention has been focused on the botanical origin because high-quality monovarietal olive oils have been recently introduced on the markets and their quality control requires the development of new and powerful analytical tools as well as new regulations to avoid fraud. CE represents a good compromise between sample throughput, sample volume, satisfactory characterization, and sustainability for the analysis of target compounds present in olive oils.

Effects of betaine supplementation and choline deficiency on folate deficiency-induced hyperhomocysteinemia in rats.

The effect of betaine status on folate deficiency-induced hyperhomocysteinemia was investigated to determine whether folate deficiency impairs homocysteine removal not only by the methionine synthase (MS) pathway but also by the betaine-homocysteine S-methyltransferase (BHMT) pathway. For this purpose, we investigated the effect of dietary supplementation with betaine at a high level (1%) in rats fed a folate-deprived 10% casein diet (10C) and 20% casein diet (20C). We also investigated the effect of choline deprivation on folate deficiency-induced hyperhomocysteinemia in rats fed 20C. Supplementation of folate-deprived 10C and 20C with 1% betaine significantly suppressed folate deprivation-induced hyperhomocysteinemia, but the extent of suppression was partial or limited, especially in rats fed 10C, the suppression of plasma homocysteine increment being 48.5% in rats fed 10C and 69.7% in rats fed 20C. Although betaine supplementation greatly increased hepatic betaine concentration and BHMT activity, these increases did not fully explain why the effect of betaine supplementation was partial or limited. Folate deprivation markedly increased the hepatic concentration of N,N-dimethylglycine (DMG), a known inhibitor of BHMT, and there was a significant positive correlation between hepatic DMG concentration and plasma homocysteine concentration, suggesting that folate deficiency increases hepatic DMG concentration and thereby depresses BHMT reaction, leading to interference with the effect of betaine supplementation. Choline deprivation did not increase plasma homocysteine concentration in rats fed 20C, but it markedly enhanced plasma homocysteine concentration when rats were fed folate-deprived 20C. This indicates that choline deprivation reinforced folate deprivation-induced hyperhomocysteinemia. Increased hepatic DMG concentration was also associated with such an effect. These results support the concept that folate deficiency impairs homocysteine metabolism not only by the MS pathway but also by the BHMT pathway.

Effects of genotype and environment on the contents of betaine, choline, and trigonelline in cereal grains.

This study examined the environmental and genetic variation in methyl donor contents and compositions of 200 cereal genotypes. Glycine betaine, choline, and trigonelline contents were determined by (1)H NMR, and significant differences were observed between cereal types (G) and across harvesting years and growing locations (E). Glycine betaine was the most abundant methyl donor in all of the 200 lines grown on a single site, and concentrations ranged from 0.43 ± 0.09 mg/g dm in oats to 2.57 ± 0.25 mg/g dm in diploid Einkorn varieties. In bread wheat genotypes there was a 3-fold difference in glycine betaine content. Choline contents, in the same lines, were substantially lower, and mean concentrations ranged from 0.17 mg/g dm in oats to 0.27 mg/g dm in durum wheat. Trigonelline was by far the least abundant of the methyl donors studied. Despite this, however, there were large differences between cereal types. Twenty-six wheat genotypes were grown in additional years at four European locations. The average glycine betaine content was highest in grains grown in Hungary and lowest in those grown in the United Kingdom. Across the six environments, there was a 3.8-fold difference in glycine betaine content. Glycine betaine levels, although moderately heritable (0.36), were found to be the most susceptible to the environmental conditions. Free choline concentrations were less variable across genotypes, but heritability of this component was the lowest of all methyl donor components (0.25) and showed a high G × E interaction. Trigonelline showed the most variation due to genotype. Heritability of this metabolite was the highest (0.59), but given that it is at a very low concentration in wheat, it is probably not attractive to plant breeders.

Determination of nonprotein amino acids and betaines in vegetable oils by flow injection triple-quadrupole tandem mass spectrometry: a screening method for the detection of adulterations of olive oils.

A novel screening method using an automated flow injection electrospray ionization tandem mass spectrometry system is proposed for the simultaneous determination of five nonprotein amino acids (ß-alanine, alloisoleucine, ornithine, citrulline, pyroglutamic acid) and three betaines (glycine betaine, trigonelline, proline betaine) after derivatization with butanolic HCl. MS/MS experiments were carried out in a triple-quadrupole instrument using multiple reaction monitoring mode in <2 min. The proposed method provided high fingerprinting power to identify the presence of five of the studied compounds in different types of vegetable oils (soybean, sunflower, corn, olive) with LODs at parts per billion levels. The method was validated, and different mixtures of extra virgin olive oil with seed oils were analyzed, achieving the typification for the detection of adulterations in extra virgin olive oils up to 2% w/w. The nonprotein amino acid ornithine was confirmed as a marker for adulteration in the olive oils analyzed.

Occurrence of pipecolic acid and pipecolic acid betaine (homostachydrine) in Citrus genus plants.

The presence of pipecolic acid and pipecolic acid betaine, also known as homostachydrine, is herein reported for the first time in Citrus genus plants. Homostachydrine was found in fruits, seeds, and leaves of orange, lemon, and bergamot (Citrus bergamia Risso et Poit). As homostachydrine was not commercially available, as a comparative source, extracts of alfalfa leaves ( Medicago sativa L.) were used, in which homostachydrine is present at high concentration. Then, the results where confirmed by comparison with an authentic standard synthesized and purified starting from pipecolic acid. The synthesized standard was characterized by a ESI-MS/MS study using a 3D ion-trap mass spectrometer. When subjected to MS/MS fragmentation in positive ion mode, homostachydrine, unlike its lower homologue proline betaine (also known as stachydrine), showed a pattern of numerous ionic fragments that allowed unambiguous identification of the compound. For the quantitation in the plant sources, high sensitivity and specificity were achieved by monitoring the transition (158 → 72), which is absent in the fragmentation patterns of other major osmolytes commonly used as markers for studies of abiotic stress. As for the metabolic origin of homostachydrine, the occurrence in citrus plants of pipecolic acid leads to the hypothesis that it could act as a homostachydrine precursor through direct methylation.

Variation of polyphenols and betaines in aerial parts of young, field-grown Amaranthus genotypes.

Amaranthus hybridus and Amaranthus mantegazzianus are commonly cultivated and the entire young fresh plants consumed as vegetables in regions of Africa and Asia. A. hybridus and A. mantegazzianus were cultivated at four sites in three climate regions of the world: Santa Rosa, Argentina; Lleida, Spain; and Prague and Olomouc, both in the Czech Republic. The contents of flavonoids (isoquercitrin, rutin, nicotiflorin), hydroxybenzoic acids (protocatechuic acid, 4-hydroxybenzoic acid, vanillic acid), hydroxycinnamic acids (caffeic acid, p-coumaric acid, ferulic acid), hydroxycinnamyl amides (N-trans-feruloyltyramine, N-trans-feruloyl-4-O-methyldopamine), and betaines (glycinebetaine, trigonelline) were determined. The variation in phytochemical content due to species and cultivation site was analyzed utilizing the multivariate statistical methods of principal component analysis (PCA) and graphical model (GM). The Argentinean samples differed from the three other locations due to higher contents of most compounds. The samples from Spain and the Czech Republic differed from each other in the content of the negatively correlated metabolites trigonelline and the flavonoids. The two amaranth species were separated primarily by a higher content of trigonelline and the two hydroxycinnamyl amides in A. mantegazzianus. The GM showed that the quantities of the different analytes within each compound group were intercorrelated except in the case of the betaines. The betaines carried no information on each other that was not given through correlations with other compounds. The hydroxycinnamic acids were a key group of compounds in this analysis as they separated the other groups from each other (i.e., carried information on all of the other groups). This study showed the contents of polyphenols and betaines in the aerial parts of vegetable amaranth to be very dependent on growth conditions, but also revealed that some of the compounds (trigonelline and the two hydroxycinnamyl amides) may be useful as features of a taxonomic classification.

Rapid NMR method for the quantification of organic compounds in thin stillage.

Thin stillage contains organic and inorganic compounds, some of which may be valuable fermentation coproducts. This study describes a thorough analysis of the major solutes present in thin stillage as revealed by NMR and HPLC. The concentration of charged and neutral organic compounds in thin stillage was determined by excitation sculpting NMR methods (double pulse field gradient spin echo). Compounds identified by NMR included isopropanol, ethanol, lactic acid, 1,3-propanediol, acetic acid, succinic acid, glycerophosphorylcholine, betaine, glycerol, and 2-phenylethanol. The concentrations of lactic and acetic acid determined with NMR were comparable to those determined using HPLC. HPLC and NMR were complementary, as more compounds were identified using both methods. NMR analysis revealed that stillage contained the nitrogenous organic compounds betaine and glycerophosphorylcholine, which contributed as much as 24% of the nitrogen present in the stillage. These compounds were not observed by HPLC analysis.