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.

HPLC and NMR quantification of bioactive compounds in flowers and leaves of Brassica rapa: the influence of aging.

Several members of the Brassicaeae family are known to possess beneficial properties which positively impact human diet, thanks to the presence of antioxidants, bioactive polyphenols and amino acids. B. rapa, one of the most widespread and economically relevant species, represents an outstanding example. The aim of this study is to investigate, at the molecular level, the effect of plant aging on the concentration of some biologically relevant compounds in different parts of the plant. Using HPLC and NMR techniques, the quantification of polyphenolic species (caffeic acid, quercetin and rutin), succinic acid and alanine was performed in flowers and leaves of young and mature B. rapa plants.

Baltic amber teething necklaces: could succinic acid leaching from beads provide anti-inflammatory effects?

BACKGROUND: Baltic amber teething necklaces have been popularized as a safe and natural alternative to conventional or pharmacological medicines for the management of teething pain. However, claims made by retailers regarding the efficacy and mechanism of action of these necklaces lack scientific or clinical basis. The claim most closely resembling science is the assertion that succinic acid will leach out of the beads and through the skin of the wearer and carry out anti-inflammatory and analgesic effects. The objective of the current research is to scientifically assess this claim. METHODS: Beads from necklaces were powdered for identification by infrared spectroscopy, and dissolved in sulfuric acid for quantification of succinic acid using HPLC. Succinic acid release from beads was assessed by long-term submersion of amber beads (separated according to light, medium and dark brown colour) in solvents relevant to human skin conditions. The potential for succinic acid to have anti-inflammatory effects was assessed by measuring the release of inflammatory cytokines IL-1α, IL-1ß, IL-8 and TNFα, and the inflammatory messenger PGE2, from THP-1 human macrophages after treatment with succinic acid and LPS. RESULTS: Amber teething necklaces were positively identified as Baltic amber, by comparison of the beads' infrared spectrum to the literature, and by their succinic acid content (1.5 mg per bead; 1.44% w/w). However, whole amber beads submerged in octanol or pH 5.5 phosphate buffered saline did not release any measurable succinic acid, except for the light-coloured beads in octanol which broke into tiny fragments. Additionally, treatment of macrophages with succinic acid did not reduce the release of any inflammatory cytokines measured, and displayed toxicity to the cells at high concentrations. CONCLUSIONS: While amber teething necklaces are genuine Baltic amber, we have found no evidence to suggest that the purported active ingredient succinic acid could be released from the beads into human skin. Additionally, we found no evidence to suggest that succinic acid has anti-inflammatory properties.

Enhancing wine ester biosynthesis in mixed Hanseniaspora uvarum/Saccharomyces cerevisiae fermentation by nitrogen nutrient addition.

The dynamic changes of wine ester production during mixed fermentation with Hanseniaspora uvarum Yun268 and Saccharomyces cerevisiae F5 was investigated at different levels and timings of nitrogen nutrient addition. Nitrogen additions were performed by supplementing yeast assimilable nitrogen (YAN) into a synthetic grape must with defined composition. Ester precursors and extracellular metabolites involved in ester synthesis were analyzed throughout the fermentation. Results showed that nitrogen additions covering 50-200 mg/L YAN at the point of yeast inoculation slightly affected yeast competition and ester profiles. Interestingly, when YAN was supplemented in the mid-stage, the survival of H. uvarum Yun268 was enhanced, resulting in more than a 2-fold increase in the levels of higher alcohol acetates compared to that at the initial stage. Furthermore, carbon fluxes may be redistributed in the central pathway, which contributed to the production of medium-chain fatty acids and eventually triggered a 1.2-fold elevation in corresponding ethyl ester levels.

Efficient succinic acid production from high-sugar-content beverages by Actinobacillus succinogenes.

This study presents the production of succinic acid (SA) by Actinobacillus succinogenes using high-sugar-content beverages (HSCBs) as feedstock. The aim of this study was the valorization of a by-product stream from the beverage industry for the production of an important building block chemical, such as SA. Three types of commercial beverages were investigated: fruit juices (pineapple and ace), syrups (almond), and soft drinks (cola and lemon). They contained mainly glucose, fructose, and sucrose at high concentration-between 50 and 1,000 g/L. The batch fermentation tests highlighted that A. succinogenes was able to grow on HSCBs supplemented with yeast extract, but also on the unsupplemented fruit juices. Indeed, the bacteria did not grow on the unsupplemented syrup and soft drinks because of the lack of indispensable nutrients. About 30-40 g/L of SA were obtained, depending on the type of HSCB, with yield ranging between 0.75 and 1.00 gSA /gS . The prehydrolysis step improved the fermentation performance: SA production was improved by 6-24%, depending on the HSCB, and sugar conversion was improved of about 30-50%.

Behavior of yeast inoculated during semi-dry coffee fermentation and the effect on chemical and sensorial properties of the final beverage.

Pulped Mundo Novo and Ouro Amarelo coffee beans were inoculated with Saccharomyces cerevisiae (CCMA 0200 and CCMA 0543) during semi-dry coffee fermentation and compared with a non-inoculated control. Samples were collected throughout the fermentation process (12days) to evaluate the persistence of the inoculum by Real-Time quantitative PCR (qPCR). Also, the chemical composition of the beans was determined by HPLC and GC-MS and the roasted beans were sensorial evaluated using the cupping test. S. cerevisiae CCMA 0543 had an average population of 5.6logcell/g (Ouro Amarelo cultivar) and 5.5logcell/g (Mundo Novo cultivar). Citric, malic, succinic and acetic acid were found in all samples, along with sucrose, fructose, and glucose. There were 104 volatile compounds detected: 49 and 55 in green and roasted coffee, respectively. All coffee samples scored over 80 points in the cupping test, indicating they were specialty-grade. Inoculation with the CCMA 0543 strain performed better than the CCMA 0200 strain. This is the first time that qPCR has been used to assess the persistence of the inoculated strains populations during coffee processing. Strain CCMA 0543 was the most suitable as an inoculant due to its enhanced persistence during the process and number of volatile compounds produced.

Polar constituents, protection against reactive oxygen species, and nutritional value of Chinese artichoke (Stachys affinis Bunge).

In the present work, we studied the chemical composition of Chinese artichoke (S. affinis tubers) by analyzing its polar constituents and its macro- and micro- nutrients. A total of nine compounds were isolated from the tuber ethanolic extract and structurally elucidated by Nuclear Magnetic Resonance (NMR) spectroscopy and mass spectrometry (MS). The marker compounds identified were oligosaccharide stachyose and the organic acid, succinic acid, as well as phenylethanoid and iridoid glycosides. The macronutrient profile was dominated by carbohydrates (36.9% dw), whereas potassium (2.36%) was the most abundant micro-nutrient. The tuber ethanolic extract was able to efficiently protect human cells (Caco-2, SHSY-5Y and K562) against t-BHP-induced oxidative damage.

Metabolic phenotyping of an adoptive transfer mouse model of experimental colitis and impact of dietary fish oil intake.

Inflammatory bowel diseases are acute and chronic disabling inflammatory disorders with multiple complex etiologies that are not well-defined. Chronic intestinal inflammation has been linked to an energy-deficient state of gut epithelium with alterations in oxidative metabolism. Plasma-, urine-, stool-, and liver-specific metabonomic analyses are reported in a naïve T cell adoptive transfer (AT) experimental model of colitis, which evaluated the impact of long-chain n-3 polyunsaturated fatty acid (PUFA)-enriched diet. Metabolic profiles of AT animals and their controls under chow diet or fish oil supplementation were compared to describe the (i) consequences of inflammatory processes and (ii) the differential impact of n-3 fatty acids. Inflammation was associated with higher glycoprotein levels (related to acute-phase response) and remodeling of PUFAs. Low triglyceride levels and enhanced PUFA levels in the liver suggest activation of lipolytic pathways that could lead to the observed increase of phospholipids in the liver (including plasmalogens and sphingomyelins). In parallel, the increase in stool excretion of most amino acids may indicate a protein-losing enteropathy. Fecal content of glutamine was lower in AT mice, a feature exacerbated under fish oil intervention that may reflect a functional relationship between intestinal inflammatory status and glutamine metabolism. The decrease in Krebs cycle intermediates in urine (succinate, α-ketoglutarate) also suggests a reduction in the glutaminolytic pathway at a systemic level. Our data indicate that inflammatory status is related to this overall loss of energy homeostasis.

[NMR-based analysis of water soluble extracts of different Astragali Radix].

Water soluble extract (WSE) is an important index for the quality evaluation of Astragali Radix (AR). In this study, the WSE of the wild AR from Shanxi province (SX) and the cultivated AR from Gansu Province (GS) were compared. The WSEs of two types of AR were determined according to the appendix of Chinese pharmacopoeia. Then the WSEs were subjected to NMR analysis, and the obtained data were analyzed using HCA, PCA, OPLS-DA, microarray analysis, and Spearman rank analysis. In addition, the Pearson correlation of differential metabolites were also calculated. The results showed that the WSE content of GS-AR (37.80%) was higher than that of SX-AR (32.13%). The main constituent of WSE was sucrose, and other 18 compounds, including amino acids, organic acids, were also detected. Multivariate analysis revealed that SX-AR contained more choline, succinic acid, citric acid, glutamate, taurine and aspartate, while GS samples contained more sucrose, arginine and fumaric acid. In addition, the Pearson correlations between different metabolites of the two types of AR also showed apparent differences. The results suggested that the WSE of two types of AR differs not only in the content, but also in the chemical compositions. Thus, the cultivation way is important to the quality of AR. This study supplied a new method for the comparison of extract of herbal drugs.

Profiling the chlorogenic acids and other caffeic acid derivatives of herbal chrysanthemum by LC-MSn.

Four samples of herbal chrysanthemum have been profiled qualitatively by LC-MS5 to identify their component chlorogenic acids and partially characterize other caffeic acid derivatives. The chlorogenic acids were minor components, and the four samples varied markedly in profile. Three p-coumaroylquinic acids, three feruloylquinic acids, four caffeoylquinic acids, six dicaffeoylquinic acids, and two tricaffeoylquinic acids were detected, 13 for the first time from this source. Partial characterization of minor components suggested the presence of five caffeoyl-hexose esters and caffeic acid-4-beta-d-glucose that have not previously been reported from this source, and eight caffeoylquinic acid glycosides and 16 dicaffeoylquinic acid glycosides that have not previously been reported in nature. Succinic acid-containing chlorogenic acids and chlorogenic acids based on epi-quinic acid, previously reported in Chrysanthemum spp., were not detected in these samples.

Chemical fingerprint analysis of rhizomes of Gymnadenia conopsea by HPLC-DAD-MSn.

A high-performance liquid chromatography-diode array detection-tandem mass spectrometry (HPLC-DAD-MS(n)) method has been firstly developed for chemical fingerprint analysis of rhizomes of Gymnadenia conopsea R. Br. and rapid identification of major compounds in the fingerprints. Comparing the UV and MS spectra with those of reference compounds, seven main peaks in the fingerprints were identified as adenosine (1), 4-hydroxybenzyl alcohol (2), 4-hydroxybenzyl aldehyde (3), dactylorhin B (4), loroglossin (5), dactylorhin A (6) and militarine (7). Compounds 4-7 were succinate derivative esters and firstly discovered from this species. The Computer Aided Similarity Evaluation System (CASES) for chromatographic fingerprint of traditional Chinese medicine was employed to evaluate the similarities of 10 samples of the rhizomes of G. conopsea collected from Sichuan, Qinghai and Hebei provinces, Tibet autonomous region of China, and Nepal. These samples from different sources had similar chemical fingerprints. This method is specific and may serve for quality identification and comprehensive evaluation of this traditional Tibetan remedy.

Molecular and sensory studies on the umami taste of Japanese green tea.

Aimed at defining the key drivers for the quality-determining umami taste of a high-grade powdered green tea, called mat-cha, a bioactivity-guided fractionation using solvent extraction, solvent precipitation, preparative chromatographic separations, and human psychophysical experiments was applied on freshly prepared mat-cha. Liquid chromatography-tandem mass spectrometry and one-/two-dimensional nuclear magnetic resonance studies on isolated fractions led to the identification of l-theanine, succinic acid, 3,4,5-trihydroxybenzoic acid (gallic acid), and (1R,2R,3R,5S)-5-carboxy-2,3,5-trihydroxycyclohexyl-3,4,5-trihydroxybenzoate (theogallin) as umami-enhancing compounds in the green tea beverage, and it can be shown by sensory studies that these compounds are able to raise the umami intensity of sodium l-glutamate proportionally.

Influence of cobalt concentration on vitamin B12 production and fermentation of mixed ruminal microorganisms grown in continuous culture flow-through fermentors.

An experiment was conducted to determine the effects of dietary concentrations of Co on vitamin B12 production and fermentation of mixed ruminal microbes grown in continuous culture fermentors. Four fermentors were fed 14 g of DM/d. The DM consisted of a corn and cottonseed hull-based diet with Co supplemented as CoCO3. Dietary treatments were 1) control (containing 0.05 mg of Co/kg of DM), 2) 0.05 mg of supplemental Co/kg of DM, 3) 0.10 mg of supplemental Co/kg of DM, and 4) 1.0 mg of supplemental Co/kg of DM. After a 3-d adjustment period, fermentors were sampled over a 3-d sampling period. This process was repeated 2 additional times for a total of 3 runs. Ruminal fluid vitamin B12 concentrations were affected by Co supplementation (P < 0.01), and there was a treatment x day interaction (P < 0.01). By sampling d 3, cultures fed the basal diet supplemented with 0.10 mg of Co/kg had greater (P < 0.05) vitamin B12 concentrations than those supplemented with 0.05 mg of Co/kg of DM, and increasing supplemental Co from 0.10 to 1.0 mg/kg of DM increased (P < 0.01) ruminal fluid vitamin B12 concentration. Ruminal fluid succinate also was affected (P < 0.10) by a treatment x day interaction. Cobalt supplementation to the control diet greatly decreased (P < 0.05) succinate in ruminal cultures on sampling d 3 but not on d 1 or 2. Molar proportions of acetate, propionate, and isobutyrate, and acetate:propionate were not affected by the addition of supplemental Co to the basal diet. However, molar proportions of butyrate, valerate, and isovalerate increased (P < 0.05) in response to supplemental Co. The majority of long-chain fatty acids observed in this study were not affected by Co supplementation. However, percentages of C18:0 fatty acids in ruminal cultures tended (P < 0.10) to be greater for Co-supplemented diets relative to the control. Methane, ammonia, and pH were not greatly affected by Co supplementation. The results indicate that a total (diet plus supplemental) Co concentration of 0.10 to 0.15 mg/kg of dietary DM resulted in adequate vitamin B12 production to meet the requirements of ruminal microorganisms fed a high-concentrate diet in continuous-flow fermentors.

[Simultaneous determination of eight organic acids in Fructus mume by RP-HPLC].

OBJECTIVE: To develop an HPLC method for the simultaneous separation and determination of oxalic acid (OA), tartaric acid(TA), malic acid(MA), vitamin C (VC), lactic acid (LA), acetic acid (AA) citric acid (CA) and succinic acid (SA) in Fructus mume. METHOD: Analytical column was Zorbax Eclipse XDB C18. Mobile phase was 0.5% (NH4) H2PO4 aqueous solution and detection wavelength was 214 nm. The flow rate of mobile phase was 0.5 mL x min(-1). RESULT: The regression equations (pH 2. 8, adjusted with phosphoric acid) of eight constituents have been established, r = 0.999 7, 0. 999 8, 0.999 2, 0.999 6, 0.999 1, 0.999 5, 0.999 8, 0.999 2 respectively. Meanwhile, the content and proportion relationship of eight organic acids in Fructus mume which yielded in Fujian (China) were investigated. CONCLUSION: This method was simple, accuracy and quick. The method can be used for the purpose of routine analysis and the quality control of a botanic (Fructus mume) containing these organic acid components.

[Study on extraction and isolation of active constituents from Sorbaria sorbifolia and antitumor effect of the constituents in vivo].

OBJECTIVE: To study the constituents from Sorbaria sorbifolia and their antitumor activities. METHODS: The constituents were isolated with silia gel column chromatography and identified by physicochemical properties and spectroscopic analysis. Thirty six mice were inoculated with sarcocarcinoma 180 according to the standard method, Sorbaria sorbifolia ethyl acetate etract was administrated for 10 days, tumor and body weight, the activities of NK cell and the contents of TNF-alpha, IL-2 in serum were detected. RESULTS: Three compounds were isolated from the ethyl acetate extract and identified as 5,2',4'-trihydroxy-6,7,5'-trimethoxyflavone(I), succinic acid(II) and p-hydroxybenzoic acid(III). The tumor of the test groups were lighter than that of control group(P < 0.05). There was no remarkable difference between the control and test groups in body weight (P > 0.05). The activity of NK cell and the contents of TNF-alpha, IL-2 in serum were higher than that of control group (P < 0.05). CONCLUSION: Compounds I and II were isolated from Sorbaria sorbifolia ethyl acetate extract for the first time. The ethyl acetate extract could improve immune function and strengthen antitumor effect in sarcocarcinoma 180.

Fermentation of pectin and glucose, and activity of pectin-degrading enzymes in the rabbit caecal bacterium Bacteroides caccae.

AIMS: To compare fermentation pattern in cultures of Bacteroides caccae supplied with pectin and glucose, and identify enzymes involved in metabolism of pectin. METHODS AND RESULTS: A strain KWN isolated from the rabbit caecum was used. Fermentation pattern, changes of viscosity and enzyme reactions products were determined. Cultures grown on pectin produced significantly more acetate and less formate, lactate, fumarate and succinate than cultures grown on glucose. Production of cell dry matter and protein per gram of substrate used was the same in pectin- and glucose-grown cultures. The principal enzymes that participated in the metabolism of pectin were extracellular exopectate hydrolase (EC 3.2.1.67), extracellular endopectate lyase (EC 4.2.2.2) and cell-associated 2-keto-3-deoxy-6-phosphogluconate (KDPG) aldolase (EC 4.1.2.14). The latter enzyme is unique to the Entner-Doudoroff pathway. Activities of pectinolytic enzymes in cultures grown on glucose were low. Activity of KDPG aldolase was similar in pectin- and glucose-grown cells. CONCLUSIONS: Metabolites and activities of pectin-degrading enzymes differed in cultures of B. caccae KWN grown on pectin and glucose. Yields of dry matter and protein were the same on both substrates. SIGNIFICANCE AND IMPACT OF THE STUDY: Information on metabolism of pectin in animal strains of Bacteroides is incomplete. This study extends the knowledge on metabolism in bacteria from the rabbit caecum.

[Analysis of volatile fragrant components in hawthorn tincture by gas chromatography-mass spectrometry].

The volatile fragrant components in hawthorn tincture were analyzed by GC/MS. 38 components were identified. The relative content of these constituents were determined with area mormalizing method and the identification ratio was 97.19%. The main components were 3-Hexen-1-ol(370.59 micrograms/g), Eugenol(320.95 micrograms/g), Butanedioic acid hydroxy, diethyl ester(191.25 micrograms/g), 2-Methyl-pentenoic acid(164.83 micrograms/g) and citric acid(80.87 micrograms/g) etc. The method is simple, rapid and highly sensitive and suitable for routine analysis and quality control.