HPLC/Q‐TOF‐MS‐Based Identification of Absorbed Constituents and Their Metabolites in Rat Serum and Urine after Oral Administration of Cistanche deserticola Extract

As a famous health food in China, Cistanche deserticola (C. deserticola) suggested an estrogenic activity according to our previous study. However, no one clarifies its active material basis to date. To find more potentially active constituents and elucidate metabolic pathways of metabolites, a method to simultaneously analyze multiple absorbed constituents and metabolites from C. deserticola in rat serum and urine was established using high‐performance liquid chromatography/quadrupole time‐of‐flight mass spectrometry (HPLC/Q‐TOF‐MS). Based on HPLC/Q‐TOF‐MS method, a total of 24 components, involving 9 prototype constituents and 15 metabolites in rat serum and urine samples, were tentatively identified based on retention time, ultraviolet spectrum, MS data, compound fragmentation laws, published literatures, and reference substances. Most of the compounds existed in the form of metabolites. The proposed metabolic pathways of main metabolites were discussed, including methylation, demethylation, hydrolysis, hydroxylation, acetoxylation, glucuronidation, dehydrogenation, sulfation, esterification, and so on. Phenylethanoid glycosides were extensively metabolized and mutually transformed in vivo. This investigation provided valuable information for further study of the active ingredients and action mechanism of C. deserticola.     

LC‐MS identification of anthocyanins in boysenberry extract and anthocyanin metabolites in human urine following dosing

The anthocyanin composition of boysenberry (Rubusloganbaccus × baileyanus Britt) extract was determined by LC‐ESI‐MS. Four anthocyanins were identified, all comprising a cyanidin‐anthocyanidin‐type skeleton. The two major components were identified as the disaccharide cyanidin‐3‐O‐sophoroside and the monosaccharide cyanidin‐3‐O‐glucoside. The two less abundant components were identified as the rutinosides, cyanidin‐3‐O‐2^G‐glucosylrutinoside and cyanidin‐3‐O‐rutinoside, respectively. These same four anthocyanins were also detected in human urine following a dosing study with boysenberry extract indicating that glycosylated anthocyanins can be absorbed from the gut and excreted intact in the urine. Several anthocyanin metabolites were also detected in the urine and were identified by LC‐ESI‐MS as monoglucuronides of peonidin, cyanidin and pelargonidin. The results suggest that anthocyanins consumed as part of a diet are bioavailable and are present as intact or metabolized forms in the body. Copyright © 2004 Society of Chemical Industry     

Chemical Profiling Using Uplc Q‐Tof/Ms and Antioxidant Activities of Fortunella Fruits

The fruits of Fortunella Swingle are widely consumed as fresh fruits and traditional medicine in China. China is the origin center and has the largest cultivated area of the genus Fortunella. In this study, the chemical compositions of ethanol extracts of the major Fortunella cultivated types including Fortunella japonica Swingle, Fortunella margarita Swingle, Fortunella crassifolia Swingle 1 (Lanshang) and Fortunella crassifolia Swingle 2 (Liuyang) were determined using ultra performance liquid chromatography coupled with quadrupole time‐of‐flight mass spectrometry (UPLC Q‐TOF/MS) method, and their antioxidant activities were evaluated. 12 compounds were identified and 5 compounds were tentatively characterized. The results showed that the chemical compositions of the ethanol extracts of 4 Fortunella cultivated types were largely the same. 3′, 5′‐di‐C‐glucopyranosylphloretin was the predominant flavonoid in Fortunella fruits, and Fortunella margarita Swingle had higher contents of flavonoids than other species. In addition, the data demonstrated high antioxidant activities of Fortunella fruits. The developed method could be available to rapidly analyze the chemical compounds in Fortunella fruits and its products. This study will provide information for further quality assessment and utilization of Fortunella resources.     

Classification of Barley Shochu Samples Produced Using Submerged Culture and Solid‐state Culture of Koji Mold by Solid‐phase Microextraction and Gas Chromatography‐Mass Spectrometry

An easy and fast method of analyzing volatile components contained in shochu, by headspace solid‐phase microextraction and gas chromatography‐mass spectrometry, was established to determine the difference in the content of the various components of barley shochu produced using a submerged culture of koji mold (submerged‐culture shochu) and a solid‐state culture of koji mold (solid‐culture shochu). The contents of the volatile components between the two types of shochu were compared. The results of the comparison revealed that the content of ethyl lactate, ethyl benzoate, 3‐methyl‐1‐pentanol, diethyl succinate, citronellol, and 2‐phenethyl acetate differed between the submerged‐culture and the solid‐culture shochu samples. In addition, the classification of barley shochu samples, including those on the market, into submerged‐culture shochu and solid‐culture shochu, was carried out by multivariate analysis using the quantitative values of the above‐mentioned six compounds, and distinct discrimination was found possible.     

Analysis and characterization of aroma‐active compounds of Schizandra chinensis (omija) leaves

Volatile components from leaves of Schizandra chinensis (omija), a native plant of Korea, were extracted by simultaneous distillation–extraction (SDE) and analyzed by gas chromatography–mass spectrometry (GC‐MS) using two types of capillary column with different polarities (DB‐5MS and DB‐Wax). The GC‐MS analysis of volatile compounds obtained by SDE revealed that germacrene D is the most abundant compound (22.6%) in omija leaves, followed by β‐elemene (17.4%), (E)‐2‐hexenal (8.7%), and (E)‐β‐ocimene (7.2%). Aroma‐active compounds were determined by gas chromatography–olfactometry (GC‐O) using the aroma‐extract‐dilution analysis method. (E,Z)‐2,6‐Nonadienal (cucumber) was the most intense aroma‐active compound due to its higher flavor‐dilution factor (243–729) than any other compound. (Z)‐3‐Hexenal (green/apple), (E)‐2‐hexenal (green/fruity), and (E)‐β‐ocimene (wither green/grass) were also identified as important aroma‐active compounds by GC‐O. In addition, the volatile compounds were extracted by solid‐phase microextraction (SPME), and the quantitative analysis of the SPME samples gave slightly different results, depending on the type of SPME fiber, compared with those from SDE, However, the aroma‐active compounds identified in SPME were similar to those in SDE. Copyright © 2004 Society of Chemical Industry     

Formation of protein adducts of 2‐amino‐1‐methyl‐6‐phenylimidazo[4,5‐b]pyridine in cooked foods

Heterocyclic amines (HCAs) are mutagenic and carcinogenic compounds found in cooked meat and fish. Although HCAs are known to form adducts with protein after metabolic activation, adduct formation during cooking has not been elucidated. In this study, we showed that 2‐amino‐1‐methyl‐6‐phenylimidazo[4,5‐b]pyridine (PhIP) is released from high molecular weight compounds by acid or enzymatic hydrolysis of cooked foods. Formation of free and protein adduct forms of PhIP was dependent on cooking temperature and time, and PhIP–protein adducts were estimated to form after formation of free PhIP. We also demonstrated that PhIP–protein adduct is formed by heating of PhIP and albumin as a model protein. A new adduct peak including [M+H]⁺ (m/z=225) of PhIP as a fragment ion was detected in the high molecular weight fraction of heat‐treated protein by LC–MS analysis. From model experiments by heating of PhIP and amino acids, the adduct was estimated to be produced by condensation of the amino group of PhIP and the carboxyl group of protein. PhIP–protein adducts were detected in several cooked meat and fish at ng/g food level as PhIP content. These results suggest that food‐borne protein adducts of HCAs may influence human HCA exposure and carcinogenic risk.     

Assessment of chemically characterised Rosmarinus officinalis L. essential oil and its major compounds as plant‐based preservative in food system based on their efficacy against food‐borne moulds and aflatoxin secretion and as antioxidant

The study explores antifungal, anti‐aflatoxigenic and antioxidant efficacy of Rosmarinus officinalis essential oil (ROEO) and its major compounds. In addition, the mode of action of ROEO and its practical efficacy as preservative have been assessed. GC‐MS analysis of ROEO identified 16 compounds; α‐pinene, 1,8‐cineole and camphor being the major compounds. The minimum concentration for inhibition of growth and aflatoxin B₁ secretion against A. flavus (LHP‐6) was found to be 1.5, >5.0, 4.0 and 3.0 μL mL^?1 and 1.25, >5.0, 3.5 and 3.0 μL mL^?1 for ROEO, α‐pinene, 1,8‐cineole and camphor, respectively. The IC₅₀ value through DPPH analysis and percentage inhibition of linoleic acid peroxidation of ROEO were 0.042 μL mL^?1 and 71.05%, respectively. The targeted site of antifungal action of ROEO was confirmed as plasma membrane through ergosterol measurement and TEM analysis. Moreover, ROEO significantly protected Piper nigrum fruits against mould infestation upto 6 months in in vivo trial.     

Identification of Bioactive Candidate Compounds Responsible for Oxidative Challenge from Hydro‐Ethanolic Extract of Moringa oleifera Leaves

Free radicals trigger chain reaction and inflict damage to the cells and its components, which in turn ultimately interrupts their biological activities. To prevent free radical damage, together with an endogenous antioxidant system, an exogenous supply of antioxidant components to the body in the form of functional food or nutritional diet helps undeniably. Research conducted by the Natl. Inst. of Health claimed that Moringa oleifera Lam possess the highest antioxidant content among various natural food sources based on an oxygen radical absorbent capacity assay. In this study, a 90% (ethanol:distilled water—90:10) gradient solvent was identified as one of the best gradient solvents for the effectual extraction of bioactive components from M. oleifera leaves. This finding was confirmed by various antioxidant assays, including radical scavenging activity (that is, 1, 1‐diphenyl‐2‐picrylhydrazyl, H₂O₂, and NO radical scavenging assay) and total antioxidant capacity (that is, ferric reducing antioxidant power and molybdenum assay). High‐performance liquid chromatography (HPLC) fingerprints of the 90% gradient extract visually showed few specific peaks, which on further analysis, using HPLC–DAD–ESI–MS, were identified as flavonoids and their derivatives. Despite commonly reported flavonoids, that is, kaempferol and quercetin, we report here for the 1st time the presence of multiflorin‐B and apigenin in M. oleifera leaves. These findings might help researchers to further scrutinize this high activity exhibiting gradient extract and its bio‐active candidates for fruitful clinical/translational investigations.     

Determination of purine alkaloids and catechins in different parts of Camellia assamica var. kucha by HPLC‐DAD/ESI‐MS/MS

BACKGROUND: Kucha (Camellia assamica var. kucha) is a novel wild tea resource grown in China and a tea plant containing a sizable amount of theacrine (1,3,7,9‐tetramethyluric acid). High‐performance liquid chromatography (HPLC) analysis of purine alkaloids and catechins in young leaves of Kucha has been reported previously. However, the compositions of purine alkaloids and catechins in other parts of the plant remain unknown, and more information about the chemical constituents of Kucha is also necessary for further research and development of this new tea resource. RESULTS: Using HPLC with diode array detection coupled with electrospray ionisation tandem mass spectrometry (HPLC‐DAD/ESI‐MS/MS), three purine alkaloids, seven catechins and four non‐catechin phenolic compounds were identified or tentatively identified in Kucha. Purine alkaloids and catechins in leaves at different developmental stages, flowers, stems, pericarps and seeds of the plant were also quantified for the first time by the HPLC method, which was fully validated. Recoveries of the quantified compounds ranged from 96.67 to 104.33%. CONCLUSION: The results showed that the total contents of purine alkaloids and catechins were highest in young leaves of Kucha. Theacrine was detected in all parts of the plant and found to be most abundant in pericarps. Copyright © 2009 Society of Chemical Industry     

Characterization of volatile compounds in Fen‐Daqu – a traditional Chinese liquor fermentation starter

Fen‐Daqu is a saccharifying agent and fermentation starter for the production of Chinese liquor Fen (alcoholic spirit) and Fen traditional vinegar. The volatile compounds produced at seven incubation steps were analysed by HS‐SPME‐GC‐MS. A total of 83 major volatile compounds were identified, including 23 esters, 8 acids, 24 alcohols, 18 ketones and aldehydes, 6 pyrazines and 4 acetals. Data obtained by HS‐SPME‐GC‐MS were subjected to principal component analysis. The trajectory plots of volatile compounds in Fen‐Daqu samples obtained during successive steps of incubation were revealed. The major compounds that contributed to discrimination were hexanal, (E)‐2‐octenal, (Z)‐2‐octen‐1‐ol, nonanoic acid, 1‐octanol, 2‐decen‐1‐ol, hexyl acetate, (E)‐2‐octen‐1‐ol, acetic acid, ethyl acetate, phenylethyl alcohol, ethyl alcohol, octanoic acid, 1‐octanol, 3‐methyl‐2‐buten‐1‐ol and pyrazines. Copyright © 2012 The Institute of Brewing & Distilling     

Bioavailability study of a polyphenol‐enriched extract from Hibiscus sabdariffa in rats and associated antioxidant status

The aqueous extracts of Hibiscus sabdariffa have been commonly used in folk medicine. Nevertheless, the compounds or metabolites responsible for its healthy effects have not yet been identified. The major metabolites present in rat plasma after acute ingestion of a polyphenol‐enriched Hibiscus sabdariffa extract were characterized and quantified in order to study their bioavailability. The antioxidant status of the plasma samples was also measured through several complementary antioxidant techniques. High‐performance liquid chromatography coupled to time‐of‐flight mass spectrometry (HPLC‐ESI‐TOF‐MS) was used for the bioavailability study. The antioxidant status was measured by ferric reducing ability of plasma method, thiobarbituric acid reactive substances assay, and superoxide dismutase activity assay. Seventeen polyphenols and metabolites have been detected and quantified. Eleven of these compounds were metabolites. Although phenolic acids were found in plasma without any modification in their structures, most flavonols were found as quercetin or kaempferol glucuronide conjugates. Flavonol glucuronide conjugates, which show longer half‐life elimination values, are proposed to contribute to the observed lipid peroxidation inhibitory activity in the cellular membranes. By contrast, phenolic acids appear to exert their antioxidant activity through ferric ion reduction and superoxide scavenging at shorter times. We propose that flavonol‐conjugated forms (quercetin and kaempferol) may be the compounds responsible for the observed antioxidant effects and contribute to the healthy effects of H. sabdariffa polyphenolic extract.     

Effects of different deodorising processes on the off‐odour compounds and gel properties of common carp surimi

In order to improve the utilisation of the common carp (Cyprinus carpio), the effects of deodorising agents [NaCl, β‐CD, malic acid (MA) and hawthorn extract (HE)] on the volatile compounds of common carp mince and its surimi gel properties were investigated. In total, thirty‐two volatile compounds were identified, seven of which were considered the main compounds of off‐odour, using HS‐SPME‐GC‐MS and OVA analyses. HS‐SPME‐GC‐MS analysis showed that MA and HE removed 84.51% and 77.18% off‐odour compounds, respectively. Three relaxation components (T₂₁, T₂₂, T₂₃) were found in surimi, and their amounts were not affected by deodorising; however, the relaxation time of major component T₂₂ was significantly increased. Surimi deodorised by HE had the highest gel strength because of its increased breaking displacement; however, a slight decrease in whiteness was observed. Thus, appropriate levels of HE could improve the palatability of surimi.     

Isolation of a free radical‐scavenging antioxidant from water spinach (Ipomoea aquatica Forsk)

Ipomoea aquatica Forsk, a green leafy vegetable that is a rich source of vitamins and amino acids with many health benefits, has been explored for the isolation and identification of its bioactive compounds. Activity‐guided repeated fractionation of a methanol extract on a silica gel column followed by an XAD column yielded a compound that exhibited antioxidant activity with an EC₅₀ value of 83 ± 1.02 µg ml^?1 reaction mixture. It also showed very strong lipid peroxidation‐inhibitory activity in a liposome model system with an EC₅₀ value of 72.2 ± 0.9 µg ml^?1. However, it showed negligible metal‐chelating activity. Based on UV, 2D nuclear magnetic resonance and gas chromatography/mass spectrometry studies, the compound was tentatively identified to be 7‐O‐β‐D ‐glucopyranosyl‐dihydroquercetin‐3‐O‐α‐D ‐glucopyranoside. This is the first report on the antioxidant properties of I aquatica leaf extracts. Copyright © 2005 Society of Chemical Industry     

GC‐MS‐Based Metabolite Profiling of Cosmos caudatus Leaves Possessing Alpha‐Glucosidase Inhibitory Activity

Cosmos caudatus, which is known as “Ulam Raja,” is an herbal plant used in Malaysia to enhance vitality. This study focused on the evaluation of the α‐glucosidase inhibitory activity of different ethanolic extracts of C. caudatus. Six series of samples extracted with water, 20%, 40%, 60%, 80%, and 100% ethanol (EtOH) were employed. Gas chromatography‐mass spectrometry (GC‐MS) and orthogonal partial least‐squares (OPLS) analysis was used to correlate bioactivity of different extracts to different metabolite profiles of C. caudatus. The obtained OPLS scores indicated a distinct and remarkable separation into 6 clusters, which were indicative of the 6 different ethanol concentrations. GC‐MS can be integrated with multivariate data analysis to identify compounds that inhibit α‐glucosidase activity. In addition, catechin, α‐linolenic acid, α‐D‐glucopyranoside, and vitamin E compounds were identified and indicate the potential α‐glucosidase inhibitory activity of this herb.     

Comparison and Characterization of Compounds with Antioxidant Activity in Lycium barbarum Using High‐Performance Thin Layer Chromatography Coupled with DPPH Bioautography and Tandem Mass Spectrometry

Methanol extracts from 50 batches of Lycium barbarum (L. barbarum, wolfberry) in China were compared and characterized using high‐performance thin‐layer chromatography coupled with 2,2‐diphenyl‐1‐picrylhydrazyl (DPPH) bioautography (HPTLC‐DPPH) and electrospray ionization quadrupole time‐of‐flight tandem mass spectrometry (ESI‐Q‐TOF‐MS/MS), respectively. Results showed that similar components occupying the major antioxidant activity existed in L. barbarum collected from different origins. However, the average antioxidant capacities of methanol extracts of L. barbarum collected in Ningxia were significantly higher than those of Qinghai, Xinjiang, Inner Mongolia, and Gansu, which may contribute to rational use of L. barbarum in China. Furthermore, the chemical structure of compound with the highest antioxidant capacity was tentatively identified as 2‐O‐β‐d ‐glucopyranosyl‐l ‐ascorbic acid using ESI‐Q‐TOF‐MS/MS analysis, which possessed high potentials to be used as an antioxidant biomarker for the quality control of L. barbarum. Results are helpful for the bioactivity‐based quality control of L. barbarum, and beneficial for the improvement of their performance in functional/health foods area, suggesting that HPTLC‐DPPH bioautography with ESI‐Q‐TOF‐MS/MS could be used as a routine approach for quality control of antioxidant components in L. barbarum.     

Characterisation of volatile compounds of farmed soft‐shelled turtle (Pelodiscus sinensis) by solid‐phase microextraction and the influence of matrix pH on the release of volatiles

Although farming and consuming of soft‐shelled turtle has been practised for centuries, the aroma‐impact compounds in the meat have not been determined. Furthermore, matrix pH, usually changing during processing and storage stage, was rarely investigated for the influence on volatile profiles. To these aims, soft‐shelled turtle meat was subjected to different pH conditions and the potential volatile compounds were analysed under different extraction conditions by headspace solid‐phase microextraction (HS‐SPME). A total of forty‐three volatiles were identified; in which, nonanal, (E, E)‐2, 4‐heptadienal, octanal, decanal, hexanal, (E)‐2‐nonenal, heptanal, 1‐octen‐3‐ol and o‐xylene were assigned as aroma‐impact compounds by high relative odour activity value (ROAV). Overly basic or acidic pH significantly (P < 0.05) facilitated the release of volatiles under moderate extraction conditions. The total volatilisation increased by 30.1% to 298% in pH‐shifted samples, while the highest one was found in pH 2 or pH 11. However, the results of principal components analysis (PCA) demonstrated that increased extraction time or temperature hindered the pH enhancement.     

Effect of Different Indigenous Yeast β‐Glucosidases on the Liberation of Bound Aroma Compounds

This study investigated β‐D‐glucosidase activity in the indigenous wine yeast present on grape berries in Yantai in Shandong Province, China. Yeast population profiles from the Yantai production area in China were examined. Among the ten species identified by RFLP analysis of the 5.8S rRNA gene, four exhibited higher β‐glucosidase activity, namely, Hanseniaspora uvarum, Trichosporon asahii, Pichia fermentans and Saccharomyces cerevisiae. The β‐glucosidases from the four representative strains were chosen to hydrolyse the glycosidic precursors of Cabernet Sauvignon. After enzymatic hydrolysis, 31 compounds were identified and quantified, including terpenes, C₁₃‐noriso‐prenoid, C₆ compounds, alcohols, aldehydes and volatile phenols. Results showed that different strains exhibited different hydrolytic abilities on the bound aroma precursors. The main variables included C₆ compounds, terpenes and alcohols. The concentration of the 14 compounds showed significant differences between enzymatic treatments, with 11 treated using the β‐glucosidase of the F6 strain (T. asahii). These findings may have some applicative value for utilizing the strains or their β‐glucosidases, which are able to complement and optimize wine quality.     

Anti‐inflammatory activity of rosemary extracts obtained by supercritical carbon dioxide enriched in carnosic acid and carnosol

The in vitro anti‐inflammatory activity of supercritical rosemary (Rosmarinus officinalis L.) extracts (rosemary A and B) is been reported in this study. To achieve that, THP‐1 macrophages were activated using lipopolysaccharide or human ox‐LDL and secretion and gene expression of TNF‐α, IL‐1β, IL‐6 and IL‐10 were evaluated, as well as COX‐2 gene expression. Results indicated that both rosemary extracts (A & B) exhibit high anti‐inflammatory activity although at a higher extent in case of rosemary B extract (5 μg mL^?1), representing a higher quantity of carnosic acid and carnosol than rosemary A. When comparing the activity of the extract to the standard itself, the anti‐inflammatory activity of standards of carnosic acid and carnosol was not as intense as that obtained with rosemary B. These data indicated that although carnosic acid content in the extracts is considered as the main anti‐inflammatory compound, a synergistic interaction with other compounds may play a significant role in enhancing its activity. Results provided the grounds for possible increase in the application of supercritical rosemary extracts in food formulations for mitigation or prevention of inflammatory diseases.     

Chemical composition and antimicrobial activity of the essential oil of Amomum tsao‐ko

BACKGROUND: The chemical composition of the essential oil obtained by hydrodistillation from the dried fruits of Amomum tsao‐ko was analysed by gas chromatography/mass spectroscopy (GC/MS). The antimicrobial activity of the oil was evaluated against 16 micro‐organisms using agar disc diffusion and broth microdilution methods. RESULTS: Seventy‐three compounds, constituting about 97.56% of the total oil, were identified. The main components were 1,8‐cineole (45.24%), ρ‐propylbenzaldehyde (6.04%), geraniol (5.11%), geranial (4.52%), α‐terpineol (3.59%) and α‐phellandrene (3.07%). The essential oil showed a broad spectrum of antimicrobial activity against all the tested micro‐organisms, including Gram‐positive and Gram‐negative bacteria, and fungi. The oil exerted the strongest bactericidal activity against Staphylococcus aureus CCTCC AB91118, with minimum inhibitory and bactericidal concentrations of 0.20 g L^?1. CONCLUSION: Our study suggests that the Amomum tsao‐ko essential oil could be one of new medicinal resources for antibacterial and antifungal agents. Copyright © 2008 Society of Chemical Industry