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.     

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.     

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     

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.     

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.     

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