Metabolites in Conocarpus erectus leaves attenuate α-amylase activity by modulating amino acid residues of α-amylase: an in vitro and docking study / Los metabolitos en las hojas de Conocarpus erectus atenúan la actividad de la α-amilasa al modular los residuos de aminoácidos de la α-amilasa: un estudio in vitro y de acoplamiento

The antioxidant activity and the inhibitory potential of α-amylase of lyophilized hydroethanolic extracts of Conocarpus erectus leaves obtained by ultrasonication were determined. The most potent extract was subjected to ultra-high performance liquid chromatography system equipped with mass spectrometer for metabolite identification. The identified metabolites were docked in α-glucosidase to assess their binding mode. The results revealed that 60% ethanolic extract exhibited highest ferric reducing antioxidant power (4.08 ± 0.187 mg TE/g DE) and α-amylase inhibition (IC50 58.20 ± 1.25 µg/mL. The metabolites like ellagic acid, 3-O-methyl ellagic acid, ferujol, 5, 2 ́-dihydroxy-6,7,8-trimethyl flavone and kaempferol glucoside were identified in the extract and subjected to molecular docking studies regarding α-amylase inhibition. The comparison of binding affinities revealed 3-O-methyl ellagic acid as most effective inhibitor of α-amylase with binding energy of -14.5911 kcal/mol comparable to that of acarbose (-15.7815 kcal/mol). The secondary metabolites identified in the study may be extended further for functional food development with antidiabetic properties.

Se determinó la actividad antioxidante y el potencial inhibidor de la α-amilasa de extractos hidroetanólicos liofilizados de hojas de Conocarpus erectus obtenidos por ultrasónicación. El extracto más potente se sometió a un sistema de cromatografía líquida de ultra alto rendimiento equipado con un espectrómetro de masas para la identificación de metabolitos. Los metabolitos identificados se acoplaron en α-glucosidasa para evaluar su modo de unión. Los resultados revelaron que el extracto etanólico al 60% exhibió el mayor poder antioxidante reductor férrico (4.08 ± 0.187 mg TE/g DE) e inhibición de la α-amilasa (IC50 58.20 ± 1.25 µg/mL. Los metabolitos como el ácido elágico, 3-O-metil elágico ácido, ferujol, 5, 2 ́-dihidroxi-6,7,8-trimetil flavona y kaempferol glucósido se identificaron en el extracto y se sometieron a estudios de acoplamiento molecular con respecto a la inhibición de la α-amilasa. La comparación de las afinidades de unión reveló 3-O-metil El ácido elágico como inhibidor más eficaz de la α-amilasa con una energía de unión de -14,5911 kcal/mol comparable a la de la acarbosa (-15,7815 kcal/mol). Los metabolitos secundarios identificados en el estudio pueden ampliarse aún más para el desarrollo funcional de alimentos con propiedades antidiabéticas.

HPLC-UV/HRMS methods for the unambiguous detection of adulterations of Ginkgo biloba leaves with Sophora japonica fruits on an extract level.

CONTEXT: Ginkgo biloba L. (Ginkgoaceae) leaf extract is one of the most frequently sold herbal extracts. There have been reports on poor quality and adulteration of ginkgo leaf extracts or the powdered plant material with extracts or powder of Styphnolobium japonicum (L.) Schott (Fabaceae) (syn. Sophora japonica L.) fruits, which is rich in flavone glycosides. OBJECTIVE: The study investigates whether ginkgo leaves genuinely contain genistein and sophoricoside and whether these two substances could be used as markers to detect adulterations with sophora fruits. MATERIALS AND METHODS: A total of 33 samples of dried ginkgo leaves were sourced from controlled plantations in China, the USA, and France. After extraction, the samples were analyzed using two high-performance liquid chromatography (HPLC) coupled with UV/HRMS methods for the detection of genistein and sophoricoside, respectively. Chromatograms were compared to standard reference materials. RESULTS: In none of the tested ginkgo samples, neither genistein nor sophoricoside could be detected. The applied method was designed to separate genistein from apigenin. The latter is a genuine compound of ginkgo leaves, and its peak may have been previously misidentified as genistein because of the same molecular mass. The method for the detection of sophoricoside allows identification of the adulteration with sophora fruit without prior hydrolysis. By both HPLC methods, it was possible to detect adulterations of ≥2% sophora fruits in the investigated ginkgo extract. CONCLUSION: The methods allow unambiguous detection of adulterations of ginkgo leaves with sophora fruits, using genistein and sophoricoside as marker compounds.

HSCCC Separation of Three Main Compounds from the Crude Extract of Dracocephalum Tanguticum by Using Dimethyl Sulfoxide as Cosolvent.

Separation of natural compounds directly from the crude extract is a challenging work for traditional column chromatography. In the present study, an efficient method for separation of three main compounds from the crude extract of Dracocephalum tanguticum has been successfully established by high-speed counter-current chromatography (HSCCC). The crude extract was directly introduced into HSCCC by using dimethyl sulfoxide as cosolvent. Ethyl acetate/n-butyl alcohol/0.3% glacial acetic acid (4: 1: 5, v/v) system was used and three target compounds with purity higher than 80% were obtained. Preparative HPLC was used for further purification and three target compounds with purity higher than 98% were obtained. The compounds were identified as chlorogenic acid, pedaliin and pedaliin-6″-acetate.

Determination of 12 herbal compounds for estimating the presence of Angelica Gigas Root, Cornus Fruit, Licorice Root, Pueraria Root, and Schisandra Fruit in foods by LC-MS/MS.

A wide variety of plant raw materials thought to promote health are used as herbal medicines as well as foods. However, there is no legal maximum or minimum concentration limit on any herbal compound when these plant raw materials are used in processed foods. Legally, these processed foods are regulated only for harmful substances, and there is no other guarantee of their contents. Therefore, the objective of this study was to determine the concentrations of 12 herbal compounds (nodakenin, decursin, decursinol angelate, morroniside, loganin, glycyrrhizic acid, liquiritigenin, puerarin, daidzin, schisandrin, gomisin A, gomisin N) in commonly used plant raw materials, such as "Angelica Gigas root", "Cornus Fruit", "Liquorice Root", "Pueraria Root", and "Schisandra Fruit"; and also in 45 processed foods, using high-performance liquid chromatography-tandem mass spectrometry (LC-MS/MS). Method validation was performed successfully using the parameters of specificity, linearity, limit of detection (LOD), limit of quantification (LOQ), accuracy, precision, matrix effect, extraction recovery, and stability. The 12 herbal compounds were determined to be present in all the foods advertised as containing each ingredient, although in very low concentrations in some cases. Three solid samples labelled as 100% pure material from one herbal species also contained herbal compounds found in others, so that intentional or unintentional adulteration was suspected.

Integrative application of licorice root extract or lipoic acid with fulvic acid improves wheat production and defenses under salt stress conditions.

Although different plant extracts and plant growth regulators are used as biostimulants to support plants grown under salt stress conditions, little information is available regarding the use of licorice root extract (LRE) or lipoic acid (LA) as biostimulants. Studies on the application of LRE or LA in combination with fulvic acid (FA) as natural biostimulants have not been performed. Therefore, in this study, two pot experiments were conducted to evaluate the potential effects of LRE (5 g L-1) or LA (0.1 mM) supplemented as a foliar spray in combination with FA (0.2 mg kg-1 soil) on osmoprotectants and antioxidants, growth characteristics, photosynthetic pigments, nutrient uptake, and yield as well as on the anatomical features of the stems and leaves of wheat plants irrigated with three levels of saline water (0.70, 7.8, and 14.6 dSm-1). Moderate (7.8 dSm-1) and high (14.6 dSm-1) levels of salinity caused a significant (p ≤ 0.05) increase in the activities of SOD, APX CAT, POX, and GR as well as in electrolyte leakage, malondialdehyde level, and reactive oxygen species (O2‒ and H2O2) levels compared to those in controls (plants irrigated with tap water). However, the leaf relative water content, membrane stability index, NPK uptake, leaf area, plant height, spike length, straw yield, grain yield, and protein content of wheat grains significantly (p ≤ 0.05) decreased. Addition of LRE or LA and/or HA to wheat plants under saline stress significantly (p ≤ 0.05) enhanced their morphological and physio-biochemical characteristics in parallel with increases in the activities of enzymatic antioxidants. Salinity stress altered (p ≤ 0.05) wheat stem and leaf structures; however, treatment with LRE + FA significantly improved these negative effects. These findings indicate that FA + LRE treatment significantly improved the antioxidant defense system of the plants, thereby reducing ROS levels and increasing wheat growth and production under saline conditions.

Characteristics and Distribution of Organic Phosphorus Fractions in the Surface Sediments of the Inflow Rivers around Hongze Lake, China.

In this study, the characteristics and distribution of the organic phosphorus (Po) fractions in the surface sediments of seven inflow rivers around Hongze Lake in China were analyzed with a soil Po fraction method, as used by Ivanoff. The relationships between the Po fractions and physiochemical features of sediments were also discussed. The results showed that, the sediments of the rivers had been moderately pollution with certain ecological risk effects except the Waste Yellow River. The relative contribution order of the Po fractions in the sediments was residual Po > HCl-Po > fulvic acid-Po > humic acid-Po > labile organic phosphorus (LOP). Moderately labile organic phosphorus (MLOP) was the main part of the Po forms in the whole sediments. The risk of phosphorus released from river sediments was the highest in the western region, followed by the southwestern region, and finally the northwestern region. There were significant correlations between Po forms and total phosphorus (TP), inorganic phosphorus (Pi), and Po. Non labile organic phosphorus (NLOP) had the strongest correlation with TP. The distribution of Po forms in each region was different due to the impact of human activities, industrial and agricultural production and the land types; the heaver polluted sediments with higher Po fractions. It is suggested that most of the sediments of the inflow rivers in the regions have certain ecological risk effects and P of them have an important contributions on the eutrophication of Hongze Lake. Po forms can provide a reliable theoretical basis for dealing with the change of water quality and should be paid more attention in the lake eutrophication investigation. There was reciprocal transformation between different Po forms, especially non-bioavailable fraction can change into bio-available ones. The results can provide a basis for the earth cycle of phosphorus and a new perspective of eutrophication control of shallow lakes.

Traditional tonifying polyherbal infusion, Jatu-Phala-Tiga, exerts antioxidant activities and extends lifespan of Caenorhabditis elegans.

BACKGROUND: The imbalance between the generation of free radicals and natural cellular antioxidant defenses, known as oxidative stress, can cause oxidation of biomolecules and further contribute to aging-associated diseases. The purpose of this study was to evaluate the antioxidant capacities of Thai traditional tonifying preparation, Jatu-Phala-Tiga (JPT) and its herbal ingredients consisting of Phyllanthus emblica, Terminalia arjuna, Terminalia chebula, and Terminalia bellirica and further assess its effect on longevity. METHOD: Antioxidant activities of various extracts obtained from JPT and its herbal components were carried out using well-established methods including metal chelating, free radical scavenging, and ferric reducing antioxidant power assays. Qualitative analysis of the chemical composition from JPT water extract was done by high-performance liquid chromatography tandem with electrospray ionisation mass spectrometry. The effect of JPT water extract on the lifespan of Caenorhabditis elegans were additionally described. RESULTS: Among the extracts, JPT water extract exerted remarkable antioxidant activities as compared to the extracts from other solvents and individual constituting plant extract. JPT water extract was found to possess the highest metal chelating activity, with an IC50 value of 1.75 ± 0.05 mg/mL. Moreover, it exhibited remarkable scavenging activities towards DPPH, ABTS, and superoxide anion radicals, with IC50 values of 0.31 ± 0.02, 0.308 ± 0.004, and 0.055 ± 0.002 mg/mL, respectively. The ORAC and FRAP values of JPT water extract were 40.338 ± 2.273 µM of Trolox/µg of extract and 23.07 ± 1.84 mM FeSO4/mg sample, respectively. Several well-known antioxidant-related compounds including amaronols, quinic acid, gallic acid, fertaric acid, kurigalin, amlaic acid, isoterchebin, chebulagic acid, ginkgolide C, chebulinic acid, ellagic acid, and rutin were found in this extract. Treatment with JPT water extract at 1 and 5 mg/mL increased C. elegans lifespan under normal growth condition (7.26 ± 0.65 vs. 10.4 0± 0.75 (p < 0.01) and 10.00 ± 0.73 (p < 0.01) days, respectively). CONCLUSIONS: The results indicated that JPT and its herbal ingredients exhibited strong antioxidant activities, in particular the water extract of the polyherbal tonic. These findings rationalize further investigation in JPT infusion as a promising agent for anti-aging and oxidative stress prevention.

In vivo pro-angiogenic effects of dracorhodin perchlorate in zebrafish embryos: A novel bioactivity evaluation platform for commercial dragon blood samples.

Dragon blood has been used in wound treatment for many years and can be obtained from several distinct plant species. Dracorhodin, the active substituent of dragon blood, is a characteristic compound of the palm tree, Daemonorops draco. At present, the only method to evaluate the quality of commercial dragon blood samples is a HPLC method which determines the amount of dracorhodin in a dragon blood sample. In this study, we used zebrafish embryos as a platform to demonstrate the in vivo pro-angiogenic activity of dracorhodin perchlorate, the chemically synthesized analog of dracorhodin. By using this platform, three different commercial dragon blood samples were also examined. Our results clearly show that even though the commercial dragon blood samples had similar amounts of dracorhodin, they showed highly variable biological activity, such as pro-angiogenic effects and toxicity. In short, an in vivo activity assay platform for rapidly examining the biological activity of commercial dragon blood samples was successfully established here, which complements the current HPLC-based assay method.

Effects of Angelica gigas Nakai on the production of decursin- and decursinol angelate-enriched eggs.

BACKGROUND: The livestock industry requires high-quality products, as well as improved productivity. There have been many studies regarding the utilization of feed additives aiming to increase productivity, enhance immune functions and prevent infectious diseases in livestock. Biofunctional feed additives would be beneficial not only for animal health, but also for consumers. In the present study, we utilized root and byproduct (stem and leaf) powders of Angelica gigas Nakai (AGN, Korean Danggui) as feed additives and examined the deposition of biofunctional compounds, such as decursin and decursinol angelate, into egg white and yolk. RESULTS: We optimized the detection system for decursin and decursinol angelate, and determined the amounts of decursin and decursinol angelate derived from AGN byproducts (stem and leaf) as well as root. In Experiment 1, laying hens were fed with the dried AGN root powder and the effective compounds were detected in egg white and yolk. Subsequently, in Experiment 2, we examined AGN byproducts as an alternative feeding supplement. Additionally, biochemical parameters were analyzed to evaluate changes in the health of the hens by feeding AGN root powder. The results obtained indicated that decursin and decursinol angelate were stably transferred into egg white and yolk by feeding AGN byproducts as well as root. Intriguingly, plasma cholesterol levels were significantly decreased in a dose-dependent manner, and those of interleukin-1ß, as an immune-related biomarker, were considerably increased in the treated hens. CONCLUSION: These results indicated that AGN root and byproducts (stem and leaf) could be utilized for the production of value-added eggs and improving the health of hens in the poultry industry. © 2018 Society of Chemical Industry.

Antimicrobial activity and active compounds of a Rhus verniciflua Stokes extract.

The Rhus verniciflua Stokes (RVS) extract is used as a traditional herbal medicine in Southeast Asian countries such as Korea and China. In the present study, one phenolic acid and six flavonoids were isolated from an 80% ethanol RVS extract to examine their antimicrobial activities. These compounds were identified as 3',4',7-trihydroxyflavone (1), methyl gallate (2), gallic acid (3), fusti (4), fisetin (5), butin (6), and sulfuretin (7) by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry and nuclear magnetic resonance spectroscopy. The antimicrobial activities of compounds 5 and 6 (at a dose of 16 µg/mL each) were superior to that of the control, cycloheximide (at a dose of 25 µg/mL), against Hypocrea nigricans; additionally, the activities of compounds 1 and 2 (at a dose of 8 µg/mL each) were superior to the control against Penicillium oxalicum. Also, chemical compounds 1 and 5 (at a dose of 16 µg/mL each) had higher activities than the control (25 µg/mL) against Trichoderma virens. Chemical compound 1 (at a dose of 8 µg/mL) had a similar activity to that of the control against Bacillus subtilis. The obtained results suggest that the RVS extract could be a promising food and nutraceutical source because of the antimicrobial properties of its phenolic compounds.

A Comparative Metabolomics Analysis Reveals the Tissue-Specific Phenolic Profiling in Two Acanthopanax Species.

Acanthopanax senticosus (Rupr. Maxim.) Harms (ASH) and Acanthopanax sessiliflorus (Rupr. Maxim.) Seem (ASS), are members of the Araliaceae family, and both are used in Asian countries. These herbals have drawn much attention in recent years due to their strong biological activity, with innocuity and little side effects. However, the common and distinct mode of compound profiles between ASH and ASS is still unclear. In this study, a high performance liquid chromatograph-mass spectrometry (HPLC-MS) method was developed to simultaneously quantify the seven major active compounds, including protocatechuate, eleutheroside B, eleutheroside E, isofraxidin, hyperoside, kaempferol and oleanolic acid. Then the targeted metabolomics were conducted to identify 19 phenolic compounds, with tight relation to the above mentioned active compounds, including nine C6C3C6-type, six C6C3-type and four C6C1-type in the two Acanthopanax species studied here. The results showed that the seven active compounds presented a similar trend of changes in different tissues, with more abundant accumulation in roots and stems for both plants. From the view of plant species, the ASH plants possess higher abundance of compounds, especially in the tissues of roots and stems. For phenolics, the 19 phenols detected here could be clearly grouped into five main clusters based on their tissue-specific accumulation patterns. Roots are the tissue for the most abundance of their accumulations. C6C3C6-type compounds are the most widely existing type in both plants. In conclusion, the tissue- and species-specificity in accumulation of seven active compounds and phenolics were revealed in two Acanthopanax species.

Establishment and application of a new HPLC qualitative and quantitative assay for Gentiana Macrophyllae Radix based on characteristic constituents of anofinic acid and its derivatives.

Gentiana Macrophylla Radix (GMR) is officially used as traditional Chinese medicine, but easily confused with Gentianae Radix et Rhizoma (GRR) and adulterants. This study aimed to establish an HPLC method for qualitative and quantitative analysis of GMR based on characteristic components, anofinic acid and its derivatives. HPLC analysis was performed on a C18 column with gradient elution using acetonitrile and 0.1% phosphoric acid as mobile phase, and detected at 240 nm by conventional methodology validation. For fingerprint analysis, RSDs of relative retention times and relative peak areas of the characteristic peaks were within 0-1.10 and 0-4.08%, respectively. For determination of 2-methoxyanofinic acid, the calibration curve showed good linearity (R2 > 0.9999) within the test range. The RSDs of precision, repeatability and stability test did not exceed 2.46, 0.83 and 1.11%, respectively. The average recoveries were between 95.08 and 103.05% with RSDs ≤2.29%. The results showed that there was no significant difference among the four species of GMR, but there were significant differences among GMR, GRR and spurious breeds by principal component analysis and hierarchical cluster analysis. Anofinic acid and its derivatives, as the characteristic markers, could be used for the identification and quality control of GMR.

Application of metabolomics profiling in the analysis of metabolites and taste quality in different subtypes of white tea.

Three subtypes of white tea, Silver Needle (SN), White Peony (WP), and Shou Mei (SM), differ in their taste, aroma, bioactivity, and commercial value. Here, a metabolomics investigation on the chemical compositions combining taste equivalent-quantification and dose-over-threshold (DoT) determination on the taste qualities were applied to comprehensively characterize the white tea subtypes for the first time. Significant differences in the contents of catechins, dimeric catechins, amino acids, phenolic acids, flavonol/flavone glycosides, and aroma precursors were observed among these 3 white teas. Metabolite content comparison and partial least-squares (PLS) analysis suggest that theanine, aspartic acid, asparagine, and AMP were positively correlated with the umami taste in white tea, and flavan-3-ols, theasinensins, procyanidin B3, and theobromine had positive correlations with higher bitterness and astringency tastes. In addition, puckering astringent (-)-epigallocatechin gallate (EGCG), (-)-epicatechin gallate (ECG) and theogallin, bitter-tasting caffeine, and the mouth-drying/velvety-like astringent γ-aminobutyric acid (GABA) were identified as key taste compounds of white tea infusion by absolute quantification and DoT factor calculations. This work provided systematic and comprehensive knowledge on the chemical components, taste qualities, and sensory active metabolites for the subtypes of white tea.

Profiling and analysis of multiple constituents in Baizhu Shaoyao San before and after processing by stir-frying using UHPLC/Q-TOF-MS/MS coupled with multivariate statistical analysis.

Baizhu Shaoyao San (BSS) is a famous traditional Chinese medicinal formula widely used for the treatment of painful diarrhea, intestinal inflammation, and diarrhea-predominant irritable bowel syndrome. According to clinical medication, three medicinal herbs (Atractylodis Macrocephalae Rhizoma, Paeoniae Radix Alba, and Citri Reticulatae Pericarpium) included in BSS must be processed using some specific methods of stir-frying. On the basis of the classical theories of traditional Chinese medicine, the therapeutic effects of BSS would be significantly enhanced after processing. Generally, the changes of curative effects mainly result from the variations of inside chemical basis caused by the processing procedure. To find out the corresponding changes of chemical compositions in BSS after processing and to elucidate the material basis of the changed curative effects, an optimized ultra-high-performance liquid chromatography-quadrupole/time-of-flight mass spectrometry in positive and negative ion modes coupled with multivariate statistical analyses were developed. As a result, a total of 186 compounds were ultimately identified in crude and processed BSS, in which 62 marker compounds with significant differences between crude and processed BSS were found by principal component analysis and t-test. Compared with crude BSS, the contents of 23 compounds were remarkably decreased and the contents of 39 compounds showed notable increase in processed BSS. The transformation mechanisms of some changed compounds were appropriately inferred from the results. Furthermore, compounds with extremely significant differences might strengthen the effects of the whole herbal formula.

An emerging strategy for evaluating the grades of Keemun black tea by combinatory liquid chromatography-Orbitrap mass spectrometry-based untargeted metabolomics and inhibition effects on α-glucosidase and α-amylase.

Quantitative analysis and untargeted liquid chromatography mass spectrum (LC-MS) based metabolomics of different grades of Keemun black tea (KBT) were conducted. Quantitative analysis did not show tight correlation between tea grades and contents of polyphenols, but untargeted metabolomics analysis revealed that high-grades KBT were distinguished from the low-grades. S-plot and Variable Importance (VIP) analysis gave 28 marker compounds responsible for the discrimination of different grades of KBT. The inhibitory effects of KBT on α-amylase and α-glucosidase were positively correlated to tea grades, and the correlation coefficient between each marker compound and inhibitory rate were calculated. Thirteen compounds were positively related to the anti-glycemic activity, and theasinensin A, afzelechin gallate and kaempferol-glucoside were confirmed as grade-related bioactive marker compounds by chemical and bioassay in effective fractions. This study suggested that combinatory metabolomics and bioactivities assay provided a new strategy for the classification of tea grades.

Variations of Escherichia coli O157:H7 Survival in Purple Soils.

Escherichia coli O157:H7 is a well-recognized cause of human illness. Survival of Escherichia coli O157:H7 in five purple soils from Sichuan Province was investigated. The dynamics of E. coli O157:H7 survival in purple soils were described by the Weibull model. Results showed that this model is suitable to fit survival curves of E. coli O157:H7 in purple soils, with the calculated td value (survival time needed to reach the detection limit of 100 CFU·g-1) ranging from 2.99 days to 26.36 days. The longest survival time of E. coli O157:H7 was observed in neutral purple soils (24.49 days), followed by alkalescent purple soil (18.62 days) and acid purple soil (3.48 days). The redundancy analysis (RDA) revealed that td values were significantly enhanced by soil nutrition (total organic carbon (OC), total nitrogen (TN), available potassium (AK) and the ratio of humic acid to fulvic acid (Ha/Fa)), but were significantly suppressed by iron and aluminum oxide.

Physical and Bioactive Properties of Muffins Enriched with Raspberry and Cranberry Pomace Powder: A Promising Application of Fruit By-Products Rich in Biocompounds.

The objective of this study was to determine the effects on quality of incorporating raspberry and cranberry pomaces into American-style muffins prepared under various baking conditions. The different baking conditions did not affect the texture or microstructure of the control muffins. The enhanced samples baked at 140 °C for 30 min were characterized by a harder texture than the control muffins and by a distributed protein matrix and distorted starch granules, while those baked at 240 °C for 15 min had a moist texture and showed incomplete starch gelatinization. The mean percent recovery of ellagic acid, flavonols, tocopherols, tocotrienols, and anthocyanins after baking were 156, 53, 48, 43, and 22 %, respectively. Lower baking temperature was better for ellagic acid and tocotrienols, but worse for flavonols, tocopherols, and anthocyanins. It seems that, for the enhanced samples, the intermediate baking conditions (180 °C for 20 min) guarantee the best microstructure and texture and the appropriate retention of phytochemicals in muffins.

Characterizing the interaction between uranyl ion and fulvic acid using regional integration analysis (RIA) and fluorescence quenching.

The development of chemometric methods has substantially improved the quantitative usefulness of the fluorescence excitation-emission matrix (EEM) in the analysis of dissolved organic matter (DOM). In this study, Regional Integration Analysis (RIA) was used to quantitatively interpret EEMs and assess fluorescence quenching behavior in order to study the binding between uranyl ion and fulvic acid. Three fulvic acids including soil fulvic acid (SFA), Oyster River fulvic acid (ORFA) and Suwannee River fulvic acid (SRFA) were used and investigated by the spectroscopic techniques. The EEM spectra obtained were divided into five regions according to fluorescence structural features and two distinct peaks were observed in region III and region V. Fluorescence quenching analysis was conducted for these two regions with the stability constants, ligand concentrations and residual fluorescence values calculated using the Ryan-Weber model. Results indicated a relatively strong binding ability between uranyl ion and fulvic acid samples at low pH (log K value varies from 4.11 to 4.67 at pH 3.50). Fluorophores in region III showed a higher binding ability with fewer binding sites than in region V. Stability constants followed the order, SFA > ORFA > SRFA, while ligand concentrations followed the reverse order, SRFA > ORFA > SFA. A comparison between RIA and Parallel Factor Analysis (PARAFAC) data treatment methods was also performed and good agreement between these two methods (less than 4% difference in log K values) demonstrates the reliability of the RIA method in this study.

[Material Composition Research on Tibetan Medicine "Brag-zhun"].

OBJECTIVE: To study the material composition of Tibetan medicine "Brag-zhun", and to provide the basis for interpretation of its source and reference for the establishment of quality standards. METHODS: Pharmacognosy routine method was used in macroscopic and microscopic identifications, inductively coupled plasma spectrometry was carried out to determine 26 kinds of mineral elements,and X-ray diffraction analysis was used for the crystalline phase detection of Brag-zhun. The content of calcium,organic matter, humic acid and fulvic acid was determined according to GB/T 14610-2008, "technical specification for soil analysis" , GB/T 11957-2001 and capacity titration, respectively. Moisture, ash content, acid insoluble ash content and water soluble extract were determined with reference to the Chinese Pharmacopoeia (2010 edition). RESULTS: Animal feces were commonly found in Brag-zhun, the main crystal phase was SiO2, 26 kinds of mineral elements content in total was about 3%, 13 batches of medicinal materials in organic matter content was 29.03%-71.79%. Humic acid content was 28.37%-58.51%; fulvic acid content was 19.69%-41.43%; moisture was 5.2% -11.7%; total ash content was 17.82%-64.39%; acid insoluble ash content was 2.10%-39.09%; and water soluble extract was 28.39%-57.40%. CONCLUSION: The traditional record of Brag-zhun from molten juice is lack of scientific basis. The mineral elements in Brag-zhun exist in amorphous. Brag-zhun contains a lot of organic matter including humic acid and fulvic acid, and inorganic matter mainly comes from sediment. The organic matter in rock has gone through a long geological evolution.

Bergenin Content and Free Radical Scavenging Activity of Bergenia Extracts. .

Our research was focused on the evaluation of bergenin content and free radical scavenging activity of extracts prepared from three different species of Bergenia - B. crassifolia (L.) Fritsch., B. ciliata (Haw.) Sternb. and B. x ornata Stein. collected during different seasons. Using an HPLC method, the highest total amount of bergenin was revealed in the leaves of B. x ornata and B. crassifolia (4.9 - 5.1 mg x g(-1)). Free radical scavenging power was determined by two methods--FRAP and NADH. The best free radical scavengers were B. crassifolia (FRAP: 6.7 - 15.9 mg GAE. 100g(-1); NADH: 20.3 - 50.9%) and B. ornata (FRAP: 13.7 - 15.2 mg GAE. 100g(-1); NADH: 29.3 - 31.1%). The lowest content of bergenin and the weakest radical scavenger was B. ciliata (bergenin: 3.1 mg x g(-1); FRAP: 5.5 - 11.0 mg GAE.100g(-1); NADH: 23.2 - 25.6%). The presence of a large percentage of bergenin is responsible for the radical scavenging activity, as shown by the results from the FRAP and NADH assays. Significant, positive correlation was found between bergenin content and radical scavenging activity in both methods.