Study on Optimization of Ethanol Reflux Extraction of Phenolic Acids from Salvia miltiorrhiza

The extraction technology of phenolic acid compounds from Salvia miltiorrhiza by ethanol reflux was studied. In this experiment, salvianolic acid B standard was used to make the standard curve. Single factor experiment and orthogonal experiment were used to study the extraction of different ethanol concentrations, reflux times and material-to-liquid ratios. The OD value of salvianolic acid compounds was measured with a spectrophotometer. The extraction rate of phenolic acid compounds under different extraction conditions was calculated through a regression equation, so as to obtain the optimal conditions for the ethanol reflux extraction process of Salvia miltiorrhiza. The experimental data can provide a reference for the ethanol reflux extraction process of salvianolic acids in the industry. According to the experiment, the extraction rate of phenolic acids in Salvia miltiorrhiza was the highest when the ethanol concentration was 60%, the reflux time was 1.5 hours, and the ratio of material-to-liquid was 1:10.

Distribution of antioxidants and phenolic compounds in flour milling fractions from hard red winter wheat

Mature wheat kernels contain three main parts:endosperm,bran,and germ.Flour milling results in multiple streams that are chemically different;however,the distribution of antioxidants and phenolic compounds has not been well documented in terms of conventional milling by-product streams.In this study,multiple analytical methods were used to investigate antioxidant activity and phenolic compound compositions of hard red winter wheat(whole ground wheat),the parts of a wheat kernel(bran,flour,germ),and wheat by-product streams(mill feed,red dog,shorts)for the first time.For each mill stream,phenolic compounds(total,flavonoid,and anthocyanin contents)were determined and antioxidant activities were evaluated with 1,1-diphenyl-2-picrylhydrazyl(DPPH)radical-scavenging activity,ferric reducing/antioxidant power(FRAP),and total antioxidant capacity assays.Significant differences(P<0.05)were observed in phenolic concentrations among fractions of bran,flour,and germ milled from the same kernels and noted that germ accounts for the majority of antioxidant properties,whereas bran contains a substantial portion of phenolic compounds and anthocyanins.Mill feed was high in phenolic content(5.29 mg FAE/g),total antioxidant capacity(866 mg/g),and antioxidant activity(up to 75% DPPH inhibition and 20.26μmol FeSO_(4)/g).The comprehensive information on distribution of antioxidants and phenolic compounds provides insights for future human consumption of commonly produced co-products from flour milling,and for selecting and using different milling fractions to make foods with improved nutritional properties.

Induced Effects of Exogenous Phenolic Acids on Allelopathy of a Wild Rice Accession(Oryza longistaminata,S37)

Four exogenous phenolic acids, including salicylic acid, fumalic acid, p-coumaric acid and p-hydroxybenzonic acid, were used to investigate the regulatory effects on allelopathy of a wild rice accession of S37 （Oryza Iongistaminata）, which is a known allelopathic rice. The four exogenous phenolic acids induced the enhancement of the allelopathic potential of wild rice S37 in target weeds though the weed-suppressive activities were low, and the inducible effects were dependent on the specific phenolic acid, concentration and treatment time. After foliar application of exogenous phenolic acids, the inhibition rates for plant height, root length and fresh weight of barnyard grass （Echinochioa crus-galli） were significantly higher than those of the control. Especially at the concentration of 100 mg/L, the inhibition rates for plant height and fresh weight of barnyard grass by fumalic acid were 38.12% and 26.31% higher than those of the control, showing that fumalic acid was more effective compared with other phenolic acids in inhibiting monocotyledon weed growth. Furthermore, the weedsuppressive activity of aqueous extract from the leaves of wild rice S37 treated with exogenous phenolic acids was increased, and it peaked at 48 h after the treatment with the aqueous extract, and then gradually declined.

The effects of phenolic acid on nitrogen metabolism in Populus 3 euramericana ‘Neva’

The declines in soil fertility and productivity in continuously cropped poplar plantations axe related to phenolic acid accumulation in the soil. Nitrogen is a vital life element for poplar and whether the accumulation of phenolic acid could influence nitrogen metabolism in poplar and thereby hinder continuous cropping is not clear. In this study, poplar cuttings of Populus × euramericana ‘Neva＇ were potted in vermiculite, and phenolic acids at three concentrations （032, 0.5X and 1.0X） were added according to the actual content （1.0X） in the soil of a second-generation poplar plantation. Each treatment had eight replicates. We measured gas exchange parameters and the activities of key enzymes related to nitrogen metabolism in the leaves. Leaf photosynthetic parameters varied with the concentration of phenolic acids. The net photosynthetic rate （PN） significantly decreased with increasing phenolic acid concentration, and non-stomatal factors might have been the primary limitation for PN- The activities of nitrate reductase （NR）, glutamine synthetase （GS） and glutamate synthase （GOGAT）, as well as the contents of nitrate nitrogen, ammonium nitrogen, and total nitrogen in the leaves decreased with increasing phenolic acid concentration. This was significantly and positively related to PN （P 〈 0.05）. The low concentration of phe- nolic acids mainly affected the transformation process of NO3- to NO2-, while the high concentration of phenolic acids affected both processes, where NO3- was transferred to NO2- and NH4＋ was transferred to glutamine （Gln）. Overall, phenolic acid had significant inhibitory effects on the photosynthetic productivity of Populus x euramericana ＇Neva＇. This was probably due to its influence on the activities of nitrogen assimilation enzymes, which reduced the amount of amino acids that were translated into protein and enzymes. Improving the absorption and utilization of nitrogen by plants could help to overcome the problems caused by continuous cropping.

Effects of Phenolic Acids on Growth and Activities of Membrane Protective Enzymes of Cucumber Seedlings

Two phenolic acids P-hydroxy benzoic acid and cinnamic acid were designated as four concentrations (0, 50 μmol/L, 100 μmol/L, 150 μmol/L) to investigate the effects of phenoic acids on the growth and the activities of membrane protective enzymes of cucumber seedlings. The results showed that both phenolic acids inhibited the seedlings growth. The inhibitory effects were increased with the concentration of phenolic acids increasing and the time of treatment prolonging. Seedlings treated with A150 (P-hydroxy benzoic acid, 150 μmol/L), B50 (cinnamic acid, 50 μmol/L), B100 (cinnamic acid, 100 μmol/L), B150 (cinnamic acid, 150 μmol/L) showed significantly shorter in plant height , smaller in leaf area. and lighter in fresh weight. The inhibitory effect of cinnamic acid was comparatively stronger than that of P-hydroxy benzoic acid. For protective enzymes system, compared to control , the POD activity increased at all concentrations of P-hydroxy benzoic acid during the treatment but increased at first then decreased before increased again at last at all concentrations of cinnamic acid .In the case of CAT, its activity increased at first, then decreased, and increased again at lower concentrations of phenolic acids. However, at higher concentrations the activities decreased at first, then increased a little, decreased continuously at last. In addition, the treatments of phenolic acids led to an increase then a decreaseof SOD activity and an increase of MDA content in the seedlings. All above indicated the accumulating of free radicalsand destruction of protective enzymes at higher concentrations of phenolic acids.

PRZEWALSKINIC ACID A,A NEW PHENOLIC ACID FROM SALVIA PRZEWALSKII MAXIM

From the root of Salvia przewalskii Maxim.a new phenolic acid, przewalskinic acid A was isolated and the structure was established by the analysis of^(13)C-NMR,~1H-NMR and two-dimensional COSY experiments.

Catalytic ozonation of phenol and oxalic acid with copper-loaded activated carbon

A novel heterogeneous catalytic ozonation process in water treatment was studied,with a copper-loaded activated carbon(Cu/AC) that was prepared by an incipient wetness impregnation method at low temperature and tested as a catalyst in the ozonation of phenol and oxalic acid.Cu/AC was characterized using XRD,BET and SEM techniques.Compared with ozonation alone,the presence of Cu/AC in the ozonation processes significantly improves the degradation of phenol or oxalic acid.With the introduction of the hydroxyl radical scavenger,i.e.,turt-butanol alcohol(t-BuOH),the degradation efficiency of both phenol and oxalic acid in the Cu/AC catalyzed ozonation process decreases by 22% at 30 min.This indicates that Cu/AC accelerates ozone decomposition into certain concentration of hydroxyl radicals.The amount of Cu(Ⅱ) produced during the reaction of Cu/AC-catalyzed ozonation of phenol or oxalic acid is very small,which shows that the two processes are both heterogeneous catalytic ozonation reactions.

Application of HPLC and ESI-MS techniques in the analysis of phenolic acids and flavonoids from green leafy vegetables (GLVs)

Diets containing high proportions of fruits and vegetables reduce the risk of onset of chronic diseases. The role of herbal medicines in improving human health is gaining popularity over the years, which also increases the need for safety and efficiency of these products. Green leafy vegetables （GLVs） are the richest source of phenolic compounds with excellent antioxidant properties. Increased consumption of diets containing phenolic compounds may give positive and better results to human health and significantly improves the immune system. Highly selective, susceptible and versatile analytical techniques are necessary for extraction, identifica- tion, and quantification of phenolic compounds from plant extracts, which helps to utilize their important biological properties. Recent advances in the pre-treatment procedures, separation techniques and spectro- metry methods are used for qualitative and quantitative analysis of phenolic compounds. The online coupling of liquid chromatography with mass spectrometry （LC-MS） has become a useful tool in the metabolic profiling of plant samples. In this review, the separation and identification of phenolic acids and flavonoids from GLVs by LC-MS have been discussed along with the general extraction procedures and other sources of mass spectrometer used. The review is devoted to the understanding of the structural configuration, nature and accumulation pattern of phenolic acids and flavonoids in plants and to highlighting the recent developments in the chemical investigation of these compounds by chromatographic and spectroscopic techniques. It concludes with the advantages of the combination of these two methods and prospects.

Temperature and Acidified Solvent Effect on Total Anthocyanins and RP-HPLC Phenolic Acids Determination in Selected Spices

Total anthocyanins of spices (Syzygium aromaticum L., Coriadrum sativum L., Cuminum cyminum L., Zingiber officinale Rosc., Elettaria cardamomum, Curcuma longa, Rhus coriaria L., Cinnamomum zeylanicum Blume, Foeniculum vulgare Mill and Laurus nobilis L.) were determined using acidified (1% HCl) solvents (methanol, ethanol and acetone) at three temperatures (20℃, 40℃ and 60℃). Also phenolic acids were separated and identified by RP-HPLC. Results showed that sumac and cinnamon had the highest levels of anthocyanins, while for the acetone the cinnamon indicated the highest amount of anthocyanins when methanol and ethanol were used as extracting solvents at 20℃. At 40℃ using ethanol, sumac showed the highest level of anthocyanins whereas acetone solvent yielded the highest anthocyanin contents for cinnamon. At 60℃, cinnamon showed the highest level of anthocyanins when methanol and acetone were the solvents, while sumac had the highest anthocyanins level using ethanol as solvent. HPLC results showed ten phenolic acids found in those spices and varied in their concentrations. Gallic acid had the highest level (1642.3 mg/100g) (cloves). Gentisic acid had the lowest level (1.2 mg/100g) in ginger. Also sumac showed the highest level of chlorogenic acid (1528.7 mg/100g). Some acids were not found in some spices, for instance, benzoic acid was not found in coriander, cumin, ginger, green cardamom, cinnamon and sweet laurel.

Phenolic Acids Content and Nutritional Quality of Conventional, Organic and Biodynamic Cultivations of the Tomato CXD271BIO Breeding Line (Solanum lycopersicum L.)

This study was undertaken to evaluate the influence of growing systems (conventional, organic and biodynamic performed with two types of manuring) on some nutritional traits of the tomato breeding line CXD271BIO (Solanum lycopersicum L.). Experimental fields were in the same geographical area and the cultivations were performed over five years. The content of macronutrients, minerals (Ca, K, Mg, P, Na), trace elements (Fe, Zn, Cu, Mn) and phenolic acids (chlorogenic, caffeic, p-coumaric and ferulic acids) was determined. The biodynamic A growing system showed the highest protein and carbohydrates content compared to the other growing systems. Data on minerals and trace elements content showed significant differences (mainly in Ca, Na, Fe and Zn content) between conventional and biodynamic tomatoes, whereas no major impact of the fertilization among organic and the two biodynamic growing systems was observed. Appreciable differences in phenolic acids biosynthesis were observed, with the year of harvest showing a marked effect especially on chlorogenic, p-cumaric and ferulic acids content. Conversely, the cultivation system did not show major influence. Principal Components Analysis (PCA), performed on mineral and phenolic acids content, substantiated the large effect of the year of harvest, also suggesting an effect of the conventional cultivation system.

The Blocking Effect of Phenolic Acid on N-Nitrosomorpholine Formation in vitro

Phenolic acids (PAs) are widely found in many daily consumed vegetables and fruits.The inhibitory effects of PAs on Nnitrosomorpholine (NMOR) formation in vitro undersimulated gastric juice condition were studied. The results showed that the inhibjtoryIX)tency of thirteen varieties of PAs differed greatly, which may be related to theirchemical structures; the blocking rate (BR) of different killds of PAs were as follows:caf feic acid, 92.5%; tannic acid, 90.0%; gallic acid, 86.8%; sinaPinic acid, 86.2%; ferulicacid, 8l.l%; chlorogenic acid, 69.4%; gentisic acid, 69.2%; syringic acid, 62.1%; protocatechuic acid, 56.0%; p-coumaric acid, 52.5%; vannilic acid, 35.4%; moreover,p-hydroxybenzoic acid and m-coumaric acid had the least blocking effect or even slightcatalyzing effect. The results also demonstrated that amounts of NMOR formed werenegatively correlated with molar ratio of PAs to nitrite and that the optimum pH forinhibition was between 2 and 3.

Research Progress on Application Effect of Biomass Charcoal and Its Restoration of Soil Phenolic Acid Pollution

Biomass charcoal has positive effects on reducing soil nutrient loss, increasing fertilizer utilization, reducing organic pollution and pesti-cide residues, inhibiting pollutant enrichment, and reducing bioavailability of pollutants. In this paper, the concept and effect of biomass charcoal were reviewed; the research situation of phenolic pollutants in soil was described; and the research progress on restoration of soil phenolic acid pol-lution by biomass charcoal application was summarized.

Addition Mechanisms of Phenol toward Formaldehyde under Acidic Condition: a Theoretical Investigation

The reaction mechanisms of phenol with formaldehyde in the first and second addition at the ortho- and para-position in acid solution were theoretically investigated at the PW91/DNP level with solvent effects included. The reaction of phenol with protonated methanediol firstly forms an adduct intermediate, via a SN2 mechanism with a water molecule as the leaving group. From the adduct intermediate, there are two reaction channels involving a proton transfer to form the addition products. One is that a proton directly transfers via a four-membered ring transition state with a notable energy barrier （Four-member mechanism）. Another mechanism involving a water molecule as catalyst to mediate the proton transfer （WCP mechanism）, is a barrierless process, indicating that the formation of the adduct intermediate, the first reaction step, is rate-limiting. The reaction products are free hydroxymethyl phenols and/or hydroxybenzy carbocation （HOC6H4CH2＋） which plays an important role in the following formation of methylene and methylene ether linkages. The second addition reactions between formaldehyde and hydroxymethyl phenol at all possible reaction sites of the phenol ring in acid solution were also investigated and discussed.

Comparison between Sulfuric Acid-phenol and Sulfuric Acid-anthrone Methods Used for Determination of Polysaccharides in Shoots of Aralia elata (Miq.) Seem.

Sulfuric acid-phenol and sulfuric acid-anthrone methods were used to detect polysaccharide content in shoots of Aralia elata（ Miq.） Seem.,and the conversion factor to glucose was measured with refined polysaccharides. Comprehensive evaluation was carried out by linear relationship,precision,reproducibility,stability and recovery rate. The results showed that the linear relationship between glucose concentration and absorbance was good when glucose concentration was0-40 μg/ml,and the average recovery rate was equal to or higher than 97. 00% with good reproducibility（ RSD 〈 1. 60%,n = 5）. It revealed that the two methods were accurate and reliable,and suitable for the determination of polysaccharide content in the shoots of A. elata. Polysaccharide content detected by sulfuric acid-phenol and sulfuric acid-anthrone methods was 19. 31% and 20. 40% respectively.

Analysis of the Fatty Acids and Phenolic Compounds in a Cereal-Based Fermented Food (Tarhana)

Tarhana is a Turkish home-made fermented food product. The mixture of wheat flour, yoghurt, yeast, cooked vegetables, salt, and spices is fermented for 1 - 7 days. Tarhana is sun dried and ground for preparation of instant Tarhana powder. Color, phenolic compounds and fatty acid composition were determined for home-made Tarhana, collected from 4 cities in the Thrace Region of Turkey. Color and phenolic content were significantly different (P < 0.05). These differences were due to the variation of ingredients used at different locations. The major unsaturated fatty acids were oleic (C18:1, 16.41% - 57.69%), and linoleic acids (C18:2, 9.74% - 56.31%). Palmitic (C16:0, 16.90% - 37.66%) and stearic acids (C18:0, 0.65% - 11.34%) were the main saturated fatty acids. This study suggested that Tarhana soup is a good source of unsaturated fatty acids. Antioxidant properties of phenolic compounds and their effect on peroxidation of lipids in Tarhana should be tested in future studies.

Phenolic Acid Content and Free Radical-Scavenging Activity of Two Differently Processed Carob Tree(Ceratonia siliqua L.)Pod

The phenolic acids in free phenolic acid (FPHA), methanol-soluble phenolic ester (MSPE), and methanol-soluble phenolic glycoside (MSPG) fractions of two carob products, natural carob-pod flour (CPF) and commercial carob-pod flour (CCPF), were identified and quantified using high-performance liquid chromatography-mass spectrometry (HPLC-MS). Six phenolic acids were identified in the carob flours (gallic acid, protocatechuic acid, gentisic acid, syringic acid, p- coumaric acid, and sinapic acid). Gallic acid was the major phenolic acid;and its concentrations in CCPF were substantially higher than in CPF (135 - 166 and 85.2 - 91.3 mg/g dw, respectively). The concentrations of p-coumaric acid and sinapic acid were also higher in CCPF than in CPF. In contrast, the concentrations of protocatechuic acid and gentisic acid were generally lower in CCPF than in CPF, with the only exception being gentisic acid glycosides. Both carob-pod flours contained almost the same amount of sinapic acid and syringic acid. The total phenolic contents of the FPHA, MSPE, and MSPG fractions of CPF were found to be 44%, 38%, and 69% that of the respective fractions of CCPF. Correspondingly, the FPHA, MSPE, and MSPG fractions of CCPF had higher free radical-scavenging activity (28.4%, 33.1%, and 26.2%, respectively) than the corresponding fractions of CPF (9.2%, 28.0%, and 9.2%, respectively). Notably, the FPHA and MSPG fractions of each sample had very similar scavenging activity while the MSPE fraction always had higher activity. The FPHA and MSPG fractions of CPF had the lowest activities.

Simultaneous Determination of Four Phenolic Acids in Radix Salviae Miltiorrhizae Formula Granules by QAMS

[Objectives]This study aimed to establish a QAMS(quantitative analysis of multi-components by single-marker)method for simultaneous determination of four phenolic acids,salvianolic acid B,tanshinol,lithospermic acid and rosmarinic acid in Radix Salviae Miltiorrhizae by HPLC and conduct methodological investigation.[Methods]Salvianolic acid B,with moderate retention time,good resolution,great response value and easy availability was selected as the internal standard,and the relative retention values and relative correction factors between tanshinol,rosmarinic acid,lithospermic acid and salvianolic acid B were established.The results of QAMS were compared with those of external standard method to verify the accuracy and applicability of QAMS.[Results]There were no significant differences between the calculated values of QAMS and the measured values of external standard method.The relative deviation was less than 3%,and the relative correction factors obtained from the experiment were credible.[Conclusions]QAMS can be used to determine the contents of phenolic acids in Radix Salviae Miltiorrhizae formula granules.