Improving Antioxidant Properties of Chitosan Films by Incorporating Extracted Fractions of Propolis

Chitosan films with propolis aqueous extract were obtained in order to improve the antioxidant capacity of the materials.Likewise,oleic acid(OA)was also incorporated to enhance the film hydrophobicity,with and without previous infusion with propolis,to extract other potentially active compounds.The propolis extracts and the chitosan film-forming dispersions were characterised as to their particle size distribution,zeta potential and rheological behaviour.Chitosan-based films with aqueous extract and with oleic acid,containing or not propolis compounds,were obtained and characterised as to their microstructure,mechanical,barrier and optical properties and antioxidant capacity in order to evaluate their ability to extend the food shelf life.The propolis compounds from the aqueous extract notably modified the chitosan conformation and charge density of chitosan molecules,giving rise to more folded chains with a milder viscous effect.These changes in the polymer gave rise to a coarser film microstructure,which exhibited greater stiffness,and less stretchability and resistance to break,but a greater barrier capacity to water vapour and oxygen.These films had a ten-fold greater antioxidant capacity,thus having great potential to preserve food from oxidative spoilage.The incorporation of OA,containing or not propolis compounds,into these films reduced the film stiffness and resistance to break,and led to a highly enhanced oxygen permeability without there being any notable changes in either the water vapour barrier capacity or antioxidant power.

Production of De-asphalted Oil and Fine Asphalt Particles by Supercritical Extraction

A continuous three-stage supercritical fluid extraction (SFE) process with a capacity of 1.0kg.h^-1 was setup to extract petroleum residue by pentane to obtain more oil for further upgrading. A discharging system integrated to the bottom of the extractor was used to recover solvent as gas while asphalt was obtained as fine particles. The influence of operating conditions on the yield and quality of extracts, i.e., deasphalted oil (DAO) and resin, was studied in the range of temperature 150-220℃, pressure of 4.0-6.0 MPa and the mass ratio of solvent to oil feed (S/O) 2.5-5.0. The particle size distribution, apparent forms and the packing density, which vary with operating pressure, were measured. The particle structures were observed by SEM as well. With the modification to conventional processes, furnace can be eliminated for solvent recovery from asphalt phase, so as to reduce energy consumption.

Optimization of Supercritical Extraction Process of Laoshan Black Tea by Response Surface Methodology

[Objectives]Laoshan black tea was subjected to supercritical CO_(2) extraction. [Methods]The extraction conditions of Laoshan black tea were studied by an orthogonal experiment and optimized by response surface methodology. [Results] The optimum extraction conditions of black tea extract by supercritical CO_(2) extraction were as follows: extraction pressure 23.53 MPa, extraction time 1.73 h, and extraction temperature 49.75 ℃, with which the extract yield could reach 5.15% theoretically. [Conclusions] Based on the traditional extraction process, a supercritical extraction method optimized by response surface methodology and a unique extraction process were formed, which enriches the extraction processes and methods of natural raw materials.

Fecal metabolomics reveals the positive effect of ethanol extract of propolis on T2DM mice

A large number of studies have shown that propolis has positive effects in the treatment of type 2 diabetes mellitus(T2DM).However,there are have only been a few reports that are based on an ultra-high performance liquid chromatography-quadrupole-time-of-flight-mass spectrometry(UPLC-Q-TOF-MS)analysis of the fecal metabolomics of ethanol extract of propolis(EEP)in the treatment of T2DM.The present investigation was designed to screen potential biomarkers of T2DM by the metabonomic method and to explain the possible anti-diabetes mechanism of EEP according to the changes in the biomarkers.The results showed that EEP improved the body weight(BW)of T2DM mice,lowered blood sugar levels,and significantly restored blood biochemical indicators related to T2DM,such as fasting insulin(FINS),homeostasis model assessment of insulin resistance(HOMA-IR),aspartate transaminase(AST),and alanine aminotransferase(ALT).Liver pathology showed that EEP reversed liver damage caused by T2DM.Metabolomics data identified 27 potential biomarkers in fecal samples.EEP effectively regulated the dysfunction in the metabolic pathways of glycerophospholipids,sphingolipids,riboflavins,and sterol lipids caused by T2DM.In summary,our research results revealed positive effects of EEP in the treatment of T2DM and provided potential candidate markers for further research and in the clinical treatment of T2DM.

Preservability of rabbit semen after chilled storage in tris based extender enriched with different concentrations of Propolis ethanolic extract (PEE)

Objective:To maintain semen quality of male rabbits during chilled storage by enrichment the tris based diluent with different concentrations of propolis ethanolic extracts.Methods: Total phenolic and total flavonoid contents, as well as antioxidant activity was determined in propolis ethanolic extract (PEE). The extract was analysed by HPLC for separation and identification of target metabolites. Semen was collected from 10 rabbit bucks, pooled, then divided into five aliquots (each of 500 μL) and diluted each in 5 mL Tris-citric acid-glucose-egg yolk extender (TCGY). The 1st aliquot served as control while PEE was added at concentration of 0.8, 1.2, 1.6 and 2.0 mg/5 mL tris extender in the aliquot 2, 3, 4 and 5 respectively. Diluted semen samples were subjected to cooling at 4℃ for 72 h. Sperm motility, sperm viability, sperm abnormality, sperm membrane integrity and acrosome integrity were evaluated in chilled semen allover the chilling period.Results:The resluts revealed presence of a considerable amount of total phenolic compounds (98.67 mg GAE/g extract) and total flavonoids (70.16 mg CE/g extract) which were parallel to an antioxidant activity assessed as ABTS, DPPH and FRAP (198.65, 180.18 and 306.17 mM TE/g extract respectively). The dominant phenolic acid was chlorogenic acids (3.959 mg/g extract). Other compounds were found in less amounts rosmarinic acid (3.959 mg/g extract), myrcetin (1.946 mg/g extract), kaempferol (1.089 mg/g extract) and apeginin-7-glucoside (1.113 mg/g extract). Obtained results clearly demonstrated that the addition of (1.2-1.6) mg PEE in the chilled extended rabbit semen proved to be beneficial for maintaining semen characteristics compared to control and the addition of 0.8 and 2 mg PEE.Conclusions: The enrichment of rabbit semen tris-basic extender with (1.2-1.6) mg PEE/5 mL tris-extender (as the best and safe concentrations) maintain the sperm characteristics in good condition all over 72 h of chilling.

The Polyphenol Content and Antimicrobial Activity of Selected Propolis’ Extracts

Propolis is a resinous natural product,produced by bees(Apis mellifera),from vegetable parts and plant secretions.Propolis’samples A,B,C and D were extracted with phosphate buffer saline(PBS)or with 70%EtOH at pH values 8.0,7.2 and 6.4 followed by:(1)reverse-phase high-performance liquid chromatography(RP-HPLC)on Purospher®Star RP-18 column,the quantity of caffeic acid,chrysin,pinocembrin and galangin was determined;(2)determination of total flavonoids in both extracts;(3)antimicrobial tests of both extracts against(a)Gram-positive bacteria:methicillin-resistant Staphylococcus aureus(MRSA),St.aureus,Streptococcus pyogenes,Str.agalactiae,(b)Gram-negative bacteria:Escherichia coli,Pseudomonas aeruginosa,Proteus mirabilis,Acinetobacter baumanii and(c)yeast:Candida albicans.The antimicrobial activity of propolis’PBS extracts against Gram-positive bacteria shows the lowest minimal inhibitory concentration(MIC,mg/mL)at pH 8.0 in sample C,followed by A,B and D.In sample C,MICs at pH 8.0 were 0.007(Str.agalactiae),0.015(MRSA),0.015(Str.pyogenes)and 0.007(St.aureus).The polyphenol content of sample C is:flavonoid content 5.47±0.62 mg/mL,caffeic acid 1.33±0.92 mg/mL,chrysin 41.02±4.22μg/mL,pinocembrin 2.93±0.33 mg/mL and galangin 41.87±4.23 mg/mL.PBS extracts against Gram-negative bacteria show the lowest MIC(mg/mL)at pH 8.0 in sample D,followed by B,C and A.In sample D,MICs at pH 8.0 were 0.003(Acin.baumanii,Pr.mirabilis,Ps.aeruginosa)and 0.007(E.coli).Polyphenol content of sample D is:flavonoids 8.28±0.92 mg/mL,caffeic acid 3.56±0.32 mg/mL,chrysin 677.42±68.42μg/mL,pinocembrin 146.49±13.89 mg/mL and galangin 59.81±5.86 mg/mL.The strongest anti C.albicans activity,with the lowest MIC(mg/mL),at pH 8.0 was in the sample C,followed by samples D,A and B.In sample C,the MIC at pH 8.0 is 0.001(PBS extract).The antimicrobial activities of selected propolis samples correlate with their polyphenol content,more precisely,flavonoid,caffeic acid,chrysin,pinocembrin and galangin content.

East Java Propolis Inhibits Cytokine Pro-inflammatory in Odontoblast-Like Cells Human Pulp

Inflammation in pulp tissue is caused by caries bacteria. Most bacteria found are Lactobacillus acidophilus. Propolis is a sticky resin material that is derived from the bees and the surrounding plants, which are reported to have several biological effects including anti-bacterial and anti-inflammatory. This study will prove the effectiveness of propolis against proinflammatory cytokines on odontoblast-like cells in human dental pulp. This study was performed on cultured odontoblast-like cells in pulp. Cell culture was derived from the pulp tissue of human M3 teeth that had been extracted. Odontoblast pulp culture was divided in three groups： group one, cultured cells with propolis; group two, cultured cells induced with inactive Lactobacillus acidophilus; group three, cultured cells induced with inactive Lactobacillus acidophilus and exposed to 3 μg/mL propolis. A measurement of TNF-α and TGF-β1 expression was done using the immunocytochemical technique to determine the effectiveness of propolis extracts from East Java Indonesia against proinfalmmatory cytokines. Data were analyzed using the analysis of variance （ANOVA） test （P = 0.05）. Propolis extract can inhibit the expression of TNF-α and increase the expression of TGF-β1 on odontoblast-like cell in human dental pulp. Anti-inflammatory effects of East Java propolis extract are associated with cytokine modulation.

Inhibitory Effects of Propolis Water Extract on the Proliferation of Cervical Cancer Cells

[Objectives]To investigate the effects of propolis extract on HeLa cells to provide theoretical guidance for facilitating clinical treatment.[Methods]The effects of propolis on inflammation,and proliferation were analyzed based on the levels of DNA transcriptional regulation and mRNA and protein expression levels.[Results]Propolis water extract(PWE)could inhibit the cell proliferation and production of DNA damage in a dosedependent manner.Moreover,the propolis water extract could significantly downregulate the expression of iNOS-Luc,PTGS-2-Luc,and IL-8-Luc,and that it was related to the expression of the NF-κB family protein.After the induction of HeLa cells by propolis,the expression of the cell cycle inhibitor gene p21 was increased,while that of the cell proliferation gene Ki67 was decreased.[Conclusions]Propolis water extract could significantly inhibit the cell proliferation and production of DNA damage,suggesting propolis as a potential candidate for the development of adjunctive therapy against cervical cancer.

Effect of Allelochemicals of Chinese-fir root extracted by supercritical CO_2 extraction on Chinese fir

Allelochemicals of Chinese-fir root was extracted by technology ofsupercritical CO_2 extraction under orthogonal experiment design, and it was used to analyzeallelopathic activity of Chinese-fir through bioassay of seed germination. The results showed thatas to the available rate of allelochemicals, the pressure and temperature of extraction were themost important factors. The allelochemicals of Chinese-fir root extracted by pure CO_2 and ethanolmixed with CO_2 have different allelopathic activities to seed germination, and the allelochemicalsextracted by ethanol mixed with CO_2 had stronger inhibitory effects on seed germination than thatextracted by pure CO_2.

Supercritical Fluid Extraction of Essential Oil from Dry Rhizome of Ligusticum chuanxiong Hort and Their Characterization by GC/MS

Supercritical fluid extraction (SFE) of essential oil from dry rhizome ofLigusticum chuanxiong Hort was developed. GC/MS was used for the determination of the composition ofessential oil. Forty-four compounds were identified. The conventional extraction method wasconducted in parallel for comparison. The extracts were qualitatively compared by GC/MS. The yieldsof SFE and steam distillation-extraction were 4.16 % ( v/w) and 0.8 % ( v/w), respectively.Application of SFE of zessential oil from dry rhizome of Ligustiaan chuanxiong Hort was preferable.

Extraction of betulin from bark of Betula platyphylla by supercritical carbon dioxide extraction

Betulin, which is a medicinal pentacyclic triterpene, is abundant in the bark of white birch (Betula platyphlly). The bark of birch was collected at Tayuan Forest Farm of Jiagedaqi, Heilongjiang Province in September 2000. Supercritical fluid extraction (SFE) that is a new separation technology has been used for the processing pharmaceutical and natural products. In this paper, the extraction of betulin from the bark of birch by supercritical CO2 extraction was studied. The authors investigated and analyzed a few parameters such as modifier dosage, extraction pressure and extraction temperature. The optimal extrac-tion conditions showed that the modifier dosage used for per gram bark powder was 1.5 mL, the extraction pressure was at 20 Mpa, and the extraction temperature was at 55 C. The velocity of flow of liquid CO2 was at 10 kg/h. The pressure and tem-perature in separation vessel were at 5.5 Mpa and 50 C, respectively.

Optimization of Supercritical Carbon dioxide Extraction of Ginger Essential Oil by Response Surface Method

[Objective] Ginger essential oil （GEO） is widely used in food production and medical field in recent years due to its prominent biological functions, and this study was conducted to obtain high-quality and high-purity ginger essential oil from the fresh ginger. [Method] GEO was extracted from ginger roots by supercritical fluid extraction （SFE） method. The effects of flow rate of CO2, mesh size of ginger powder and volume of entrainer were investigated by single-factor experiments and response surface method. The content and extraction rate of 6-gingerol represented the extraction index of GEO. [Result] The conditions were optimized as follows： flow rate of CO2 at 25 L/h, mesh size of ginger power of 80 mesh, and volume of anhydrous ethanol as entrainer of 92.46 ml. The optimal extraction rate of 6-gingerol was 3.21%, which was predicted by RSM. [Conclusion] The optimal process of supercritical carbon dioxide extraction of ginger essential oil was identified by singlefactor experiments and response surface method. The present study provides a satisfactory method for purifying GEO from ginger for industrial purpose.

Combination of Supercritical Fluid Extraction with Ultrasonic Extraction for Obtaining Sex Hormones and IGF-1 from Antler Velvet

Supercritical CO2 (SC-CO2) extraction technology and ultrasonic technology were used to extract two active sex hormones, estradiol and progesterone, and insulin-like growth factor-1 (IGF-1) from antler velvet. The effects of SC-CO2 extraction condition on the extraction yield and content of sex hormones, the ultrasonic extrac-tion condition on the content of IGF-1 and the SC-CO2 extraction condition on the activity remaining of IGF-1 were studied. The optimal conditions were obtained. The experimental results showed that, in presence of 75% ethanol as the co-solvent, the mean yield and content of estradiol and progesterone were 87.67 pg·g-1 and 1224.10 pg·g-1, 12.38 ng·g-1 and 354.06 ng·g-1, respectively, with extraction pressure of 30 MPa, temperature of 35°C, extraction time of 30 min and CO2 consumption of 15 L·g-1 at the flow rate of 2.0 L·min-1. The highest content of IGF-1 was 7425.75 ng·g-1 antler velvet residue, when the pH10 ammonia-ammonium chloride buffer solution was used as the solvent, the ratio of solvent to sample was 20/1 (volume/mass), the extraction temperature was 0-35°C, and the ex-traction time was 4×15 min. Under these conditions, 93.68% activity remaining of IGF-1 in the residue was ob-tained, while little IGF-1 activity exists in traditional residue. The experimental results indicate that the technology of SC-CO2 with co-solvent is of advantage for getting high content sexual hormones and keeping high activity of IGF-1 in the residue, which can not be achieved by traditional extraction methods.

A Comparative Study on Chemical Composition of Supercritical CO_2 Extraction Products in Peels of Trichosanthes kirilowii Maxim. from Changqing District

[Objective] This study aimed to investigate the differences in chemical composition of supercritical CO2 extraction products in peels of Trichosanthes kirilowii Maxim. from Changqing district. [Method] Supercritical fluidextraction （SFE） and GCMS method were applied to determine and analyze the chemical components of the extracts in peels of three strains of Trichosanthes kirilowii Maxim. [Result] The chemical components of supercritical CO2 extraction products in peels of three strains of Trichosanthes kirilowii Maxim. varied., and the number of chemical components with normalized percentage content higher than 1% was 5, 7 and 8, respectively. There are 14 kinds of common components, and the relative content of hexadecanoic acid was the highest. [Conclusion] Supercritical CO2 extracts in peels of different strains of Trichosanthes kirilowii Maxim. contain different chemical components, providing scientific basis for breeding excellent varieties and the development and utilization of Trichosanthes kirilowii Maxim.

Effect of Allelochemicals of Chinese-fir root extracted by supercritical CO2 extraction on Chinese fir

Allelochemicals of Chinese-fir root was extracted by technology of supercritical CO2 extraction under orthogonal experiment design, and it was used to analyze allelopathic activity of Chinese-fir through bioassay of seed germination. The results showed that as to the available rate of allelochemicals, the pressure and temperature of extraction were the most important factors. The allelochemicals of Chinese-fir root extracted by pure CO2 and ethanol mixed with CO2 have different allelopathic activities to seed germination, and the allelochemicals extracted by ethanol mixed with CO2 had stronger inhibitory effects on seed germination than that extracted by pure CO2.

Evaluating bioactivity and bioaccessibility properties of the propolis extract prepared with L-lactic acid:An alternative solvent to ethanol for propolis extraction

Ethanol readily solubilizes the active components of propolis.Due to certain disadvantages of ethanolic extracts,developing new non-ethanolic extraction methods is highly desirable.This study aimed to determine the content of phenolic compounds and antioxidant activity of propolis by using lactic acid extraction compared to ethanol.Various concentrations of ethanol and lactic acid were utilized to produce extracts,and an in vitro digestion procedure was applied to the extracts to compare the bioaccessibility of phenolic compounds by using LC-MS/MS in extracts and HPLC-DAD in digested samples.The compounds detected in the lactic acid-based propolis so-lution had similar values to those in the ethanol solution at a propolis concentration of 10%.The propolis-lactic acid solution had a significantly higher antioxidant activity against DPPH radical than the one prepared with ethanol at the same concentration(p<0.05).Chrysin and naringenin were found as the most abundant phenolic compounds in the initial samples during in vitro digestion experiments.Pinocembrin was the most bioaccessible component among the dialyzed IN fractions of ethanol and lactic acid-based solutions,whereas some compounds were not detected.The results suggest that lactic acid may be used as an alternative to ethanol for propolis extraction.

Extraction of Perilla Seed Oil Using Supercritical Carbon Dioxide and GC-MS Analysis

[Objective] This study was aimed to determine the optimal parameters for the extraction of perilla seed oil to obtain high-quality perilla seed oil and analyze its compositions. [Method] In this study, perilla seed oil was extracted using supercritical CO2 （SC-CO2）. The effects of extraction time, temperature and pressure were investigated by single-factor experiments and orthogonal array testing （ORT）. The chemical compositions of extracted perilla seed oil were investigated by GC-MS. [Result] The optimal conditions for the extraction of perilla seed oil using SC-CO2 were extraction time of 4 h, extraction temperature at 40 ℃, and extraction pressure at 23 MPa. Under these conditions, the extraction yield of perilla seed oil was maximized to 12.43%. GC-MS analysis revealed that perilla seed oil was a complex mixture containing 76.183% α-linolenic acid. [Conclusion] Supercritical CO2 extraction was proven to be an effective technology to extract oil from perilla seed, and GCMS was also a satisfactory method for analyzing the compositions of perilla seed oil.

Isolation and Extraction of Total Flavonoids from Epimedium Koreanum Nakai by Supercritical Fluid Extraction

In the present paper is reported the method for the isolation and extraction of total flavonoids of Epimedium Koreanum Nakai by means of supercritical fluid extraction(SFE). By examining pressure, temperature, amounts of modifier and extraction time, the optimized condition of SFE is confirmed as 30 MPa and 60 ℃, with 70% ethanol as the modifier. The samples were statically extracted for 30 min, followed by dynamic extraction for 120 min at a flow rate of 6 mL/min. The quantitative analysis of total flavonoids was performed by UV-Vis spectrophotometry. Compared with the conventional method, the SFE method is more efficient, more rapid and more friendly environmentally.

Supercritical Fluid Extraction of Total Flavonoids from Leaves of Acanthopanax Senticosus Harms

The herbal plant Acanthopanax Senticosus Harms is natural herb of Changbaishan in Jilin Province of China, which belongs to the Araliaceae family. As the ingredients of folk medicine, it has long been used to treat a variety of human diseases, such as cerebrovascular disease, diabetes, tumor, isochemic heart diseases, hypertension, rheumatic arthritis, etc. E2,33. Flavonoids, a class of constituent compounds, which have a broad distribution in the nature and are found in Acanthopanax Senticosus Harms, have gained particular attention. A number of studies have shown that flavonoid compounds have wide biological activities, such as anti-aging, anticancer, anti-HIV,

Experimental optimization and mathematical modeling of supercritical carbon dioxide extraction of essential oil from Pogostemon cablin

The supercritical carbon dioxide extraction was applied to obtain essential oil from Pogostemon cablin in this work.Effect of extraction parameters including temperature,pressure,extraction time and particle size on extraction yield was investigated,and the response surface methodology with a Box–Behnken Design was used to achieve the optimized extraction conditions.The maximum yield of essential oil was 2.4356%under the conditions of extraction temperature 47°C,pressure 24.5 MPa and extraction time 119 min.Moreover,based on the Brunauer–Emmett–Teller theory of adsorption,a mathematical modeling was performed to correlate the measured data.The model shows a function relationship between extraction yield and time by a simple equation with three significantly adjustable parameters.These model parameters have been optimized through simulated annealing algorithm.The predicted data from the mathematical model show a good agreement with the experimental data of the different extraction parameters.