Analysis of anticancer compound,indole-3-carbinol,in broccoli using a new ultrasound-assisted dispersive-filter extraction method based on poly(deep eutectic solvent)-graphene oxide nanocomposite

Indole-3-carbinol(I3C),an important anticancer compound found in broccoli,has attracted considerable attention.The rapid extraction and accurate analysis of I3C in the pharmaceutical industry in broccoli is challenging as I3C is unstable at low pH and high temperature.In this study,a rapid,accurate,and lowcost ultrasound-assisted dispersive-filter extraction(UADFE)technique based on poly(deep eutectic solvent)-graphene oxide(PDES-GO)adsorbent was developed for the isolation and analysis of I3C in broccoli for the first time.PDES-GO with multiple adsorption interactions and a fast mass transfer rate was synthesized to accelerate adsorption and desorption.UADFE was developed by combining dispersive solid-phase extraction(DSPE)and filter solid-phase extraction(FSPE)to realize rapid extraction and separation.Based on the above two strategies,the proposed PDES-GO-UADFE method coupled with high-performance liquid chromatography(HPLC)allowed the rapid(15-16 min),accurate(84.3%-96.4%),and low-cost(adsorbent:3.00 mg)analysis of I3C in broccoli and was superior to solid-phase extraction,DSPE,and FSPE methods.The proposed method showed remarkable linearity(r=0.9998;range:0.0840-48.0 mg/g),low limit of quantification(0.0840 mg/g),and high precision(relative standard deviation<5.6%).Therefore,the PDES-GO-UADFE-HPLC method shows significant potential in the field of pharmaceutical analysis for the separation and analysis of anti-cancer compounds in complex plant samples.

A Review on Sources,Extractions and Analysis Methods of a Sustainable Biomaterial:Tannins

Condensed and hydrolysable tannins are non-toxic natural polyphenols that are a commercial commodity industrialized for tanning hides to obtain leather and for a growing number of other industrial applications mainly to substitute petroleum-based products.They are a definite class of sustainable materials of the forestry industry.They have been in operation for hundreds of years to manufacture leather and now for a growing number of applications in a variety of other industries,such as wood adhesives,metal coating,pharmaceutical/medical applications and several others.This review presents the main sources,either already or potentially commercial of this forestry by-materials,their industrial and laboratory extraction systems,their systems of analysis with their advantages and drawbacks,be these methods so simple to even appear primitive but nonetheless of proven effectiveness,or very modern and instrumental.It constitutes a basic but essential summary of what is necessary to know of these sustainable materials.In doing so,the review highlights some of the main challenges that remain to be addressed to deliver the quality and economics of tannin supply necessary to fulfill the industrial production requirements for some materials-based uses.

Online identification and extraction method of regional large-scale adjustable load-aggregation characteristics

This article introduces the concept of load aggregation,which involves a comprehensive analysis of loads to acquire their external characteristics for the purpose of modeling and analyzing power systems.The online identification method is a computer-involved approach for data collection,processing,and system identification,commonly used for adaptive control and prediction.This paper proposes a method for dynamically aggregating large-scale adjustable loads to support high proportions of new energy integration,aiming to study the aggregation characteristics of regional large-scale adjustable loads using online identification techniques and feature extraction methods.The experiment selected 300 central air conditioners as the research subject and analyzed their regulation characteristics,economic efficiency,and comfort.The experimental results show that as the adjustment time of the air conditioner increases from 5 minutes to 35 minutes,the stable adjustment quantity during the adjustment period decreases from 28.46 to 3.57,indicating that air conditioning loads can be controlled over a long period and have better adjustment effects in the short term.Overall,the experimental results of this paper demonstrate that analyzing the aggregation characteristics of regional large-scale adjustable loads using online identification techniques and feature extraction algorithms is effective.

Optimization of Ultrasound-assisted Extraction Conditions for Polysaccharides from Pinus tabulaeformis Seed Shells

[Objective] This study aimed to investigate the optimal technological condi- tions for ultrasouqd-assisted extraction of polysaccharides from Pinus tabulaeformis seed shells. [Melhod] Polysaccharides were extracted from P. tabulaeformis seed shells by ultrasound-assisted hot water extraction method and detected by phenol- concentrated sulfuric acid method. The effect of extraction temperature, extraction duration, solid-liquid ratio and extraction frequency on the extraction of polysaccha- rides was investigated using single-factor gradient experiment and the extraction conditions for polysaccharides from P. tabulaeformis seed shells were optimized us- ing L9（34） orthogonal design. [Result] The content of polysaccharides extracted twice at 60 ℃ for 25 min with solid-liquid ratio of 1：3.5 reached the maximum. The re- covery rate （n=-5） ranged from 97.71% to 100.67% and RSD was 1.56%, suggesting high stability. [Conclusion] A simple and efficient method for ultrasound-assisted ex- traction of polysaccharides from P. tabulaeformis seed shells was established in this study, which laid solid foundation for the development and utilization of abandoned pine nut shell resources.

Optimization of ultrasound-assisted extraction of camptothecin from Camptotheca acuminata seeds

Naturally occurring camptothecin（CPT） is an important source of chemotherapeutic agents.The extraction from Camptotheca acuminata is still the main approach to obtain CPT compared with total synthesis.In the present study,ultrasound-assisted extractions（UAE） of CPT from C.acuminata seeds with alkaline solutions were investigated and CPT yield were determined by High Performance Liquid Chromatography.The conditions of alkaline species and concentrations,extraction time,extraction temperature and ultrasonic power were optimized.Results show that both Na3PO4 and Na2CO3 solutions gain good extraction yields,whereas Na3PO4 solution has stronger basicity and need higher concentration than Na2CO3 solution does,thus aqueous Na2CO3 is more beneficial for the extraction.The optimal condition was ultrasonically extracted with 0.5% aqueous Na2CO3 at 50°C and ultrasonic power of 400 W for 60 min.Comparing with UAE with ethanol,the extraction with 0.5% Na2CO3 solution achieves higher yield.Moreover,aqueous Na2CO3 as a solvent has various advantages including non-toxicity,inflammable,non-corrosive and low cost,which ensure this UAE method is a superior method with high utilizing prospect.

Ultrasound-assisted Enzymatic Extraction of SDF on Passion Fruit Seeds

Ultrasound-assisted enzymatic extraction of soluble dietary fiber(SDF) from passion fruit seeds was carried out in order to shorten the extraction time and increase the yield of SDF. The effects of cellulase concentration, ultrasonic power, extraction temperature, solid-liquid ratio and water bath time on the SDF yield of passion fruit were studied by single factor analysis. And the orthogonal experiment was designed to optimize the extraction conditions. The results showed that the optimum conditions for the extraction of SDF from passion fruit by ultrasoundassisted enzymatic method were cellulase concentration 1.5%, ultrasonic power 120 W, solid-liquid ratio(g/mL) 1 ∶ 15, extraction temperature 55 ℃, and water bath time 160 min. The SDF extraction rate was determined to be 5.12%.

Effects of Ethanol Condensation Reflux Extraction and Ultrasound-assisted Extraction on the Content of Total Flavonoids of Pueraria edulis

With Pueraria edulis from different habitats in Yunnan Province,Guangdong Province and Guangxi Zhuang autonomous region as materials,the medicinal materials of P.edulis were extracted by ethanol condensation reflux extraction and ultrasound-assisted extraction,and the effects of different extraction methods the contents of total flavonoids in P.edulis from different areas were compared.The results showed that the best extraction time for P.edulis in Yunnan Province was 20 min,and that for Guangdong and Guangxi Zhuang Autonomous Region was 30 min.The yield of total flavonoids from P.edulis extracted by the condensation reflux extraction method was relatively better.The content of total flavonoids in P.edulis in Yunnan was the highest with an average content of 0.2557%,while the average content of total flavonoids in P.edulis in Guangdong was the lowest with an average content of 0.2108%.

Pectinmethylesterase extraction from orange solid wastes: Optimization and comparison between conventional and ultrasound-assisted treatments

During orange juice production, a half of fresh oranges weight is considered as production waste (peels, pulp, seeds, orange leaves and damaged orange fruits). An alternative for the management of these wastes is their treatment by addition of lime and a latter pressing, obtaining a press cake and a press liquor rich in sugars (10&#176;Brix) and citric acid, protein, pectin and ethanol. For non-thermal concentration of press liquor to obtain citruss molasses (65&#176;-70&#176;Brix), the removal of pectin is necessary. Traditionally, depectinization of juices has been done by using pectinmethylesterase (PME) enzymes from external sources. In this work it performed the extraction of PME enzymes from orange peels to obtain the optimum extraction conditions. Two different methods of solventextraction were compared (conventional andultrasound-assisted methods). For the conventional extraction experiments, a central composite design with three variables ([NaCl], pH and time) and five replicates of the center point was used. For ultrasound-assisted extraction, experiments were done at pH = 5.5 and [NaCl] = 1.25M), varying extraction time (1-30 min). Response variables were PME activity, protein content and a ratio between them, named PME effectiveness (ηPME). At the same experimental conditions (pH =5.5, [NaCl] = 1.25 M, t = 15 min) it was found that conventional extractions led to slightly better results in terms of ηPME than ultrasound-assisted extraction method.

Effect of Ultrasound-Assisted Extraction of Phenolic Compounds on the Characteristics of Walnut Shells

Walnuts are one of nature’s more waste-heavy products. 67% of the nut is shell and husk, low value by-products that are rich in phenolic compounds. The phenolic compounds extracted from walnut shells are potentially good natural sources of antioxidants for the food and pharmaceutical industries. In this study, phenolic compounds were extracted using an ultrasonic bath, an ultrasonic probe and a standard shaking method. The extraction yield achieved with an ultrasonic probe was 51.2 mg GAE/g DW, two times higher than both the shaking method and the ultrasonic bath method which were 20.6 mg GAE/g DW and 25.8 mg GAE/g DW, respectively. Phenolic extraction was further improved by a size reduction of the walnut shells. The best extraction yield of 52.8 mg GAE/g DW was attained when the particle size was between 45 - 100 mesh. The ultrasonic probe treatment is the best method for extraction of phenolic compounds from walnut shells. Scanning electron microscopy (SEM) imaging indicated that the ultrasonic probe treatment could better rupture the hard structure of the cells, increasing the penetration of solvents and thus the extraction yield.

Isolation of Chlorella vulgaris and Its DNA Extraction Methods

[Objective] The aim of this study is to isolate Chlorella vulgaris(chlorella)and extract its genomic DNA.[Method] Both the dilution method and drip method were employed to isolate chlorella from lake water samples;the conditions for culturing chlorella were optimized and its genomic DNA was extracted by improved CTAB method and SDS method.[Result] The proper conditions for chlorella culture were as following:temperature 20-25 ℃,illumination 4.39-5.86 W/m2 and rotational speed 100-150r/min;improved CTAB method was suitable for extracting genomic DNA from chlorella.[Conclusion] The study is helpful to study the chlorella at molecular level and promote the exploitation and utilization of chlorella resources.

Comparative Study on Four Methods for Quick Extraction of Sorghum Genomic DNA

[Objective] This study was to find out a quick,simple,and low-cost method for the extraction of sorghum genomic DNA.[Method] Four plant genomic DNA extraction methods based on CTAB,including liquid nitrogen grinding method（method I）,buffer grinding method（method II）,drying grinding method（method III）and directly grinding method（method IV）,were used to extract the sorghum genomic DNA from leaves;further the quantity and quality of the yielded DNA were detected by gel electrophoresis,SSR-PCR and SRAP-PCR.[Result] These four methods performed no remarkable difference in DNA product.The method I and method II produced DNA with higher purity and better integrity,which,especially from method I,is effective for SRAP-PCR and SSR-PCR.While the DNA extracted via method III and method IV had less integrality and lower purity,and only effective in SSR-PCR.[Conclusion] Enough amount of sorghum genomic DNA to perform tens of PCR could be quickly extracted using all these four methods.The DNA obtained via method I and method II had a broader application spectrum（SRAP,RAPD,ISSR and SSR）than that via method III and method IV which is only proper for PCR targeting small DNA fragments（SSR）.

Comparison of RNA Extraction Methods of Sugarcane Stem

[Objective] The study was to explore the most effective and feasible method for sugarcane stalks RNA extraction.[Method] SDS extraction method,kit extraction method and GHCL extraction method were used to extract the sugarcane RNA in stalks,and the quality of the extracted RNA was compared.[Result] RNA extracted by kit extraction method had a high-yield,the bands were clear and RNA had a good integrity,there was no significant degradation of RNA,and the OD260 nm/OD280 nm value was closed to 2.0.[Conclusion] Kit extraction method was the effectively method to extract sugarcane RNA,and this study had provided a theoretical basis for the molecular biology study of sugarcane.

Comparison on Four Extraction Methods of Genomic DNA from Clematis fasciculiflora Franch

[Objective] This study aimed at comparing the four extraction methods of genomic DNA from Clematis fasciculiflora Franch and determining the optimal extraction method for extracting the genomic DNA from Clematis fasciculiflora Franch.[Method] Leavies of Clematis fasciculiflora Franch were used as materials for comparing the purity and concentration of extracted DNA and extracting time among the four extraction methods of genomic DNA including improved CTAB method Ⅰ,improved CTAB method Ⅱ,improved CTAB method Ⅲ and improved SDS method.[Result] The four extraction methods could all be successfully used for extracting the genomic DNA from Clematis fasciculiflora Franch.The purity of genomic DNA was the highest using improved CTAB method Ⅰ,with the longest extracting time;while the concentration of genomic DNA was the maximum using the improved SDS method,with the shortest extracting time and relatively low purity;the extracting time of improved CTAB method Ⅲ was the shortest.[Conclusion] This study had established the optimal extraction method for extracting the genomic DNA from Clematis fasciculiflora Franch and supported for the further research using molecular biological methods.

Comparison of Genomic DNA Extraction Methods for Chenopodium quinoa Willd

To rapidly obtain high-quality genomic DNA from Chenopodium quinoa Willd, the genomic DAN in different tissues （leaves, stems and roots） of Chenopodi- um quinoa Willd was extracted by modified CTAB method, SDS method and high- salt Iow-pH method, respectively. The quality and yield of extracted DNA was deter- mined using agarose gel electrophoresis and UV spectrophotometry. At the same time, the PCR-SSR and SSCP molecular detection was also performed. The results showed that the gel test strips, without obvious decomposition, of all the extraction methods were relatively obvious; the genomic DNA yield extracted by modified CTAB method was highest, followed by that by SDS method, and the genomic DNA extracted by high-salt Iow-pH method was lowest： the genomic DNA yields extracted by different methods from Chenopodium quinoa Wiltd leaves were all high- er than those from roots and stems; the quality of Chenopodium quinoa Willd ge- nomic DNA extracted by modified CTAB method and high-salt Iow-pH method was better, and polyphenols, polysaccharides and other impurities were removed more completely. The PCR-SSR and SSCP detection results showed that the genomic DNA extracted by different methods from different tissues of Chenopodium quinoa Willd all could be better amplified, and high-quality strips could be obtained. So the Chenopodium quinoa Willd genomic DNA extracted by the three methods all can be used for subsequent molecular biology research.

Extraction of Pumpkin Polysaccharide by Complex Enzyme Method and Its Antioxidant Research

[Objective] This paper aims to study a new method of extracting pumpkin polysaccharide from pumpkin. Single factor experiments were conducted to examine the effects of extracting time,temperature,the solid-liquid ratio and pH value on the extraction yield of polysaccharide from pumpkin. [Method] The best enzyme ratio and extraction conditions for complex enzymes extraction were determined through orthogonal tests. Scavenging ·OH and O-2 activities of pumpkin polysaccharides were also investigated by salicylic acid and improved self-oxidation of o-pheno methods respectively. [Results] The results showed that the biggest extraction yield of polysaccharide from pumpkin can be got when adding 1% cellulose enzyme,1.5% pectinase,1.0% papain and Na2HPO4-citric acid buffer solution (pH was 4.6),and oscillating for 30 min under water at 40 ℃ with the solid-liquid ratio of 1:30. In addition,pumpkin polysaccharides had a strong activity of eliminating ·OH,but very weak activity to scavenge O-2. [Conclusion] This study provided basic data for research and application of Pumpkin polysaccharide.

A New Rapid and Batch-oriented Crushing Method for DNA Extraction from Maize Leaves

The crushing of leaves is the most time and effort-consuming part in DNA extraction which is a fundamental step in the study of molecular biology. In this study, a new rapid and batch-oriented crushing method for DNA extraction from maize leaves was developed. In addition, the practicability of the developed method in molecular marker-assisted breeding was verified using SSR molecular maker technology so as to provide a rapid, batch-oriented, low-cost and non-toxic leafcrushing method for a large number of molecular marker tests, improving test efficiency.

An Improved Method for DNA Extraction from the Faeces of Red Deer

[Objective] To introduce an improved method for DNA extraction from the faeces of red deer. [Method] Based on the traditional method of CTAB lysis, we proposed an improved DNA extraction method according to the characteristics of red deer faeces. [Result] This improved method extracted high-quality fecal DNA from Tianshan red deer and amplified the mitochondrial cytochrome b gene. With the muscle and fur DNA of red deer as the control, the sequencing results further con- firmed the reliability of the method. [Conclusion] The method requires no proteinase K in the process of extraction, and the extracted DNA can be used for PCR ampli- fication directly without the purification of DNA purification kit, thus, it is cost-saving.

A Mass and Rapid Method for DNA Extraction of Beauveria bassiana

[Objective] The aim was to optimize the mass and rapid method for DNA extraction of Beauvena bassiana. [Method] Boiling water DNA extraction method was improved, DNA extraction liquid was heated by PCR instrument and the extraction process was finished rapidly. [ Resuit] The quality of DNA obtained through mass and rapid extraction of fungal genomic DNA could meet the requirement of RAPD amplification analysis. The clear bands were amplified from 22 tested strains, the number of clear bands were different in the range of 2 -6 and the size of band were mainly concentrated in 450 -800 bp. The DNA extracted by this method also could completely meet the requirement of SCAR amplification. The amplified specific DNA bands used to mark the strain F263 were very clear. [Conclusion] This research provided relatively perfect method for mass and rapid extraction of fungal clenomic DNA.

A Comparative Study on Three Methods for the Extraction of Total RNA from Pinus bungeana

[Objective] The aim of this study was to compare three RNA extraction methods and thus find out the suitable one for isolating intact and high quality total RNA from Pinus bungeana.[Method] Employing three extraction methods of Trizol,RNeasy Mini Kit,LiCl precipitation,total RNAs of P.bungeana were extracted from pine leaf samples,and their integrity and purity were detected via agarose gel electrophoresis and spectrophotometry for a comparative study.[Result] Among the three extraction methods,LiCl precipitation method demonstrated higher yield and better integrity of total extracted RNA,with OD260/OD280 ratio between 1.8-2.0 and clear 28 S and 18 S bands in electrophoresis pattern.[Conclusion] LiCl precipitation method could be used to extract highly pure and intact total RNA from P.bungeana.

DNA extraction of birch leaves by improved CTAB method and optimization of its ISSR system

The basic method of DNA extraction （CTAB） was improved as the multi-times STE-CTAB extraction method and used to extract the DNA of birch leaved in this experiment. Results showed that the improved method is suitable not only for genomic DNA extraction of birch but also for that of other plants. The purity of genornic DNA extracted by the.multi-times STE-CTAB extraction method is higher than that by one time STE-CTAB method, and it does not need the process of RNase. The factors of influencing ISSR system were explored based on the genomic DNA of birch extracted by the two methods. The optimal conditions for ISSR system were determined as follows： Mg2＋ concentration is 1.5-3.0 mmol·L^-1, dNTP concentration 0.104）.25 mmol·L^-1, the quantity of Taq polymerase 0.5-2.0 U, template DNA 30-100 ng, and the concentration of primer is 0.2-0.4 pmmol·L^-1, and the reaction program was as： initial denaturation for 5 min at 94℃, 30 cycles of denaturation for 30 s at 94℃,annealing for 30 s at 51℃, extension for 30 s at 72℃, and a final 7 min extension at 72℃.