Effective Extraction Procedure Research Articles

Exploring Varied (Green) Extraction Methods to Optimize Galia Melon Peel Antioxidant Potential

Cucumis melo L. (C. melo), commonly known as the melon, is a widely cultivated tropical fruit associated with nutritional benefits and bioactive properties. With global production reaching 40 million tons annually, the fruit processing industry generates significant waste, primarily peels, totaling 8 to 20 million tons yearly. These organic by-products are rich in bioactive compounds such as antioxidants, offering health benefits such as a reduced risk of cancer and cardiovascular diseases, as well as of diabetes and neurogenerative diseases, offering an opportunity for sustainable utilization. C. melo by-products have demonstrated various health benefits, including anti-inflammatory, analgesic, and antioxidant properties, attributed mainly to polyphenols. Recognizing the potential of melon waste, this study systematically explored different extraction methods, including stirring (ST), ultrasound (US), and pulsed electric field (PEF) methods, while considering factors such as extraction time, temperature, and solvent composition. The primary goal was to identify the most effective extraction procedures and optimal conditions for maximizing the yield of total polyphenols and antioxidant capacity (using the FRAP and DPPH methods) from C. melo peel by-products. According to the results, the optimum conditions include ST as the extraction method, an ethanolic solvent with a strength of 50%, a 150 min extraction duration, and an 80 °C extraction temperature. The maximum values of total polyphenols that can be observed are 3.75 mg gallic acid equivalents (GAE)/g of dry weight (dw) and 25.77 μmol ascorbic acid equivalents (AAE)/g dw and 34.44 μmol AAE/g dw from FRAP and DPPH antioxidant assays, respectively. The polyphenols identified were the following: gallic acid, neochlorogenic acid, catechin, chlorogenic acid, epicatechin, and kaempferol. By securing the maximum isolation of bioactive content and antioxidant activity, the research will contribute to sustainable waste management by reducing waste and developing value-added products.

Optimization of the QuEChERS Extraction of Mycotoxins in Maize Using Response Surface Methodology (RSM)

AbstractSeveral methods have been developed for the analysis and detection of mycotoxins in food; however, most do not make use of critical statistics and mathematical tools for precise optimization. This study developed, optimized, and validated a modified quick, easy, cheap, effective, rugged, and safe (QuEChERS) extraction procedure for the extraction of multiple mycotoxins in maize and subsequent validation using liquid chromatography tandem mass spectrometry (LC–MS/MS). Central composite design (CCD) was used to optimize extraction conditions. Data analysis of full factorial screening experiments revealed that MeCN (%), FA (%), and extraction time significantly affected the mycotoxins recovery. Assessment of the statistical significance of the generated model using analysis of variance (ANOVA), coefficient tables, and surface plots showed the relative interactions of factors and the adequacy of the model. Thus, P values from the lack of fit (LOF) test ranged from 0.137– 0.467 and a composite desirability function of 0.91 was obtained. Using the optimum extraction conditions of 0.1% (v/v) FA in 80.2% MeCN for 74 min, 10 mycotoxins were effectively extracted with satisfactory recoveries (85–114%), coefficients of regression (R2 > 0.98), coefficients of variation (CVs < 15%), limit of quantifications (LOQs) (0.33–60.45 µg/kg), and other associated method validation parameters. The method validation was carried out according to Commission Implementing Regulation 2021/808 and Commission Regulation (EC) No 401/2006 of 23 February 2006. Application of this method to 20 maize samples collected from markets in Botswana showed detectable mycotoxins in 13 samples, with 2 exceeding the European Union (EU) maximum aflatoxin B1 (AFB1) limit, suggesting potential exposure to high levels of toxic mycotoxins in Botswana.

Extraction, isolation, and standardization of anthocyanin from Vaccinium myrtillus to evaluate their potential in glaucoma

In order to preserve the structural integrity and bioactivity of anthocyanins while improving their extraction, identification, and isolation from Vaccinium myrtillus berries is the goal of this study. The following three goals were pursued: · Creating effective extraction procedures, · Using cutting-edge chromatographic techniques for identification, and · Isolating certain anthocyanin chemicals for possible bioactivity research. Three extraction techniques (A, B, and C) were contrasted with a fresh berry control for Objective 1. The maximum anthocyanin production (14.2 mg/g) and antioxidant activity (90%) were produced by Method C, which can be attributable to the high concentration of Petunidin-3-glucoside (55%). Method A produced anthocyanins with a concentration of 12.5 mg/g, predominantly made up of cyanidin-3-glucoside (45%), compared to Method B's 9.8 mg/g, primarily made up of delphinidin-3-glucoside (30%). The structural integrity of each approach was kept to a satisfactory level, as shown by the absorbance tests. Objective 2 was to identify anthocyanins using sophisticated chromatographic methods. Cyanidin-3-glucoside (5.21 min, 12500), Delphinidin-3-glucoside (7.46 min, 9800), and Petunidin-3-glucoside (9.83 min, 14200) were measured for retention times and peak areas. Anthocyanin molecules were identified using purification methods to work towards Objective 3. Three fractions were collected, and the anthocyanin chemicals in each fraction were identified: Fraction 1, 2.5 mg, 95% purity of Cyanidin-3-glucoside; Fraction 2, 1.8 mg, 90% purity of Delphinidin-3-glucoside; and Fraction 3, 3.2 mg, 92% purity of Petunidin-3-glucoside. Analysing UV-Vis spectra showed the purity, and chromatographic retention durations matched benchmark values. Further confirmation of chemical identities was obtained by mass spectrometry and NMR investigations, with detected peaks matching recognised references. This thorough investigation contributes to our understanding of how to extract and characterise anthocyanins from V. myrtillus berries. The findings demonstrate that Method C is a better extraction technique, producing high anthocyanin content while maintaining bioactivity.

From Aroma Chemistry via Bioactives to Toxicology – Remembering the Broad Expertise of Michael Granvogl in Food Science

On March 23, 2022, the field of food science lost a pioneering and impassioned scientist through the tragic and untimely passing of Prof. Michael Granvogl. Born on December 21, 1974 in Munich, Germany Michael’s inquisitive nature led him to study food chemistry at the Technical University of Munich (TUM). After graduating in 1998, he remained at TUM to pursue a doctoral degree (conferred in 2007), then habilitation (2016), qualifying him as Associate Professor in Food Chemistry. In 2019, Michael Granvogl was appointed Full Professor at the Institute of Food Chemistry, University of Hohenheim, Germany where he managed a young and dynamic team of enthusiastic scientists in their pursuits to push the boundaries of food chemistry research in their specialized disciplines. Indeed, Michael was a renowned international subject-matter expert on multiple facets of food chemistry, foremost on aroma-active compounds and food-borne toxicants, with a deep knowledge of their formation and degradation mechanisms, and with skilled application of effective extraction procedures and comprehensive analytical techniques to explore and elucidate their nature. Beyond his teaching and research, Michael was a staunch and dedicated member of the Division of Agricultural and Food Chemistry (AGFD) of the American Chemical Society (ACS), taking active and prominent roles on the executive committee to promote the field and support the next generation of scientists. As a native of the city of Munich, it might be noted that his passion for science, evidenced in lecture theaters and laboratories, was mirrored in his private life through his enthused and loyal support of his native soccer team, FC Bayern Munich. That team must now move forward with the loss of one dedicated fan, but the field of food chemistry – in particular the sub-discipline food bioactives – must overcome the void previously filled with Michael Granvogl’s stalwart presence and must cope with the absence of discoveries that will no longer materialize. There are many researchers in the field who lost a dedicated mentor, an esteemed colleague and a dear friend on that fateful day in spring 2022.

Comparison of Different Extraction Procedures for Isolating Bacteriostatic and Anti-pseudorabies Virus Components from Rodgersia Sambucifolia (Hemsl)

Medicinal plants are considered a repository of bioactive ingredients. Rodgersia sambucifolia (Hemsl), a herbaceous perennial originating from East Asia, has demonstrated bacteriostatic and antiviral properties. The aim of this study was to identify an effective extraction procedure for isolating potential bacteriostatic and antiviral compounds from the plant. The crude extracts were obtained using ethanol, methanol or acetone, and stepwise extracted with ethyl acetate, n-butanol or water. The minimum inhibitory concentration (MIC), minimum bactericidal concentration (MBC), and diameters of the inhibition zone were evaluated for bacteriostatic efficacy. These tests were carried out with Enterococcus faecalis, Enterococcus faecium, Shigella dysenteriae, Streptococcus lactis, Staphylococcus aureus, and Streptococcus lactis. The antiviral activities of prevention, treatment, and neutralization, utilizing pseudorabies virus, were evaluated using swine testicular cell viability. The active ingredients from the best bioactivity extraction procedure were analyzed using liquid chromatography-mass spectrometry (LC-MS). When ranked highest to lowest, the bacteriostatic and antiviral effects were obtained in the order of the crude extracts prepared from ethanol, methanol, and acetone. Bacteriostatic and antiviral effects were highest in the stepwise extraction of ethyl acetate, followed by the extraction of n-butanol and water. The main compounds identified by LC-MS in the ethyl acetate phase in descending order of content were: bergenin, palmitic acid, baicalein, linoleic acid, chrysin, γ-linolenic acid, catechin gallate, catechin, ursolic acid, and baicalin. This study provides a research basis for the development of bacteriostatic and antiviral drugs from traditional Chinese herbal medicines.

Development of a sensitive LC-APCI-MS/MS method for simultaneous determination of eleven nitrosamines in valsartan and irbesartan with a simple extraction approach

Nitrosamines (NAs) are potent genotoxic agents (GAs) in several animal species, and some are classified as probable or possible human carcinogens by the International Agency for Research on Cancer (IARC). In July 2018, angiotensin II receptor blockers (ARBs) which are used to treat high blood pressure have been recalled owing to contamination with NAs. In this study, a simple and sensitive method for the determination of eleven NAs in a single analysis was developed, using atmospheric pressure chemical ionisation source coupled liquid chromatography tandem mass spectrometer (LC-APCI-MS/MS). By performing the 17 min-run in dynamic multiple reaction monitoring (dMRM) mode, eleven NAs were separated on a Poroshell HPH C18 (4.6 × 150 mm, 2.7 µm) column with gradient elution implementing mobile phase A consisting of 0.2 % formic acid in water and mobile phase B consisting of methanol. The developed analytical method was successfully applied in both active pharmaceutical ingredients (APIs) and finished products (FPs) of valsartan and irbesartan with straightforward and effective extraction procedures. Good linearity with a correlation coefficient (R2) > 0.996 was achieved over the concentration in a range of 0.5–50 ng/mL. The limits of detection (LODs) ranged in 0.001–0.008 ppm and limits of quantitation (LOQs) ranged in 0.008–0.05 ppm of the method fulfilled thresholds of US Food and Drug Administration (US-FDA) and European Medicines Agency (EMA) for testing of GAs in valsartan and irbesartan. The accuracy of the proposed method ranged from 73.1 % to 115.2 % for APIs and the relative standard deviation (RSD %) was ≤11.3 while these validation parameters were in the range of 80.2–128.5 % and ≤ 10.6 for FPs, respectively.

Comparative Study of Various Procedures for Extracting Doxorubicin from Animal Tissue Samples

This article presents a comparative study of selected deproteinization-, liquid–liquid-extraction- (LLE), and solid-phase-extraction (SPE)-based procedures for the isolation of doxorubicin (DOX) and daunorubicin (DAU) as an internal standard (IS) from rat tissue samples. During the experiments, all samples were analyzed via liquid chromatography coupled with fluorescence detection (LC-FL), with analytes being monitored at excitation and emission wavelengths of 487 and 555 nm, respectively. The absolute recoveries of the sample-preparation procedure were then calculated and compared, and the advantages and disadvantages of each approach were considered in depth. Ultimately, SPE with hydrophilic–lipophilic balanced (HLB) sorbents was selected as the most effective extraction procedure as it enabled the absolute recovery of DOX from tissue samples at a level of 91.6 ± 5.1%. Next, the selected HLB-SPE protocol was coupled with LC-FL separation and the resultant method was validated according to FDA and ICH requirements. The validation data confirmed that the developed procedure met all required criteria for bioanalytical methods, with a limit of detection (LOD) and limit of quantification (LOQ) of 0.005 µg/g and 0.01 µg/g, respectively. Finally, the developed protocol was successfully tested on various rat tissues enriched with DOX, confirming its potential as an interesting alternative to previously reported protocols for pharmacokinetic studies and clinical investigations aimed at analysis of the level and biodistribution of DOX in tissue samples after systemic administration of this drug.

Deep Learning-Based Image Extraction

The development of the web and advancements in computation and multimedia technologies have led to an increase in the variety of photo databases and the collection of hundreds of images that include medical images, e-libraries, and art galleries. The necessary images from that kind of big collection may demand a lengthy period to retrieve using traditional image extraction techniques like Textual Based Images Retrieval. It is essential to develop an effective image extraction procedure that can handle such vast amounts of data at once. The main objective is to develop a trustworthy tool that effectively generates, uses, and responds to data. It employs a strategy for creating an effective picture retrieval application that enables individuals to ask questions about the software and extract it from a huge database.

Quality by Design in optimizing the extraction of (poly)phenolic compounds from Vaccinium myrtillus berries

Quality by Design was adopted for developing an effective extraction procedure of (poly)phenolic compounds from bilberry (Vaccinium myrtillus L.) fruits, using a pooled sample of berries from different regions of Ukraine. Mechanical solvent extraction, microwave-assisted extraction (MAE) and ultrasonic-assisted extraction (UAE) were investigated by screening matrices. Extraction time (Time, from 5 to 15 min), organic solvent type (OS type, methanol, ethanol and acetone), organic solvent percentage (OS%, from 50% to 90%), sample/extractant ratio (S/E ratio, from 0.025 to 0.1 g mL−1), and, only for MAE, extraction temperature (T, from 30 to 60°C), were selected as critical method parameters (CMPs). The spectrophotometric assays total soluble polyphenols (TSP), total monomeric anthocyanins (TMA), and radical scavenging activity (evaluated by the 2,2-diphenyl-1-picrylhydrazyl radical (DPPH), the 2,2′-azino-di-(3-ethylbenzthiazoline sulfonic acid), and the ferric reducing antioxidant power methods) were chosen as critical method attributes (CMAs). The screening procedure allowed for selecting UAE and methanol, while the other CMPs underwent further optimization through Response Surface Methodology. Target values for TSP, TMA and DPPH were selected and the method operable design region (MODR) was defined by means of Monte-Carlo simulations. The optimized conditions, with the corresponding MODR intervals in bracket, were the following: (i) Time, 17 min (15–23 min); OS%, 56% (44–59%); S/E ratio, 0.030 (0.022–0.034) g mL−1. Under these experimental conditions, CMAs values of the pooled sample were the following (n = 3): TSP=4433±176 mg (+)-catechin eq/100 g dry weight (d.w.); TMA=3575±194 mg cyanidin-3-glucoside eq/100 g d.w.; DPPH=273±5 μg DPPH inhib./mg d.w. The optimized extraction method was tested for matrix effect (ME%) in the UHPLC-MS/MS analysis of 15 anthocyanins and 20 non-anthocyanins individual (poly)phenols commonly found in bilberries, as well as for luteolin, sinapic acid, and pelargonidin-3-glucoside, absent in this fruit and therefore added to the extracts as surrogate standards for evaluating apparent recovery (AR%). |ME%| was in any case ≤ 23% and AR% of the surrogate standards in the range 91–95%, confirming the very good performances of the optimized extraction method.

Rapid screening of 84 pesticide residues in dendrobium by Sin-QuEChERS Nano purification column with gas chromatography-tandem mass spectrometry

利用Sin-QuEChERS Nano净化柱结合气相色谱-串联质谱(GC-MS/MS)分析,建立了石斛基质中84种不同极性农药残留的快速筛查方法。比较了采用不同的提取溶剂(1%乙酸乙腈、丙酮)和不同的提取方式(加水浸泡和不加水浸泡)下目标物的提取效率。利用金钗石斛样品系统比较了Sin-QuEChERS Nano法与经典的基质固相分散法(dSPE)、固相萃取法(SPE)、QuEChERS法的净化效果及提取回收率,以及净化效果较好的Sin-QuEChERS Nano法与dSPE法基质效应的差异。目标物经DB-1701MS石英毛细管色谱柱(30 m×0.25 mm×0.25 μm)程序升温分离,GC-MS/MS多反应监测(MRM)模式检测,基质匹配溶液外标法定量。通过GC-MS/MS检测方法对金钗石斛和铁皮石斛中的84种代表性农药进行了方法学验证。结果表明:各目标物在不同范围内呈良好的线性相关,相关系数(r2)均>0. 990,方法的检出限(LOD, S/N=3)为1.5~5.8 μg/kg,方法的定量限(LOQ, S/N=10)为5.0~15.0 μg/kg。在两个水平下,目标农药的加标回收率为68.7%~116.2%,相对标准偏差(RSD, n=6)均低于15%。与其他经典的前处理方法相比,Sin-QuEChERS Nano法在净化效果方面表现更好,该法不仅可以有效去除色素、有机酸、碱性干扰物等物质,还可以节省样品制备时间,避免溶剂转移造成的损失,无需进一步涡旋或离心,是一种简单而有效的提取物纯化程序。该方法灵敏、快速、简便、可靠,有效地提高了石斛中农药快速筛查时的检测效率,具有较强的实际应用价值。此外,所开发的方法可以进一步扩展目标农药的类型,并可以用于检测其他更多食品及中药材中的农药残留。

Development of dispersive solid-liquid extraction method based on organic polymers followed by deep eutectic solvents elution; application in extraction of some pesticides from milk samples prior to their determination by HPLC-MS/MS

An effective extraction procedure based dispersive solid phase extraction was developed for the extraction of different pesticides (dimethoate, imidacloprid, pirimicarb, carbaryl, fenitrothion, hexythiazox, and phosalone) from milk samples. The analytes were determined by high performance liquid chromatography-tandem mass spectrometry. In this method, an organic polymer (polystyrene) was dissolved in a water-miscible organic solvent (tetrahydrofuran) and it was injected into the aqueous phase obtained from milk sample after deprotonation. By this action the sorbent (polymer) was precipitated and dispersed in the solution as tiny particles. By doing so, the pesticides were adsorbed onto the sorbent. After that, the sorbent was separated by centrifugation and the adsorbed analytes were eluted by tetrabutylammonium chloride: dichloroacetic acid deep eutectic solvent. After optimization, the method validation was followed according to Food and Drug Administration and International Research Council guidelines and the results showed that wide linear ranges (0.93–500 ng/mL) and low limits of detection (0.09–0.27 ng/mL) and quantification (0.31–0.93 ng/mL) were obtained. Extraction recoveries and extraction and enrichment factors were in the ranges of 81–94% and 162–188, respectively. Relative standard deviations were less than 9% for intra– (n = 6) and inter–day (n = 6) precisions at three concentration of 2, 5, and 25 ng mL−1 of each analyte. The developed method was successfully applied to twenty cow milk samples for determination of the pesticides.

Determination of Avermectins Residues in Soybean, Bean, and Maize Using a QuEChERS-Based Method and Ultra-High-Performance Liquid Chromatography Coupled to Tandem Mass Spectrometry

Soybean, maize, and bean are crops of great economic importance, but in recent years have suffered with infestations of the caterpillar Helicoverpa armigera, with the main reason being the resistance of this pest to most pesticides. Avermectin emamectin benzoate was recently released to control this pest. Other avermectins, like abamectin, doramectin, eprinomectin, and ivermectin are used in large scale because they potent acaricidal, anthelmintic, and insecticidal activities. Thus, a simple and fast method for the determination of avermectins in these crops based on a quick, easy, cheap, effective, rugged, and safe (QuEChERS) extraction procedure and ultra-high performance liquid chromatography with tandem mass spectrometry (UHPLC-MS/MS) analysis was developed and validated. For extraction, water followed by acetonitrile:isopropanol and a partition step with salts was stablished. With the clean-up step using activated EMR-Lipid, limits of detection of 1.2 μg kg−1 for abamectin, doramectin, emamectin benzoate, and ivermectin, and of 2.4 μg kg−1 for eprinomectin were achieved. The validation showed satisfactory results and the method was successfully applied to commercial samples, indicating that it is suitable for routine analysis.

The Bark of the Spruce Picea jezoensis Is a Rich Source of Stilbenes.

Stilbenes are plant phenolic secondary metabolites that show beneficial effects on human health and possess high antifungal activity against plant pathogens. Currently, a search for plant sources with high stilbene levels is relevant, since stilbene content in various plant species can vary substantially and is often at a low level. In this paper, the bark and wood of Picea jezoensis were analyzed for the content and composition of stilbenes and compared with other known stilbene sources. The HPLC-MS analysis of P. jezoensis bark and wood extracted with different solvents and at different temperatures revealed the presence of 11 and 5 stilbenes, respectively. The highest number of stilbenes of 171 and 229 mg per g of the dry weight (mg/g DW) was extracted from the bark of P. jezoensis using methanol or ethanol at 60 °C for 2 h. Trans-astringin, trans-piceid, and trans-isorhapontin prevailed over other stilbenoids (99% of all detected stilbenes). The most abundant stilbene was trans-isorhapontin, reaching 217 mg/g DW or 87% of all stilbenes. An increase in the extraction time from 2 to 6 h did not considerably increase the detected level of stilbenes, while lower extraction temperatures (20 and 40 °C) significantly lowered stilbene yield. The content of stilbenes in the P. jezoensis bark considerably exceeded stilbene levels in other stilbene-producing plant species. The present data revealed that the bark of P. jezoensis is a rich source of stilbenes (primarily trans-isorhapontin) and provided effective stilbene extraction procedures.

Screening of multi-mycotoxins in fruits by ultra-performance liquid chromatography coupled to ion mobility quadrupole time-of-flight mass spectrometry

A modified quick, easy, cheap, effective, rugged, and safe extraction procedure combined with ultra-performance liquid chromatographic separation and ion mobility quadrupole time-of-flight mass spectrometry was developed to determine the presence of 20 mycotoxins (i.e., Alternaria toxins, ochratoxin, patulin, aflatoxin and trichothecenes) in fruit samples from Xinjiang. A complete platform, including screening via an in-house library, confirmation and quantification using reference standards, was established, which provided accurate MS data and complete spectra containing the fragment ions for each analyte. To evaluate the performance of the developed method, satisfactory validation parameters, such as linearity (R2 ≥ 0.9992), precision (RSDs ≤ 9.8%), recovery (81.2–99.2%), LOD (0.06–2.22 µg kg−1), and LOQ (0.2–7.39 µg kg−1), were obtained. The analysis of 130 fruit samples revealed nonnegligible contamination with mycotoxins; specifically, the highest levels of three Alternaria toxins were detected in jujube, wolfberries and raisins.

Accurate mass screening of pesticide residues in wine by modified QuEChERS and LC-hybrid LTQ/Orbitrap-MS

In this research, a quick, easy, cheap, effective, rugged, and safe (QuEChERS) extraction procedure and Ultra-High Performance Liquid Chromatography-Orbitrap-Mass Spectrometry (UHPLC-Orbitrap-MS), were combined to obtain a sensitive and rapid method for the determination of multiclass pesticides in white and red wines. The optimization strategy involved the selection of buffering conditions, by applying different QuEChERS procedures and sorbents for the cleanup step in order to achieve acceptably high recoveries and low co-extractives in the final extracts. Identification was based on both accurate mass and retention time, while further confirmation was achieved by MS fragmentation. The method was evaluated in terms of linearity, recovery, precision, limit of detection (LOD) and quantification (LOQ), matrix effects (ME) and expanded uncertainty. The validated method was successfully applied to real samples (home-made and commercial) revealing the presence of two selected fungicides, in relatively low levels compared to the MRLs defined by the EU for vinification grapes.

Optimization of batch extraction of functional ingredients from stevia leaves with response surface methodology

The aim of the study was to reveal an effective extraction procedure for maximization of the yield of stevioside, flavonoid, and total phenolic compounds as well as antioxidant activity in stevia extract. Extraction was carried out at temperature variations of 75 ÂΩC, 80 ÂΩC, 90 ÂΩC and variations of water solvents 280 mL/g, 290 mL/g, 300 mL/g. Stevioside and flavonoid analysis used spectrophotometer method, while phenol analysis used the Folin-Cioccalteau reagent method. The result is a change in temperature and solvent ratio affect the changes in stevioside, flavonoid, and phenol levels. Optimal conditions of stevioside, flavonoid, and phenol using response surface methodology (RSM) occur at a temperature range of 75 ÂΩC with a solvent ratio of 280-290 mL/g.

Extraction Methods of Cyanotoxins Aqueous Media and Sediments

Cyanotoxins are chemical compounds produced by cyanobacterial mats grown in aquatic ecosystems. These may threaten human health and aquatic organisms. Extraction of these toxins is usually associated with many difficulties due to their concentration in aquatic ecosystems. This study is designed to provide suitable and effective extraction procedures that can effectively extract low concentration cyanotoxin from water and bacterial cells. The methodology is based on collecting raw material of cyanobacterial mats from naturally growing sites such as Wadi Gaza along with 16 liters of aquatic surrounding media. The materials were left in the Lab for 24 - 48 h for stabilization of the mats. The floating mats were collected using special funnel and allowed to air drying. The aqueous phase was extracted by liquid/liquid extraction using solvent mixture (hexane + ethylacetate 10% w:w), and by liquid solid extraction using several types of organoclays complexes. The solid phase was extracted by acetone and ultrasonic device. Results showed some difficulties were associated with liquid/liquid extraction whereas effective and easy extraction procedures were obtained by liquid solid extraction using either organoclay complex or activated charcoal. In contrast combination of both solid materials did not show improvement in the extracted cyanotoxin. Thus we recommend the use of organoclays or activated charcoal separately for extracting cyanotoxin. Further improvement of extraction can be tailored by using a specific organoclay complex that has some similarity in the chemical structure between the pre-adsorbed organic cation to the clay mineral and the chemical structure of cyanotoxin.

Extraction and analysis of oleanolic acid and ursolic acid from apple processing waste materials using ultrasound-assisted extraction technique combined with high performance liquid chromatography

The aim of the present study was to develop a is simple, effective, eco-friendly, reproducible and high-yield two-stage ultrasound-assisted extraction (UAE) procedure combined with quantitative determination high performance liquid chromatographic (HPLC) method for obtaining isomeric triterpene acids - oleanolic acid (OA) and ursolic acid (UA) in the crystalline dried powdered form from apple processing agro-industrial waste material. A rapid, sensitive and specific HPLC method was developed and validated with respect to robustness, specificity, linearity-range, accuracy, precision and sensitivity. The effect of the nature and the volume of the extraction solvent, the extraction time and the sample size on the extraction efficiency were investigated. The optimal conditions for high-yield extraction were found.

Optimization of QuEChERS Method for Simultaneous Determination of Neonicotinoid Residues in Pollinator Forage.

Consistent with the large-scale use of pesticide seed treatments in U.S. field crop production, there has been an increased use of neonicotinoid-treated corn and soybean seed over the past decade. Neonicotinoids can move downwind to adjacent off-field pollinator habitats in dust from planting and/or move downslope to habitats in surface water. The extent of potential neonicotinoid exposure to pollinators from neonicotinoid movement into these adjacent pollinator habitats is unclear. Pollen and leaf tissue extractions were completed using a quick, easy, cheap, effective, rugged, and safe (QuEChERS) extraction procedure. Samples were subjected to a clean-up step using dispersive solid-phase extraction (dSPE) techniques prior to analysis. The compounds in the extracts were separated on a reversed-phase column with gradient elution and confirmed with tandem mass spectrometry. The extraction method showed acceptable recoveries of analytes ranging from 78.4 to 93.6% and 89.4 to 101% for leaf tissue and pollen, respectively. The method’s detection limits ranged from 0.04 to 0.3 ng/g in milkweed leaf tissue and 0.04 to 1.0 ng/g in pollen. The method is currently being employed in ongoing studies surveying pollen from a diversity of forbs and milkweed leaves obtained from habitat patches established within fields with a history of using neonicotinoid-treated seeds.

Evaluation of effective protein extraction procedure to profile petiole of Moringa oleifera

Moringa oleifera is widely known as multipurpose tree since all of its parts confer multiple functions. The leaf is highly favourable among consumers while the petiole is mostly wasted. There are numerous studies on the flavonoid and antioxidant property of the stem and twig. However, study on the petiole has never been done. There-upon, this study was conducted to develop protein profiling of the petiole. In this study, 6 different protein extraction methods were tested on the fresh petiole before its protein quantity and quality were checked via Bradford assay and Sodium Dodecyl Sulfate Polyacrylamide Gel Electrophoresis (SDS-PAGE) respectively. The in-solution digestion was then done prior to LC-MS/MS analysis. The protein electrophoretic pattern from the SDS-PAGE proves that method 6 using Tris HCl buffer with incorporation of dithiothreitol (DTT) and phenylmethylsulfonyl fluoride (PMSF) confers the best quality of protein. It produced the highest number of visible individual bands compared to other methods. Meanwhile, 93 proteins were successfully identified via LCMS analysis where the protein, signal response and carbohydrate metabolism categories confer the highest percentage. High quality and content of the protein extracted from the petiole including the antioxidant, anticancer and antidiabetic protein identified suggested that consuming this part of the plant could enhance nutrients of human body.