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Background: Tangerine peel is rich in flavonoids, particularly hesperidin, which has anti-inflammatory, antioxidant and anticancer biological activities. However, it is often wasted during citrus processing. The current common extraction method for hesperidin is solvent extraction, which has the characteristics of low extraction rate and high contamination. The aim of this study was to investigate the effect of pulsed electric field-assisted alkali dissolution extraction, followed by an acidification precipitation method, on the extraction rate and structure of hesperidin from tangerine peel., Results: The results showed that the selected factors (material/liquid ratio, electric field intensity and pulse number) had a significant effect on the extraction yield. An optimum condition of 66.00 mL g -1 , 4.00 kV cm -1 and 35.00 pulses gave the maximum amount (669.38 μg mL -1 ), which was consistent with the theoretically predicted value by software (672.10 μg mL -1 ), indicating that the extraction process was feasible. In addition, the purified extract was further identified as hesperidin from UV and NMR spectra., Conclusion: An appropriate strength of pulsed electric field-assisted alkali dissolution extraction followed by an acidification precipitation method can effectively improve the extraction rate of orange peel, and the purity of the extracted orange peel is higher. Compared with the traditional extraction, the pulsed electric field-assisted extraction method may be a potential technology for hesperidin extraction, which is beneficial for the high-value utilization of citrus resources. © 2024 Society of Chemical Industry., (© 2024 Society of Chemical Industry.)

The demand for polyphenols and essential oils (EOs) on the food market is high and grows every year. Its partially the result of the fact that these compounds can be used in formulation of clean label foods, a fast growing food sector. A significant share of polyphenols and EOs are extracted from herbs. The quality of the extracts is determined mainly by the extraction method. Conventional extraction techniques of phytochemicals are time-consuming, operate at high temperatures, and require usage of organic solvents and energy in large quantities. According to the United Nations Sustainability Development Plan, chemical processes should be replaced by green alternatives that would reduce the use of solvents and energy. Ultrasound-Assisted Extraction (UAE), Microwave-Assisted Extraction (MAE) and Cold Plasma-Assisted Extraction (CPAE) meets these criteria. The review shows that each of these techniques seems to be a great alternative for conventional extraction methods ensuring higher yields of bioactive compounds., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 The Authors. Published by Elsevier Ltd.. All rights reserved.)

One of the principal byproducts of coffee roasting is the coffee parchment. It is abundant in bioactive substances, including derivatives of chlorogenic acids, which are well-known for their exceptional antioxidant effects. It is advantageous to use environmentally friendly extraction techniques on such residues since it adds value to the entire coffee production process supply chain. The aim of this work was to assess and enhance the ability of non-conventional extraction techniques to extract derivatives of chlorogenic acid from coffee parchment. A central composite design was used to maximize the recovery of those phenolic compounds. The optimized extraction conditions were with 5 min extraction period, at a temperature of 70 °C, and 80% ethanol in the extractor solvent. In this conditions extraction recovery of chlorogenic acids was of 0.8% by the use of microwave-aided extraction (MAE). The optimized conditions are practical, economical, and ecologically friendly method to extract phenolic compounds and, consequently, underscores the potential for sustainable utilization of coffee parchment, offering a valuable contribution to the development of environmentally conscious strategies within the coffee industry., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

Background: Extracts of fresh wine grape seeds/skin or of grape pomace seeds were used to prepare antioxidant natural toothpastes., Results: Ethanol extracted twice more polyphenols than water; ultrasound did not provide any improvement in the extraction. The addition of freeze-dried ethanol extracts of seeds or skin, at 2% and 10%, to the commercial toothpaste significantly increased the polyphenol content, both from white grape seeds and skin and from red grape seed pomace. The evaluation of time stability (shelf life) revealed a decrease, after 4 months, of 3.9% and 9.4% in total polyphenol content, in 5% and 10% water extracts, but not for ethanol extracts. 1,1-Diphenyl-2-picrilhydrazil 1 antiradical activity was the highest in 10% of seed water extract toothpaste and, after 4 months, the activity was stable., Conclusion: Ethanol and water are efficient and safe solvents to create natural toothpaste with grape or pomace seed extract with antioxidant activity. © 2021 The Authors. Journal of The Science of Food and Agriculture published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry., (© 2021 The Authors. Journal of The Science of Food and Agriculture published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.)

The objective of this study was to assess the biological activity of essential oils (EOs) of four Juniperus species obtained via two different distillation methods and their potential as biopesticides. The studied factors were juniper species ( Juniperus communis L., J. oxycedrus L., J. pygmaea C. Koch., and J. sibirica Burgsd), plant sex (male (M) and female (F)), and distillation method (hydrodistillation via a standard Clevenger apparatus (ClevA) and semi-commercial (SCom) steam distillation). The hypothesis was that the EO will have differential antioxidant, antimicrobial, and insecticidal activities as a function of plant species, plant sex, and distillation method. The two distillation methods resulted in similar EO composition within a given species. However, there were differences in the EO content (yield) due to the sex of the plant, and also differences in the proportions of some EO components. The concentration of α-pinene, β-caryophyllene, δ-cadinene and δ-cadinol was dissimilar between the EO of M and F plants within all four species. Additionally, M and F plants of J. pygmaea , and J. sibirica had significantly different concentrations of sabinene within the respective species. The EOs obtained via ClevA extraction showed higher antioxidant capacity within a species compared with those from SCom extraction. All of the tested EOs had significant repellent and insecticidal activity against the two aphid species Rhopalosiphum padi (bird cherry-oat aphid) and Sitobion avenae (English grain aphid) at concentrations of the EO in the solution of 1%, 2.5%, and 5%. The tested EOs demonstrated moderate activity against selected pathogens Fusarium spp., Botrytis cinerea , Colletotrichum spp., Rhizoctonia solani and Cylindrocarpon pauciseptatum . The results demonstrate that the standard ClevA would provide comparable EO content and composition in comparison with SCom steam distillation; however, even slight differences in the EO composition may translate into differential bioactivity.

A self-standing, robust bacterial cellulose (BC)-based multifunctional composite membrane embedded with desirable nano-adsorbents has been successfully fabricated via a facile versatile strategy. As expected, the developed BC-based composite membrane enables the simultaneous and efficient removal of multiple co-existing pollutants.

Effective carbon dioxide (CO 2 ) separation by nominal energy utilization is the factual attempt in the present era of energy scarcity and environmental calamity. In this perspective, the membrane- based gas separation technology is a budding endeavour owing to its cost -effectiveness, ease of operational maintenance and compact modular design. Among various membrane materials, bio-based polymers are of interest as they are abundant and can be obtained from renewable resources, and can also reduce our dependency on exhaustible fossil fuel-based sources. In this review, the structure-property relationship of chitosan and some of its film-forming derivatives has been critically studied for the first time in view of the fundamental properties required for gas separation applications. Various factors affecting the gas permeation performance of chitosan-based membranes have been highlighted along with prospects and propositions for the design of a few novel bio-based membranes based on the exhaustive analyses., (Copyright © 2021 Elsevier Ltd. All rights reserved.)

Background: α-lactalbumin (α-La) is of great interest to the industry as a result of its excellent functional properties and nutritional value. Aqueous two-phase flotation (ATPF) of thermo-sensitive polymer poly (ethylene glycol-ran-propylene glycol) monobutyl ether (UCON) and KH 2 PO 4 was applied to directly separate and purify α-La from milk whey, which was purposed to simplify the production process and reduced cost of production., Results: The effect of ATPF composition and operating parameters on the flotation efficiency (E) and purity of α-La were investigated. The optimal conditions included 2 min of premixing time, 30 mL min -1 flow velocity and 20 min of flotation time, whereas the composition conditions comprised 35.0 mL 0.18 g mL -1 phosphate solution (containing 10% (cow milk whey/salt solution, v/v) cow milk whey, 50 ppm defoamer and 2 g NaCl) and 5.0 mL of 40% (w/w) UCON solution. Under the optimal conditions, E of α-La was 95.67 ± 1.04% and purity of α-La was 98.78 ± 1.19%. UCON was recovered by a thermally-induced phase separation and reused in next ATPF process without reducing E of α-La. Purified α-La was characterized by several key technologies. The results indicated that α-La in cow milk whey could be directly separated and purified by the ATPF and the purity was satisfactory. Moreover, it was suggested there was no obvious structure difference between the α-La separated by ATPF and the α-La standard., Conclusion: The present study enabled the recycling of UCON, providing an effective, economically viable and environmentally friendly approach for the separation and purification of protein. © 2021 Society of Chemical Industry., (© 2021 Society of Chemical Industry.)

Different parts of a plant (seeds, fruits, flower, leaves, stem, and roots) contain numerous biologically active compounds called "phytoconstituents" that consist of phenolics, minerals, amino acids, and vitamins. The conventional techniques applied to extract these phytoconstituents have several drawbacks including poor performance, low yields, more solvent use, long processing time, and thermally degrading by-products. In contrast, modern and advanced extraction nonthermal technologies such as pulsed electric field (PEF) assist in easier and efficient identification, characterization, and analysis of bioactive ingredients. Other advantages of PEF include cost-efficacy, less time, and solvent consumption with improved yields. This review covers the applications of PEF to obtain bioactive components, essential oils, proteins, pectin, and other important materials from various parts of the plant. Numerous studies compiled in the current evaluation concluded PEF as the best solution to extract phytoconstituents used in the food and pharmaceutical industries. PEF-assisted extraction leads to a higher yield, utilizes less solvents and energy, and it saves a lot of time compared to traditional extraction methods. PEF extraction design should be safe and efficient enough to prevent the degradation of phytoconstituents and oils.

The nicotine addiction problem is of great concern, particularly in adolescents. Notably, nicotine addiction drives humans to continue smoking. Notably, several diseases and disorders are caused by smoking. To date, various adsorbents have been proposed to develop a functionalization filter tip for reducing nicotine content in mainstream smoke. However, the nicotine adsorption efficiencies of most of the reported functionalization filter tips were not satisfactory, and their preparation process was complex and time-consuming. Herein, we demonstrate a highly active and adsorbing filter tip for cigarettes, fabricated by decorating polydopamine (PDA) on the surface of a commercial filter tip in situ. The PDA coating on the filter tip was obtained by the self-polymerization of dopamine (DA) within 16 h, which was quicker and easier than the preparation processes of other reported functionalized filter tips. Significantly, the PDA-decorated filter tip had a nicotine adsorption efficiency as high as ∼95%, which was much higher than most of the commercial filter tips.

Isolating high quality RNA is a limiting factor in molecular analysis, since it is the base for transcriptional studies. The RNA extraction method can directly affect the RNA quality and quantity, as well as, its overall cost. The industrial importance of the yeast genus Candida in several sectors comes from their capacity to produce Lipases. These enzymes are one of the main metabolites produced by some Candida species, and it has been shown that Candida yeast can biodegrade petroleum hydrocarbons and diesel oil from biosurfactants that they can produce, a feature that turns these organisms into potential combatants for bioremediation techniques. Thus, this study aimed to determine an efficient method for isolating high quality RNA from Candida viswanathii biomass. To achieve this aim, three different RNA extraction methods, TRIzol, Hot Acid Phenol, and CTAB (Cetyltrimethylammonium Bromide), were tested. The three tested methods allowed the isolation of high-quality RNA from C. viswanathii biomass and yielded suitable RNA quantity for carrying out RT-qPCR studies. In addition, all methods displayed high sensitivity for the expression analysis of the CvGPH1 gene through RT-qPCR, with TRIzol and CTAB showing the best results and the CTAB method displaying the best cost-benefit ratio (US$0.35/sample)., (Copyright © 2021 Elsevier B.V. All rights reserved.)

Background: Volatile organic compounds are present at very low concentration but exhibit an important influence on flavor and aroma of tobacco leaves and products. During tobacco processing, at different stages, tobacco wastes occur. Since they are delivered directly from the tobacco plant, they are expected to have a similar aroma profile., Results: The volatile composition of three types of tobacco waste (scrap, dust, and midrib) was characterized for the first time and compared with tobacco leaves' volatile composition. Ultrasound-assisted extraction with hexane followed by gas chromatography-mass spectrometry was successfully applied. Different ultrasound-assisted extraction parameters (temperature, time, and solvent:solid ratio) showed a significant influence on the volatile profiles of the extracts obtained. The most important compounds in tobacco leaves, scrap, and dust with the highest abundance were nicotine (up to 87.5%), 4,8,13-duvatriene-1,3-diol (up to 16.2%), and neophytadiene (up to 9.4%). In midrib, only nicotine was present in all extracts. The most abundant compounds in the extracts were quantified and subjected to optimization using response surface methodology., Conclusion: Regression analysis showed that 83-98% of the variation was explained by the models obtained. The experimentally obtained values agreed with those predicted, thus indicating the suitability of the model employed and the success of response surface methodology in optimizing the extraction conditions. © 2020 Society of Chemical Industry., (© 2020 Society of Chemical Industry.)

Background: Red beet plants are cultivated worldwide for the consumption of their roots, generating large amounts of unexploited by-products. In particular, beet leaves (BLs) represent about 50% of the whole plant and are usually discarded as waste. This constitutes not only an economic issue, since multiple resources invested in the production will be wasted, but also an environmental problem because of the pollution associated with their disposal. However, BLs comprise an important source of functional compounds (polyphenols and betalains) that could be recovered from the raw material, representing a sustainable solution for the underutilization of this by-product. This study proposes the recovery of polyphenols and betalains using an aqueous ultrasound-assisted extraction (UAE) process at different powers (35, 50, and 100 W) that was characterized and optimized., Results: UAE significantly enhanced the recovery of bioactive compounds and shortened the time required for extraction in comparison with traditional macerations (35 < 50 < 100 W). During UAE, the temperature of the systems increased as a function of the power applied, favouring the recovery of these phytochemicals. Additionally, a Box-Behnken design and response surface methodology were employed to optimize UAE conditions (90 W ultrasound power, 1:20 solid:liquid ratio, 16 min extraction time), under which the yields were 14.9 mg g -1 (polyphenols), 949.1 μg g -1 (betaxanthins), and 562.2 μg g -1 (betacyanins), consistent with the values predicted by the models., Conclusion: This study enabled the development of a green-solvent UAE process that constitutes an effective post-harvest by-products strategy to minimize losses and increase biomass utilization through the recovery of bioactive compounds from BLs, promoting sustainability in the agri-food chain. © 2020 Society of Chemical Industry., (© 2020 Society of Chemical Industry.)

The study was an attempt to compare batch and circulating processes for polyphenols extraction from pomelo peels by liquid-phase pulsed discharge (LPD) in order to assess the extraction efficiency of the two processes. Response surface methodology was used to optimize batch (8-12 kV discharge voltage, 30-50 mL/g liquid to solid ratio and 2-4 min extraction time) and circulating (8-12 kV discharge voltage, 30-50 mL/g liquid to solid ratio and 20-40 mL/min flow rate) extractions. The highest polyphenols yield was 2.50 ± 0.02% at 42.2 mL/g, 12 kV and 4 min in batch extraction, while circulating extraction produced the most polyphenols (2.42 ± 0.01%) at 43.7 mL/g, 10.4 kV and 27.6 mL/min. The results showed that batch extraction achieved much greater yields than circulating extraction with lower-cost equipment. Therefore, batch extraction was a promising technology for the separation of high value-added products from pharmaceuticals and fine chemicals., (Copyright © 2020 Elsevier Ltd. All rights reserved.)

A weak ion-exchange membrane (P-COOH) was synthesized by alkaline hydrolysis of a polyacrylonitrile nanofiber membrane prepared by electrospinning process. The P-COOH membrane was characterized for its physical properties and its application for purification of lysozyme from chicken egg white was investigated. The lysozyme adsorption efficiency of the P-COOH membrane operating in a stirred cell contactor (Millipore, Model 8010) was evaluated. The effects of key parameters such as the feed concentration, the rotating speed, the flow rate of feed and the operating pressure were studied. The results showed successful purification of lysozyme with a high recovery yield of 98% and a purification factor of 63 in a single step. The purification strategy was scaled-up to the higher feedstock loading volume of 32.7 and 70 mL using stirred cell contactors of Model 8050 and 8200, respectively. The scale-up processes achieved similar purification results, proving linear scalability of the purification technique adopted., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2020 Elsevier Ltd. All rights reserved.)

The dried Ganoderma lucidum (GL) has been widely used for its pharmacological properties and bioactive ganoderic acids (GAs). Herein, extraction procedures combining ultra-sonication and heating were optimized using response surface methodology based on four variables (antioxidant activity, anti-diabetic activity, total GAs content, and total polysaccharide content) and principal component analysis. The extraction of freeze-dried GL at temperatures between 64.2 and 70 °C for 1.2 h maximized the antioxidant activity and GA content, whereas the polysaccharide content and anti-diabetic activity were maximized by extraction between 66.8 and 70 °C for more than 2.8 h. Heat-dried GL extracted at 50 °C for 3 h provided the greatest anti-inflammatory activity against HaCaT cells by suppressing the response to inflammation related cytokines at mRNA levels. These results suggest that extraction conditions might be a limiting factor for target-oriented investigations, and optimized extraction methods may improve the potential effect and quality of harvested GL products., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2020 Elsevier Ltd. All rights reserved.)

Production of inulin from yam bean tubers by ultrasonic assisted extraction (UAE) was optimized by using response surface methodology (RSM) and genetic algorithms (GA). Yield of inulin was obtained between 11.97%-12.15% for UAE and 11.21%-11.38% for microwave assisted extraction (MAE) using both the methodologies, significantly higher than conventional method (9.9 %) using optimized conditions. Under such optimized condition, SEM image of root tissues before and extraction showed disruption and microfractures over surface. UAE provided a shade better purity of extracted inulin than other two techniques. Degree of polymerization in inulin was also recorded to be better, might be due lesser degradation during extraction. Significant prebiotic activity was recorded while evaluation using Lactobacillus fermentum and it was 36 % more than glucose treatment. Energy density by UAE was few fold lesser than MAE. Carbon emission was far more less in both these methods than the conventional one., (Copyright © 2020 Elsevier Ltd. All rights reserved.)

1-Deoxynojirimycin (1-DNJ), a polyhydroxylated alkaloid, is a highly selective and potent glycosidase inhibitor that has garnered great interest as a tool to study cellular recognition and as a potential therapeutic agent. The development of analytical methods for the quantification polyhydroxylated alkaloids in natural products requires a multifaceted approach. Many publications over the past five decades have described analytical methods for this compound. However, recently more advanced techniques have come to prominence for sample extraction, purification, detection, and identification. This review provides an updated, extensive overview of the available methods for the extraction, purification, identification or detection of 1-DNJ. The review highlights different strategies for the design of 1-DNJ detection methods, which we analyzed in light of recent detection data. Finally, we conclude with perspectives on possible strategies for increasing the efficiency of identification and quantification of 1-DNJ in the future.

Global health and food security constantly face the challenge of emerging human and plant diseases caused by bacteria, viruses, fungi, and other pathogens. Disease outbreaks such as SARS, MERS, Swine Flu, Ebola, and COVID-19 (on-going) have caused suffering, death, and economic losses worldwide. To prevent the spread of disease and protect human populations, rapid point-of-care (POC) molecular diagnosis of human and plant diseases play an increasingly crucial role. Nucleic acid-based molecular diagnosis reveals valuable information at the genomic level about the identity of the disease-causing pathogens and their pathogenesis, which help researchers, healthcare professionals, and patients to detect the presence of pathogens, track the spread of disease, and guide treatment more efficiently. A typical nucleic acid-based diagnostic test consists of three major steps: nucleic acid extraction, amplification, and amplicon detection. Among these steps, nucleic acid extraction is the first step of sample preparation, which remains one of the main challenges when converting laboratory molecular assays into POC tests. Sample preparation from human and plant specimens is a time-consuming and multi-step process, which requires well-equipped laboratories and skilled lab personnel. To perform rapid molecular diagnosis in resource-limited settings, simpler and instrument-free nucleic acid extraction techniques are required to improve the speed of field detection with minimal human intervention. This review summarizes the recent advances in POC nucleic acid extraction technologies. In particular, this review focuses on novel devices or methods that have demonstrated applicability and robustness for the isolation of high-quality nucleic acid from complex raw samples, such as human blood, saliva, sputum, nasal swabs, urine, and plant tissues. The integration of these rapid nucleic acid preparation methods with miniaturized assay and sensor technologies would pave the road for the "sample-in-result-out" diagnosis of human and plant diseases, especially in remote or resource-limited settings., (Copyright © 2020 Elsevier B.V. All rights reserved.)

Four extraction techniques (that is, hot water extraction [HWE], alkaline-acidic extraction [AAE], ultrasound assisted extraction [UAE], and microwave assisted extraction [MAE]) were compared for flaxseed gum extraction and their influence on the yield, purity, structural characterization (monosaccharide composition, molecular weight distribution, and microstructure by transmission electron microscope), and antioxidant activity (in terms of scavenging ability of 2,2-diphenyl-1-picrylhydrazyl [DPPH], 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulphonic acid [ABTS], and reducing power) was investigated. The HWE achieved higher yield (8.96%) than UAE (7.84%) followed by MAE (7.01%) and AAE (6.44%). Moreover, the four flaxseed gum (FSG) samples exhibited the identical monosaccharide composition, but slight difference was observed in the content, whereas the molecular weight ratios exhibited significant difference. All samples displayed concentration-dependent manner for all antioxidant assays. UAE-FSG showed significant higher scavenging ability on DPPH free radical, ABTS free radical, reducing power, and β-carotene bleaching assay followed by HWE-FSG, MAE-FSG, and AAE-FSG. Overall results showed that UAE was favorable to the purity of FSG, whereas HWE was more advantageous to improve the extraction yield and facile as it requires no special equipment. PRACTICAL APPLICATION: Extraction methods directly affect the recovery and purity of the extracted compound. Thus, this study could help in selection of appropriate extraction method for FSG. The results suggested that FSG possesses potential healthcare application in food industry because of their nutrition composition and antioxidant activities, and thus, it can be used for formulation of functional food as a natural antioxidant agent., (© 2020 Institute of Food Technologists®.)

Rationale: A viable and accurate method based on high-power ultrasound-assisted microextraction and inductively coupled plasma mass spectrometry was developed to determine metals in human serum from patients with bipolar disorder and schizophrenia., Methods: A simple and rapid sample preparation method using a cup-horn sonoreactor was developed. The acid concentration of HNO 3 (10, 20, and 40% v/v) and HCl (1, 5, 15, and 30% v/v) of the extraction solution, the sonication time (1, 3, 6, and 10 min), and the sonication amplitude (20, 40, 60, and 80%) were evaluated. Cd, Cu, Fe, Li, Pb, and Zn were determined in serum samples from patients with bipolar disorder and schizophrenia, and from healthy controls. Quantitative metal recoveries using the proposed method were compared under the same conditions using an ultrasonic bath, magnetic stirring, and microwave-assisted digestion., Results: Optimum extraction conditions were obtained using HNO 3 (40% v/v) + HCl (30% v/v) as the extraction solution with 3 min sonication time and 60% sonication amplitude. Significant differences were observed among the methods compared. On application of the sample preparation method based on high-power ultrasound-assisted microextraction coupled with inductively coupled plasma mass spectrometry, Pb and Cd in all the studied samples were below the limit of detection of our method. Compared with healthy controls, the concentration of Cu, Li, Fe, and Zn was found to be significantly higher for the bipolar disorder group, while these metals and Li were found at a lower level for the group diagnosed with schizophrenia., Conclusions: Principal component analysis showed a significant separation for the groups studied based on their ionomic profiles after the application of high-power ultrasound-assisted microextraction as a sample preparation strategy., (© 2020 John Wiley & Sons, Ltd.)

Sedimentation velocity analytical ultracentrifugation is a powerful classical method to study protein self-association processes in solution based on the size-dependent macromolecular migration in the centrifugal field. This technique can elucidate the assembly scheme, measure affinities ranging from picomolar to millimolar K d , and in favorable cases provide information on oligomer lifetimes and hydrodynamic shape. The present step-by-step protocols detail the essential steps of instrument calibration, experimental setup, and data analysis. Using a widely available commercial protein as a model system, the protocols invite replication and comparison with our results. A commentary discusses principles for modifications in the protocols that may be necessary to optimize application of sedimentation velocity analysis to other self-associating proteins. ©2020 Wiley Periodicals LLC. Basic Protocol 1: Measurement of external calibration factors Basic Protocol 2: Sedimentation velocity experiment for protein self-association Basic Protocol 3: Sedimentation coefficient distribution analysis in SEDFIT and isotherm analysis in SEDPHAT., (©2020 Wiley Periodicals LLC. This article has been contributed to by US Government employees and their work is in the public domain in the USA.)

Bioactive components possess various functionalities and are most interested for different food, nutraceutical and pharmaceutical formulations. The current review will discuss the preparation methods and fabrication techniques to design microemulsions (MEs) for the solubilization, separation, encapsulation and purification of various agro-food bioactive compounds. ME systems have shown suitable potential in enhancing oil recovery, protein extraction, and isolation of bioactive compounds. Moreover, the capability of ME based systems as drug and nutraceutical delivery cargos, and synthesis of various organic and inorganic nanoparticles, especially using biopolymers, will be investigated. ME liquid membranes are also developed as nano-extractor/nano-reactor vehicles, capable of simultaneous extraction, encapsulation or even synthesis of hydrophilic and lipophilic bioactive compounds for food, nutraceutical and drug applications., Competing Interests: Declaration of Competing Interest All authors declare that there is no conflict of interest., (Copyright © 2020. Published by Elsevier B.V.)

The flavonoids are compounds synthesized by plants, and they have properties such as antioxidant, anticancer, anti-inflammatory, and antibacterial, among others. One of the most important bioactive properties of flavonoids is their antioxidant effect. Synthetic antioxidants have side toxic effects whilst natural antioxidants, such as flavonoids from natural sources, have relatively low toxicity. Therefore, it is important to incorporate flavonoids derived from natural sources in several products such as foods, cosmetics, and drugs. For this reason, there is currently a need to extract flavonoids from plant resources. In this review are described the most important parameters involved in the extraction of flavonoids by unconventional methods such as ultrasound, pressurized liquid extraction, mechanochemical, high hydrostatic pressure, supercritical fluid, negative pressure cavitation, intensification of vaporization by decompression to the vacuum, microwave, infrared, pulsed electric field, high-voltage electrical discharges, and enzyme-assisted extraction. There are no unified operation conditions to achieve high yields and purity. Notwithstanding, progress has been achieved in the development of more advanced and environmentally friendly methods of extraction. Although in literature are found important advances, a complete understanding of the extraction process in each of the unconventional techniques is needed to determine the thermodynamic and kinetic mechanisms that govern each of the techniques., Competing Interests: The authors declare that they have no conflicts of interest., (Copyright © 2020 Sara Luisa Rodríguez De Luna et al.)

In the present study, a fast multiresidue method determining three novel fungicides fenpicoxamid, isofetamid, and mandestrobin in cereals was developed and validated for the first time using ultrahigh-performance liquid chromatography coupled to tandem mass spectrometry (UHPLC-MS/MS). Samples were extracted by QuEChERS (Quick, Easy, Cheap, Effective, Rugged, and Safe) methodology, and cleaned up using the disposable pipette extraction (DPX) tips containing primary secondary amine (PSA) and silica gel modified with zirconium oxide (Z-Sep) in less than 1 min. Linearity (r > 0.99) of three fungicides in the calibration range of 0.001-0.1 µg mL -1 was satisfactory. Mean recoveries (n = 15) from all matrices were between 84.8% and 100.3% as the corresponding intra-day and inter-day relative standard deviations (RSDs) were less than 10.6%. Limits of quantitation (LOQs) of all analytes in different matrices were defined at 0.01 mg kg -1 . The results indicate this method can serve as a sensitive and rapid approach to monitoring contents of fenpicoxamid, isofetamid, and mandestrobin in cereals., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2020 Elsevier B.V. All rights reserved.)

The high consumer demand for cosmetic products has caused the authorities and the industry to require rigorous analytical controls to assure their safety and efficacy. Thus, the determination of prohibited compounds that could be present at trace level due to unintended causes is increasingly important. Furthermore, some cosmetic ingredients can be percutaneously absorbed, further metabolized and eventually excreted or bioaccumulated. Either the parent compound and/or their metabolites can cause adverse health effects even at trace level. Moreover, due to the increasing use of cosmetics, some of their ingredients have reached the environment, where they are accumulated causing harmful effects in the flora and fauna at trace levels. To this regard, the development of sensitive analytical methods to determine these cosmetic-related compounds either for cosmetic control, for percutaneous absorption studies or for environmental surveillance monitoring is of high interest. In this sense, (micro)extraction techniques based on nanomaterials as extraction phase have attracted attention during the last years, since they allow to reach the desired selectivity. The aim of this review is to provide a compilation of those nanomaterial-based (micro)extraction techniques for the determination of cosmetic-related compounds in cosmetic, biological and/or environmental samples spanning from the first attempt in 2010 to the present.

In this work, a kinetic model for protein extraction from Camellia oleifera seed cake using high voltage electrical discharge extraction (HVED) was built with discharge energy inputs as primary variables. The results showed that both the equilibrium yields and the mass transfer coefficient of HVED were highly dependent on the HVED specific energy input per pulse (kJ/kg). After linear and nonlinear fitting with five different basic functions, the best model satisfied the power function relationship through optimizing the influence of specific energy input per pulse on the equilibrium yields and the mass transfer coefficients. Based on the observations, a predictive model that correlates the energy input, mass of raw material and kinetics of HVED extraction was proposed. The validity of the predictive model was verified, and the observed deviation was found to be less than 10%. This could provide a model basis for optimization of HVED at different processing capacities., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2020 Elsevier Ltd. All rights reserved.)

Loose nanofiltration (NF) membranes with diverse selectivity can meet the great demands in various bioseparation applications. Thus, a facile strategy to tune the properties such as pore size, surface charge, and hydrophilicity of the NF membrane is required to produce tailor-made loose NF membranes without changing the existing production line. Herein, we systematically investigated the post-treatment of the nascent poly(piperazine amide) NF membranes using different reagents (organic acids, weak bases, organic solvents and ionic liquid (IL)). Various characterizations revealed that the skin/separation layer became looser and permeance was promoted with the decrease of salt rejection in varying degrees. It was found that the O/N ratio did not rigorously represent the cross-linking degree of the skin layer, because besides the hydrolysis of the residual acyl chloride impeding the amido bond formation, the breaking of existing amido bonds and the grafting of free trimesoyl chloride molecules on the nascent membranes could also increase the O/N ratio during post-treatments. Then three mechanisms including hydrolysis, swelling rearrangement and capping reaction effects were proposed to better understand the membrane properties variations. All these effects resulted in larger pore size of the NF membrane, and the hydrolysis/capping effect might increase negative charge and hydrophilicity on the membrane, while the swelling rearrangement could produce less defective skin structure. These three effects might be involved together during a single treatment. Finally, the NF membrane post-treated by N-hexane could efficiently separate antibiotics and NaCl with the highest permeate flux, whereas the one post-treated by ionic liquid outperformed others for the decoloration of cane molasses (much more efficient than NF270, DL, and NTR7450 membranes). The long-term operating stability of the post-treated membranes selected was also confirmed by a continuous crossflow filtration for 15 h with regular alkaline cleaning.

Phycocyanin, a natural blue colorant, is typically extracted from liquid biomass of Arthrospira platensis, a blue-green algae called spirulina. In this study, we developed a scalable process to extract phycocyanin from dried spirulina biomass. First, we established the optimal ionic strength and pH for the extraction buffer. The results showed that a minimum ionic strength (>5 g/L NaCl) must be maintained to minimize the co-extraction of the green chlorophyll. The optimal pH of the phosphate buffer (100 mM) for phycocyanin extraction is 7.5; however, the pH should be immediately adjusted to 6.0 to 6.5 after the extraction to keep phycocyanin stable. Second, we also investigated three processing techniques, that is, high-pressure processing (HPP), pulsed electric field (PEF), and ultrasonication, to break the cell walls of spirulina and facilitate the release of phycocyanins into extraction buffers. HPP and PEF do not lead to the release of phycocyanin into the extraction buffer. However, ultrasonication breaks down the spirulina into fine particles and releases most of the phycocyanin, along with other impurities, immediately after the treatment. Last, it has been revealed that most of the phycocyanin can be extracted from biomass within 3 hr by phosphate buffer only (pH 7.5, 100 mM) at room temperature. It is concluded that there is no need to treat the rehydrated biomass solution by HPP, PEF, or ultrasonication due to the minimal benefits they brought for the extraction. Based on these results, we proposed an extraction process for the plant production of phycocyanin starting from dried spirulina biomass. PRACTICAL APPLICATIONS: Limited information can be found on the extraction of phycocyanin from dried spirulina biomass, especially how to better preserve the natural blue color of phycocyanin during extraction. We have investigated the method and presented a different view from previous processes. Pulsed electric field, high-pressure processing, and ultrasonication were employed to accelerate the extraction of phycocyanin from dried biomass. However, it was found that, unlike the extraction from live wet biomass, these techniques do not help with the extraction from dried biomass. Based on investigations, we have proposed a process that can be easily scaled up for the commercial production of phycocyanin from dried spirulina biomass., (© 2020 Institute of Food Technologists®.)

Hydrosoluble trehalose lipid (a biosurfactant) was employed for the first time as a green extraction solution to extract the main antioxidant compounds (geniposidic acid, chlorogenic acid, caffeic acid, and rutin) from functional plant tea (Eucommia ulmoides leaves). Single-factor tests and response surface methodology were employed to optimize the extraction conditions for ultrasound-assisted micellar extraction combined with ultra-high-performance liquid chromatography in succession. A Box-Behnken design (three-level, three-factorial) was used to determine the effects of extraction solvent concentration (1-5 mg/mL), extraction solvent volume (5-15 mL), and extraction time (20-40 min) at a uniform ultrasonic power and temperature. In consequence, the best analyte extraction yields could be attained when the trehalose lipid solution concentration was prepared at 3 mg/mL, the trehalose lipid solution volume was 10 mL and the extraction time was set to 35 min. In addition, the recoveries of the antioxidants from Eucommia ulmoides leaves analyzed by this analytical method ranged from 98.2 to 102%. These results indicated that biosurfactant-enhanced ultrasound-assisted micellar extraction coupled with a simple ultra-high-performance liquid chromatography method could be effectively applied in the extraction and analysis of antioxidants from Eucommia ulmoides leaf samples., (© 2019 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.)

Globe artichoke (Cynara cardunculus var. scolymus L.) is not only used as a vegetable and ornamental plant, but also as important medicinal plant for the treatment of dyspeptic disorders. The European Pharmacopoeia describes a method for the quality assessment of dry artichoke leaves, which is time-consuming and requires huge amounts of organic solvents. In this study, an ultrasound-assisted extraction method was studied, which proved to be more efficient than the standard protocol of the European Pharmacopoeia, since it led to comparable results, was faster and easier to handle, and was more sustainable due to a reduced need for organic solvents., (Copyright © 2019 Elsevier B.V. All rights reserved.)

Fish-scale waste is rich in biocompatible hydroxyapatite (HAp). In the present study, an environmentally friendly method of extracting HAp from fish-scale waste was developed in an effort to promote environmental sustainability. Deep eutectic solvents (choline chloride/glycerol, 1/2) were used to extract HAp from bighead carp (Aristichthys nobilis) scales. A relatively high extraction rate of 47.67% ± 1.81% was obtained under optimum conditions (70 °C, a solid/liquid ratio of 1/15 g/g and a 2.5 hr extraction time). The obtained HAp was characterized and its purity was determined using Fourier transform infrared spectroscopy and X-ray diffraction, respectively. The chemical composition was performed by energy-dispersive X-ray spectrometry and inductively coupled plasma-optical emission spectroscopy. Its morphology and particle size were observed using scanning electron microscopy and particle size distribution analysis. Thermogravimetric analysis was used to determine its thermal stability. Blood compatibility was determined using a hemolytic test. The results showed that this extraction yielded HAp with the irregular morphology, the higher Ca/P ratio, good thermal stability, and blood compatibility, indicating that the proposed method is an excellent alternative for the improved utilization of fish scale waste. PRACTICAL APPLICATION: Biocompatible hydroxyapatite (HAp) was extracted from fish scale (FS) waste by using an environmentally friendly deep eutectic solvent. The optimized extraction and structure characterization of extracted HAp were investigated in this study. The results showed that the extracted HAp had the irregular morphology, the higher Ca/P ratio, good thermal stability, and blood compatibility, which indicated that the proposed method was an excellent alternative to improving the utilization of FS waste., (© 2019 Institute of Food Technologists®.)

Urinary microRNAs show promise as noninvasive biomarkers in renal disease. Here, we describe a detailed protocol for the column-based extraction and quantification of miRNAs by reverse-transcription quantitative polymerase chain reaction (RT-qPCR) from urine samples.

This work reports the development of a very-simple-to-construct stir-bar extraction device so called "a dumbbell-shaped stainless steel stir-bar." The extraction device was assembled from a rolled up stainless steel net filled with an XAD-2 sorbent and a metal rod to allow the use of a magnetic stirrer during extraction. The dumbbell-shaped stainless steel stir-bar was used to extract diethyl phthalate (DEP), dibutyl phthalate (DBP), and di(2-ethylhexyl) phthalate (DEHP) before analysis by a gas chromatograph equipped with an electron capture detector (GD-ECD). Under the optimal conditions, the developed method provided a good linearity from 10.0 to 1,000.0 ng mL -1 for all three compounds. Limits of detection and limits of quantification were 9.37 ± 0.29 ng mL -1 and 31.22 ± 0.95 ng mL -1 for DEP, 5.73 ± 0.31 ng mL -1 and 19.1 ± 1.0 ng mL -1 for DBP and 3.30 ± 0.06 ng mL -1 and 11.0 ± 0.19 ng mL -1 for DEHP, respectively. Good recoveries in the range of 81.89 ± 0.17 to 109.5 ± 2.0% were achieved when the method was used to extract phthalate esters in five instant noodle and two rice soup samples.

We describe the development of a high-resolution, noncontact fraction collector for liquid chromatography (LC) separations, allowing high-resolution fractionation in high-density well plates. The device is based on a low-dead-volume solenoid valve operated at 1-30 Hz for accurate collection of fractions of equal volume. The solenoid valve was implemented in a modified autosampler resulting in the so-called FractioMate fractionator. The influence of the solenoid supply voltage on solvent release was determined and the effect of the frequency, flow rate, and mobile phase composition was studied. For this purpose, droplet release was visually assessed for a wide range of frequencies and flow rates, followed by quantitative evaluation of a selection of promising settings for highly accurate, repeatable, and stable fraction collection. The potential of the new fraction collector for LC-based bioactivity screening was demonstrated by fractionating the LC eluent of a mixture of estrogenic and androgenic compounds, and a surface water sample (blank and spiked with bioactives) combining mass spectrometric detection and two reporter gene assays for bioactivity detection of the fractions. Additionally, a mixture of two compounds was repeatedly LC separated and fractionated to assess the feasibility of the system for analyte isolation followed by nuclear magnetic resonance analysis.

In this work, a monolithic polymer based on poly(methacrylic acid-co-ethylene glycol dimethacrylate) (MAA-co-EDMA) was prepared inside 200 μL pipette tips for the extraction of drug of abuse from oral fluid samples. After an appropriate surface tip modification, several polymerization mixtures with different monomer/cross-linker ratios, and percentage of porogen were studied. The most appropriate monolith to easily flow organic solvents and oral fluid samples was prepared with a MAA/EDMA ratio of 8:92 wt/wt and dodecanol containing 10 wt% toluene, as porogenic solvent. Parameters affecting the extraction procedure were evaluated and the monolith was characterized in terms of binding capacity, reusability, and precision, using α-pyrrolidinovalerophenone as model compound. Cocaine, diazepam, methamphetamine and 20 new psychoactive substances were determined in oral fluids, using the synthesized poly(MAA-co-EDMA) monolith in-tip on an eight-channel micropipette extraction and ultra-high performance liquid chromatography tandem mass spectrometry. Appropriate recoveries were obtained, ranging from 64 to 115%, with limit of detection values from 0.03 to 0.6 μg L -1 , and a high precision with relative standard deviation values lower than 10% for all the evaluated drugs., (Copyright © 2019 Elsevier B.V. All rights reserved.)

Background: The ultrasound-assisted extraction of pectic polysaccharides from orange by-products was investigated. Kinetics of mechanical agitation (0.2 × g) and acoustic (US1: 542 W L -1 and US2: 794 W L -1 ) extractions, were obtained and modelled at different pH values (1.5 and 2.0). All extractions were carried out at 25 °C, using citric acid as the extraction solvent., Results: Higher pectic polysaccharides extraction yields were obtained with ultrasonic assistance, in comparison with the results obtained using mechanical agitation. Moreover, yield increases were significantly higher using the more acidic pH. Thus, at pH 1.5, pectin yield increased from ∼19%, obtained with agitation, to ∼47%, applying ultrasound; whereas, at pH 2.0, this increase was from ∼10%, with agitation, to ∼18%, applying ultrasound. A considerable decrease of the galacturonic acid proportion was observed on the extracts when ultrasound were applied for 60 min under pH 2.0. High methoxyl pectins were extracted at pH 1.5 whereas at pH 2.0, pectins exhibited a low methylation degree. Curves of acoustic and mechanical agitation extractions were properly represented by a second-order rate model (average mean relative error ≤ 7.4%). The extraction rate constant, initial extraction rate and maximum yield were determined for all experimental conditions., Conclusion: Overall, the results clearly indicated that the effect of ultrasound was highly dependent on the pH. Therefore, adequate acidic conditions must be applied in order to improve the efficiency of ultrasound on the pectin extraction process. © 2019 Society of Chemical Industry., (© 2019 Society of Chemical Industry.)

Digital PCR is a powerful amplification method for absolute quantification of nucleic acids. The systems that integrated the nucleic acid extraction and amplification can reduce detection time, improve accuracy, and reduce labor costs. However, current nucleic acid extraction systems cannot be integrated well with integrated fluidic circuit (IFC) dPCR or droplet digital PCR chips perfectly and limit the application of digital PCR. In this study, a polytetrafluoroethylene (PTFE)-based nucleic acid extraction (PNE) system, which was able to achieve fully closed extraction for micro samples and was able to be integrated with IFC dPCR or droplet digital dPCR (ddPCR) chips perfectly is proposed. For this system, PTFE tubing with an inner diameter of 1 mm was used to load the reagents and superparamagnetic particles (PMPs) were used to extract nucleic acids. The system can extract nucleic acids from cells and blood in 5 minutes. Meanwhile, when nucleic acid extraction was completed, PNE was able to be directly combined with IFC dPCR or ddPCR chips without any intermediate steps. Therefore, the PNE system can realize sample-in-digital-answer-out. It will be highly useful in point-of-care (POC) and promote the development and application of dPCR.

Byproducts such as orange peel have potential uses because of their bioactive compounds, which are important for their potential to reduce the risk factors of diseases caused by aging. The lack of effective techniques and the high levels of pollution produced by the conventional extraction of bioactive compounds using organic solvents have highlighted the need to enhance the 'green chemistry' trend. This study evaluates the use of ultrasound to extract bioactive compounds from orange peel. The antioxidant capacity, phenolic content, ascorbic acid, total carotenoids, and HPLC profile of phenolic compounds from orange peel extracts were obtained by a physicochemical evaluation. The results demonstrate that the optimal conditions for the ultrasound-assisted extraction of bioactive orange peel compounds were a power of 400 W, a time of 30 min, and 50% ethanol in water. These conditions were used to obtain a total carotenoid concentration of 0.63 mg ß-carotene/100 g, vitamin C concentration of 53.78 mg AA/100 g, phenolic concentration of 105.96 mg GAE/100 g, and antioxidant capacity of ORAC = 27.08 mM TE and TEAC = 3.97 mM TE. The major phenolic compound identified in all orange peel extracts was hesperidin, with a maximum concentration of 113.03 ± 0.08 mg/100 g.

Salmonella enterica serovar typhimurium (S. typhimurium) causes food poisoning in human and animals. Its infection rate is the highest among all salmonella serotypes. Metabolomics is a potential way to study the pathogenesis of S. typhimurium via analysis of various small molecular substances. Due to the lack of a uniform protocol for the extraction of metabolites, we evaluated five commonly used extraction methods including cold methanol (CM), hot ethanol (HE), chloroform-methanol cocktail (CMC), perchloric acid (PCA), and alkali (AL) for their efficacy in extracting the intracellular metabolites of S. typhimurium. Samples were quenched in 60% methanol at - 40 °C, and then the five methods were used to extract the metabolites. After derivatization, all samples were analyzed on a gas chromatography-triple quadrupole mass spectrometry (GC-MS/MS). Our results suggest that CM and HE extraction methods provide the best compromise allowing identification of 98 and 95 metabolites in a single analysis. For targeted metabolome analysis, the optimal extraction method for alcohols and organic acids is HE. CMC preferentially extracted lipid metabolites. PCA is suitable for extraction of small molecular carbohydrates. The optimal extraction method for macromolecular carbohydrates is the CM method.

Accelerated solvent extraction (ASE) and solid-phase extraction (SPE) conditions were optimized by a high-performance liquid chromatography-fluorescence detector (HPLC-FLD) method for the detection of piperazine in chicken tissues and pork. Piperazine residues were determined by precolumn derivatization with trimethylamine and dansyl chloride. Samples were extracted with 2% formic acid in acetonitrile using an ASE apparatus and purified using a Strata-X-C SPE column. The monosubstituted product of the reaction of piperazine with dansyl chloride was 1-dansyl piperazine (1-DNS-piperazine). Chromatographic separations were performed on an Athena C 18 column (250 × 4.6 mm, id: 5 μm) with gradient elution using ultrapure water and acetonitrile (5:95, V/V) as the mobile phase. The calibration curves showed good linearity over a concentration range of LOQ-200.0 μg/kg with a coefficient of determination (R 2 ) ≥ .9992. The recoveries and relative standard deviations (RSD values) ranged from 78.49% to 97.56% and 1.19% to 5.32%, respectively, across the limit of quantification (LOQ) and 0.5, 1, and 2.0 times the maximum residue limit (MRL; μg/kg). The limits of detection (LODs) and LOQs were 0.96 to 1.85 μg/kg and 3.20 to 5.50 μg/kg, respectively. The method was successfully applied for the validation of animal products in the laboratory., (© 2019 Wiley Periodicals, Inc.)

The extraction of bioanalytes is the first step in many diagnostic and analytical assays. However, most bioanalyte extraction methods require extensive dilution-based washing processes that are not only time-consuming and laborious but can also result in significant sample loss, limiting their applications in rare sample analyses. Here, we present a method that enables the efficient extraction of multiple different bioanalytes from rare samples (down to 10 cells) without washing-centrifugation-assisted immiscible fluid filtration (CIFF). CIFF utilizes centrifugal force to drive the movement of analyte-bound glass microbeads from an aqueous sample into an immiscible hydrophobic solution to perform an efficient, simple, and nondilutive extraction. The method can be performed using conventional polymerase chain reaction (PCR) tubes with no requirement of specialized devices, columns, or instruments, making it broadly accessible and cost-effective. The CIFF process can effectively remove approximately 99.5% of the aqueous sample in one extraction with only 0.5% residual carryover, whereas a traditional "spin-down and aspirate" operation results in a higher 3.6% carryover. Another unique aspect of CIFF is its ability to perform two different solid-phase bioanalytes extractions simultaneously within a single vessel without fractionating the sample or performing serial extractions. Here we demonstrate efficient mRNA and DNA extraction from low-input samples (down to 10 cells) with slightly higher to comparable recovery compared to a traditional column-based extraction technique and the simultaneous extraction of two different proteins in the same tube using CIFF.

Proteome is extremely complex as many proteins with a large dynamic range are involved. Nano-liquid chromatography/mass spectrometry-based proteomics has made it possible to separate and identify thousands of proteins in one shot. Although the number of identified proteins in proteomics has significantly improved, it is necessary to increase detection sensitivity to clearly identify low-abundant proteins. In this study, we developed meter-long monolithic columns with a small inner diameter and applied them to selected reaction monitoring-based proteomics for improving proteomic detection sensitivity. Bovine serum albumin tryptic digests were analyzed with optimized selected reaction monitoring methods, and separation efficiency and detection sensitivity in each monolithic column were evaluated. As a result, peak capacity increased by about 1.8-fold and peak area of peptide levels increased by about 2.3-fold. Although flow rate was reduced during analysis with columns of a smaller inner diameter, the peak area reproducibility was maintained. These data displayed the value of meter-long monolithic columns with small inner diameter for selected reaction monitoring-based proteomics., (Copyright © 2019 The Society for Biotechnology, Japan. Published by Elsevier B.V. All rights reserved.)

Although magnesium is the second most abundant cation present in the cell, the transport mechanism of Mg 2+ across membranes is poorly understood. Importantly, the prokaryotic MgtE Mg 2+ channel is related to mammalian SLC41A1 transporters and, therefore, biochemical and biophysical characterization of MgtE and its orthologs assumes significance. To date, the purification and structure determination of MgtE from Thermus thermophilus has been carried out using the widely used nonionic detergent, n-dodecyl-β-d-maltopyranoside (DDM). However, DDM is an expensive detergent and alternative methods to produce high-quality proteins in stable and functional form will be practically advantageous to carry out structural studies in a cost-effective manner. In this work, we have utilized 'dual-detergent strategy' to successfully purify MgtE channel in a stable and functional form by employing relatively inexpensive detergents (Triton X-100 and Anzergent 3-14) for membrane solubilization and subsequently changed to DDM during purification. Our results show that Triton X-100 and Anzergent 3-14 extract MgtE well and the quality of purified protein is comparable to DDM-extracted MgtE. Interestingly, addition of high concentration of salt and glycerol during solubilization does not significantly affect the quantity and quality of MgtE. Importantly, limited proteolysis assay, circular dichroism spectroscopy and ensemble tryptophan fluorescence strongly support the use of Triton X-100, in particular, as an inexpensive, alternative detergent for the purification of MgtE without compromising the structural integrity of the channel and Mg 2+ -induced gating-related conformational dynamics. Overall, these results are relevant for the cost-effective purification of stable and functional membrane proteins in general, and magnesium channels, in particular., (Copyright © 2019 Elsevier Inc. All rights reserved.)

High-throughput screening platforms for the identification of bioactive compounds in mixtures have become important tools in the drug discovery process. Miniaturization of such screening systems may overcome problems associated with small sample volumes and enhance throughput and sensitivity. Here we present a new screening platform, coined picofractionation analytics, which encompasses microarray bioassays and mass spectrometry (MS) of components from minute amounts of samples after their nano liquid chromatographic (nanoLC) separation. Herein, nanoLC was coupled to a low-volume liquid dispenser equipped with pressure-fed solenoid valves, enabling 50-nL volumes of column effluent (300 nL/min) to be discretely deposited on a glass slide. The resulting fractions were dried and subsequently bioassayed by sequential printing of nL-volumes of reagents on top of the spots. Unwanted evaporation of bioassay liquids was circumvented by employing mineral oil droplets. A fluorescence microscope was used for assay readout in kinetic mode. Bioassay data were correlated to MS data obtained using the same nanoLC conditions in order to assign bioactives. The platform provides the possibility of freely choosing a wide diversity of bioassay formats, including those requiring long incubation times. The new method was compared to a standard bioassay approach, and its applicability was demonstrated by screening plasmin inhibitors and fibrinolytic bioactives from mixtures of standards and snake venoms, revealing active peptides and coagulopathic proteases.

The research of fungi is of great importance in a number of fields, such as environmental and healthcare studies. While there are a large number of optical and molecular methods available for characterization and identification of fungi and their spores, their isolation is still conducted using slow and labor-intensive methods. Here, we develop a microfluidic device for the continuous separation of fungal spores from other eukaryotic cells. The spores were separated through the microfluidic device by expanding pinched flow fractionation (PFF) containing the spores, achieving a spatial separation perpendicular to the flow direction according to the spore size. Further branch flow fractionation (BFF) and co-flow of a Newtonian and viscoelastic fluid were used to enhance the separation performance. Using this microfluidic device, we demonstrated the separation of two different types of fungal spores and further separation of fungal spores from eukaryotic cells with a separation efficiency of above 90%. Compared to the existing conventional methods, our microfluidic flow focusing device requires little manual handling and uses small amounts of samples without any pre-treatment steps of the samples.

The conversion of lignocellulosic biomass into biofuels and bio-products leads to oxygenated matrices having a wide range of polarities and molecular weights. A complete analytical characterization of these complex mixtures is necessary to improve conversion processes. In this study, an innovative centrifugal partition chromatography (CPC) protocol was developed to fractionate aqueous biomass samples with a MTBE-water solvent system, by mixing elution, displacement and extrusion modes in the same run. This new protocol was validated on model molecules and applied to the water soluble phase of a fast pyrolysis bio-oil. It demonstrated a promising separation with a relevant selectivity on the most significant chemical families of biomass samples: carbohydrates, furans, carboxylic acids and phenols. CPC fractions of the sample were collected and analyzed comprehensively by HPLC-UV/MS (with ESI negative and positive ionization modes). This CPC x LC approach allowed more accurate attributions on the 217 peaks detected. The use of different detection modes gave a complete view of the water soluble phase of a fast pyrolysis bio-oil through 2D maps. Molecular characterization was enhanced by independent information: CPC retention time, LC retention time, UV and MS spectra. Concomitance of these different chemical information is of precious help for unambiguous identification., (Copyright © 2019 Elsevier B.V. All rights reserved.)

In this study, a novel core-shell structured Fe 3 O 4 @TA@P(NVP-co-NIPAM) magnetic microsphere has been successfully developed for polyphenol extraction employing tannic acid (TA) directed copolymerization. The TA directing strategy guided copolymerization preferentially on the surface of the magnetic core and facilitated formation of the shell for coating the magnetic core. It was unveiled that the amount of TA, the ratio of the two monomers, and the amount of surfactant were critical for the preparation of the magnetic microspheres. The identity and proportion of the shell were analyzed by FT-IR and TGA. Magnetic hysteresis loop measurement confirmed the superparamagnetic nature and no loss of magnetization after coating. The prepared magnetic microspheres demonstrated outstanding absorption ability to typical polyphenols. Kinetic studies suggested that the adsorption behavior followed Langmuir absorption model. The developed magnetic microspheres should be useful for biological studies. The TA directing strategy would be valuable for synthesis of core-shell structured particles., (Copyright © 2019 Elsevier Ltd. All rights reserved.)

Liquid chromatography-electrospray ionization tandem mass spectrometry (LC-ESI-MS/MS) with a triple quadrupole (QqQ) is proposed for determining the vitamin K homologues, phylloquinone (PK), menaquinone-4 (MK) and menadione (MD), in vegetables. The analytes were isolated from the samples (1-1.5 g) by ultrasound assisted extraction using acetonitrile (2 mL), and the liquids were submitted to microwave assisted cloud point extraction with Triton X-45. The enrichment factors were between 20 and 50, depending on the vitamin homologue in question. The analytes were determined by LC-ESI-QqQ-MS/MS in the multiple reaction monitoring (MRM) mode, providing unequivocal identification and quantification, with limits of detection of 0.8, 1.0, and 16 ng/g for MK, PK, and MD, respectively. Recovery assays for samples spiked at two concentration levels, between 40 and 600 ng/g depending on the compound, provided recoveries in the 90-114% range. Only PK was detected in the samples analyzed, at concentrations in the 90-2350 ng/g range.

Silver nanoparticles were coated on a glass stir bar by overnight exposure of the bar to the nanoparticle solution, using 3-aminopropyltriethoxysilane as a binding agent and were used for the extraction of heavy metal Fe 3+ , Cu 2+ , Pb 2+ and Zn 2+ cations from water samples after their complexation with the ligand 1-(2-pyridylazo)-2-naphthol, followed by their determination by high performance liquid chromatography. Various parameters affecting the extraction were evaluated by one variable-at-a-time and Box-Behnken design methods and optimized. Six independent variables, the pH and ionic strength of the solution, the Triton X-114 (surfactant) concentration, amount of ligand, and adsorption and desorption times, were coded as A, B, C, D, E and F at three various levels (-1, 0, and 1). The linear ranges of the technique were 0.5-250.0 μg L -1 for all four analytes. Under the optimum conditions, the limits of detection were calculated as 0.08, 0.11, 0.12 and 0.17 μg L -1 for copper, iron, lead and zinc, respectively. The technique was used for the determination of these cations in drinking water samples.