Impact of extraction method on the lipids of Himalayan marmot oil with ultrahigh-performance liquid chromatography Q-Exactive Orbitrap mass spectrometry analysis.

RATIONALE: Himalayan marmot oil (SPO) has been used for pharmaceutical purposes for centuries, but its composition is still unclear. The bioactivity of SPO highly depends on the techniques used for its processing. This study focused on the comprehensive lipidomics of SPO, especially on the ones derived from dry rendering, wet rendering, cold pressing, and ultrasound-assisted solvent extraction. METHODS: We performed lipid profiling of SPO acquired by different extraction methods using ultrahigh-performance liquid chromatography Q-Exactive Orbitrap mass spectrometry, and 17 classes of lipids (2 BMPs, 12 LysoPCs, 9 LysoPEs, 41 PCs, 24 PEs, 23 Plasmenyl-PCs, 10 Plasmenyl-PEs, 10 MGs, 63 DGs, 187 TGs, 2 MGDGs, 3 Cer[NDS]s, 22 Cer[NS]s, 2 GlcCer[NS]s, 14 SMs, 14 CEs, and 6 AcylCarnitines) were characterized. RESULTS: Fifty-five lipids were differentially altered (VIP > 1.5, p < 0.05) between the extraction techniques, which can be used as potential biomarkers to differentiate SPO extracted by various methods. Additionally, the contents of oleic acid and arachidic acid were abundant in all samples that may suggest their medicinal values and are conducive to in-depth research. CONCLUSIONS: These findings reveal the alterations of lipid profile and free fatty acid composition in SPO obtained with different extraction methods, providing a theoretical foundation for investigating its important components as functional factors in medicines and cosmetics.

A comprehensive review on pulse protein fractionation and extraction: processes, functionality, and food applications.

The increasing world population requires the production of nutrient-rich foods. Protein is an essential macronutrient for healthy individuals. Interest in using plant proteins in foods has increased in recent years due to their sustainability and nutritional benefits. Dry and wet protein fractionation methods have been developed to increase protein yield, purity, and functional and nutritional qualities. This review explores the recent developments in pretreatments and fractionation processes used for producing pulse protein concentrates and isolates. Functionality differences between pulse proteins obtained from different fractionation methods and the use of fractionated pulse proteins in different food applications are also critically reviewed. Pretreatment methods improve the de-hulling efficiency of seeds prior to fractionation. Research on wet fractionation methods focuses on improving sustainability and functionality of proteins while studies on dry methods focus on increasing protein yield and purity. Hybrid methods produced fractionated proteins with higher yield and purity while also improving protein functionality and process sustainability. Dry and hybrid fractionated proteins have comparable or superior functionalities relative to wet fractionated proteins. Pulse protein ingredients are successfully incorporated into various food formulations with notable changes in their sensory properties. Future studies could focus on optimizing the fractionation process, improving protein concentrate palatability, and optimizing formulations using pulse proteins.

Deep eutectic solvents as sustainable extraction media for extraction of polysaccharides from natural sources: Status, challenges and prospects.

Deep eutectic solvents (DES) emerge as promising alternatives to conventional solvents, offering outstanding extraction capabilities, low toxicity, eco-friendliness, straightforward synthesis procedures, broad applicability, and impressive recyclability. DES are synthesized by combining two or more components through various synthesis procedures, such as heat-assisted mixing/stirring, grinding, freeze drying, and evaporation. Polysaccharides, as abundant natural materials, are highly valued for their biocompatibility, biodegradability, and sustainability. These versatile biopolymers can be derived from various natural sources such as plants, algae, animals, or microorganisms using diverse extraction techniques. This review explores the synthesis procedures of DES, their physicochemical properties, characterization analysis, and their application in polysaccharide extraction. The extraction optimization strategies, parameters affecting DES-based polysaccharide extraction, and separation mechanisms are comprehensively discussed. Additionally, this review provides insights into recently developed molecular guides for DES screening and the utilization of artificial neural networks for optimizing DES-based extraction processes. DES serve as excellent extraction media for polysaccharides from different sources, preserving their functional features. They are utilized both as extraction solvents and as supporting media to enhance the extraction abilities of other solvents. Continued research aims to improve DES-based extraction methods and achieve selective, energy-efficient processes to meet the demands of this expanding field.

Rice proteins: A review of their extraction, modification techniques and applications.

Rice protein is highly nutritious and easy to digest and absorb. Its hydrolyzed peptides have significant effects on lowering blood pressure and cholesterol. First, a detailed and comprehensive explanation of rice protein extraction methods was given, and it was found that the combination of enzymatic and physical methods could improve the extraction rate of rice protein, but it was only suitable for laboratory studies. Second, the methods for improving the functional properties of rice protein were introduced, including physical modification, chemical modification, and enzymatic modification. Enzymatic modification of the solubility of rice protein to improve its functional properties has certain limitations due to the low degree of hydrolysis, the long time required, the low utilization of the enzyme, and the possible undesirable taste of the product. Finally, the development and utilization of rice protein was summarized and the future research direction was suggested. This paper lists the advantages and disadvantages of various extraction techniques, points out the shortcomings of existing extraction techniques, aims to fill the gap in the field of rice protein extraction, and then provides a possible improvement method for the extraction and development of rice protein in the future.

Green and efficient method to acquire high-value phycobiliprotein from microalgal biomass involving deep eutectic solvent-based ultrasound-assisted extraction.

Phycoerythrin (PE) is a phycobiliprotein holding great potential as a high-value food colorant and medicine. Deep eutectic solvent (DES)-based ultrasound-assisted extraction (UAE) was applied to extract B-PE by disrupting the resistant polysaccharide cell wall of Porphyridium purpureum. The solubility of cell wall monomers in 31 DESs was predicted using COSMO-RS. Five glycerol-based DESs were tested for extraction, all of which showed significantly higher B-PE yields by up to 13.5 folds than water. The DES-dependent B-PE extraction efficiencies were proposedly associated with different cell disrupting capabilities and protein stabilizing effects of DESs. The DES-based UAE method could be considered green according to a metric assessment tool, AGREEprep. The crude extract containing DES was further subjected to aqueous two-phase system, two-step ammonium sulfate precipitation, and ultrafiltration processes. The final purified B-PE had a PE purity ratio of 3.60 and a PC purity ratio of 0.08, comparable to the purity of commercial products.

Differences in physicochemical properties of pectin extracted from pomelo peel with different extraction techniques.

In order to obtain high yield pomelo peel pectin with better physicochemical properties, four pectin extraction methods, including hot acid extraction (HAE), microwave-assisted extraction (MAE), ultrasound-assisted extraction, and enzymatic assisted extraction (EAE) were compared. MAE led to the highest pectin yield (20.43%), and the lowest pectin recovery was found for EAE (11.94%). The physicochemical properties of pomelo peel pectin obtained by different methods were also significantly different. Pectin samples obtained by MAE had the highest methoxyl content (8.35%), galacturonic acid content (71.36%), and showed a higher apparent viscosity, thermal and emulsion stability. The pectin extracted by EAE showed the highest total phenolic content (12.86%) and lowest particle size (843.69 nm), showing higher DPPH and ABTS scavenging activities than other extract methods. The pectin extracted by HAE had the highest particle size (966.12 nm) and degree of esterification (55.67%). However, Fourier-transform infrared spectroscopy showed that no significant difference occurred among the different methods in the chemical structure of the extracted pectin. This study provides a theoretical basis for the industrial production of pomelo peel pectin.

Ultrasonic-assisted extraction of polysaccharide from Paeoniae Radix alba: Extraction optimization, structural characterization and antioxidant mechanism in vitro.

Paeoniae Radix alba is used in Traditional Chinese Medicine for the treatment of gastrointestinal disorders, immunomodulatory, cancer, and other diseases. In the current study, the yield of Paeoniae Radix alba polysaccharide (PRP) was significantly increased with optimal ultrasound-assisted extraction compared to hot water extraction. Further, an acidic polysaccharide (PRP-AP) was isolated from PRP after chromatographic separation and was characterized as a typical pectic polysaccharide with side chains of arabinogalactans types I and II. Moreover, it showed antioxidant effects on LPS-induced damage on IPEC-J2 cells determined by qRT-PCR and ELISA, including decreasing the pro-inflammatory factors' expressions and increasing the antioxidant enzymes activities, which was shown to be related to the Nrf2/Keap1 pathway modulated by PRP-AP. The metabolites change (such as itaconate, cholesterol sulfate, etc.) detected by untargeted metabolomic analysis in cells was also shown to be modulated by PRP-AP, and these metabolites were further utilized and protected cells damaged by LPS. These results revealed the cellular active mechanism of the macromolecular PRP-AP on protecting cells, and supported the hypothesis that PRP-AP has strong benefits as an alternative dietary supplement for the prevention of intestinal oxidative stress by modulating cellular metabolism.

Characterization and prebiotic potential of polysaccharides from Rosa roxburghii Tratt pomace by ultrasound-assisted extraction.

In this study, polysaccharides (RRTPs) were extracted from Rosa roxburghii Tratt pomace by hot water or ultrasound (US)-assisted extraction. The structural properties and potential prebiotic functions of RRTPs were investigated. Structural characterization was conducted through HPAEC, HPGPC, GC-MS, FT-IR and SEM. Chemical composition analysis revealed that RRTPs extracted by hot water (RRTP-HW) or US with shorter (RRTP-US-S) or longer duration (RRTP-US-L) all consisted of galacturonic acid, galactose, glucose, arabinose, rhamnose and glucuronic acid in various molar ratio. US extraction caused notable reduction in molecular weight of RRTPs but no significant changes in primary structures. Fecal fermentation showed RRTPs could reshape microbial composition toward a healthier balance, leading to a higher production of beneficial metabolites including total short-chain fatty acids, curcumin, noopept, spermidine, 3-feruloylquinic acid and citrulline. More beneficial shifts in bacterial population were observed in RRTP-HW group, while RRTP-US-S had stronger ability to stimulate bacterial short-chain fatty acids production. Additionally, metabolic profiles with the intervention of RRTP-HW, RRTP-US-S or RRTP-US-L were significantly different from each other. The results suggested RRTPs had potential prebiotic effects which could be modified by power US via molecular weight degradation.

Seaweed-derived phenolic compounds as diverse bioactive molecules: A review on identification, application, extraction and purification strategies.

Seaweed, a diverse group of marine macroalgae, has emerged as a rich source of bioactive compounds with numerous health-promoting properties. Among these, phenolic compounds have garnered significant attention for their diverse therapeutic applications. This review examines the methodologies employed in the extraction and purification of phenolic compounds from seaweed, emphasizing their importance in unlocking the full potential of these oceanic treasures. The article provides a comprehensive overview of the structural diversity and biological activities of seaweed-derived phenolics, elucidating their antioxidant, anti-inflammatory, and anticancer properties. Furthermore, it explores the impact of extraction techniques, including conventional methods and modern green technologies, on the yield and quality of phenolic extracts. The purification strategies for isolating specific phenolic compounds are also discussed, shedding light on the challenges and advancements in this field. Additionally, the review highlights the potential applications of seaweed-derived phenolics in various industries, such as pharmaceuticals, cosmetics, and functional foods, underscoring the economic value of these compounds. Finally, future perspectives and research directions are proposed to encourage continued exploration of seaweed phenolics, fostering a deeper understanding of their therapeutic potential and promoting sustainable practices in the extraction and purification processes. This comprehensive review serves as a valuable resource for researchers, industry professionals, and policymakers interested in harnessing the untapped potential of phenolic compounds from seaweed for the betterment of human health and environmental sustainability.

Ultrasonic-assisted extraction to enhance the recovery of bioactive phenolic compounds from Commiphora gileadensis leaves.

The "ultrasonic-assisted extraction (UAE)" method was utilized in this work to assess how different process parameters affected the yield and recovery of phenolic compounds from the leaf of Commiphora gileadensis, which is one of the medicinal plants with a variety of biological functions. Its leaf is used for a various of therapeutic applications, such as the treatment of bacterial infections, inflammation, and wound healing. The "One-Factor-At-a-Time (OFAT)" approach was employed to examine the impacts of various UAE process parameters on the process of extraction, which include time of extraction, sample/solvent ratio, ultrasonic frequency, and solvent (ethanol) concentration. The extracts were then investigated for the presence of several phytochemicals using analytical techniques such as "Gas Chromatography-Mass Spectroscopy (GC-MS)" and "Fourier Transform Infrared Spectroscopy (FTIR)" studies. The findings showed that the maximum extraction yield, the total phenolic content (TPC), and the total flavonoids content (TFC) of the ethanolic extract of the leaves of C. gileadensis using the UAE method were at 31.80 ± 0.41 %, 96.55 ± 2.81 mg GAE/g d.w. and 31.66 ± 2.01 mg QE/g d.w. accordingly under a procedure duration of 15 min, ultrasonic frequency of 20 kHz, solvent/sample ratio of 1:20 g/mL, and solvent concentration of 40 % v/v. The leaves extract of C. gileadensis included 25 phenolic compounds that were previously unreported, and GC-MS analysis confirmed their presence. Hence, it follows that the UAE technique can successfully extract the phytochemicals from C. gileadensis for a variety of therapeutic uses.

Sustainable green extraction of anthocyanins and carotenoids using deep eutectic solvents (DES): A review of recent developments.

Recently, deep eutectic solvents (DES) have been extensively researched as a more biocompatible and efficient alternative to conventional solvents for extracting pigments from natural resources. The efficiency of DES extraction for the anthocyanin and carotenoid can be enhanced by microwave-assisted extraction (MAE) and/or ultrasound-assisted extraction (UAE) techniques. Apart from the extraction efficiency, the toxicity and recovery of the pigments and their bioavailability are crucial for potential applications. A plethora of studies have explored the extraction efficiency, toxicity, and recovery of pigments from various natural plant-based matrices using DES. Nevertheless, a detailed review of the deep eutectic solvent extraction of natural pigments has not been reported to date. Additionally, the toxicity, safety, and bioavailability of the extracted pigments, and their potential applications are not thoroughly documented. Therefore, this review is designed to understand the aforementioned concepts in using DES for anthocyanin and carotenoid extraction.

Fibrillization of lentil proteins is impacted by the protein extraction conditions and co-extracted phenolics.

Legume proteins can be induced to form amyloid-like fibrils upon heating at low pH, with the exact conditions greatly impacting the fibril characteristics. The protein extraction method may also impact the resulting fibrils, although this effect has not been carefully examined. Here, the fibrillization of lentil protein prepared using various extraction methods and the corresponding fibril morphology were characterized. It was found that an acidic, rather than alkaline, protein extraction method was better suited for producing homogeneous, long, and straight fibrils from lentil proteins. During alkaline extraction, co-extracted phenolic compounds bound proteins through covalent and non-covalent interactions, contributing to the formation of heterogeneous, curly, and tangled fibrils. Recombination of isolated phenolics and proteins (from acidic extracts) at alkaline pH resulted in a distinct morphology, implicating a role for polyphenol oxidase also in modifying proteins during alkaline extraction. These results help disentangle the complex factors affecting legume protein fibrillization.

Ultrasonic-assisted extraction of grape seed procyanidins, preparation of liposomes, and evaluation of their antioxidant capacity.

The residue remaining after oil extraction from grape seed contain abundant procyanidins. An ultrasonic-assisted enzyme method was performed to achieve a high extraction efficiency of procyanidins when the optimal extraction conditions were 8 U/g of cellulase, ultrasound power of 200 W, ultrasonic temperature of 50 ℃, and ultrasonic reaction time of 40 min. The effects of free procyanidins on both radical scavenging activity and thermal stability at 40, 60, and 80 ℃ of the procyanidins-loaded liposomal systems prepared by the ultrasonic-assisted method were discussed. The presence of procyanidins at concentrations ranging from 0.02 to 0.10 mg/mL was observed to be effective at inhibiting lipid oxidation by 15.15 % to 69.70 % in a linoleic acid model system during reaction for 168 h, as measured using the ferric thiocyanate method. The procyanidins-loaded liposomal systems prepared by the ultrasonic-assisted method were characterized by measuring the mean particle size and encapsulation efficiency. Moreover, the holographic plots showed that the effect-response points of procyanidins combined with α-tocopherol in liposomes were lower than the addition line and 95 % confidence interval limits. At the same time, there were significant differences between the theoretical IC50add value and the experimental IC50mix value. The interaction index (γ) of all combinations was observed to be less than 1. These results indicated that there was a synergistic antioxidant effect between procyanidins combined with α-tocopherol, which will show promising prospects in practical applications. In addition, particle size differentiation and morphology agglomeration were observed at different time points of antioxidant activity determination (0, 48, 96 h).

Fractionation of oil palm fronds using ethanol-assisted deep eutectic solvent: Influence of ethanol concentration on enhancing enzymatic saccharification and lignin ß-O-4 content.

Numerous fractionation methods have been developed in recent years for separating components such as cellulose, hemicellulose, and lignin from lignocellulosic biomass wastes. Deep eutectic solvents (DES) have recently been widely investigated as captivating green solvents for biomass fractionation. However, most acidic-based deep eutectic solvent fractionation produces condensed lignin with low ß-O-4 content. Besides, most DESs exhibit high viscosity, which results in poor mass transfer properties. This study aimed to address the challenges above by incorporating ethanol into the deep eutectic solvent at various concentrations (10-50 wt%) to fractionate oil palm fronds at a mild condition, i.e., 80 °C, 1 atm. Cellulose residues fractionated with ethanol-assisted deep eutectic solvent showed a maximum glucose yield of 85.8% when 20 wt% of ethanol was incorporated in the deep eutectic solvent, significantly higher than that achieved by pure DES (44.8%). Lignin extracted with ethanol-assisted deep eutectic solvent is lighter in color and higher in ß-O-4 contents (up to 44 ß-O-4 per 100 aromatic units) than pure DES-extracted lignin. Overall, this study has demonstrated that incorporating ethanol into deep eutectic solvents could enhance the applicability of deep eutectic solvents in the complete valorization of lignocellulosic biomass. Highly enzymatic digestible cellulose-rich solid and ß-O-4-rich lignin attained from the fractionation could serve as sustainable precursors for the production of biofuels.

Toward well-defined colloidal particles: Efficient fractionation of lignin by a multi-solvent strategy.

Colloidal lignin particles (CLPs) have sparked various intriguing insights toward bio-polymeric materials and triggered many lignin-featured innovative applications. Here, we report a multi-solvent sequential fractionation methodology integrating green solvents of acetone, 1-butanol, and ethanol to fractionate industrial lignin for CLPs fabrication. Through a rationally designed fractionation strategy, multigrade lignin fractions with variable hydroxyl group contents, molecular weights, and high purity were obtained without altering their original chemical structures. CLPs with well-defined morphology, narrow size distribution, excellent thermal stability, and long-term colloidal stability can be obtained by rational selection of lignin fractions. We further elucidated that trace elements (S, N) were reorganized onto the near-surface area of CLPs from lignin fractions during the formation process in the form of -SO42- and -NH2. This work provides a sustainable and efficient strategy for refining industrial lignin into high-quality fractions and an in-depth insight into the CLPs formation process, holding great promise for enriching the existing libraries of colloidal materials.

Efficient Fast Fractionation of Biomass Using a Diol-Based Deep Eutectic Solvent for Facilitating Enzymatic Hydrolysis and Obtaining High-Quality Lignin.

Current DES pretreatment is often performed under relatively severe conditions with high temperature, long time, and high DES usage. This work studied a short-time diol DES (deep eutectic solvent) pretreatment under mild conditions to fractionate the bamboo, facilitate enzymatic hydrolysis, and obtain high-quality lignin. At an optimized condition of 130 °C for only 10 min, lignin and xylan removal reached 61.34 % and 84.15 %, with residual glucan showing a ~90 % enzymatic hydrolysis yield. Equally important, the dissolved lignin could be readily recovered with 97.51 % yield, exhibiting 96.65 % ß-O-4 preservation. The fractionation and lignin protection mechanisms were unveiled by XRD, FTIR, cellulose-DP, 2D HSQC NMR, 31P NMR and GPC analysis. This study highlighted that short-time fractionation of bamboo can be achieved by a diol-based DES which is an ideal strategy to upgrade the lignocellulose biomass for high enzymatic hydrolysis yields and high-quality lignin stream.

Ultrasound assisted wall-breaking extraction and primary structures, bioactivities, rheological properties of novel Exidia yadongensis polysaccharide.

New natural multifunctional polysaccharide and its innovatory extraction technology may be urgently needed for food industries. Our aims were to establish new extraction method and investigate the primary structures, bioactivities and rheological properties of novel E. yadongensis polysaccharide (EYP). Ultrasound assisted mechanical wall-breaking extraction (MAUE) was successfully established for the EYP extraction from a new E. yadongensis. Based on the MAUE with RSM, the polysaccharide yield of 17.92 ± 0.56 % with the optimal parameters of five extraction factors were obtained, and current MAUE was characterized by its high yield, low extraction temperature and short ultrasound time. After the isolation and purification, the EYP as a protein-bound polysaccharide was obtained. FT-IR and NMR analysis showed that the main backbone of the EYP comprised of (1 â†’ 4)-ß-D-glucopyranosyl and (1 â†’ 6)-ɑ-D-mannopyranosyl groups; EYP exhibited significant antioxidant, antibacterial, antitumor, antidiabetic activities, and good viscoelastic properties in low pH solutions (P < 0.05). The EYP may be used as a natural functional and cohesive agent in food industries.

Dry fractionation process operations in the production of protein concentrates: A review.

The market for plant proteins is expanding rapidly as the negative impacts of animal agriculture on the environment and resources become more evident. Plant proteins offer competitive advantages in production costs, energy requirements, and sustainability. Conventional plant-protein extraction is water and chemical-intensive, posing environmental concerns. Dry fractionation is an energy-efficient and environmentally friendly process for protein separation, preserving protein's native functionality. Cereals and pulses are excellent sources of plant proteins as they are widely grown worldwide. This paper provides a comprehensive review of the dry fractionation process utilized for different seeds to obtain protein-rich fractions with high purity and functionality. Pretreatments, such as dehulling and defatting, are known to enhance the protein separation efficiency. Factors, such as milling speed, mill classifier speed, feed rate, seed type, and hardness, were crucial for obtaining parent flour of desired particle size distribution during milling. The air classification or electrostatic separation settings are crucial in determining the quality of the separated protein. The cut point in air classification is targeted based on the starch granule size of the seed material. Optimization of these operations, applied to different pulses and seeds, led to higher yields of proteins with higher purity. Dual techniques, such as air classification and electrostatic separation, enhance protein purity. The yield of the protein concentrates can be increased by recycling the coarse fractions. Further research is necessary to improve the quality, purity, and yield of protein concentrates to enable more efficient use of plant proteins to meet global protein demands.

Reverse flow enhanced inertia pinched flow fractionation.

Particle separation plays a critical role in many biochemical analyses. In this article, we report a method of reverse flow enhanced inertia pinched flow fractionation (RF-iPFF) for particle separation. RF-iPFF separates particles by size based on the flow-induced inertial lift, and in the abruptly broadened segment, reverse flow is utilized to further enhance the separation distance between particles of different sizes. The separation performance can be significantly improved by reverse flow. Generally, compared with the case without reverse flow, this RF-iPFF technique can increase the particle throughput by about 10 times. To demonstrate the advantages of RF-iPFF, RF-iPFF was compared with traditional iPFF through a control experiment. RF-iPFF consistently outperformed iPFF across various conditions we studied. In addition, we use tumor cells spiked into the human whole blood to evaluate the separation performance of RF-iPFF.

A gas phase fractionation acquisition scheme integrating ion mobility for rapid diaPASEF library generation.

Data independent acquisition (DIA/SWATH) MS is a primary strategy in quantitative proteomics. diaPASEF is a recent adaptation using trapped ion mobility spectrometry (TIMS) to improve selectivity/sensitivity. Complex DIA spectra are typically analyzed with reference to spectral libraries. The best-established method for generating libraries uses offline fractionation to increase depth of coverage. More recently strategies for spectral library generation based on gas phase fractionation (GPF), where a representative sample is injected serially using narrow DIA windows that cover different mass ranges of the complete precursor space, have been introduced that performed comparably to deep offline fractionation-based libraries. We investigated whether an analogous GPF-based approach that accounts for the ion mobility (IM) dimension is useful for the analysis of diaPASEF data. We developed a rapid library generation approach using an IM-GPF acquisition scheme in the m/z versus 1/K0 space requiring seven injections of a representative sample and compared this with libraries generated by direct deconvolution-based analysis of diaPASEF data or by deep offline fractionation. We found that library generation by IM-GPF outperformed direct library generation from diaPASEF and had performance approaching that of the deep library. This establishes the IM-GPF scheme as a pragmatic approach to rapid library generation for analysis of diaPASEF data.