Alternative Extraction Method Research Articles

Ionic liquid–based aqueous biphasic system as an alternative tool for enhanced bacterial DNA extraction

BackgroundDeveloping an alternative and benign method for DNA extraction is imperative due to the high cost and potential harms associated with conventional techniques. Investigation of Ionic liquid (IL) as a solvent for DNA storage and stability revealed the ability of IL to assist DNA processes. IL–based aqueous biphasic system emerges as a comprehensive extraction platform capitalizing on the task–specificity of ILs and the wide applicability of ABS for biomolecule extractions. Therefore, it is beneficial to optimize an IL–based ABS specifically for DNA extraction, taking into account the fundamental interactions between the IL and DNA. ResultsThe primary objective was to design ABS consisting of Ammonium based ILs, and Potassium phosphate buffer as the salting–out agent for the partitioning of salmon sperm DNA. The analysis focused on optimizing biocompatible anions for the extraction. Moreover, the stability of the DNA in the IL rich phases was analysed to validate the method. The proposed process was then employed for extracting plasmid DNA from bacteria, demonstrating results comparable to those obtained with a commercially available kit. Further validation using agarose gel electrophoresis and transformation of the extracted DNA into E.coli were conducted, producing promising outcomes. Although there is room for improvement in terms of recovery of DNA and reusability of ABS, the described approach is comparable with the conventional one while being cost–effective, and showcases a noticeable and convincing link to eco–friendly processes. SignificanceThere is limited literature on IL–based ABS for DNA extraction, and the existing studies predominantly concentrate on systems derived from Cholinium ILs. However, their high hydrophilicity limits the choice of the second–phase forming component to polymers for the formation of ABS. Ammonium ILs efficiently form biphasic systems with various available salting–out agents, and biocompatible anions are introduced to mitigate the toxicity of the ILs.

Microalgae as a New Source of Oxylipins: A Comprehensive LC-MS-Based Analysis Using Conventional and Green Extraction Methods.

Microalgae are promising sources of essential lipids, including omega-3 and omega-6 polyunsaturated fatty acids (n-3 and n-6 PUFA) and novel lipid metabolites like oxylipins. However, limited data exist on the oxylipin profile, its characterization, and the potential impact of the extraction process on these metabolites in microalgae. Thus, our study aimed to investigate the fatty acid and oxylipin profile of four microalgal species of interest (Microchloropsis gaditana, Tisochrysis lutea, Phaeodactylum tricornutum, and Porphyridium cruentum) while also examining the impact of the extraction method, with a focus on developing a greener process using ultrasound-assisted extraction (UAE) and ethanol. The UAE method showed similar oxylipin profiles, generally yielding concentrations comparable to those of the conventional Folch method. In total, 68 oxylipins derived from n-3 and n-6 PUFA were detected, with the highest concentrations of n-3 oxylipins found in P. tricornutum and T. lutea and of n-6 oxylipins in P. cruentum. This study provides the most extensive oxylipin characterization of these microalgae species to date, offering insights into alternative extraction methods and opening new avenues for further investigation of the significance of oxylipins in microalgae.

Extraction method combining saponin and trehalose useful for analyzing fragile intermolecular association

Immunoprecipitation (IP) and co-immunoprecipitation (co-IP) are well-established methodologies to analyze protein expression and intermolecular interaction. Composition of extraction and washing buffer for preparing protein is important to accomplish experimental purpose. Various kinds of detergents are included in buffer to adjust extraction efficiency and washing effect. Among them, Triton X-100 (Tx-100), Nonidet P-40 (NP40), deoxycholic acid (DOC) and SDS are generally used according to experimental purpose and characteristic features of protein of interest. In some cases, general detergents disrupt intermolecular interaction and make it impossible to analyze molecular relation of protein of interest with its binding partners. In this study, we propose saponin, a natural detergent, is useful for co-immunoprecipitation when analyzing fragile intermolecular interactions, in which dystrophin and dystroglycan are used as a representative interaction. One of the most notable findings in this report is that intermolecular association between dystrophin and dystroglycan is maintained in saponin buffer whereas general detergents, such as Tx-100, NP40 and DOC, dissociate its binding. Furthermore, supplementation of trehalose, which has been shown to act as a molecular chaperone, facilitates efficient detection of dystrophin-dystroglycan macromolecular complex in co-IP assay. Importantly, the extraction buffer comprising 3 % saponin, 0.5 M trehalose and 0.05 % Tx-100 (we named it STX buffer) is applicable to co-IP for another molecular interaction, N-cadherin and β-catenin, indicating that this methodology can be used for versatile proteins of interest. Thus, STX buffer emerges as an alternative extraction method useful for analyzing fragile intermolecular associations and provides opportunity to identify complex interactomes, which may facilitate proteome-research and functional analysis of proteins of interest.

Improvement and prediction of the extraction parameters of lupeol and stigmasterol metabolites of Melia azedarach with response surface methodology

BackgroundMelia azedarach is known as a medicinal plant that has wide biological activities such as analgesic, antibacterial, and antifungal effects and is used to treat a wide range of diseases such as diarrhea, malaria, and various skin diseases. However, optimizing the extraction of valuable secondary metabolites of M. azedarach using alternative extraction methods has not been investigated. This research aims to develop an effective, fast, and environmentally friendly extraction method using Ultrasound-assisted extraction, methanol and temperature to optimize the extraction of two secondary metabolites, lupeol and stigmasterol, from young roots of M. azedarach using the response surface methodology.MethodsBox-behnken design was applied to optimize different factors (solvent, temperature, and ultrasonication time). The amounts of lupeol and stigmasterol in the root of M. azedarach were detected by the HPLC-DAD. The required time for the analysis of each sample by the HPLC-DAD system was considered to be 8 min.ResultsThe results indicated that the highest amount of lupeol (7.82 mg/g DW) and stigmasterol (6.76 mg/g DW) was obtained using 50% methanol at 45 °C and ultrasonication for 30 min, and 50% methanol in 35 °C, and ultrasonication for 30 min, respectively. Using the response surface methodology, the predicted conditions for lupeol and stigmasterol from root of M. azedarach were as follows; lupeol: 100% methanol, temperature 45 °C and ultrasonication time 40 min (14.540 mg/g DW) and stigmasterol 43.75% methanol, temperature 34.4 °C and ultrasonication time 25.3 min (5.832 mg/g DW).ConclusionsThe results showed that the amount of secondary metabolites lupeol and stigmasterol in the root of M. azedarach could be improved by optimizing the extraction process utilizing response surface methodology.

Research progress and latest application of ultrasonic treatment on protein extraction and modification

SummaryThe advancement of the modern food industry has prompted a shift away from traditional protein extraction methods due to their inherent limitations. As a result, there is a growing interest in exploring alternative methods for protein extraction and modification such as ultrasonic (US) treatment. This shift towards more sustainable and efficient protein extraction methods is a key research focus in obtaining safe proteins in an environmentally friendly manner. Furthermore, enhancing the functional properties of proteins through modification can expand their application. This paper reviews recent research progress and applications of US treatment on protein extraction and modification as well as the impact on protein structure and functional properties. In addition, the latest applications of ultrasound‐modified‐proteins, including nano‐proteins, 3D printing and interactions between proteins and other food components, are reviewed. This review may provide some inspiration for the potential application of US technologies in food fields.

Depollution of Polymeric Leather Waste by Applying the Most Current Methods of Chromium Extraction.

The leather industry is one of the most polluting industries in the world due to the large amounts of waste following raw hide processing but also due to the high content of chemical substances present in leather waste. The main problem with chromium-tanned leather solid waste is related to the storage, due to the ability of chromium to leach into soil or water, and also owing to the high ability of trivalent chromium to oxidize to its toxic form, hexavalent chromium. The purpose of this article is to present the most current methods of chromium extraction from solid tanned leather waste in order to obtain non-polluting leather, which can constitute secondary raw material in new industrial processes. The extraction methods identified in the present study are based on acid/basic/enzymatic hydrolysis and substitution with the help of organic chelators (organic acids and organic acid salts). In addition, this study includes a comparative analysis of the advantages and disadvantages of each identified extraction method. At the same time, this study also presents alternative chromium extraction methods based on the combination of conventional extraction methods and ultrasound-assisted extraction.

Optimizing ultrasound-assisted extraction of bioactive compounds from Canthium horridum blume leaves utilizing polyols: A study on skin-related activities

The focus on reducing organic solvent usage, owing to their negative environmental and health impacts, is driving a search for innovative green alternative solvents in academia and industry. Canthium horridum Blume (CH) exhibits many therapeutic activities, including antioxidant and anti-inflammatory efficacy. The objective of this study is to evaluate the optimal solvent concentration using a simplex-lattice design with an aqueous-polyols mixture and to optimize the parameters for extracting bioactive compounds and antioxidant activities from ultrasound-assisted extraction (UAE) of CH leaves through central composite design (CCD) in response surface methodology (RSM). This study examines the total phenolic content (TPC), antioxidant activities, comparison of different extraction conditions, identification of bioactive compounds, cell cytotoxicity, cellular antioxidant activity, and melanin content reduction efficacy of the extracts. According to the findings from the simplex-lattice model, the ideal solvent composition consisted of 32.57%w/w butylene glycol, 32.92%w/w glycerine, and 34.51%w/w water. Furthermore, based on the response model, optimal extraction conditions were identified as a 15-min extraction time and a solvent-to-sample ratio of 32.94:1. In comparison to alternative extraction methods, ultrasonic-assisted extraction using the aqueous-glycerine-butylene glycol (GB-UAE) extract resulted in notably elevated TPC and antioxidant responses (p < 0.05). Major antioxidant bioactive compounds included 4-(Butoxymethyl) phenol, 3-O-Caffeoyl-4-O-methylquinic acid, Quercetin 3-(2G-glucosylrutinoside), 2,4-Dihydroxybenzoic acid and other bioactive compounds. The GB-UAE extract revealed greater cell viability than UAE using ethanol (EtOH-UAE) extract in both cytotoxicity and cellular antioxidant assays at the same concentration. Additionally, it exhibited comparable melanin content reduction efficacy at a higher concentration compared to that of EtOH-UAE extract. The researcher anticipates that the current study will advance the utilization of an aqueous-polyols system for extracting bioactive compounds extending beyond CH leaves. Although the potential applications of CH leaves in cosmetics and pharmaceutical formulations have been identified, further comprehensive mechanistic and clinical studies are required to fully understand their effects.

Cranberry (Vaccinium macrocarpon Aiton) seeds: An exceptional source of tocotrienols

Cranberries (Vaccinium macrocarpon Aiton) are rich in beneficial phytochemicals and are valuable ingredients in the formulation of functional foods and pharmaceutical products, contributing to improved human health and well-being. Nevertheless, cranberry seed chemical composition is still scarcely investigated. Therefore, in the current study, tocochromanol (tocopherol (T) and tocotrienol (T3)) profile variance with respect to genotypes and harvest seasons were investigated. Tocochromanols were extracted by using a standard saponification protocol, as well as an environmentally friendlier and more rapid alternative extraction method, in the seeds of eight cranberry cultivars from two harvest seasons. γ-T3 constituted 87 % of the tocochromanols found in cranberry seeds, while the remaining 13 % were as follows: α-T3 (7 %), δ-T3 (3 %), and γ-T (3 %). Year, cultivar, and their interaction have a significant impact on the content of tocochromanols in cranberry seeds. Tocochromanol concentration varies significantly between cultivars, but is relatively stable over different harvest seasons. Concentration did not significantly differ between the standard saponification protocol and the rapid, environmentally friendlier method. Thereby, the present study shows that the simple and environmentally conscious/friendlier approach is feasible for the extraction of tocochromanols from cranberry seeds with comparable recovery, repeatability and reproducibility, to the saponification protocol.

SOH Estimation for Lithium-Ion Batteries Based on Health Indicators Extraction and MKRVR

Aiming at the problems of difficult feature extraction and low State of Health (SOH) prediction accuracy of lithium-ion batteries, this paper proposes a SOH estimation approach that involves extracting Health Indicators (HIs) and utilizing mixed kernel function relevance vector regression (MKRVR). In order to accurately and efficiently extract battery HIs, the paper first examines the limitations of extracting HIs based on Incremental Capacity (IC) curves and puts forward an alternative method of HIs extraction based on voltage-capacity (V-C) curves. Following correlation analysis of the HIs, the paper establishes the MKRVR model that combines a range of kernel functions to estimate SOH. To determine the hyper-parameters and weight coefficients of the MKRVR model, the paper integrates Differential Evolution (DE) and Levy flight into the Gray Wolf Optimizer (GWO) to enhance the population diversity and random search ability of GWO. Finally, the paper conducts experimental validation utilizing three distinct battery datasets. The results indicate that the proposed approach outperforms Support Vector Regression (SVR) and Relevance Vector Regression (RVR), with evaluation index values of MAE, MSE, and RMSE all below 1%. Moreover, the coefficients of determination exceed 0.95, which demonstrates the superiority of this approach over other methods.

Characterization of the Chemical Composition and Biological Activities of Bog Bilberry (Vaccinium uliginosum L.) Leaf Extracts Obtained via Various Extraction Techniques.

This investigation aimed to assess the chemical composition and biological activities of bog bilberry (Vaccinium uliginosum L.) leaves. Hydroethanolic extracts were obtained using four extraction techniques: one conventional (CE) and three alternative methods; ultrasound (UAE), microwave (MAE) and high-pressure (HPE) extractions. Spectrophotometric analysis was conducted to determine their chemical content, including the total phenolic content (TPC) and total flavonoid content (TFC). Furthermore, their antioxidative and antimicrobial properties were evaluated. HPLC (high performance liquid chromatography) analysis identified and quantified 17 phenolic compounds, with chlorogenic acid being the predominant compound, with the lowest level (37.36 ± 0.06 mg/g) for the bog bilberry leaf extract obtained by CE and the highest levels (e.g., HPE = 44.47 ± 0.08 mg/g) for the bog bilberry leaf extracts obtained by the alternative methods. Extracts obtained by HPE, UAE and MAE presented TPC values (135.75 ± 2.86 mg GAE/g; 130.52 ± 1.99 mg GAE/g; 119.23 ± 1.79 mg GAE/g) higher than those obtained by the CE method (113.07 ± 0.98 mg GAE/g). Regarding the TFC values, similar to TPC, the highest levels were registered in the extracts obtained by alternative methods (HPE = 43.16 ± 0.12 mg QE/g; MAE = 39.79 ± 0.41 mg QE/g and UAE = 33.89 ± 0.35 mg QE/g), while the CE extract registered the lowest level, 31.47 ± 0.28 mg QE/g. In the case of DPPH (1,1-diphenyl-2-picrylhydrazyl) antioxidant activity, the extracts from HPE, UAE and MAE exhibited the strongest radical scavenging capacities of 71.14%, 63.13% and 60.84%, respectively, whereas the CE extract registered only 55.37%. According to Microbiology Reader LogPhase 600 (BioTek), a common MIC value of 8.88 mg/mL was registered for all types of extracts against Staphylococcus aureus (Gram-positive bacteria) and Salmonella enterica (Gram-negative bacteria). Moreover, the alternative extraction methods (UAE, HPE) effectively inhibited the growth of Candida parapsilosis, in comparison to the lack of inhibition from the CE method. This study provides valuable insights into bog bilberry leaf extracts, reporting a comprehensive evaluation of their chemical composition and associated biological activities, with alternative extraction methods presenting greater potential for the recovery of phenolic compounds with increased biological activities than the conventional method.

Extraction of silica from sugarcane bagasse ash and its utilization in zeolite 4A synthesis for CO2 adsorption.

Sugarcane bagasse ash (SCBA) is a solid waste containing a high amount of silica (SiO2) and is suitable to utilize as a silica source for synthesizing zeolite NaA. SCBA is typically calcined at high temperatures before silica extraction. The method is not environmentally friendly because it consumes energy and produces CO2. This work demonstrates an alternative extraction method of SiO2 from SCBA by treating it with hydrochloric (HCl) and sodium hydroxide (NaOH) solution. The obtained mixture was separated by paper filter No. 1 (P) and a combination of paper filter and syringe filter (PS). The solution was neutralized by HCl solution, producing silica (SiO2-P and SiO2-PS) with a purity of 98 wt%. Both SiO2 samples and SCBA were utilized to synthesize zeolite NaA for CO2 adsorption. The CO2 adsorption capacities of NaA-P and NaA-PS were 4.30 and 4.10 mmol gadsorbent -1, in the same range as commercial NaA. The capacity is influenced by the total basicity of zeolite. The CO2 adsorption behavior of all samples correlates well with the Toth model. The CO2 adsorption kinetics agrees well with the pseudo-second-order kinetic model. Overall, this work shows the successful extraction of silica via using a direct NaOH solution, yielding high-purity silica sufficient for synthesizing zeolite NaA, a promising adsorbent of CO2.

Optimizing Neurodegenerative Disease Classification with Canny Segmentation and Voting Classifier: An Imbalanced Dataset Study

This study explores the efficacy of a Voting Classifier, combining Logistic Regression, Random Forest, and Gaussian Naive Bayes, in the classification of neurodegenerative diseases, focusing on Alzheimer's Disease (AD), Parkinson’s Disease (PD), and control groups. Utilizing a dataset pre-processed with Canny segmentation and Hu Moments feature extraction, the research aimed to address the challenges posed by imbalanced datasets in medical image classification. The classifier's performance was evaluated through a 5-fold cross-validation approach, with metrics including accuracy, precision, recall, and F1-Score. The results revealed a consistent recall rate of approximately 46% across all folds, indicating the model's effectiveness in identifying cases of neurodegenerative diseases. However, the precision and F1-Score were notably lower, averaging around 22% and 29%, respectively, underscoring the difficulties in achieving accurate classification in imbalanced datasets. The study contributes to the understanding of machine learning applications in medical diagnostics, specifically in the challenging context of neurodegenerative disease classification. It highlights the potential of using advanced image processing techniques combined with machine learning ensembles in enhancing diagnostic accuracy. However, it also draws attention to the inherent challenges in such approaches, particularly regarding precision in imbalanced datasets. Recommendations for future research include exploring data balancing techniques, alternative feature extraction methods, and different machine learning algorithms to improve the precision and overall performance. Additionally, applying the model to a broader and more diverse dataset could provide more generalizable and robust findings. This study is significant for researchers and practitioners in medical imaging and machine learning, offering insights into the complexities and potential of automated disease classification

Protein composition and nutritional aspects of pea protein fractions obtained by a modified isoelectric precipitation method using fermentation.

Pea albumins are promising for their nutritional, biological, and techno-functional properties. However, this fraction is usually discarded in the industry due to its low protein content compared to globulin fraction and the presence of some anti-nutritional compounds. In the present study, we used an alternative method of pea protein extraction based on alkaline solubilization/isoelectric precipitation in which the reduction of pH was achieved by lactic acid fermentation using specific starters instead of mineral acids. Hence, the main objective of this study was to examine the protein profile and the content of anti-nutritional and nutritional active compounds in pea albumin-rich fractions obtained by the isoelectric extraction method without (control) or with fermentation with different lactic acid bacteria (Streptococcus thermophilus, Lactiplantibacillus plantarum, and their co-culture). Different pea cultivars (Cartouche, Ascension, and Assas) were used here for their differences in protein profile. The results revealed a higher total nitrogen content in albumin-rich fraction for fermented samples and, in particular, for co-culture. The majority of total nitrogen was determined as non-protein (~50%), suggesting the degradation of proteins by LAB to small peptides and amino acids, which were solubilized in the soluble fraction (albumin) as confirmed by size exclusion chromatography (SEC-HPLC) analysis. Moreover, the higher antioxidant activity of fermented albumin samples was attributed to the production of small peptides during extraction. Lactic acid fermentation also resulted in a significant reduction of trypsin inhibitor activity, α-galactoside, and phytic acid content of this fraction compared to control.

Gelatin extraction through acetic acid using tannery waste as raw material

The present study proposes an alternative gelatin extraction method from waste materials generated by tanneries, including trimmings, offcuts, and leather scraps. These industrial byproducts yield substantial monthly volumes that are managed by an environmental entity and disposed of in a sanitary landfill. To harness the potential of these waste materials, experimental techniques were employed, involving protein analysis, assessment of response to varying concentrations of acetic acid, and the application of various treatments to individual samples. The objective was to maximize yield and minimize ecological impact. The most effective treatment was found to be T16 (1 mole of acid per liter) for trimmings, T12 (0.8 moles of acid per liter) for offcuts, and T8 (0.7 moles of acid per liter) for leather scraps. Approximately 90% of the obtained protein values correspond to collagen, with a more pronounced presence in the trimmings samples.

The optimization of oil extraction by surfactant-assisted aqueous extraction process of rice bran (Oryza sativa L.) using Box-Behnken design

Surfactant-assisted aqueous extraction process (SAAEP) is used as an environmentally friendly alternative method for oil extraction. Rice (Oryza sativa L.) is one of the most widely produced and consumed foods, and it produces bran as a by-product. Therefore, this study aims to determine the optimum conditions for SAAEP from rice bran. The response surface methodology (RSM) technique was used to determine the optimum conditions. Furthermore, the Box-Behnken design (BBD) was selected as the study design with three factors and three levels. The independent variables used were surfactant and oil ratio (SOR) (3, 3.5, and 4), ultrasonication amplitude (60%, 70%, and 80%), and ultrasonication contact time (15, 20, and 25 mins. Meanwhile, the dependent variable used was oil content. The extraction was carried out based on SAEEP. Also, the rice bran used was the IR 32 SR variety which passed the stabilization stage with a moisture content of 4.25%, extracted oil of 20.64%, and 2.04 meqO2/kg FFA levels. The optimal point was predicted to produce an extracted oil of 19.326%. Furthermore, the prediction results were evaluated by experimental extraction at the optimal level. The results showed an extracted oil of 18.52±0.04%, which is a difference of about 0.81% from the predicted values. It was concluded that the rice bran oil content can be successfully optimized using the RSM technique. Therefore, the optimum conditions for extracting rice bran oil are within the specified range of factors

Effect of foliar application of cow dung extract on growth and yield of waterleaf (Talinum triangulare Jacq.) in an ultisol

The research was conducted at the Umuowa Ibu 1 community via National Horticultural Research Institute, Mbato, Okigwe Imo State, Nigeria, where waterleaf production and consumption is predominant. The objective of this study was to evaluate the effect of foliar application of cow dung extract on the performance of water leaf. The experiment was laid out in a randomized complete block design. Treatments were four levels of cow dung extract (3.75, 7.50, 11.25, and 15.00 L/ha) and three controls (5t/ha cow dung, 200kg/ha NPK fertilizer, and no soil amendment). Growth and yield data were subjected to analysis of variance. Significant means were compared with the least significant difference at 5%. The result showed a significant increase in waterleaf growth and yield parameters, with an increase in extract application. The foliar yield obtained from the 11.25 L/ha treatment (13.92 and 14.50 t/ha) and 15L/ha (14.46 and 14.75 t/ha) were not significantly different. The lowest foliar yields, 4.31 and 4.26 t/ha, were recorded in no soil amendment. Treatment of 15 L/ha had a significant foliar yield of 3-70% and 1-71% in 2021 and 2022. The result showed liquid manure had lower nutrient content than solid cow dung. In order to reduce frequent application due to its low nutrient content, this study suggests an alternative method of extraction instead of water. Therefore, applying 11.25L/ha to waterleaf was recommended in the interim.

Comparison of 3 DNA extraction methods for extracting DNA from an adult Culex quinquefasciatus (Diptera: Culicidae).

The aim of this study was to compare 3 DNA extraction methods: the PureLink Genomic DNA kit, DNAzol Direct reagent, and a microwave-based method, for extracting DNA from an adult Culex quinquefasciatus by focusing on the quantity and purity of DNA, cost, and time required. Ten mosquitoes were individually used for DNA extraction by each method. Based on the results obtained, DNA was extracted from each method using specific primers, resulting in a polymerase chain reaction (PCR) product with a length of 274 bp. The DNA quantity extracted using the DNAzol Direct (179.08 ± 3.77 ng/µl) differs significantly from that of the commercial kit (115.98 ± 4.57 ng/µl) and a microwave-based method (119.26 ± 3.06 ng/µl). The absorbance ratio of DNA extracted using the PureLink Genomic DNA kit, the DNAzol Direct, and the microwave-based methods was 1.92 ± 0.02, 1.79 ± 0.01, and 1.87 ± 0.01, respectively. Among the 3 methods evaluated, the microwave-based method is simpler, less expensive, and more time efficient. This is the first evaluation of the microwave-based method for extracting DNA from an adult mosquito. This study provides a useful guide for alternative DNA extraction methods for PCR-based assays, especially in low-resource settings.

Alternative methods for RuBisCO extraction from sugar beet waste: A comparative approach of ultrasound and high voltage electrical discharge

Ultrasound (US) and high voltage electric discharge (HVED) with water as a green solvent represent promising novel non-thermal techniques for protein extraction from sugar beet (Beta vulgaris subsp. vulgaris var. altissima) leaves. Compared to HVED, US proved to be a better alternative method for total soluble protein extraction with the aim of obtaining high yield of ribulose-1,5-bisphosphate carboxylase-oxygenase enzyme (RuBisCO). Regardless of the solvent temperature, the highest protein yields were observed at 100% amplitude and 9 min treatment time (84.60 ± 3.98 mg/gd.m. with cold and 96.75 ± 4.30 mg/gd.m. with room temperature deionized water). US treatments at 75% amplitude and 9 min treatment time showed the highest abundance of RuBisCO obtained by immunoblotting assay. The highest protein yields recorded among HVED-treated samples were observed at a voltage of 20 kV and a treatment time of 3 min, disregarding the used gas (33.33 ± 1.06 mg/gd.m. with argon and 34.89 ± 1.59 mg/gd.m. with nitrogen as injected gas), while the highest abundance of the RuBisCO among HVED-treated samples was noticed at 25 kV voltage and 3 min treatment time. By optimizing the US and HVED parameters, it is possible to affect the solubility and improve the isolation of RuBisCO, which could then be purified and implemented into new or already existing functional products.

Optimization of pH for extracting sinapates from mustard varieties using green technology

Mustard seeds contain higher levels of phenolic compounds, especially sinapine, compared to many other oilseeds. Due to their rich bioactive composition, mustard seeds have considerable potential in food and pharmaceutical industries as antimicrobials, antioxidants, and chemoprotective agents. The yields of phenolic compounds, particularly the formation of canolol, could be substantially enhanced by modifying the pre-treatment conditions and extraction techniques. To achieve this, there is a need for innovative green technology methods to minimize environmental damage. This novel approach involved pre-heating by sautéing, and adjusting the pH of the extract to acid, alkaline or neutral conditions using safe household ingredients, such as vinegar and sodium bicarbonate. The treated whole or crushed seeds were then extracted by pressurized heating at 115 °C at 10.2 psi, and the extractability of the phenolic compounds was assessed.Statistical analysis of selected mustard varieties showed an interesting trend in which pH affected the mustard endogenous phenolic compound profile, in which neutral pH proved to be superior to acid and alkaline treatment Among the selected varieties, both yellow and black mustard had competitive amounts of sinapine, sinapic acid, and canolol. Sinapic acid and canolol were predominant in crushed black mustard seeds (2.71± 0.0.8 and 2.61± 0.35 mg/g DW, respectively) (p < 0.05). Meanwhile, crushed yellow mustard prevailed in sinapine concentration (385 ± 49.47 mg/g DW).Overall, on the basis of electrostatic interactions between the solvent and the extractants, using water with the combination of preheat treatment and pressure was an alternative green extraction method to optimize the extraction of phenolic compounds from Oriental, black, and yellow mustard seeds.

Evaluation of DNA extraction methods for the detection of Shiga toxin producing Escherichia coli in food by polymerase chain reaction

Reported food-borne outbreaks of Shiga toxin producing Escherichia coli (STEC) have involved a very diverse range of foods. Contemporary analytical methods for the detection of STEC in foods typically include PCR screening of enrichment media. However, PCR inhibitors present in food enrichments can produce false negative results when screening for DNA sequences associated with the pathogen. To avoid false negative results in enrichment screening, it is advantageous to have DNA extraction methods that are effective at removing PCR inhibitors from a wide range of foods. The standard Canadian STEC method MFLP-52 uses Bio-Rad Instagene Matrix for DNA extraction. In this study, three DNA extraction protocols using commercial kits (Instagene Matrix with Beckman Coulter Ampure XP Beads; Qiagen Gentra Puregene Yeast/Bact. Kit; Qiagen DNeasy Blood & Tissue) were assessed as alternative DNA extraction methods for the detection of the Shiga toxin gene by PCR in enrichments from sixteen different foods inoculated with STEC O157. The inoculated foods were bean sprouts, blackberries, blue cheese, cilantro, cocoa powder, coleslaw, cream of mushroom dried soup mix, cream of vegetable dried soup mix, flaxseed, guacamole, peanut butter, soft cheese, soy butter, spinach, walnut, and wheat flour. Two of the protocols, Instagene Matrix with Ampure XP Beads, and Gentra Puregene Yeast/Bact, produced no false-negative or false positive results in the analysis of triplicate enrichment samples from sixteen inoculated foods.