Reverse micellar extraction (RME) has emerged as a versatile and efficient tool for downstream processing (DSP) of various biomolecules, including structural proteins and enzymes, due to the substantial advantages over conventional DSP methods. However, the RME system is a complex dependency of several parameters that influences the overall selectivity and performance of the RME system, hence this justifies the need for optimization to obtain higher possible extraction results. For the last two decades, many experimental design strategies for screening and optimization of RME have been described in literature. The objective of this article is to review the use of different experimental designs and response surface methodologies that are currently used to screen and optimize the RME system for various types of biomolecules. Overall, this review provides the rationale for the selection of appropriate screening or optimization techniques for the parameters associated with both forward and backward extraction during the RME of biomolecules.
Matched MeSH terms: Chemical Fractionation/methods*
Biomolecules, specifically proteins, polysaccharides, and secondary metabolites are potential lead compounds due to their remarkable pharmacological properties. However, the complex molecular structure of the biomolecules makes their separation processes of great challenges. The conventional downstream processes require multistep protocols that are less efficient, high solvent consumption, expensive, time-consuming, and laborious. Hence, aqueous two-phase system (ATPS) is a reliable technique for the extraction and purification of biomolecules from a complex mixture. ATPS is an environmentally friendly, simple, cost effective, and easily scalable process. It requires a short processing time to separate biomolecules of industrial values simultaneously in a single process. Modifications have also been performed by introducing deep eutectic solvents, ionic liquids, carbohydrates, amino acids or copolymers to enhance the process efficiency with an increased yield, purity and bioactivity of recovered biomolecules. This review attempts to review the recent developed ATPSs and their efficiency to extract, isolate, and purify biomolecules such as proteins, polysaccharides, secondary metabolites and other biological substances. The review provides insights into the feasibility and reliability of ATPS for biomolecule recovery.
Matched MeSH terms: Chemical Fractionation/methods
This study optimized subcritical water extraction (SWE) conditions to maximize pectin yield from cocoa pod husk (CPH) and compared the characteristics of CPH pectin extracted through SWE with those of CPH pectin obtained through conventional extraction (CE) with citric acid. The Box-Behnken experimental design was employed to optimize SWE and examine the influence of process parameters, including temperature (100 °C-120 °C), extraction time (10-30 min), and solid:liquid ratio (SLR) (1:30-2:30 g/mL), on pectin yield. The maximum pectin yield of 6.58% was obtained under the optimal extraction conditions of 120 °C for 10 min with 1:15 g/mL SLR and closely corresponded with the predicted value of 7.29%. Compared with CE, SWE generated a higher yield and resulted in a higher degree of esterification, methoxyl content, and anhydrouronic acid value but a lower equivalent weight. The extracted pectin was pure, had low-methoxyl content, and similar melting and degradation temperatures.
Matched MeSH terms: Chemical Fractionation/methods
Oil palm empty fruit bunch (EFB) was pretreated by Formiline process to overcome biomass recalcitrance and obtain hemicellulosic syrup and lignin. Higher formic acid concentration led to more lignin removal but also higher degree of cellulose formylation. Cellulose digestibility could be well recovered after deformylation with a small amount of lime. After digested by enzyme loading of 15 FPU+10 CBU/g solid for 48 h, the polysaccharide conversion could be over 90%. Simultaneous saccharification and fermentation (SSF) results demonstrated that ethanol concentration reached 83.6 g/L with approximate 85% of theoretic yield when performed at an initial dry solid consistency of 20%. A mass balance showed that via Formiline pretreatment 0.166 kg of ethanol could be produced from 1 kg of dry EFB with co-production of 0.14 kg of high-purity lignin and 5.26 kg hemicellulosic syrup containing 2.8% xylose. Formiline pretreatment thus can be employed as an entry for biorefining of EFB.
Matched MeSH terms: Chemical Fractionation/methods
The aim of this study was to determine the best parameter for extracting phenolic-enriched kenaf (Hibiscus cannabinus L.) seeds by a pulsed ultrasonic-assisted extraction. The antioxidant activities of ultrasonic-assisted kenaf seed extracts (KSE) were determined by a 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging capacity assay, 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulphonic acid) (ABTS) radical scavenging assay, β -carotene bleaching inhibition assay, and ferric reducing antioxidant power (FRAP) assay. Total phenolic content (TPC) and total flavonoid content (TFC) evaluations were carried out to determine the phenolic and flavonoid contents in KSE. The KSE from the best extraction parameter was then subjected to high performance liquid chromatography (HPLC) to quantify the phenolic compounds. The optimised extraction condition employed 80% ethanol for 15 min, with the highest values determined for the DPPH, ABTS, and FRAP assay. KSE contained mainly tannic acid (2302.20 mg/100 g extract) and sinapic acid (1198.22 mg/100 g extract), which can be used as alternative antioxidants in the food industry.
Matched MeSH terms: Chemical Fractionation/methods*
A series of experiments were carried out to determine the best medium for the recovery of cobalt by means of an electrogenerative system. Use of the electrogenerative system with a chloride medium had shown promising performance with the highest free energy of -389.8 kJ mol(-1) compared to that with sulphate and nitrate media. Subsequently, the influence of catholyte concentrations on cobalt recovery using the electrogenerative process was carried out by varying the initial cobalt concentration and sodium chloride concentration. The results showed that almost 100% recovery was attained within 1-4 h of the recovery process. Influence of pH was investigated where the electrogenerative system performed best between pH 5.0 and 7.0. Maximum cell performance of 83% with 99% cobalt removal was obtained at 90 min when 100 mg L(-1) of Co(2+) in 0.5 M NaCl was taken as catholyte solution. The values of ΔH(o) and ΔS(o) of the process were evaluated as 33.41 kJ mol(-1) and 0.13 kJ mol(-1), respectively.
Matched MeSH terms: Chemical Fractionation/methods*
Hollow fibre liquid-phase microextraction with in situ derivatization using dansyl chloride has been successfully developed for the high-performance liquid chromatography-ultraviolet (HPLC-UV) determination of the biogenic amines (tryptamine, putrescine, cadaverine, histamine, tyramine, spermidine) in food samples. Parameters affecting the performance of the in situ derivatization process such as type of extraction solvent, temperature, extraction time, stirring speed and salt addition were studied and optimized. Under the optimized conditions (extraction solvent, dihexyl ether; acceptor phase, 0.1M HCl; extraction time, 30 min; extraction temperature, 26 degrees C; without addition of salt), enrichment factors varying from 47 to 456 were achieved. Good linearity of the analytes was obtained over a concentration range of 0.1-5 microg mL(-1) (with correlation coefficients of 0.9901-0.9974). The limits of detection and quantification based on a signal-to-noise ratio of 3-10, ranged from 0.0075 to 0.030 microg mL(-1) and 0.03 to 0.10 microg mL(-1), respectively. The relative standard deviations based on the peak areas for six replicate analysis of water spiked with 0.5 microg mL(-1) of each biogenic amine were lower than 7.5%. The method was successfully applied to shrimp sauce and tomato ketchup samples, offering an interesting alternative to liquid-liquid extraction and solid phase extraction for the analysis of biogenic amines in food samples.
Matched MeSH terms: Chemical Fractionation/methods*
There are relatively limited studies on the protein of honey samples mainly because of the low amount of protein in honey (0.1-0.5 %), the difficulty in extracting honey protein from the sugar-rich environment, and the hindrance of protein characterization by conventional approaches. Several protein extraction methods such as mechanical (ultrafiltration and ultracentrifugation) and chemical (precipitation) techniques have been applied to different types of honey samples. Most of these studies reported the quantity and molecular size of honey protein from gel electrophoresis, but were unable to identify and characterize the protein. This limitation might be due to the low capacity of analytical equipment in those days. Although different precipitants have also been used, not all them are compatible with mass spectrometric methods during downstream analysis. As a result, the sample preparation step is essential in order to confidently characterize the low and varied amount of honey protein. Nowadays, honey protein is getting attention from researchers because of its potential activity in pharmacological applications. Therefore, honey protein extraction and determination by mass spectrometry are critically reviewed in order to stimulate further honey protein research.
Matched MeSH terms: Chemical Fractionation/methods*
The effect of physicochemical treatment on pectin yield, degree of esterification, along with the kinetics and thermodynamics characteristics was investigated in the present study. Several extraction parameters were observed to have impacted the yield and degree of esterification significantly, and the best extraction condition was as follows: agitation rate of 250 rpm, temperature of 70 °C, extraction time of 120 min, pH 2, and liquid to solid ratio of 10 v/w which has resulted in 28.20% of pectin yield, with DE (degree of esterification) of 57.00%. A theoretical model which describes the extractability, dissolution and degradation rate of pectin to predict the maximal yield at the maximal time was established to study the extraction kinetics of pectin from HPP. The kinetic analysis from Panchev's model shows the extraction rate was found highest at LSR 10 with ymax 30.85%. The calculated activation energy for pectin dissolution and degradation was found to be 4.532 kJ/mol and 28.054 kJ/mol, respectively. The thermodynamic study has suggested that the process was endothermic, spontaneous and reversible. These results suggest that the physical and chemical treatment applied could be an efficient technique for the extraction of pectin from Hylocereus polyrhizus peels.
Matched MeSH terms: Chemical Fractionation/methods*
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.
Matched MeSH terms: Chemical Fractionation/methods*
Optimization process is an important aspect in the natural product extractions. Herein, an alternative approach is proposed for the optimization in extraction, namely, the Generalized Likelihood Uncertainty Estimation (GLUE). The approach combines the Latin hypercube sampling, the feasible range of independent variables, the Monte Carlo simulation, and the threshold criteria of response variables. The GLUE method is tested in three different techniques including the ultrasound, the microwave, and the supercritical CO2 assisted extractions utilizing the data from previously published reports. The study found that this method can: provide more information on the combined effects of the independent variables on the response variables in the dotty plots; deal with unlimited number of independent and response variables; consider combined multiple threshold criteria, which is subjective depending on the target of the investigation for response variables; and provide a range of values with their distribution for the optimization.
Matched MeSH terms: Chemical Fractionation/methods*
Extraction of DNA, RNA, and protein is the basic method used in molecular biology. These biomolecules can be isolated from any biological material for subsequent downstream processes, analytical, or preparative purposes. In the past, the process of extraction and purification of nucleic acids used to be complicated, time-consuming, labor-intensive, and limited in terms of overall throughput. Currently, there are many specialized methods that can be used to extract pure biomolecules, such as solution-based and column-based protocols. Manual method has certainly come a long way over time with various commercial offerings which included complete kits containing most of the components needed to isolate nucleic acid, but most of them require repeated centrifugation steps, followed by removal of supernatants depending on the type of specimen and additional mechanical treatment. Automated systems designed for medium-to-large laboratories have grown in demand over recent years. It is an alternative to labor-intensive manual methods. The technology should allow a high throughput of samples; the yield, purity, reproducibility, and scalability of the biomolecules as well as the speed, accuracy, and reliability of the assay should be maximal, while minimizing the risk of cross-contamination.
Matched MeSH terms: Chemical Fractionation/methods*
The present study aimed to investigate the effects of ultrasound-assisted extraction (UAE) condition on the yield, antioxidant activity and stability of the oil from papaya seed. The studied ultrasound variables were time, temperature, ultrasound power and solvent to sample ratio. The main goal was to optimise UAE condition providing the highest recovery of papaya seed oil with the most desirable antioxidant activity and stability. The interaction of ultrasound variables had the most and least significant effects on the antioxidant activity and stability, respectively. Ultrasound-assisted extraction provided a relatively high oil recovery (∼ 73%) from papaya seed. The strongest antioxidant activity was achieved by the extraction at the elevated temperature using low solvent to sample ratio. The optimum ultrasound extraction was set at the elevated temperature (62.5 °C) for 38.5 min at high ultrasound power (700 W) using medium solvent to sample ratio (∼ 7:1 v/w). The optimum point was practically validated.
Matched MeSH terms: Chemical Fractionation/methods*
The main goal of this study was to investigate the effect of extraction conditions on the enzymatic properties of thermoacidic amylase enzyme derived from dragon peel. The studied extraction variables were the buffer-to-sample (B/S) ratio (1:2 to 1:6, w/w), temperature (-18°C to 25°), mixing time (60 to 180 seconds), and the pH of the buffer (2.0 to 8.0). The results indicate that the enzyme extraction conditions exhibited the least significant (P < 0.05) effect on temperature stability. Conversely, the extraction conditions had the most significant (P < 0.05) effect on the specific activity and pH stability. The results also reveal that the main effect of the B/S ratio, followed by its interaction with the pH of the buffer, was significant (P < 0.05) among most of the response variables studied. The optimum extraction condition caused the amylase to achieve high enzyme activity (648.4 U), specific activity (14.2 U/mg), temperature stability (88.4%), pH stability (85.2%), surfactant agent stability (87.2%), and storage stability (90.3%).
Matched MeSH terms: Chemical Fractionation/methods*
The purification of thermo-acidic amylase enzyme from red pitaya (Hylocereus polyrhizus) peel for the first time was investigated using a novel aqueous two-phase system (ATPS) consisting of a thermo-separating copolymer and an organic solvent. The effectiveness of different parameters such as molecular weight of the thermo-separating ethylene oxide-propylene oxide (EOPO) copolymer and type and concentration of organic solvent on the partitioning behavior of amylase was investigated. In addition, the effects of phase components, volume ratio (VR), pH and crude load of purification factor and yield of amylase were evaluated to achieve the optimum partition conditions of the enzyme. In the novel ATPS method, the enzyme was satisfactorily partitioned into the polymer-rich top phase in the system composed of 30% (w/w) EOPO 2500 and 15% (w/w) 2-propanol, at a volume ratio of 1.94 and with a crude load scale of 25% (w/w) at pH 5.0. Recovery and recycling of components was also measured in each successive step of the ATPS process. The enzyme was successfully recovered by the method with a high purification factor of 14.3 and yield of 96.6% and copolymer was also recovered and recycled at a rate above 97%, making the method was more economical than the traditional ATPS method.
Matched MeSH terms: Chemical Fractionation/methods*
A rapid reversed-phase high performance liquid chromatographic method using a monolithic column for the determination of eight catechin monomers and caffeine was developed. Using a mobile phase of water:acetonitrile:methanol (83:6:11) at a flow rate of 1.4 mL min(-1), the catechins and caffeine were isocratically separated in about 7 min. The limits of detection and quantification were in the range of 0.11-0.29 and 0.33-0.87 mg L(-1), respectively. Satisfactory recoveries were obtained (94.2-105.2 ± 1.8%) for all samples when spiked at three concentrations (5, 40 and 70 mg L(-1)). In combination with microwave-assisted extraction (MAE), the method was applied to the determination of the catechins and caffeine in eleven tea samples (6 green, 3 black and 2 oolong teas). Relatively high levels of caffeine were found in black tea, but higher levels of the catechins, especially epigallocatechin gallate (EGCG) were found in green teas.
Matched MeSH terms: Chemical Fractionation/methods*
Different extraction conditions were applied to investigate the effect of temperature, extraction time and substrate-extractant ratio on pectin extraction from cocoa husks. Pectin was extracted from cocoa husks using water, citric acid at pH 2.5 or 4.0, or hydrochloric acid at pH 2.5 or 4.0. Temperature, extraction time and substrate-extractant ratio affected the yields, uronic acid contents, degrees of methylation (DM) and degrees of acetylation (DA) of the extracted pectins using the five extractants differently. The yields and uronic acid contents of the extracted pectins ranged from 3.38-7.62% to 31.19-65.20%, respectively. The DM and DA of the extracted pectins ranged from 7.17-57.86% to 1.01-3.48%, respectively. The highest yield of pectin (7.62%) was obtained using citric acid at pH 2.5 [1:25 (w/v)] at 95 °C for 3.0 h. The highest uronic acid content (65.20%) in the pectin was obtained using water [1:25 (w/v)] at 95 °C for 3.0 h.
Matched MeSH terms: Chemical Fractionation/methods*
A modeling technique based on absorbed microwave energy was proposed to model microwave-assisted extraction (MAE) of antioxidant compounds from cocoa (Theobroma cacao L.) leaves. By adapting suitable extraction model at the basis of microwave energy absorbed during extraction, the model can be developed to predict extraction profile of MAE at various microwave irradiation power (100-600 W) and solvent loading (100-300 ml). Verification with experimental data confirmed that the prediction was accurate in capturing the extraction profile of MAE (R-square value greater than 0.87). Besides, the predicted yields from the model showed good agreement with the experimental results with less than 10% deviation observed. Furthermore, suitable extraction times to ensure high extraction yield at various MAE conditions can be estimated based on absorbed microwave energy. The estimation is feasible as more than 85% of active compounds can be extracted when compared with the conventional extraction technique.
Matched MeSH terms: Chemical Fractionation/methods*
A new halogen-free and environmental-friendly method using water and ethanol is developed as an alternative for the recovery of polyhydroxyalkanoates (PHA) from recombinant Cupriavidus necator in comparison to the established chloroform extraction method. After optimisation, our results showed that the halogen-free method produced a PHA with 81% purity and 96% recovery yield, in comparison to the chloroform extraction system which resulted in a highly pure PHA with 95% yield. Although the purity of the PHA using the new method is lower, the molecular weight of the extracted PHA is not compromised. This new method can be further developed as an alternative and more environmental-friendly method for industrial application.
Matched MeSH terms: Chemical Fractionation/methods
A headspace single-drop microextraction (HS-SDME) procedure is optimized for the analysis of organochlorine and organophosphorous pesticide residues in food matrices, namely cucumbers and strawberries by gas chromatography with an electron capture detector. The parameters affecting the HS-SDME performance, such as selection of the extraction solvent, solvent drop volume, extraction time, temperature, stirring rate, and ionic strength, were studied and optimized. Extraction was achieved by exposing 1.5 microL toluene drop to the headspace of a 5 mL aqueous solution in a 15-mL vial and stirred at 800 rpm. The analytical parameters, such as linearity, correlation coefficients, precision, limits of detection (LOD), limits of quantification (LOQ), and recovery, were compared with those obtained from headspace solid-phase microextraction (HS-SPME) and solid-phase extraction. The mean recoveries for all three methods were all above 70% and below 104%. HS-SPME was the best method with the lowest LOD and LOQ values. Overall, the proposed HS-SDME method is acceptable in the analysis of pesticide residues in food matrices.
Matched MeSH terms: Chemical Fractionation/methods*