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Fulltext Microwave-Assisted Brine Extraction for Enhancement of the Quantity and Quality of Lipid Production from Microalgae Nannochloropsis sp

Zghaibi N, Omar R, Kamal SMM, Biak DRA, Harun R

Molecules, 2019 Oct 04;24(19).
PMID: 31590304 DOI: 10.3390/molecules24193581

Toward attaining a sustainability and eco-friendly process, a green and low-cost solvent-brine (NaCl solution) is proposed, as microwave-assisted extraction (MAE) technique solvent to extract lipids from microalgae Nannochloropsis sp. The effect of NaCl concentration on the quantity and quality of the extracted lipid was assessed, while MAE parameters were optimized using response surface methodology (RSM). The content of fatty acid methyl esters (FAMEs) in the lipid was analyzed by using a gas chromatography-flame ionization detector (GC/FID). The highest lipid yield (16.1%) was obtained using 10% (w/v) brine at optimum extraction parameters of 5% (w/v) solid loading, 100 °C, and 30 min. The lipid extraction yield via optimized MAE-brine technique was thrice better than that Soxhlet extraction did and only 2% less than Bligh and Dyer (B&D) lipid extraction, which utilized harmful solvents. The proposed MAE-brine technique offered better quality lipids containing the highest amount of polyunsaturated fatty acids (PUFA) (44.5%) and omega-3 fatty acids (FAs) (43%). Hence, the MAE-brine solvent technique appears to be a promising extraction method for cheaper, greener, and faster extraction of a high-quality lipid for specialty food applications.

Matched MeSH terms: Chemical Fractionation/methods*
Generation, Fractionation, and Characterization of Iron-Chelating Protein Hydrolysate from Palm Kernel Cake Proteins

Zarei M, Ghanbari R, Tajabadi N, Abdul-Hamid A, Bakar FA, Saari N

J Food Sci, 2016 Feb;81(2):C341-7.
PMID: 26720491 DOI: 10.1111/1750-3841.13200

Palm kernel cake protein was hydrolyzed with different proteases namely papain, bromelain, subtilisin, flavourzyme, trypsin, chymotrypsin, and pepsin to generate different protein hydrolysates. Peptide content and iron-chelating activity of each hydrolysate were evaluated using O-phthaldialdehyde-based spectrophotometric method and ferrozine-based colorimetric assay, respectively. The results revealed a positive correlation between peptide contents and iron-chelating activities of the protein hydrolysates. Protein hydrolysate generated by papain exhibited the highest peptide content of 10.5 mM and highest iron-chelating activity of 64.8% compared with the other hydrolysates. Profiling of the papain-generated hydrolysate by reverse phase high performance liquid chromatography fractionation indicated a direct association between peptide content and iron-chelating activity in most of the fractions. Further fractionation using isoelectric focusing also revealed that protein hydrolysate with basic and neutral isoelectric point (pI) had the highest iron-chelating activity, although a few fractions in the acidic range also exhibited good metal chelating potential. After identification and synthesis of papain-generated peptides, GGIF and YLLLK showed among the highest iron-chelating activities of 56% and 53%, whereas their IC50 were 1.4 and 0.2 μM, respectively.

Matched MeSH terms: Chemical Fractionation
High methoxyl pectin extracts from Hylocereus polyrhizus's peels: Extraction kinetics and thermodynamic studies

Zaid RM, Mishra P, Tabassum S, Wahid ZA, Sakinah AMM

Int J Biol Macromol, 2019 Dec 01;141:1147-1157.
PMID: 31494156 DOI: 10.1016/j.ijbiomac.2019.09.017

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*
Isolation of Wheat Genomic DNA for Gene Mapping and Cloning

Yu G, Hatta A, Periyannan S, Lagudah E, Wulff BBH

Methods Mol Biol, 2017;1659:207-213.
PMID: 28856653 DOI: 10.1007/978-1-4939-7249-4_18

DNA is widely used in plant genetic and molecular biology studies. In this chapter, we describe how to extract DNA from wheat tissues. The tissue samples are ground to disrupt the cell wall. Then cetyltrimethylammonium bromide (CTAB) or sodium dodecyl sulfate (SDS) is used to disrupt the cell and nuclear membranes to release the DNA into solution. A reducing agent, β-mercaptoethanol, is added to break the disulfide bonds between the cysteine residues and to help remove the tanins and polyphenols. A high concentration of salt is employed to remove polysaccharides. Ethylenediaminetetraacetic acid (EDTA) stops DNase activity by chelating the magnesium ions. The nucleic acid solution is extracted with chloroform-isoamyl alcohol (24:1) or 6 M ammonium acetate. The DNA in aqueous phase is precipated with ethanol or isopropanol, which makes DNA less hydrophilic in the presence of sodium ions (Na+).

Matched MeSH terms: Chemical Fractionation/methods*
Fulltext Low dose triterpene-quinone fraction from Ardisia crispa root precludes chemical-induced mouse skin tumor promotion

Yeong LT, Abdul Hamid R, Saiful Yazan L, Khaza'ai H, Mohtarrudin N

BMC Complement Altern Med, 2015;15(1):431.
PMID: 26638207 DOI: 10.1186/s12906-015-0954-3

Drastic increment of skin cancer incidence has driven natural product-based chemoprevention as a promising approach in anticancer drug development. Apart from its traditional usages against various ailments, Ardisia crispa (Family: Myrsinaceae) specifically its triterpene-quinone fraction (TQF) which was isolated from the root hexane extract (ACRH) was recently reported to exert antitumor promoting activity in vitro. This study aimed at determining chemopreventive effect of TQF against chemically-induced mouse skin tumorigenesis as well as elucidating its possible pathway(s).

Matched MeSH terms: Chemical Fractionation
Vasorelaxant Action of the Chloroform Fraction of Orthosiphon stamineus via NO/cGMP Pathway, Potassium and Calcium Channels

Yam MF, Tan CS, Ahmad M, Ruan S

Am J Chin Med, 2016;44(7):1413-1439.
PMID: 27785939

Orthosiphon stamineus Benth. (Lamiaceae) is an important plant in traditional folk medicine that is used to treat hypertension and kidney stones. In humans, this plant has been tested as an addition regiment for antihypertensive treatment. Among the treatments for hypertension, O. stamineus had been to have diuretic and vasorelaxant effects in animal models. There is still very little information regarding the vasorelaxant effect of O. stamineus. Therefore, the present study was designed to investigate the vasorelaxant activity and mechanism of action of the fractions of O. stamineus. The vasorelaxant activity and the underlying mechanisms of the chloroform fraction of the 50% methanolic extract of O. stamineus (CF) was evaluated on thoracic aortic rings isolated from Sprague Dawley rats. CF caused relaxation of the aortic ring pre-contracted with phenylephrine in the presence and absence of endothelium, and pre-contracted with potassium chloride in endothelium-intact aortic ring. In the presence of endothelium, both indomethacin (a nonselective cyclooxygenase inhibitor) and [Formula: see text]-[1,2,4]Oxadiazolo[4,3-[Formula: see text]]quinoxalin-1-one (ODQ, selective soluble guanylate cyclase inhibitor) had a small effect on the vasorelaxation response. On the other hand, in the presence of Nω-nitro-L-arginine methyl ester (L-NAME, nitric oxide synthase inhibitor), methylene blue (cyclic guanosine monophosphate lowering agent), tetraethylammonium ([Formula: see text], nonselective calcium activator [Formula: see text] channel blocker), 4-aminopyridine (4-AP, voltage-dependent [Formula: see text] channel blocker), barium chloride ([Formula: see text], inwardly rectifying [Formula: see text] channel blocker), glibenclamide (nonspecific ATP-sensitive [Formula: see text] channel blocker), atropine (muscarinic receptor blocker) and propranolol (β-adrenergic receptor blocker), the vasorelaxant effect significantly reduced the relaxation stimulated by CF. CF was also found to be active in reducing [Formula: see text] release from the sarcoplasmic reticulum and blocking calcium channels.

Matched MeSH terms: Chemical Fractionation
Fulltext Binary solvent extraction system and extraction time effects on phenolic antioxidants from kenaf seeds (Hibiscus cannabinus L.) extracted by a pulsed ultrasonic-assisted extraction

Wong YH, Lau HW, Tan CP, Long K, Nyam KL

ScientificWorldJournal, 2014;2014:789346.
PMID: 24592184 DOI: 10.1155/2014/789346

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*
Inhibition of Staphylococcus aureus by crude and fractionated extract from lactic acid bacteria

Wong CB, Khoo BY, Sasidharan S, Piyawattanametha W, Kim SH, Khemthongcharoen N, et al.

Benef Microbes, 2015 Mar;6(1):129-39.
PMID: 25213027 DOI: 10.3920/BM2014.0021

Increasing levels of antibiotic resistance by Staphylococcus aureus have posed a need to search for non-antibiotic alternatives. This study aimed to assess the inhibitory effects of crude and fractionated cell-free supernatants (CFS) of locally isolated lactic acid bacteria (LAB) against a clinical strain of S. aureus. A total of 42 LAB strains were isolated and identified from fresh vegetables, fresh fruits and fermented products prior to evaluation of inhibitory activities. CFS of LAB strains exhibiting a stronger inhibitive effect against S. aureus were fractionated into crude protein, polysaccharide and lipid fractions. Crude protein fractions showed greater inhibition against S. aureus compared to polysaccharide and lipid fractions, with a more prevalent effect from Lactobacillus plantarum 8513 and L. plantarum BT8513. Crude protein, polysaccharide and lipid fractions were also characterised with glycine, mannose and oleic acid being detected as the major component of each fraction, respectively. Scanning electron microscopy revealed roughed and wrinkled membrane morphology of S. aureus upon treatment with crude protein fractions of LAB, suggesting an inhibitory effect via the destruction of cellular membrane. This research illustrated the potential application of fractionated extracts from LAB to inhibit S. aureus for use in the food and health industry.

Matched MeSH terms: Chemical Fractionation
Optimisation of the batch reactor for the removal of cobalt ions from chloride media

Wen Min S, Hasnat MA, Rahim AA, Mohamed N

Chemosphere, 2013 Jan;90(2):674-82.
PMID: 23063484 DOI: 10.1016/j.chemosphere.2012.09.048

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*
Novel IEF Peptide Fractionation Method Reveals a Detailed Profile of N-Terminal Acetylation in Chemotherapy-Responsive and -Resistant Ovarian Cancer Cells

Weiland F, Arentz G, Klingler-Hoffmann M, McCarthy P, Lokman NA, Kaur G, et al.

J Proteome Res, 2016 11 04;15(11):4073-4081.
PMID: 27569743

Although acetylation is regarded as a common protein modification, a detailed proteome-wide profile of this post-translational modification may reveal important biological insight regarding differential acetylation of individual proteins. Here we optimized a novel peptide IEF fractionation method for use prior to LC-MS/MS analysis to obtain a more in depth coverage of N-terminally acetylated proteins from complex samples. Application of the method to the analysis of the serous ovarian cancer cell line OVCAR-5 identified 344 N-terminally acetylated proteins, 12 of which are previously unreported. The protein peptidyl-prolyl cis-trans isomerase A (PPIA) was detected in both the N-terminally acetylated and unmodified forms and was further analyzed by data-independent acquisition in carboplatin-responsive parental OVCAR-5 cells and carboplatin-resistant OVCAR-5 cells. This revealed a higher ratio of unacetylated to acetylated N-terminal PPIA in the parental compared with the carboplatin-resistant OVCAR-5 cells and a 4.1-fold increase in PPIA abundance overall in the parental cells relative to carboplatin-resistant OVCAR-5 cells (P = 0.015). In summary, the novel IEF peptide fractionation method presented here is robust, reproducible, and can be applied to the profiling of N-terminally acetylated proteins. All mass spectrometry data is available as a ProteomeXchange repository (PXD003547).

Matched MeSH terms: Chemical Fractionation/methods
Advanced analytical techniques for the extraction and characterization of plant-derived essential oils by gas chromatography with mass spectrometry

Waseem R, Low KH

J Sep Sci, 2015 Feb;38(3):483-501.
PMID: 25403494 DOI: 10.1002/jssc.201400724

In recent years, essential oils have received a growing interest because of the positive health effects of their novel characteristics such as antibacterial, antifungal, and antioxidant activities. For the extraction of plant-derived essential oils, there is the need of advanced analytical techniques and innovative methodologies. An exhaustive study of hydrodistillation, supercritical fluid extraction, ultrasound- and microwave-assisted extraction, solid-phase microextraction, pressurized liquid extraction, pressurized hot water extraction, liquid-liquid extraction, liquid-phase microextraction, matrix solid-phase dispersion, and gas chromatography (one- and two-dimensional) hyphenated with mass spectrometry for the extraction through various plant species and analysis of essential oils has been provided in this review. Essential oils are composed of mainly terpenes and terpenoids with low-molecular-weight aromatic and aliphatic constituents that are particularly important for public health.

Matched MeSH terms: Chemical Fractionation
Stereoselective method development and validation for determination of concentrations of amphetamine-type stimulants and metabolites in human urine using a simultaneous extraction-chiral derivatization approach

Wan Raihana WA, Gan SH, Tan SC

J Chromatogr B Analyt Technol Biomed Life Sci, 2011 Jan 1;879(1):8-16.
PMID: 21147046 DOI: 10.1016/j.jchromb.2010.10.037

Amphetamine-type stimulants (ATS) are a group of chiral amine drugs which are commonly abused for their sympathomimetic and stimulant properties. ATS are extensively metabolised by hepatic cytochrome P450 enzymes. As metabolism of ATS has been shown to be highly stereospecific, stereoselective analytical methods are essential for the quantitative determination of ATS concentrations for both in vivo and in vitro studies of ATS metabolism. This paper describes a new stereoselective method for the simultaneous determination of amphetamine (AM), methamphetamine (MA), 3,4-methylenedioxymethamphetamine (MDMA), 3,4-methylenedioxyamphetamine (MDA), 4-hydroxy-3-methoxymethamphetamine (HMMA), 4-hydroxy-3-methoxyamphetamine (HMA), 3,4-hydroxymethamphetamine (HHMA) and 3,4-hydroxyamphetamine (HHA) in human urine samples validated according to the United States Food and Drug Administration guidelines. In this method, analytes are simultaneously extracted and derivatized with R-(-)-α-methoxy-α-(trifluoromethyl)phenylacetyl chloride (R-MTPCl) as the chiral derivatization reagent. Following this, the analytes were subjected to a second derivatization with N-methyl-N-trimethylsilyltrifluoroacetamide (MSTFA) which targets the hydroxyl groups present in HMMA, HMA, HHMA and HHA. The derivatized analytes were separated and quantified using gas chromatography-mass spectrometry (GC-MS). The method was evaluated according to the established guidelines for specificity, linearity, precision, accuracy, recovery and stability using a five-day protocol. Intra-day precision ranged from 0.89 to 11.23% RSD whereas inter-day precision was between 1.03 and 12.95% RSD. Accuracy values for the analytes ranged from -5.29% to 13.75%. Limits of quantitation were 10 μg/L for AM, MA, MDMA, HMA and HMMA and 2μg/L for MDA, HMA and HHA. Recoveries and stability values were also within accepted values. The method was applied to authentic ATS-positive samples.

Matched MeSH terms: Chemical Fractionation/methods*
Effect of functional groups in acid constituent of deep eutectic solvent for extraction of reactive lignin

Tan YT, Ngoh GC, Chua ASM

Bioresour Technol, 2019 Jun;281:359-366.
PMID: 30831515 DOI: 10.1016/j.biortech.2019.02.010

In this study, acidic deep eutectic solvents (DES) synthesized from various organic carboxylic acid hydrogen bond donors were applied to lignocellulosic oil palm empty fruit bunch (EFB) pretreatment. The influence of functional group types on acid and their molar ratios with hydrogen bond acceptor on lignin extraction were evaluated. The result showed presence of hydroxyl group and short alkyl chain enhanced biomass fractionation and lignin extraction. Choline chloride:lactic acid (CC-LA) with the ratio of 1:15 and choline chloride:formic acid (CC-FA) with 1:2 ratio extracted more than 60 wt% of lignin. CC-LA DES-extracted lignin (DEEL) exhibited comparable reactivity with technical and commercial lignin based on its phenolic hydroxyl content (3.33-3.72 mmol/glignin). Also, the DES-pretreated EFB comprised of enriched glucan content after biopolymer fractionation. Both DES-pretreated EFB and DEEL can be potential feedstock for subsequent conversion processes. This study presented DES as an effective and facile pretreatment method for reactive lignin extraction.

Matched MeSH terms: Chemical Fractionation
Optimization of protein extraction for proteomic analyses of fresh and frozen "Musang King" durian pulps

Tan XY, Misran A, Daim LDJ, Lau BYC

Food Chem, 2021 May 01;343:128471.
PMID: 33143964 DOI: 10.1016/j.foodchem.2020.128471

Four different methods were evaluated to extract proteins from "Musang King" durian pulps and subsequently proteins with different abundance between fresh and long term frozen storage were identified using two-dimensional polyacrylamide gel electrophoresis coupled with matrix-assisted laser desorption/ionization time-of-flight mass spectrometer analyses. The acetone-phenol method was found to produce good protein yields and gave the highest gel resolution and reproducibility. Differential protein analyses of the durian pulp revealed that 15 proteins were down-regulated and three other proteins were up-regulated after a year of frozen storage. Isoflavone reductase-like protein, S-adenosyl methionine synthase, and cysteine synthase isoform were up-regulated during frozen storage. The down-regulation of proteins in frozen durian pulps indicated that frozen storage has affected proteins in many ways, especially in their functions related to carbohydrate and energy metabolisms, cellular components, and transport processes. This study will enable future detailed investigations of proteins associated with quality attributes of durians to be studied.

Matched MeSH terms: Chemical Fractionation/methods*
Fulltext DNA, RNA, and protein extraction: the past and the present

Tan SC, Yiap BC

J Biomed Biotechnol, 2009;2009:574398.
PMID: 20011662 DOI: 10.1155/2009/574398

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*
An effective placental cotyledons proteins extraction method for 2D gel electrophoresis

Tan NJ, Daim LD, Jamil AA, Mohtarrudin N, Thilakavathy K

Electrophoresis, 2017 03;38(5):633-644.
PMID: 27992069 DOI: 10.1002/elps.201600377

Effective protein extraction is essential especially in producing a well-resolved proteome on 2D gels. A well-resolved placental cotyledon proteome, with good reproducibility, have allowed researchers to study the proteins underlying the physiology and pathophysiology of pregnancy. The aim of this study is to determine the best protein extraction protocol for the extraction of protein from placental cotyledons tissues for a two-dimensional gel electrophoresis (2D-GE). Based on widely used protein extraction strategies, 12 different extraction methodologies were carefully selected, which included one chemical extraction, two mechanical extraction coupled protein precipitations, and nine chemical extraction coupled protein precipitations. Extracted proteins were resolved in a one-dimensional gel electrophoresis and 2D-GE; then, it was compared with set criteria: extraction efficacy, protein resolution, reproducibility, and recovery efficiency. Our results revealed that a better profile was obtained by chemical extraction in comparison to mechanical extraction. We further compared chemical extraction coupled protein precipitation methodologies, where the DNase/lithium chloride-dense sucrose homogenization coupled dichloromethane-methanol precipitation (DNase/LiCl-DSH-D/MPE) method showed good protein extraction efficiency. This, however, was carried out with the best protein resolution and proteome reproducibility on 2D-gels. DNase/LiCl-DSH-D/MPE was efficient in the extraction of proteins from placental cotyledons tissues. In addition, this methodology could hypothetically allow the protein extraction of any tissue that contains highly abundant lipid and glycogen.

Matched MeSH terms: Chemical Fractionation
Benchtop Isolation and Characterisation of Small Extracellular Vesicles from Human Mesenchymal Stem Cells

Tan KL, Chia WC, How CW, Tor YS, Show PL, Looi QHD, et al.

Mol Biotechnol, 2021 Sep;63(9):780-791.
PMID: 34061307 DOI: 10.1007/s12033-021-00339-2

The objective of this study is to develop a simple protocol to isolate and characterise small extracellular vesicles (sEVs) from human umbilical cord-derived MSCs (hUC-MSCs). hUC-MSCs were characterised through analysis of morphology, immunophenotyping and multidifferentiation ability. SEVs were successfully isolated by ultrafiltration from the conditioned medium of hUC-MSCs. The sEVs' size distribution, intensity within a specific surface marker population were measured with zetasizer or nanoparticle tracking analysis. The expression of surface and internal markers of sEVs was also assessed by western blotting. Morphology of hUC-MSCs displayed as spindle-shaped, fibroblast-like adherent cells. Phenotypic analysis by flow cytometry revealed that hUC-MSCs expressed MSC surface marker, including CD90, CD73, CD105, CD44 and exhibited the capacity for osteogenic, adipogenic and chondrogenic differentiation. Populations of sEVs with CD9, CD63 and CD81 positive were detected with size distribution in the diameter of 63.2 to 162.5 nm. Typical sEVs biomarkers such as CD9, CD63, CD81, HSP70 and TSG101 were also detected with western blotting. Our study showed that sEVs from hUC-MSCs conditioned medium were successfully isolated and characterised. Downstream application of hUC-MSCs-sEVs will be further explored.

Matched MeSH terms: Chemical Fractionation/methods*
A combined microwave pretreatment/solvent extraction process for the production of oil from palm fruit: optimisation, oil quality and effect of prolonged exposure

Tan JC, Chuah CH, Cheng SF

J Sci Food Agric, 2017 Apr;97(6):1784-1789.
PMID: 27470073 DOI: 10.1002/jsfa.7975

BACKGROUND: Conventional palm oil milling involves multiple stages after fruit collection; in particular, oil clarification introduces water into the pressed oil, which results in a large quantity of wastewater.
RESULTS: A combined process of microwave pretreatment and solvent extraction to mill crude palm oil, without introducing water or steam, is described. An excellent yield (up to 30%) of oil was obtained with pretreatment in a 42 L, 1000 W and 2450 MHz microwave oven followed by hexane extraction. The optimum conditions (10 min microwave pretreatment and 12 h solvent extraction) yielded an oil with a low free fatty acid content (<1.0%) and an acceptable anisidine value (<3.0 meq kg(-1) ). The oil had a fatty acid composition not resembling those of conventional crude palm oil and crude palm kernel oil. In the pretreatment, the leached oil had 6.3% lauric acid whereas the solvent extracted oil had only 1.5% lauric acid. Among the factors affecting the oil quality, microwave pretreatment affected the oil quality significantly; however, an optimised duration that would ensure high efficiency in solvent extraction also resulted in ruptured fruitlets, although not to the extent of causing excessive oxidation. In fact, microwave pretreatment should exceed 12 min; after only 15 min, the oil had 1-methylcyclopentanol (12.96%), 1-tetradecanol (9.44%), 1-nonadecene (7.22%), nonanal (7.13%) and 1-tridecene (5.09%), which probably arose from the degradation of fibres.
CONCLUSION: Microwave pretreatment represents an alternative milling process for crude palm oil compared with conventional processes in the omission of wet treatment with steam. © 2016 Society of Chemical Industry.

Matched MeSH terms: Chemical Fractionation/methods*
A comparative study of physicochemical characteristics and functionalities of pinto bean protein isolate (PBPI) against the soybean protein isolate (SPI) after the extraction optimisation

Tan ES, Ying-Yuan N, Gan CY

Food Chem, 2014;152:447-55.
PMID: 24444960 DOI: 10.1016/j.foodchem.2013.12.008

Optimisation of protein extraction yield from pinto bean was investigated using response surface methodology. The maximum protein yield of 54.8 mg/g was obtained with the optimal conditions of: temperature=25 °C, time=1 h and buffer-to-sample ratio=20 ml/g. PBPI was found to obtain high amount of essential amino acids such as leucine, lysine, and phenylalanine compared to SPI. The predominant proteins of PBPI were vicilin and phytohemagglutinins whereas the predominant proteins of SPI were glycinin and conglycinins. Significantly higher emulsifying capacity was found in PBPI (84.8%) compared to SPI (61.9%). Different isoelectric points were found in both PBPI (4.0-5.5) and SPI (4.0-5.0). Also, it was found that PBPI obtained a much higher denaturation temperature of 110.2 °C compared to SPI (92.5 °C). Other properties such as structural information, gelling capacity, water- and oil-holding capacities, emulsion stability as well as digestibility were also reported.

Matched MeSH terms: Chemical Fractionation
Application of experimental designs and response surface methods in screening and optimization of reverse micellar extraction

Sy Mohamad SF, Mohd Said F, Abdul Munaim MS, Mohamad S, Azizi Wan Sulaiman WM

Crit Rev Biotechnol, 2020 May;40(3):341-356.
PMID: 31931631 DOI: 10.1080/07388551.2020.1712321

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*

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