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An important technique which allows purification of mixture components is chromatography based on interaction between a stationary and mobile phase. The mixture components redistribute themselves between the phases either adsorption, partition, ion exchange or size exclusion. Here, we presented a review of applications of column, paper, thin layer...
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... chromatography is an analytical method used to separate colored chemicals or substances using a paper as the stationery phase ( Figure 4). In paper chromatography support material consists of a layer of cellulose highly saturated with water. ...
Citations
... Purification of protease by Ion exchange chromatography: The next step for protease purification was ion exchange chromatography which is a technique for separating organic molecules based on size, shape, charge, and solubility (19) . There are mobile and stationary phases in this procedure, CM-Cellulose is a type of weak cation exchanger; it can bind to the reverse charge of the target protein (20) . ...
hABSTRACT Background: Escherichia coli is one of the significant bacteria that belongs to Enterobacteriaceae bacteria family, which found in human intestinal tracts. Several Escherichia coli clone have been known to be extremely virulent and multidrug resistant. Escherichia coli poses a significant public health challenge for Iraq. Objectives: The current study aimed to cleavage the mucin protein by partial purified protease enzyme produced by pathogenic Escherichia coli bacteria. Materials and methods: This study was conducted on isolates of Escherichia coli bacteria that isolated from stool of people with gastroenteritis and diarrhoea. These isolates were examined on skim milk agar medium in order to screening of protease enzyme. Escherichia coli A29 was efficient isolate for protease production. The protease enzyme was purified by ion exchange (CM-Cellulose column) and gel filtration chromatography (Sephacryl S-300) after precipitated by ammonium sulphite saturation of 80%. Characterization for protease enzyme was done for effect of pH and temperature on activity and stability. The next step was treatment of protease (19250 U/mg) with mucin (0.11 mg/ml) and passed across the Sephacryl S-300 column. Results: The results showed that the purification of protease by these chromatography techniques was given specific activity of 19250 U/mg, purification fold 4.94 and yield 49. The maximum activity for purified protease was at pH 6.0 (77.527 U/ml) and pH stability for enzyme activity was between 5.5 and 9, while the optimum temperature was 37˚C (77.7 U/ml) and the stability for activity was kept between 15 and 50°C. Conclusion: The protease was cleavage of mucin for 3 peaks which represent fragments of mucin.
... The IEX media, which are commonly available on the market, include Capto, MacroCap, MiniBeads, Monobeads, Sephadex, Sepharose, and SOURCE. The disadvantage of IEX is the dilemma in selecting the IEX media and column hardware as they have great influence on the lateral outcomes [79]. ...
Peptides are short sequences of proteins consisting of two or more amino acids that are linked by peptide bonds. Peptide-based designs and drug deliveries can offer several advantages, such as antioxidant, antimicrobial, antihypertensive activities, along with immunomodulatory and antithrombotic properties, with hormone or drug-like potential. Peptide-based therapeutic formulations are used as drug candidates for the treatment of various diseases. However, there are several concerns associated with the efficacy of peptides in pharmaceutical design and delivery, including rapid degradation, limited solubility, and poor permeability. The nanoformulation of peptides has been identified as a promising approach for improving the stability of peptides and providing metabolic stability and bioavailability. This article provides an overview of the advances in the development of peptides for drug design and formulation applications. It discusses various peptide nanoformulation approaches as well as recent developments in the in vitro and in vivo analyses of nanoformulated peptides for pharmaceutical applications.
... However, during this method, the mixture was first dropped in the bottom regions of the plate with varying flow rates using a pipette. [160] Hence, the separation of analytes is accomplished. Additionally, TLC is a basic, cost-effective, and simple-to-use analytical chemistry technology with several applications, including the discovery of novel drugs and various types of medicinal plant compositions. ...
Galacto-oligosaccharides (GOS) are the prebiotic constituents with various functional qualities that promote the optimal proliferation of gut microbiota and are consequently beneficial to human health. GOS are not absorbed by humans and animals, but they play numerous health-related positive roles in the human body. Currently, their production has occupied much attention due to the increasing demand for functional foods. It could be produced by enzymatic and chemical approaches. Many microbial enzymes have been utilized for GOS production but the main source of these enzymes is lactose. Moreover, characterization of GOS structures is also a necessary and critical effort for understanding their mechanism of action. Hence, this review is written to understand the extensive but related features for the synthesis of GOS which includes: the enzymatic production of GOS through various enzymes and the types of enzymes such as free, immobilized, recombinant, crude, and purified enzymes that are involved in their synthesis, the techniques used in the purification of GOS, the analysis and quantification of GOS through various practices, and the functional properties of GOS are also
discussed in this review. It also aims to combine new findings with current information in order to improve understanding of GOS formation and other factors related to its functionality.
... Chromatography is defined as the separation of a combination of compounds into specific entities by using two phases; one is mobile and the other one is stationary [2,3]. This technique was first invented in 1903 by Mikhail Semyorivich Tswett, an Italian born Russian botanist, and was later considered the 'Father of Chromatography' [4,5]. Chromatography combines two Greek words, i.e., chromo means 'color' and graphene means 'to write' [6]. ...
Background: Chromatography is defined as a set of techniques that are used for the separation of constituents in a mixture. Introduction: High-Pressure Liquid Chromatography or High-Performance Liquid Chromatography (HPLC) is known as a specialized technique in which columns as well as liquid chromatography are used in the separation, characterizationand investigation of the active moieties existing in the mixture. Objective: Current review focuses on the HPLC technique, including its principles, instrumentation, types, applications, advancements, and patents. Result: HPLC technique is important both for quantitative as well as qualitative analysis and is used for the evaluation of biological and
... Each component's velocity is determined by its chemical composition, the stationary phase's nature (column) and the mobile phase's composition. The retention time of a particular analyte is the time at which it elutes (emerges from the column) [46]. Numerous column varieties are available, each with a unique combination of adsorbents with variable particle sizes, porosity and surface chemistry. ...
Nanoplastics (NPs) are a rapidly developing subject that is relevant in environmental and food research, as well as in human toxicity, among other fields. NPs have recently been recognized as one of the least studied types of marine litter, but potentially one of the most hazardous. Several studies are now being reported on NPs in the environment including surface water and coast, snow, soil and in personal care products. However, the extent of contamination remains largely unknown due to fundamental challenges associated with isolation and analysis, and therefore, a methodological gap exists. This article summarizes the progress in environmental NPs analysis and makes a critical assessment of whether methods from nanoparticles analysis could be adopted to bridge the methodological gap. This review discussed the sample preparation and preconcentration protocol for NPs analysis and also examines the most appropriate approaches available at the moment, ranging from physical to chemical. This study also discusses the difficulties associated with improving existing methods and developing new ones. Although microscopical techniques are one of the most often used ways for imaging and thus quantification, they have the drawback of producing partial findings as they can be easily mixed up as biomolecules. At the moment, the combination of chemical analysis (i.e., spectroscopy) and newly developed alternative methods overcomes this limitation. In general, multiple analytical methods used in combination are likely to be needed to correctly detect and fully quantify NPs in environmental samples.
... Chromatographic methods have made it possible, in the last 70 years, the separation of complex mixtures, both on an analytical and preparative scale. Column and thin-layer chromatography (Enyoh et al. 2019), based on adsorptive processes, uses solid supports as a stationary phase, while that based on the partition between two liquid phases, the liquid stationary phase is adsorbed on a support. In both cases, the solute-solid interaction can cause irreversible adsorption and/or chemical modifications in the components of the mixtures subjected to separation. ...
Alkaloids are nitrogen-containing compounds of the secondary metabolism of plants and microorganisms, which can also be found in animals, insects, and marine organisms. Because of the presence of one or more nitrogen atoms in the molecule, these compounds can form salts in the presence of acids, which are soluble in water and not in organic solvents. This characteristic is useful for their extraction from the matrix source and is explored in liquid–liquid partitioning techniques like countercurrent chromatography. Solvent systems used in classic purifications of alkaloids by this separation technique consist of an organic solvent such as chloroform or dichloromethane and a water buffer where the pH varies along the purification. The addition of other organic solvents like methanol and other alcohols to the solvent system is discussed, as well as the use of less polar systems. Recently developed techniques in countercurrent separations, such as pH-zone-refining countercurrent chromatography, are also presented. This comprehensive review covers the early work on separation of alkaloids with the Craig and Post apparatus and the evolution in the use of modern equipment for the isolation and purification of this class of bioactive natural products.
... Vast combinations of diverse bioactive compounds are contained in medicinal plants, and therefore expertise is required in their isolation, identification and characterization procedures. The frequently used methods for the isolation of bioactive compounds, which have been shown to be efficient, include immunoassay, phytochemical screening assay, and chromatographic techniques such as column chromatography, high-pressure liquid chromatography (HPLC), thin-layer chromatography (TLC), flash chromatography and Sephadex chromatography [56,57]. After the isolation of bioactive compounds, they are identified, quantified and characterized using Fourier transform infrared (FTIR) spectroscopy and gas chromatography-mass spectrometry (GC-MS) [58][59][60][61][62][63][64]. ...
The therapeutic actions of plants rely solely on the bioactive compounds contained in such plants. Bioactive compounds are synthesized basically from plant primary metabolites such as amino acids, carbohydrates and lipids. Bioactive compounds can be isolated, identified and characterized using standard analytical protocols. Edible leaves of Vernonia amygdalina, Ocimum gratissimum, Piper guineense and Gongronema latifolium are used traditionally in various parts of the world, especially in Africa and Asia, for the alleviation of pathologic conditions. The present review highlighted the medicinal usefulness of edible leaves of V. amygdalina, O. gratissimum, P. guineense and G. latifolium in connection with the diverse bioactive compounds that are linked to their therapeutic potencies. Herbal therapeutics are preferred to synthetic drugs by many people worldwide due to their ease of accessibility, low cost of usage and low incidences of side effects associated with the use of herbs as well as cultural consideration. Thus, the leaves of V. amygdalina, O. gratissimum, P. guineense and G. latifolium should be subjected to more extensive and rigorous medicinal evaluation as well as consider the use of bioactive compounds from these plant materials for the design and development of novel drugs.
... The technique is similar with the AF4 except that, AF4 contains only one permeable wall so the cross-flow is caused only by the carrier liquid. In GE, NPs are separated as they move through a gel with an electric field [140,141]. The separation is possible because NPs have charges on its surfaces [11,81]. ...
... The CE technique adopts similar mechanism as GE, except that the particle travels through a column containing an electrolyte solution [140,141]. The SEC is applied to separate particles in a sample by size and sometimes on molecular weight. ...
... SEC is not plastic-specific just like GE and CE. The sample is passed through a column of porous beads [141]. Smaller particles are slowed since they have to pass through the porous matrix of the beads while larger particles pass around the beads. ...
Air pollutants are perhaps the largest cause of diseases and death in the world today. Increasing urbanization and industrialization have caused an increase in number of diverse forms and types of new pollutants, which are difficult to detect and characterize due to their stench behaviour and complex sources of production. Such pollutants have been called emerging pollutants (EPs) and their list is ever increasing. Therefore, the understanding of the method of analysis and health implication of (EPs) in air is critical to providing a more robust understanding of exposure routes, regulations and mitigation. EPs in air discussed in this study are not in any way exhaustive but limited to emerging VOCs (including acrylonitrile, 1-3-butadiene, chloroform, dichloromethane, ethylene oxides, formaldehyde, toluene, trichloroethylene, 1,4-Dioxane) and metals (arsenic, manganese, and vanadium), ultrafine particles, micro-and nano-plastics, engineered nanoparticles, diesel/black carbon and bioaerosols. Occurrence, detection and health implications of these EPs in air are still unfolding due to limited monitoring studies, lack of standard methodology and regulations. To address this knowledge gap, authors conducted an in-depth review of available information. Their spatial distribution, analytical methods and health implications are discussed including the novel coronavirus (COVID-19) as a potential EP in air. The study concluded with highlights of gaps in knowledge and suggestions to key areas for future research. This information is of general interest to environmental scientists and of specific interest to both health and sanitation workers and policymakers at private, government and international organizations.
... Subsequently adding dichloromethane (DCM) to the extract, dichloromethane (DCM) soluble fraction was removed and the remaining portion was methanolic fraction of the extract. The methanolic fraction was subjected to column chromatography to separate their constituents based on adsorption using wet packing method [12]. Later these column fractions were analyzed by thin layer chromatography [13] and compounds of interest were isolated using preparative layer chromatography (PLC) [13]. ...
Methanolic extract of the leaves of Bridelia stipularis was studied. From this study, we have
isolated three flavones. They are identified as 7-O-methyl luteolin, apigenin and 5, 7, 2’, 5’
tetrahydroxyflavone by NMR spectroscopic studies. All of them are first time documented for this
plant. Different solvent fractions were subjected to in vitro antioxidant and cytotoxicity studies.
Both apigenin and ethyl acetate soluble fraction of Bridelia stipularis showed strong antioxidant
activity having IC 50 value of 8.005, 8.77 µg/mL respectively. Chloroform soluble fraction of
Bridelia stipularis exerted highest toxicity to brine shrimp and petroleum ether soluble fraction
showed moderate toxicity having LC 50 value of 1.05, 1.71 µg/mL respectively .
Keywords: Bridelia stipularis, flavone, luteolin, apigenin, tetrahydroxyflavone, antioxidant,
cytotoxicity
... Henceforth they flow along with the mobile phase at different time intervals [21]. Finally, analysis of the eluted fractions can be done by using different techniques including TLC, HPLC, HPTLC, LC-MS, GC-MS etc. [22]. ...
In the present study, the antimicrobial activity of fractions and sub-fractions of methanol extract of Acacia nilotica L. twig was done, and bioactive compounds were identified by GC-MS. Fractionation was done by column chromatography using different solvents, and their antimicrobial potential was checked by the agar well diffusion method. Minimum inhibitory concentration (MIC) was performed by the micro broth dilution method. Oral pathogens, including Enterococcus faecalis, Staphylococcus aureus, and Candida albicans were selected for the study. Results indicated that fraction (Fr-III) of A. nilotica methanol extract showed a significant zone of inhibition (ZOI) in the range of 14-15 mm against selected pathogens. Further sub-fraction, Fr-IIIf & Fr-IIIg of Fr-III exhibited maximum ZOI in a range of 38-40 mm at P<0.05. MIC of sample fractions was in the range of 80-210 µg/mL. GC-MS analysis represented that Piperidine,2,2,6,6-tetramethyl-was the major phytochemical in Fr-III. In the case of Fr-IIIg, n-Hexadecanoic acid was the main component, whereas behenic alcohol was in Fr-IIIf. Therefore, these bioactive compounds may be used as a potential therapeutic agent for oral health.
