Figure 4 - uploaded by Enyoh Christian Ebere
Content may be subject to copyright.
Source publication
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...
Contexts in source publication
Context 1
... 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. ...
Similar publications
High‐purity octafluoropropane (C3F8) electronic specialty gas is a key chemical raw material in semiconductor and integrated circuit manufacturing industry, while selective removal of hexafluoropropylene (C3F6) impurity for C3F8 purification is essential but a challenging task. Here we report a fluorinated cage‐like MOF Zn‐bzc‐CF3 (bzc=5‐(trifluoro...
Citations
... Ion-exchange chromatography (IEC) is an optional step generally performed after hydrolysis to purify biomolecules by removing inorganic contamination, particularly in mineral-bound matrices Vane et al., 2018). IEC uses interactions between charged molecules in the stationary phase (e.g., Na + , Ca 2+ , or OH -) and any interfering counterions in an analyte solution (Cummings et al., 2010;Ebere et al., 2019). Although both anion-and cation-exchange columns are available, biological and geochemical samples see anionic impurities more frequently, and the pH dependency of ionization typically generates a positively charged solution (due to the HCl in which it is dissolved) (Macko et al., 1987;Cummings et al., 2010). ...
... The system contains counter ions that are in an equilibrium state between the two phases, resulting in two IEC formats: anion and cation exchange. Catex resin is used to exchange cations, while Annex resin is used to exchange anions (Ebere et al. 2019). Other examples of anionic exchangers are Diethyl Aminoethyl (DEAE), Quaternary Aminoethyl (QAE) and Quaternary ammonium (Q) are examples of Catex while Carboxymethyl (CM), Sulfopropyl (S) and Methyl sulfonate (M). ...
... Applications This technique is used for the separation and purification of charged biomolecules like polypeptides, proteins, polynucleotides nucleic acids, blood components such as albumin, recombinant growth factors enzymes and other charged biomolecules (Ebere et al. 2019). Proteins, polypeptides, nucleic acids, polynucleotides, other charged biomolecules can also be separated and purified by using IEX. ...
Background
Proteomics is an analytical technique employed for the identification and quantitative analysis of total protein content in a cell, tissue or organism using various methods and tools. Proteomics succeeds in genomics (study of set of DNA) and transcriptomics (study of set of RNA) of biological systems. A proteome is an array of proteins generated by a particular organism, system, or biological condition. Though, an organism’s genome is more or less constant, a proteome is more complicated than a genome as cells generate a variety of sets of protein at different times or under different situations, such as cellular development and differentiation, cell cycle or carcinogenesis. Therefore, new approaches for capturing crucial biological information have been developed. Proteomics-based technologies are used for several research purposes including detection of various diagnostic markers, vaccine candidates as well as understanding pathogenicity mechanisms, changes in expression patterns in response to various signals, and interpretation of functional protein pathways in various diseases. Analytical methods like two-dimensional gel electrophoresis, mass spectrometry and various hyphenated techniques have been developed.
Main body
This review highlights various analytical tools used in qualitative and quantitative analysis of proteomics and novel approaches for the use of proteomics in disease diagnostics, as well as their other applications.
Conclusion
This review shall help academicians and researchers as a reference guide to update their knowledge on analytical tools for proteomics as well as the application of proteomics.
... We prove that the process described in this manuscript also works for detecting pharmacological agents in saliva samples, providing a simple, efficient way to isolate charged small molecules from protein-rich matrices. Future efforts can automate and simplify the current manual process either by incorporating directly into a staged LFA, by modifying the column to operate using pressure instead of centrifugation (either positive or negative), or by leveraging paper-based ion-exchange matrices [36]. In conclusion, the simplicity of the method described in this manuscript will enable more POC assays to operate in urine, saliva, and other biofluids at a higher degree of confidence, enabling more assays to be performed at the site of care; the end result should be a decrease in the time and cost to analyze samples, resulting in improved health outcomes. ...
The current COVID-19 pandemic has highlighted the power, speed, and simplicity of point-of-care (POC) diagnostics. POC diagnostics are available for a wide range of targets, including both drugs of abuse as well as performance-enhancing drugs. For pharmacological monitoring, minimally invasive fluids such as urine and saliva are commonly sampled. However, false positives or negatives caused by interfering agents excreted in these matrices may confound results. For example, false positives have, in most cases, prevented the use of POC diagnostics for pharmacological agent detection; the consequence is that centralized labs are instead tasked to perform these screenings, resulting in significant delays between sampling and testing. Thus, a rapid, simple, and inexpensive methodology for sample purification is required for the POC to reach a field-deployable tool for the pharmacological human health and performance assessments. Buffer exchange is a simple, rapid approach to remove interfering agents, but has traditionally been difficult to perform on small pharmacological molecules. Therefore, in this communication, we use salbutamol, a performance-enhancing drug, as a case example to demonstrate the efficacy of ion-exchange chromatography as a technique to perform buffer exchange for charged pharmacological agents. This manuscript demonstrates the efficacy of this technique leveraging a commercial spin column to remove interfering agents found in simulant urines, such as proteins, creatinine, and urea, while retaining salbutamol. The utility and efficacy of the method was then confirmed in actual saliva samples. The eluent was then collected and run on the lateral flow assays (LFAs), improving the reported limit of detection by over 5× (new lower limit of detection of 10 ppb compared to reported 60 ppb by the manufacturer) while simultaneously removing noise due to background interfering agents.
... 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.
... 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.
