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Journal of Food and Drug Analysis Dec 2020In recent years, carbon dots (C-dots) have gained appreciable interest owing to their unique optical properties, including tunable fluorescence, stability against... (Review)
Review
In recent years, carbon dots (C-dots) have gained appreciable interest owing to their unique optical properties, including tunable fluorescence, stability against photobleaching and photoblinking, and strong fluorescence. Simple and low-cost hydrothermal and electrochemical approaches have been widely used in the preparation of biocompatible and high-quality C-dots. Various C-dots have been used for the quantitation of small analytes, mostly based on analyte induced fluorescence quenching. Depending on the nature of precursors, synthetic conditions (such as reaction temperature and time), and surface conjugation, multi-function C-dots can be prepared and used in diagnostics and therapeutics. Their strong fluorescence and photostability, enables use in cell imaging. Their biological activity from the surface residues and capability of generating reactive oxygen species, have allowed many C-dots to become candidates as antibacterial and anticancer reagents. After suitable conjugation, biocompatible and fluorescent C-dots can be used for diagnostics and therapeutics, thus, showing their great potential in the area of theranostics.
PubMed: 35696139
DOI: 10.38212/2224-6614.1154 -
Biointerphases Mar 2020Nowadays, high-resolution imaging techniques are extensively applied in a complementary way to gain insights into complex phenomena. For a truly complementary analytical...
Nowadays, high-resolution imaging techniques are extensively applied in a complementary way to gain insights into complex phenomena. For a truly complementary analytical approach, a common sample carrier is required that is suitable for the different preparation methods necessary for each analytical technique. This sample carrier should be capable of accommodating diverse analytes and maintaining their pristine composition and arrangement during deposition and preparation. In this work, a new type of sample carrier consisting of a silicon wafer with a hydrophilic polymer coating was developed. The robustness of the polymer coating toward solvents was strengthened by cross-linking and stoving. Furthermore, a new method of UV-ozone cleaning was developed that enhances the adhesion of the polymer coating to the wafer and ensures reproducible surface-properties of the resulting sample carrier. The hydrophilicity of the sample carrier was recovered applying the new method of UV-ozone cleaning, while avoiding UV-induced damages to the polymer. Noncontact 3D optical profilometry and contact angle measurements were used to monitor the hydrophilicity of the coating. The hydrophilicity of the polymer coating ensures its spongelike behavior so that upon the deposition of an analyte suspension, the solvent and solutes are separated from the analyte by absorption into the polymer. This feature is essential to limit the coffee-ring effect and preserve the native identity of an analyte upon deposition. The suitability of the sample carrier for various sample types was tested using nanoparticles from suspension, bacterial cells, and tissue sections. To assess the homogeneity of the analyte distribution and preservation of sample integrity, optical and scanning electron microscopy, helium ion microscopy, laser ablation inductively coupled plasma mass spectrometry, and time-of-flight secondary ion mass spectrometry were used. This demonstrates the broad applicability of the newly developed sample carrier and its value for complementary imaging.
Topics: Animals; Hydrophobic and Hydrophilic Interactions; Imaging, Three-Dimensional; Nanoparticles; Polymers; Pseudomonas putida; Rabbits; Skin; Surface Properties; Temperature; Water; Zea mays
PubMed: 32212739
DOI: 10.1116/1.5143203 -
Methods in Molecular Biology (Clifton,... 2022The separation of complex mixtures is ubiquitous throughout molecular biology, and techniques such as gel-based electrophoresis are common laboratory practice. Such...
The separation of complex mixtures is ubiquitous throughout molecular biology, and techniques such as gel-based electrophoresis are common laboratory practice. Such methods are not without their drawbacks, however, which include non-specific interactions between analyte and the separation matrix, poor yields in purification and non-continuous analyte throughput. Microfluidic techniques, which exploit physical phenomena unique to the microscale, promise to improve many aspects of traditional laboratory procedures. These methods offer a quantitative, solution-based alternative to traditional gel electrophoresis, with rapid measurement times enabling the analysis of transient or weak biomolecular interactions that would be challenging to observe with traditional methods. Here, we present a protocol for the lithographic fabrication and operation of microfluidic chips capable of free-flow electrophoretic (FFE) fractionation and analysis of biological analytes. We demonstrate the efficacy of our approach through a protein-sensing methodology based on FFE fractionation of DNA-protein mixtures. In addition, the FFE technique described here can be readily adapted to suit a variety of preparative and analytical applications, providing information on the charge, zeta-potential, and interactions of analytes.
Topics: Electrophoresis; Electrophoresis, Microchip; Proteins
PubMed: 35094333
DOI: 10.1007/978-1-0716-1811-0_16 -
Metabolites Sep 2023Volumetric absorptive microsampling (VAMS) has arisen as a relevant tool in biological analysis, offering simplified sampling procedures and enhanced stability. Most of... (Review)
Review
Volumetric absorptive microsampling (VAMS) has arisen as a relevant tool in biological analysis, offering simplified sampling procedures and enhanced stability. Most of the attention VAMS has received in the past decade has been from pharmaceutical research, with most of the published work employing VAMS targeting drugs or other exogenous compounds, such as toxins and pollutants. However, biomarker analysis by employing blood microsampling has high promise. Herein, a comprehensive review on the applicability of VAMS devices for the analysis of endogenous metabolites/biomarkers was performed. The study presents a full overview of the analysis process, incorporating all the steps in sample treatment and validation parameters. Overall, VAMS devices have proven to be reliable tools for the analysis of endogenous analytes with biological importance, often offering improved analyte stability in comparison with blood under ambient conditions as well as a convenient and straightforward sample acquisition model.
PubMed: 37887363
DOI: 10.3390/metabo13101038 -
Analytical and Bioanalytical Chemistry May 2022Chromatographic retention times are usually modeled considering only one analyte at a time. However, it has certain limitations as no information is shared between the...
Chromatographic retention times are usually modeled considering only one analyte at a time. However, it has certain limitations as no information is shared between the analytes, and consequently the model predictions poorly generalize to out-of-sample analytes. In this work, a publicly available dataset was used to illustrate the benefits of pooling the individual data and analyzing them simultaneously utilizing Bayesian hierarchical approach. Statistical analysis was carried out using the Stan program coupled with R, which enables full Bayesian inference with Markov chain Monte Carlo sampling. This methodology allows (i) incorporating prior knowledge about the likely values of model parameters, (ii) considering the between-analyte variability and the correlation between the model parameters, (iii) explaining the between-analyte variability by available predictors, and (iv) sharing information across the analytes. The latter is especially valuable when only limited information is available in the data about certain model parameters. The results are obtained in the form of posterior probability distribution, which quantifies uncertainty about the model parameters and predictions. Posterior probability is also directly relevant for decision-making. In this work, we used the Neue model to describe the relationship between retention factor and acetonitrile content in the mobile phase for 1026 analytes. The model was parametrized in terms of retention factor in 100% water, retention factor in 100% acetonitrile, and curvature coefficient, and considered log P and pK as predictors. From this analysis, we discovered that the analytes formed two clusters with different retention depending on the degree of analyte dissociation. The final model turned out to be well calibrated with the data. It gives insight into the behavior of analytes in the chromatographic column and can be used to make predictions for a structurally diverse set of analytes if their log P and pK values are known.
Topics: Bayes Theorem; Chromatography, High Pressure Liquid; Water
PubMed: 35347353
DOI: 10.1007/s00216-022-03968-x -
RSC Advances Sep 2020We report a novel bioluminescent aptasensor, which consists of 2'-F-RNA aptamer modules joined into a bi-specific aptamer construct. One aptamer module binds the...
We report a novel bioluminescent aptasensor, which consists of 2'-F-RNA aptamer modules joined into a bi-specific aptamer construct. One aptamer module binds the analyte, then after structural rearrangement the second module recruits non-covalently Ca-dependent photoprotein obelin from the solution, thus providing a bioluminescent signal. This concept allows using free protein as a reporter, which brings such advantages as no need for aptamer-protein conjugation, a possibility of thermal re-folding of aptamer component with no harm to a protein, and simpler detection protocol. We developed the new 2'-F-RNA aptamer for obelin, and proposed the strategy for engineering structure-switching bi-modular aptamer constructs which bind the analyte and the obelin in a sequential manner. With the use of hemoglobin as a model analyte, we showed the feasibility of utilizing the aptasensor in a fast and straightforward bioluminescent microplate assay. With a proper design of a secondary structure, this strategy of aptasensor engineering might be further extended to bi-specific aptamer-based bioluminescent sensors for other analytes of interest.
PubMed: 35516485
DOI: 10.1039/d0ra05117a -
Nanoscale Jul 2023Nanoscience shows promise for scientific advancement in many sectors, such as biology, energy, materials, environment, and manufacturing. Nanocomposites are mixtures of... (Review)
Review
Nanoscience shows promise for scientific advancement in many sectors, such as biology, energy, materials, environment, and manufacturing. Nanocomposites are mixtures of two or more materials, one of which is nanosized particles. The composites are expected to show combined features resulting in general enhancements in their physical and chemical properties. Metal-organic frameworks (MOFs) are coordination polymers that have attracted attention from researchers in recent years due to their porosity and controllable functionality. Another example of interesting nanomaterials is carbon nanotubes (CNTs) which are also known for their mechanical and thermal properties. Incorporation of both these materials into a nanocomposite has shown an enhancement in properties and conquered challenges in the defects of construction. This mini-review sheds light on the recent synthetic approaches and characterization of MOF-CNT nanocomposites in order to access porous selective nanocomposites that can improve analyte detection in environmental matrixes and biological systems. A summary of the chemical composition of nanocomposites, analytes in the target, and analytical techniques used is provided.
PubMed: 37378958
DOI: 10.1039/d3nr01074k -
Scandinavian Journal of Clinical and... Apr 2022The analytical stability of laboratory tests relies mostly on internal and external quality control procedures. Summarized patient data has in several studies been shown...
The analytical stability of laboratory tests relies mostly on internal and external quality control procedures. Summarized patient data has in several studies been shown to be a good supplement for monitoring analytical stability. In our present investigation, we evaluate a datamining method for retrospective evaluation and assessment of analyte stability in whole blood. Results from the laboratory information system were used as the basis for the datamining approach. Blood tests were requested by the general practitioners and drawing of the blood sample was either at the general practitioner's or at the hospital outpatient clinics. We were able to split data into groups based on sample collection place and time to analysis. The datamining approach was compared to experiments where samples were incubated at a single temperature as well as an experiment where the temperatures were changed during incubation. To demonstrate the method, we selected three laboratory tests considered representative: potassium, phosphate, and lactate dehydrogenase. The datamining approach showed results similar to the reference experiment. Furthermore, our results show that the analytes phosphate and potassium were not stable after short storage at a lower temperature.
Topics: Blood Specimen Collection; Humans; Phosphates; Potassium; Retrospective Studies; Specimen Handling; Temperature
PubMed: 35112638
DOI: 10.1080/00365513.2022.2031280 -
Journal of Analytical Toxicology Sep 2021Analytical performance of stable isotope-labeled internal standardization (SIL-IS) and threshold accurate calibration (TAC) methods of matrix normalization are compared...
Analytical performance of stable isotope-labeled internal standardization (SIL-IS) and threshold accurate calibration (TAC) methods of matrix normalization are compared for quantitation of 51 drugs and metabolites (analytes) in urine with analysis by ultra performance liquid chromatography with tandem mass spectrometry (UPLC-MS-MS). Two SIL-IS methods of analysis were performed, one method using analyte-specific internal standardization (ASIL-IS) and another method using a shared stable isotope from another analyte for internal standardization (SSIL-IS). Variance in inter-specimen matrix effect, without the use of a matrix normalization method, was studied by UPLC-MS-MS analysis of 338 urine donor samples and showed >200% variation in ion response for some analytes. Matrix normalization methods were evaluated for precision, accuracy, calibration, multi-matrix recovery and positive casework quantitation. Acceptable calibration and quality control criteria were achieved for all methods when calibrators and controls were prepared from the same urine matrix pool. Quantitative accuracy, determined by the addition of analytes to multi-donor urine pools at two concentration levels, resulted in acceptable percent relative standard deviation (%RSD) and bias for TAC and ASIL-IS methods. SSIL-IS method quantitations in analyte-supplemented donor pools revealed a %RSD ranging from 20% to 60% for >30% of the analytes and a method bias that ranged up to 87%, with a differential matrix effect on analyte and shared internal standard accounting for the imprecision and bias. Analyte quantitation in 162 authentic case samples showed close agreement for TAC and ASIL-IS methods, with greater variance in the SSIL-IS method. The study demonstrates effective matrix normalization by ASIL-IS and TAC methods and a matrix-caused bias in the SSIL-IS method.
Topics: Chromatography, High Pressure Liquid; Chromatography, Liquid; Pharmaceutical Preparations; Substance Abuse Detection; Tandem Mass Spectrometry
PubMed: 34013336
DOI: 10.1093/jat/bkab052 -
Methods in Molecular Biology (Clifton,... 2020The greatest advantage of frontal affinity chromatography (FAC) is that the analyte concentration does not need to be taken into consideration, and this renders FAC an... (Review)
Review
The greatest advantage of frontal affinity chromatography (FAC) is that the analyte concentration does not need to be taken into consideration, and this renders FAC an extremely favorable analytical tool for weak interactions. In this short review, we propose a straightforward explanation of the underlying mechanism. When FAC is performed using analyte solutions at relatively high concentrations, concentration-dependent retardation is observed due to competition among analyte molecules, and the elution volume changes depending on the degree of saturation of the immobilized ligand.However, when the analyte concentration is very low, no competition occurs among the analytes, and the elution volume reaches a constant value, which reflects the proportion of bound state to free state of a single analyte molecule. Therefore, the binding strength can be determined using a minimum analyte concentration.
Topics: Biomedical Research; Chromatography, Affinity; Lectins; Ligands; Polysaccharides; Protein Binding
PubMed: 32306312
DOI: 10.1007/978-1-0716-0430-4_4