A systematic review of various pKa determination techniques.

The pKa values of functional groups is crucial in determining the pharmacokinetic properties of a drug, affecting its absorption and thus bioavailability. This physicochemical property is also vital for the designing of drug excipients and vehicles. There are currently 13 known methods of determining a pKa value, namely: potentiometric titration, spectrometry, fluorometry, NMR, HPLC, conductometry, electrophoresis, voltammetry, solubility, partition coefficient, calorimetry, computational, and surface tension. Some of these techniques are more widely utilized and well-established compared to others, with each having their inherent advantages and disadvantages. This review discusses each of the aforementioned techniques with emphasis on their pros and cons.

Comparative characterization of sugar beet fibers to sugar beet pectin and octenyl succinic anhydride modified maltodextrin in aqueous solutions using viscometry, conductometry, tensiometry and component analysis.

BACKGROUND: Knowledge about specific functional characteristics, such as viscosimetric, conductometric, tensiometric and structural properties of polysaccharide aqueous solutions is highly important in the successful and adequate application in food emulsion formulation. For the first time detailed characterization of sugar beet fibers aqueous solutions in comparison to high molecular weight (sugar beet pectin) and low molecular weight [octenyl succinic anhydride (OSA) maltodextrin] hydrocolloids/stabilizers was performed through viscometry, conductometry, tensiometry and component analysis. RESULTS: Sugar beet fibers and its water-soluble fraction were investigated. All sugar beet fiber samples showed substantial surface-active properties but different effect on the viscosity values of aqueous solutions. Sugar beet pectin had higher impact on aqueous solutions viscosity values compared to sugar beet fiber samples. Structural bonding between investigated polysaccharides were evaluated through conductometric measurements. Intermolecular linking and probable embedding of OSA maltodextrin molecules into the sugar beet fiber complex structure was detected in conductometric studies. The increased concentration of sugar beet fibers in the presence of sugar beet pectin led to the accelerated increase in specific conductivity values indicating effects of 'macromolecular crowding', intermolecular and intramolecular conformation changes and charge formation. CONCLUSIONS: Detailed characterization of sugar beet fibers provided scientific insight towards fundamental characteristics of sugar beet fiber aqueous solutions. The presented characteristics are particularly applicable in the field of food emulsion stabilization due to the presented surface-active properties of sugar beet fibers as well as specific characteristics of investigated multi-polysaccharide systems. © 2022 Society of Chemical Industry.

New trends in methyl salicylate sensing and their implications in agriculture.

Methyl salicylate (MeSal) is an organic compound present in plants during stress events and is therefore a key marker for early plant disease detection. It has usually been detected by conventional methods that require bulky and costly equipment, such as gas chromatography or mass spectrometry. Currently, however, chemical sensors provide an alternative for MeSal monitoring, showing good performance for its determination in the vapour or liquid phase. The most promising concepts used in MeSal determination include sensors based on electrochemical and conductometric principles, although other technologies based on mass-sensitive, microwave, or spectrophotometric principles also show promise. The receptor elements or sensitive materials are shown to be part of the key elements in these sensing technologies. A literature survey identified a significant contribution of bioreceptors, including enzymes, odourant-binding proteins or peptides, as well as receptors based on polymers or inorganic materials in MeSal determination. This work reviews these concepts and materials and discusses their future prospects and limitations for application in plant health monitoring.

Recent advancements in urea biosensors for biomedical applications.

The quick progress in health care technology as a recurrent measurement of biochemical factors such as blood components leads to advance development and growth in biosensor technology necessary for effectual patient concern. The review wok of authors present a concise information and brief discussion on the development made in the progress of potentiometric, field effect transistor, graphene, electrochemical, optical, polymeric, nanoparticles and nanocomposites based urea biosensors in the past two decades. The work of authors is also centred on different procedures/methods for detection of urea by using amperometric, potentiometric, conductometric and optical processes, where graphene, polymer etc. are utilised as an immobilised material for the fabrication of biosensors. Further, a comparative revision has been accomplished on various procedures of urea analysis using different materials-based biosensors, and it discloses that electrochemical and potentiometric biosensor is the most promise one among all, in terms of rapid response time, extensive shelf life and resourceful design.

Rapid and Selective Biomarker Detection with Conductometric Sensors.

The biomarker detection in human body fluids is crucial as biomarkers are important in diagnosing diseases. Conventional invasive techniques for biomarker detection are associated with infection, tissue damage, and discomfort. Non-invasive devices are an attractive alternative. Here, metal oxide (oxygen-deficient zinc oxide, ZnO) based conductometric sensors with two-terminal electrodes for rapid detection of biomarkers in real-time, are presented. This platform can be engineered for non-invasive, sensitive, and on-demand selective detection of biomarkers based on surface functionalization. The three novelties in this biosensing technique include an on-demand target selection device platform, short (<10 min) incubation times, and real-time monitoring of the biomarker of interest by electrical (resistance change) measurements. Cardiac inflammatory biomarkers interleukin 6 (IL-6) and C-reactive protein (CRP) are used as the model antigens. The devices can detect 100× lower concentration of IL-6 than healthy levels in human saliva and sweat and 1000× and ≈50× lower CRP concentrations than healthy levels in human saliva and sweat, respectively. The devices show high selectivity for IL-6 and CRP antigens when tested with a mixture of biomarkers. This sensor platform can be extended to selective measurements for viruses or DNA screening, which enables a new category of compact and rapid point-of-care medical devices.

Affordable automated phenotypic antibiotic susceptibility testing method based on a contactless conductometric sensor.

User-friendly phenotypic antibiotic susceptibility testing (AST) methods are urgently needed in many fields including clinical medicine, epidemiological studies and drug research. Herein, we report a convenient and cost-effective phenotypic AST method based on online monitoring bacterial growth with a developed 8-channel contactless conductometric sensor (CCS). Using E. coli and V. parahaemolyticus as microorganism models, as well as enoxacin, florfenicol, ampicillin, kanamycin and sulfadiazine as antibiotic probes. The minimum inhibitory concentration (MIC) determination was validated in comparison with standard broth microdilution (BMD) assay. The total essential agreements between the CCS AST assays and the reference BMD AST assays are 68.8-92.3%. The CCS has an approximate price of $9,000 (USD). Requiring neither chemical nor biotic auxiliary materials for the assay makes the cost of each sample < $1. The MICs obtained with the automated CCS AST assays are more precise than those obtained with the manual BMD. Moreover, in 72 percent of the counterpart, the MICs obtained with the CCS AST assays are higher than that obtained with the BMD AST assays. The proposed CCS AST method has advantages in affordability, accuracy, sensitivity and user-friendliness.

Conductometric determination of trospium chloride by silver nitrate and phosphomolybidic acid.

Two simple, accurate, sensitive and precise conductometric methods were developed for determination of trospium chloride in pure form and in pharmaceutical formulations. It is based on using two precipitating reagents; Phosphomolybdic acid (PMA) and Silver nitrate (AgNO3). The mean recovery for Silver nitrate is in the range (98-100.95%) and for Phosphomolybdic acid in the range (98-101.69%). A molar ratio has been determined conductometrically for the two reagents, revealed (1/1) for (drug/reagent). The proposed methods were validated and successfully applied for the determination of the studied drug in pure form and in its pharmaceutical preparation. The results of the proposed methods were compared to the results of reported method with no significant difference between them.

Voltammetric kanamycin aptasensor based on the use of thionine incorporated into Au@Pt core-shell nanoparticles.

A signal-on aptasensor is described for the voltammetric determination of kanamycin (KANA). Au@Pt core-shell nanoparticles with large surface and good electrical conductivity were synthetized and act as both a conductive material and as the carrier for complementary strands (CS2) and thionine (TH). In the presence of KANA, the electrochemical response of TH changes due to hybridization between CS1 immobilized on the electrode and the Au@Pt-CS2/TH system. The peak current increases linearly with the logarithm of the KANA concentration in the range from 1 pM to 1 µM, and the limit of detection is 0.16 pM. The sensor was characterized in terms of selectivity, reproducibility and stability, and satisfactory results were obtained. It was also utilized for the determination of KANA in (spiked) chicken samples. The recoveries (95.8-103.2%) demonstrate the potential of the method for KANA detection in real samples. Graphical abstract A signal-on aptasensor for kanamycin (KANA) was developed by using Au@Pt core-shell nanoparticles as nanocarrier for probe aptamer and as a sensing probe.

An Evaluation of the Glass Vial Hydrolytic Resistance Method.

The European Pharmacopoeia (Ph. Eur.) hydrolytic resistance method for glass vials is routinely used to screen pharmaceutical glass vial supply. In an effort to better understand and control the factors affecting method precision and robustness, several potential sources of variability in the Ph. Eur. alkalinity method have been studied for 3 cc glass vials. Method parameters including vial rinsing, vial covering, autoclave cycle execution, sample hold times, and titration procedure were evaluated in this study. The results of this study indicate the method parameters which require stringent control in order to achieve acceptable method precision and robustness.LAY ABSTRACT: The European Pharmacopoeia (Ph. Eur.) hydrolytic resistance method for glass vials is routinely used to screen biopharmaceutical glass vial supply. The method was studied to assess contributions to its variability and to potentially improve its reliability. The results of this study indicate which method parameters require stringent control in order to generate reliable data using the Ph. Eur. hydrolytic resistance method.

Simultaneous Determination of Inorganic Anions and Cations in Water and Biological Samples by Capillary Electrophoresis with a Capacitive Coupled Contactless Conductivity Detector Using Capillary Filling Method.

An analytical method for concurrent analysis of inorganic anions and cations has been developed using a capillary electrophoresis (CE)-capacitively coupled contactless conductivity detector (C4D) system. Although hydrodynamic and electrokinetic injection techniques have been widely used in CE, we employed a capillary filling method (CFM) for the analysis of inorganic ions. The procedure is relatively simple and has the advantage that CMF does not require pressure control and vial exchange. Three anions (chloride, sulfate, nitrate) and five cations (ammonium, potassium, sodium, magnesium, calcium) were successfully separated and detected at ppm levels within 80 s using a 9 mM histidine/15 mM malic acid (pH 3.6) containing 50 mM N-dodecyl-N,N-dimethyl-3-ammonio-1-propanesulfonate as background electrolyte. Applying this analytical condition, the electroosmotic flow is negligible and anions and cations were migrated concurrently to different polarities according to their electrophoretic mobility. Obtained raw data showed stepwise increases in detected conductivity due to the migration of sample components, which expresses as peak profiles by differentiation of electropherograms. The RSD values of the peak area and migration times for the anions and cations were satisfactory and were less than 5.15 and 2.04%, respectively. The developed method was applied for the analysis of inorganic anions and cations in commercial mineral waters, tap water, urine, and exhaled breath condensate. These results indicate that the CE-C4D system with CFM is suitable for the rapid analysis of inorganic anions and cations in various samples.

Conductometric biosensor for arginine determination in pharmaceutics.

A new conductometric biosensor based on coimmobilized urease and arginase has been developed for arginine determination in pharmaceutics. First, the main parameters of the selected method of immobilization (concentrations of arginase, urease, and glutaraldehyde, time of incubation) were optimized. An influence of the solution parameters (buffer ionic strength, capacity, pH, Mn2+ concentration) on the biosensor operation was studied, working conditions were optimized. After biosensor optimization, the main analytical characteristics were as follows. The limit of detection - 2.5 µM, the linear range - 2.5-500 µM, the sensitivity to arginine 13.4 ±â€¯2.4 µS/mM, the response time - 20 s. The signals repeatability and operational stability in continuous exploitation were studied over one working day and during one week. Additionally, the selectivity of the developed biosensor towards arginine was essayed relative to other amino acids. The developed biosensor has been used to measure arginine concentrations in some drugs. The results obtained were in high correlation with the characteristics declared by producers.

Laser based thermo-conductometry as an approach to determine ribbon solid fraction off-line and in-line.

Ribbon solid fraction is one of the most important quality attributes during roll compaction/dry granulation. Accurate and precise determination is challenging and no in-line measurement tool has been generally accepted, yet. In this study, a new analytical tool with potential off-line as well as in-line applicability is described. It is based on the thermo-conductivity of the compacted material, which is known to depend on the solid fraction. A laser diode was used to punctually heat the ribbon and the heat propagation monitored by infrared thermography. After performing a Gaussian fit of the transverse ribbon profile, the scale parameter σ showed correlation to ribbon solid fraction in off-line as well as in-line studies. Accurate predictions of the solid fraction were possible for a relevant range of process settings. Drug stability was not affected, as could be demonstrated for the model drug nifedipine. The application of this technique was limited when using certain fillers and working at higher roll speeds. This study showed the potentials of this new technique and is a starting point for additional work that has to be done to overcome these challenges.

Enzyme- and label-free electrochemical aptasensor for kanamycin detection based on double stir bar-assisted toehold-mediated strand displacement reaction for dual-signal amplification.

It is critically important to detect antibiotic residues for monitoring food safety. In this study, an enzyme- and label-free electrochemical aptasensor for antibiotics, with kanamycin (Kana) as a typical analyte, was developed based on a double stir bar-assisted toehold-mediated strand displacement reaction (dSB-TMSDR) for dual-signal amplification. First, we modified two gold electrodes (E-1 and E-2) with different DNA probes (S1/S2 hybrid probe in E-1 and DNA fuel strand S3 in E-2). In the presence of Kana, an S1/S2 probe can be disassembled from E-1 to form an S2/Kana complex in supernatant. The S2/Kana could react with S3 on E-2 to form S2/S3 hybrid and release Kana through TMSDR. After then, the target recycling was triggered. Subsequently, the formed S2/S3 hybrid can also trigger a hybridization chain reaction (HCR). Consequently, the dual-signal amplification strategy was established, which resulted in many long dsDNA chains on E-2. The chains can associate with methylene blue (MB) as redox probes to produce a current response for the quantification of Kana. The assay exhibited high sensitivity and specificity with a detection limit at 16 fM Kana due to the dual-signal amplification. The double stir bars system can both increase phase separation and prevent leakage of DNA fuel to reduce background interference. Moreover, it allows flexible sequence design of the TMSDR probes. The assay was successfully employed to detect Kana residues in food and showed potential application value in food safety detection.

An ultrasensitive hollow-silica-based biosensor for pathogenic Escherichia coli DNA detection.

A novel electrochemical DNA biosensor for ultrasensitive and selective quantitation of Escherichia coli DNA based on aminated hollow silica spheres (HSiSs) has been successfully developed. The HSiSs were synthesized with facile sonication and heating techniques. The HSiSs have an inner and an outer surface for DNA immobilization sites after they have been functionalized with 3-aminopropyltriethoxysilane. From field emission scanning electron microscopy images, the presence of pores was confirmed in the functionalized HSiSs. Furthermore, Brunauer-Emmett-Teller (BET) analysis indicated that the HSiSs have four times more surface area than silica spheres that have no pores. These aminated HSiSs were deposited onto a screen-printed carbon paste electrode containing a layer of gold nanoparticles (AuNPs) to form a AuNP/HSiS hybrid sensor membrane matrix. Aminated DNA probes were grafted onto the AuNP/HSiS-modified screen-printed electrode via imine covalent bonds with use of glutaraldehyde cross-linker. The DNA hybridization reaction was studied by differential pulse voltammetry using an anthraquinone redox intercalator as the electroactive DNA hybridization label. The DNA biosensor demonstrated a linear response over a wide target sequence concentration range of 1.0×10-12-1.0×10-2 µM, with a low detection limit of 8.17×10-14 µM (R2 = 0.99). The improved performance of the DNA biosensor appeared to be due to the hollow structure and rough surface morphology of the hollow silica particles, which greatly increased the total binding surface area for high DNA loading capacity. The HSiSs also facilitated molecule diffusion through the silica hollow structure, and substantially improved the overall DNA hybridization assay. Graphical abstract Step-by-step DNA biosensor fabrication based on aminated hollow silica spheres.

Determination of apple juice authenticity using gas chromatography-mass spectrometry, volt-ampere and new conductometric methods.

BACKGROUND: Organoleptic properties and positive health effect on consumers both justify the importance of determining apple juice authenticity with respect to retail and wholesale trade networks worldwide. RESULTS: Adulteration of apple juice through dilution with water, as well as with ascorbic acid, has been determined. It has been demonstrated that polarization curves of a platinum electrode in apple juice are useful for undeclared antioxidant determination, as a result of the movement of the polarization curve to the left, by 200 mV, when ascorbic acid was added. A difference between the equivalent electrical conductivity of suspended solids of undiluted and diluted (1:1) juices has been substantiated as a new quantitative criterion for determining adulteration through dilution. The criterion values range from 0.030 to 0.034 S m-1 for juices of different apple varieties. Gas chromatography-mass spectrometry results have shown the presence of (E)-2-hexenal and n-hexanal in all juices tested obtained from different apple varieties. CONCLUSION: A new conductometric method for the determination of diluted apple juices has been substantiated. It has been established that the volt-ampere method is useful for the investigation of undeclared ascorbic acid. © 2018 Society of Chemical Industry.

Ultrasensitive photoelectrochemical biosensor for the detection of HTLV-I DNA: A cascade signal amplification strategy integrating λ-exonuclease aided target recycling with hybridization chain reaction and enzyme catalysis.

Sensitive and specific detection of DNA is of great significance for clinical diagnosis. In this paper, an effective cascade signal amplification strategy was introduced into photoelectrochemical (PEC) biosensor for ultrasensitive detection of human T-cell lymphotropic virus type I (HTLV-I) DNA. This proposed signal amplification strategy integrates λ-exonuclease (λ-Exo) aided target recycling with hybridization chain reaction (HCR) and enzyme catalysis. In the presence of target DNA (tDNA) of HTLV-I, the designed hairpin DNA (h1DNA) hybridized with tDNA, subsequently recognized and cleaved by λ-Exo to set free tDNA. With the λ-Exo aided tDNA recycling, an increasing number of DNA fragments (output DNA, oDNA) were released from the digestion of h1DNA. Then, triggered by the hybridization of oDNA with capture DNA (cDNA), numerous biotin-labeled hairpin DNAs (h2DNA and h3DNA) could be loaded onto the photoelectrode via the HCR. Finally, avidin-labeled alkaline phosphatase (avidin-ALP) could be introduced onto the electrode by specific interaction between biotin and avidin. The ALP could catalyze dephosphorylation of phospho-L-ascorbic acid trisodium salt (AAP) to generate an efficient electron donor of ascorbic acid (AA), and thereby greatly increasing the photocurrent signal. By utilizing the proposed cascade signal amplification strategy, the fabricated PEC biosensor exhibited an ultrasensitive and specific detection of HTLV-I DNA down to 11.3 aM, and it also offered an effective strategy to detect other DNAs at ultralow levels.

A sensitive Potentiometric resolved ratiometric Photoelectrochemical aptasensor for Escherichia coli detection fabricated with non-metallic nanomaterials.

In this work, a sensitive potentiometric resolved ratiometric photoelectrochemical aptasensor for Escherichia coli (E. coli) detection was successfully fabricated with non-metallic nanomaterials. To avoid the use of precious metals or heavy metals, three-dimensional graphene hydrogel-loaded carbon quantum dots (C-dots/3DGH) and graphene-like carbon nitride (g-C3N4) with excellent PEC activity and matched potential were prepared. These two materials were modified onto two adjacent areas on the ITO electrode. By applying different bias voltage, the cathodic current generated by C-dots/3DGH and the anodic current generated by g-C3N4 can be clearly distinguished and would not interfere with one another. Then E. coli aptamer was modified onto the surface of C-dots/3DGH. In the presence of targets, the binding of E. coli with aptamer lead to the steric hindrance greatly increased and the cathodic current decreased significantly. Meanwhile, the anodic current generated by g-C3N4 was not influenced and it can serve as a stable reference to evaluate the environmental factors. Therefore, the concentration of E. coli can be quantified by the ratio of cathodic current to anodic current, which can effectively eliminate these analyte-independent factors and provide a more precise analysis. In addition, this ratiometric PEC biosensor also showed a good sensitivity and a wide linear range (2.9 cfu/mL to 2.9 × 106 cfu/mL).

Thermal Scanning Conductometry (TSC) as a General Method for Studying and Controlling the Phase Behavior of Conductive Physical Gels.

The thermal scanning conductometry protocol is a new approach in studying ionic gels based on low molecular weight gelators. The method is designed to follow the dynamically changing state of the ionogels, and to deliver more information and details about the subtle change of conductive properties with an increase or decrease in the temperature. Moreover, the method allows the performance of long term (i.e. days, weeks) measurements at a constant temperature to investigate the stability and durability of the system and the aging effects. The main advantage of the TSC method over classical conductometry is the ability to perform measurements during the gelation process, which was impossible with the classical method due to temperature stabilization, which usually takes a long time before the individual measurement. It is a well-known fact that to obtain the physical gel phase, the cooling stage must be fast; moreover, depending on the cooling rate, different microstructures can be achieved. The TSC method can be performed with any cooling/heating rate that can be assured by the external temperature system. In our case, we can achieve linear temperature change rates between 0.1 and approximately 10 °C/min. The thermal scanning conductometry is designed to work in cycles, continuously changing between heating and cooling stages. Such an approach allows study of the reproducibility of the thermally reversible gel-sol phase transition. Moreover, it allows the performance of different experimental protocols on the same sample, which can be refreshed to initial state (if necessary) without removal from the measuring cell. Therefore, the measurements can be performed faster, in a more efficient way, and with much higher reproducibility and accuracy. Additionally, the TSC method can be also used as a tool to manufacture the ionogels with targeted properties, like microstructure, with an instant characterization of conductive properties.

Determination of the Critical Micelle Concentration of Neutral and Ionic Surfactants with Fluorometry, Conductometry, and Surface Tension-A Method Comparison.

Micelles are of increasing importance as versatile carriers for hydrophobic substances and nanoprobes for a wide range of pharmaceutical, diagnostic, medical, and therapeutic applications. A key parameter indicating the formation and stability of micelles is the critical micelle concentration (CMC). In this respect, we determined the CMC of common anionic, cationic, and non-ionic surfactants fluorometrically using different fluorescent probes and fluorescence parameters for signal detection and compared the results with conductometric and surface tension measurements. Based upon these results, requirements, advantages, and pitfalls of each method are discussed. Our study underlines the versatility of fluorometric methods that do not impose specific requirements on surfactants and are especially suited for the quantification of very low CMC values. Conductivity and surface tension measurements yield smaller uncertainties particularly for high CMC values, yet are more time- and substance consuming and not suitable for every surfactant.

A sandwich-type electrochemical immunoassay for ultrasensitive detection of non-small cell lung cancer biomarker CYFRA21-1.

Many studies confirm that the aberrant expression of Cytokeratin 19 fragment 21-1 (CYFRA21-1) is highly correlated with non-small cell lung cancer (NSCLC), especially for squamous cell carcinoma. Herein, we report a sandwich-type electrochemical immunosensor based on signal amplification strategy of multiple nanocomposites to test CYFRA21-1 selectively and sensitively. The proposed immunosensor fabricated by three-dimensional graphene (3D-G), chitosan (CS) and glutaraldehyde (GA) composite on the glass carbon electrode (GCE) with a large surface area is prepared to immobilize primary antibodies (Ab1) and provide excellent conductivity. To further amplify the electrochemical signal, the trace tag on the foundation of gold nanoparticles (AuNPs) is coated with amino-functionalized carbon nanotube (MWCNT-NH2) nanocomposite through thionine linking, which provides more amino groups to capture more horseradish peroxidase-labeled antibodies (HPR-Ab2) and enhances the conductivity. Under optimal conditions, the developed immunosensor exhibits excellent analytical performance for the determination of CYFRA21-1 with a wide linear range from 0.1 to 150ng·mL-1 and a low detection limit (LOD) of 43pg·mL-1. Furthermore, satisfactory results are obtained for the determination of CYFRA21-1 in real clinical serum samples, indicating the potential of the immunoassay to be applied in clinical analysis.