Concentration Of Titrant Research Articles

Protein corona of SiO2 nanoparticles with grafted thermoresponsive copolymers: Calorimetric insights on factors affecting entropy vs. enthalpy-driven associations

The mechanism of protein adsorption on various surfaces of tailored functionality is often accessed by isothermal titration calorimetry (ITC), the powerful tool yielding the full set of thermodynamic parameters in one label-free experiment. In this work, the non-ionic polymer brushes with thermo-switchable hydrophilic-hydrophobic balance, based on diethylene glycol methacrylate (DEGMA) and 4-vinyl pyridine (4VP) copolymers, grafted to SiO2 nanoparticles, were titrated with individual blood proteins, their mixture, and diluted human plasma. The concentration of adsorption sites was calculated from the sizes of protein molecules, assuming the formation of a monolayer. For the titrations with the protein mixture and diluted plasma, the concentration of titrant was an estimate, limiting the accuracy of thermodynamic parameters. The particles with grafted polymer brushes showed negative ξ-potentials similar to the blood proteins and their interactions occurred against the Coulomb forces. Two cases of exothermic hydrophobic and endothermic polar interactions were distinguished. The strength of adsorption, expressed with the affinity constant Ka, was dependent on brush composition and temperature. The adsorption of proteins from blood plasma was always exothermic and, therefore, the dominating hydrophobic interactions were assumed.

Toward quantification of active sites and site-specific activity for polyaromatics hydrogenation on transition metal sulfides

Quantification of active sites is indispensable for understanding a catalytic material, ultimately enabling meaningful comparisons across different catalysts and discerning the underlying intrinsic kinetics. At present, however, few methods are available for quantitatively probing the catalytic surface of transition metal sulfides (TMS) during catalysis. TMS catalysts expose Brønsted acidic (sulfhydryls, SH) and Lewis acidic sites (coordinatively unsaturated sites, CUS) at their surfaces and, thus, are conceptually amenable to in situ acid-base titration with judicious choices of bases that strongly bind to the acidic entities. In this proof-of-concept study, in situ titration using several N-containing organic bases allowed us to determine the surface concentration of active sites and their specific activities for polyaromatics hydrogenation on unsupported TMS catalysts. Ni-promoted sulfide catalysts were more active than the unpromoted ones, because of a modest increase in the site density and a 2- to 3-fold enhancement in the TOF averaged over all titrated sites. By further discriminating between the titrated CUS and SH with site-selective titrants, a substantial difference in the TOF was found between CUS and SH among the titrated pool of sites. In addition to the titrated sites that contributed to the majority (80–95%) of the hydrogenation activity, some active sites remained unoccupied by the studied base molecules. The contribution of these residual sites, likely inaccessible to large-sized and less mobile titrants, depended on the temperature, H2 pressure and the catalyst, but was unaffected by the space time, titrant concentration, and the type or concentration of the polyaromatic reactant, for a given catalyst. This work leads to several emerging opportunities to better understand complex TMS catalysts.

An acidic-groups detection method and its application to analysis of Chinese humic acid samples.

The method of non-aqueous conductivity titration (NACT) of organic weak acids was applied to quickly and accurately determine the phenolic-hydroxyl and carboxyl-groups contents in humic acid. By varying the pH of the humic-acid sample, the concentration of the titrant, and the nitrogen-gas flow rate, the optimal titration conditions were determined to be a sample pH of 4, titrant concentration of 0.05 mol/L, and nitrogen-gas flow rate of 80 mL/min. Applying the detection method to p-hydroxybenzoic acid showed that its phenolic-hydroxyl content was 758.82±111.76 cmol/kg and carboxyl content was 744.44±51.11 cmol/kg. The theoretical phenolic-hydroxyl and carboxyl-groups contents of the p-hydroxybenzoic acid were 723.96 cmol/kg respectively, indicating that the method can accurately quantify the carboxyl and phenolic-hydroxyl groups in the sample. The NACT was used to measure the phenolic-hydroxyl and carboxyl-groups contents in humic acid quickly and accurately. In addition, 29 humic acid samples from 8 provinces of China covering the main humic-acid producing areas were collected and analyzed for acidic-groups content using the reported method.

Determination of Sodium Dodecyl Sulfate via Turbidimetric Titration with Poly(Diallyldimethylammonium Chloride)

AbstractTitration of sodium dodecyl sulfate (SDS) solution with poly(diallyldimethylammonium chloride) (PDDA) results in the formation of colloidal dispersion and gradual increase of turbidity. The titration end point can be determined in different ways, depending on both titrant concentration and optical detector used. The maximum value of turbidity depends on concentration of the titrant solution. An immersion probe is applicable to measure turbidity changes only at low concentrations of the colloidal dispersion. An external optical sensor is well suitable at both low and high concentrations of PDDA titrant. Near to the equimolar ratio of the analyte and titrant, large floccules are formed and turbidity values undergo chaotic fluctuations. Calibration graphs for SDS determination are straight lines. Determination range is from 0.2 to 40 mg L−1.

SPECTROPHOTOMETRIC STUDY OF ACID-BASE PROPERTIES OF SPATIAL-DISTORTED DERIVATIVES OF PORPHYRINS IN ACETONITRILE

Spectrophotometric titration method was used to study the acidic properties of: 5,10,15,20-tetrakis(thienyl-2-yl)-2,3,7,8,12,13,17,18-octaethylporphyrin (I), 5,10,15,20-tetrakis(4’-t-butylphenyl)-2,3,7,8,12,13,17,18-octaethylporphyrin (II), 5,10,15,20-tetrakis(3,5-di-t-butylphe-nyl)-2,3,7,8,12,13,17,18-octaethylporphyrin (III), 5,10,15,20-tetraphenyl-2,3,7,8,12,13,17,18-octaethylporphyrin (IV), 5,15-bis(4’-methoxyphenyl)-10,20-bis(4-nitro-phenyl)-2,8,12,18-tetrame-thyl-3,7,13,17-tetraarylporphyrin (V), 5,15-bis(4’-methoxyphenyl)-10,20-diphenyl-2,3,7,8,12,13, 17,18-octaethylporphyrin (VI), 5,15-bis(4’-methoxyphenyl)-2,8,12,18-tetramethyl-3,7,10,13,17,20-hexaethylporphin (VII), 5,10,15,20-tetrakis(4’-methoxyphenyl)-2,3,7,8,12,13,17,18-octaethylpor-phyrin (VIII) in the presence of a deprotonated agent, 1,8-diazabicyclo[5.4.0] undec-7-ene, in acetonitrile. Under deprotonation two families of spectral curves in the absorption spectra, having its own system of isobestic points, are formed. Electron absorption spectrum of molecular form transforms gradually into the spectrum of the final form of dianion with titrant concentration increase. Determination of the coordinates of the inflection (and the corresponding concentration of DBU) on the titration curve allowed us to distinguish two areas in the electron absorption spectra of the reaction system, which are likely belong to the first and second steps of deprotonation, namely, to the formation of mono- and dianionic forms (HP– and P2–) of the compound. Spectral characteristics of the ionized forms and combined ionization constant for the first and second steps (lgКа1 and lgКа2) were revealed. At the first step in the order of decreasing acidic properties of the ligands the compounds form a series of: III< V< VIII< VI < VII< II< IV < I; at the second step in decreasing order of the acidic properties of the ligands the compounds form a series of:.III < V < VI < II <VIII< VII < IV < I. The effect of structural and electronic properties of substituents on acid-base properties of porphyrins is analyzed. Thus, chemical modification of the macrocycle can lead to direct changes in acid-base properties of supramolecular macroheterocyclic ligands and can be a powerful tool to control the reactivity of compounds of the porphyrins class.

The Determination of Lower Acidity in Several Coloured Oils by Catalyzed Thermometric Titration

A novel method of catalytic thermometric titration has been developed for the determination of the acidity of oils using titration with KOH in isopropanol as titrant. The amounts of oil, concentration and delivery rate of titrant, volumes and types of thermometric end-point indicator were investigated. The results show that mixture of acetone and chloroform used as indicator exhibits strongly exothermic effects to reflect end point obviously. The results obtained from the thermometric titration have good agreement with standard potentiometric and visual titration methods. The procedure is fast, easy to use, accurate, and highly reproducible to measure lower acidity in coloured oils. It is very suitable for the routine process and quality control of many types of oils.

Thermometric Titration for Rapid Determination of Trace Water in Jet Fuel

Water content in jet fuels is detected by thermometric titration (TMT), and the optimal detected system is 2,2-dimethoxypropane as titrant, cyclohexane and isopropanol as titration solvents, and methanesulfonic acid as catalyst in this method. The amounts of oil, concentration and delivery rate of titrant, volumes, and the reliability and accuracy of thermometric titration were emphasized. The results show that the accuracy, validity, and reliability of TMT are excellent by different indicated spiked water contents. The obtained results between TMT and Karl Fischer titration have been proven to be in accord. But, the duration of titration merely spends 3–5 min in the whole process, greatly shortening the detected time. Therefore, rapid and accurate determination of trace water in a jet fuel can be realized by TMT.

Semiautomated pH gradient ion-exchange chromatography of monoclonal antibody charge variants.

A new approach using a chromatography system equipped with isocratic pumps and an electrolytic eluent generator (EG) is introduced, replacing external pH gradient delivery using conventional gradient systems, in which bottled buffers with preadjusted pH are mixed using a gradient pump. The EG is capable of generating high purity base or acid required for online preparation of the buffer at the point of use, utilizing deionized water as the only carrier stream. Typically, the buffer was generated from online titration of a reagent composed of low molecular weight amines. The reagent was delivered isocratically into a static mixing tee, where it was titrated to the required pH with electrolytically generated base or acid. The required pH gradient was thus conveniently generated by electrically controlling the concentration of titrant. Also, since the pH was adjusted at the point of use, this approach offered enhanced throughput in terms of eluent preparation time and labor, and with a more reproducible pH profile. The performance of the system was demonstrated by running pH gradients ranging from pH 8.2 to 10.9 on a polymer monolith cation-exchange column for high throughput profiling of charge heterogeneity of intact, basic therapeutic monoclonal antibodies. A high degree of flexibility in modulating the key parameters of the pH gradient, including the buffer concentration, the pH gradient slope and the operating pH range was demonstrated. This enabled fine-tuning of the separation conditions for each individual antibody in order to enhance the chromatographic resolution.

Quantitative determination of anionic surfactants using optical sensors with microtiter plate reader

In this study, it is shown that anionic surfactants can be readily determined by using polymer film-based optodes containing an appropriate chromoionophore. The optodes are prepared with poly(vinyl chloride) (PVC), polyurethane (PU), bis(2-ethylhexyl) sebacate (DOS), and 2′,7′-dichlorofluorescein octadecylester (DCFOE). These optodes exhibit reproducible and sensitive absorbance changes in response to the varying titrant concentrations. Three different cationic surfactants, cetyltrimethylammonium bromide (CTAB), benzyldimethyltetradecylammonium chloride (zephiramine, Zeph), and benzyldimethylhexadecylammonium chloride (CDB), are employed as titrants to determine anionic surfactant concentrations. In addition, membrane electrodes were also employed for estimating the interactions between the cationic and anionic species. The reaction stoichiometries are compared based on the electrode measurements and Scatchard plots are prepared to estimate the strength of binding interaction of titrants with the target surfactants.

Practical application and statistical analysis of titrimetric monitoring of water and sludge samples

Titrimetry offers the possibility of simultaneous measurement at low cost of several (buffering) components. A first step in the study towards practical application of the titrimetric technique was the titrimetric analysis by up- or down-titration of standard solutions, standard mixtures, solids digester samples and water samples coming from autotrophic nitrogen-removal reactors. The resulting raw data were further processed with an Excel-based program. This program first converts the raw data into a buffer curve upon which a linear buffer capacity model is fitted to the experimental data by estimating the (buffer) concentrations and corresponding pKa values. As such the type of component and the concentration can be determined. As a second step the resulting calculated concentrations were analysed statistically to assess the accuracy and precision of the titrimetric technique. For this purpose, the data were paired, i.e. the difference between the concentration obtained with titrimetry and the concentration obtained with another technique such as colorimetry or gas chromatography was calculated. First the normality of the paired data was assessed. Then, a paired t-test (normal data) or a paired Wilcoxon test (normal data) was used to statistically compare the results obtained with the titrimetric technique to either the stock solution concentration or measurements with another method (colorimetry or gas chromatography). The statistical tests showed that, depending on the titrant concentration, concentrations from 50 mg/. to 3 000 mg/. could adequately be measured with the titrimetric technique.

Application of dc and mark-space bias differential electrolytic potentiometry for determination of cyanide using a programmable syringe pump

This paper describes a rapid sequential injection titration method for the determination of cyanide in aqueous solutions. Mercuric nitrate was used as a titrant and a pair of gold-amalgam electrodes as an indicating system. The technique of differential electrolytic potentiometry using both mark-space bias (m.s.b.) and dc current for polarization was employed. The optimum values of current and percentage bias were 5 μA and 13%, respectively. The effects of dispense time, volume of analyte, supporting electrolyte, and the concentration of titrant were investigated. The results obtained are in agreement with those of the standard method (APHA), with a relative standard deviation of 1.43%, t = 0.783, F = 1.713. A sampling rate of about 20 samples per hour was achieved with good reproducibility and lower consumption of reagents.

Spectrophotometric determination of various polyanions with polymeric film optodes using microtiter plate reader

Polycation-sensitive membrane optodes based on the chromoionophore 2′,7′-dichlorofluorescein octadecylester (DCFOE) have previously been developed and used for determination of heparin via a titrimetric method. In this study, it is shown that some other important polyanions such as PPS (pentosan polysulfate), DNA, xanthan, Na-alginate, and carrageenan (food additive) can also be readily determined by using DCFOE-based microtiter plate-format optodes (MPOs) and polycationic titrants that bind these polyanionic species. The optical sensors are prepared with poly(vinyl chloride) (PVC), polyurethane (PU), bis(2-ethylhexyl)sebacate (DOS), and 2′,7′-dichlorofluorescein octadecylester (DCFOE) and exhibit reproducible and sensitive absorbance changes in response to the varying polycationic titrant concentrations. Three different polycations; protamine, poly- l-lysine and poly- l-arginine, are employed as titrants. The method has a detection limit of 1 μg mL −1, and a dynamic range of 1–40 μg mL −1. After the quantitative determinations are successfully demonstrated in buffered solutions, similar titrations are also performed in real samples. The method is validated by recovery studies in these samples. The average polyanion recoveries were quantitative [99.7(±1.3) % for pastry cream with vanillin (protamine titrant); 100.4 (±3.3) % for pastry gel with strawberry(PLA titrant), and 102.9(±2.0) % for pastry gel with strawberry (PLL titrant)].

Explicit formulation of titration models for isothermal titration calorimetry

Isothermal titration calorimetry (ITC) produces a differential heat signal with respect to the total titrant concentration. This feature gives ITC excellent sensitivity for studying the thermodynamics of complex biomolecular interactions in solution. Currently, numerical methods for data fitting are based primarily on indirect approaches rooted in the usual practice of formulating biochemical models in terms of integrated variables. Here, a direct approach is presented wherein ITC models are formulated and solved as numerical initial value problems for data fitting and simulation purposes. To do so, the ITC signal is cast explicitly as a first-order ordinary differential equation (ODE) with total titrant concentration as independent variable and the concentration of a bound or free ligand species as dependent variable. This approach was applied to four ligand–receptor binding and homotropic dissociation models. Qualitative analysis of the explicit ODEs offers insights into the behavior of the models that would be inaccessible to indirect methods of analysis. Numerical ODEs are also highly compatible with regression analysis. Since solutions to numerical initial value problems are straightforward to implement on common computing platforms in the biochemical laboratory, this method is expected to facilitate the development of ITC models tailored to any experimental system of interest.

Comparative Biophysical Characterization of p53 with the Pro-apoptotic BAK and the Anti-apoptotic BCL-xL

The p53 transcription-independent apoptosis in mitochondria, mediated by its interaction with the pro-apoptotic and the anti-apoptotic members of the Bcl2 family of proteins, has been described in vivo, especially in radiosensitive tissues. We have characterized the interaction of p53 with both the pro-apoptotic Bak and the anti-apoptotic Bcl-x(L) proteins, comparing their affinity and their interaction surfaces, using biophysical techniques such as fluorescence anisotropy, analytical ultracentrifugation, and NMR. We have shown that both proteins interact with only the p53 core domain and not with its N- and C-terminal regions. Further, p53 has a higher affinity for Bcl-x(L) than for Bak, which is consistent with the previously described sequential binding of Bcl-x(L) and Bak by p53. Interestingly, although the interaction with both proteins is electrostatic in character, they have different binding sites. Using NMR spectroscopy, we have determined that Bcl-x(L) interacts with the DNA binding site of p53, but Bak does not interact with this site. A new potential interaction surface for Bak is proposed.

Sensitive Potentiometric Method for Determination of Micromolar Level of Polyethoxylated Nonionic Surfactants in Effluents

Abstract The Metrohm NIO surfactant electrode has been used as end-point indicator for potentiometric titration of low concentration level of polyethoxylated nonionic surfactants. This can be achieved by using of a diluted titrant concentration, thus reducing the amount of precipitate formed during titration and preventing the electrode deterioration. The solutions of low levels (down to 10−6 mol/L) of selected nonionic surfactants containing 5 to 23 EO groups were successfully titrated with diluted (as low as 10−4 mol/L) sodium tetraphenylborate as standard anionic titrant, increasing up to 20 times the sensitivity of the method. The low surfactant concentration has been determined in synthetic formulations of widely used detergent products and industrial waste waters. The titration end-point has been determined by applying extended Savitzky-Golay least-squares regression. The accuracy and precision has been evaluated by using the standard addition method. Relative standard deviation within results was between 3.4 and 12.8 % depending on the sample complexity and the surfactant concentration level.

Interaction of “extremely diluted solutions” with aqueous solutions of hydrochloric acid and sodium hydroxide: A calorimetric study at 298 K

The “extremely diluted solutions” (EDS) have revealed a really intriguing behaviour, characterized by multiple independent variables. Because of their behaviour, EDS can be described as far-from-equilibrium systems, capable of auto-organizing as a consequence of little perturbations. We measured the heats of mixing of basic and acid solutions with such EDS and their electrical conductivity, comparing with the analogous heats of mixing and electrical conductivity of the untreated solvent. In particular, calorimetric titrations have been performed with NaOH or HCl solutions at various concentrations. Plots of the excess heat as a function of the concentration of titrant reveal differences and similarities between the two. By analysing these plots we were able to formulate new hypotheses about the supramolecular organization of the solvent water, when subject to mechanical perturbations stemming from the EDS preparation protocol, that is comprised of an iterative process of successive dilutions and succussions.

FIA acid–base titrations with a new flow-through pH detector

A pH-sensitive detector for flow-through potentiometry based on a graphite/quinhydrone composite electrode was applied for flow-injection analysis (FIA) titrations. Hydrochloric acid and acetic acid were titrated by injection of samples into a sodium hydroxide carrier solution. System conditions were optimised by variation of flow rate, injection volume and titrant concentration. The parameters sampling frequency, residence time and dispersion coefficient were determined. The evaluation of peak width (time between the two inflection points on each side of the peak), peak area and slope of the raising edge of the peak lead were studied with respect to their use for calibration. Hydrochloric acid and acetic acid could be titrated down to a concentration of 2 x 10(-4) mol L(-1) using 150-microL injection volumes, which is almost ten times lower than can be achieved using colour indicators and a spectrophotometric detection.

A program for the simultaneous potentiometric determination of protonation constants and electrode calibration parameters

A program for the potentiometric determination of the protonation constants of mononuclear polyprotic substances is described. A maximum of twelve parameters can be determined simultaneously, including up to six protonation constants, four electrode calibration parameters, the protolysis constant of the solvent and the titrant concentration. Optimization is carried out by using the non-simplified Newton-Raphson method, which is potentiated by the Marquardt algorithm and a distance speeding-up coefficient. A direct search method is also used to improve the initial set of values. Variances are calculated very accurately, since the real Hessian function is used. Statistical weights and ionic strength corrections are also considered. The program has been tested by using simulated titration curves of polyprotic acids with close constants.

A novel methodology for evaluation of formation constants of complexes: example of lanthanide–Arsenazo III complexes

A novel method is proposed for the computation of formation constants of complexes. It makes use essentially of the data from the first stages of the titration and is based on the Taylor expansion of the concentration of the ligand as a function of the titrant concentration. The applicability of the methodology is examplified by the computation of the complexation constants of lanthanides (Ln) by 3,6-bis[(2-arsonophenyl)azo]-4,5-dihydroxy-2,7-naphtalenedisulphonic acid disodium salt (Arsenazo III). Arsenazo III is known to form stable complexes with a large variety of ions. The spectrophotometric study of these complexes in the visible domain is facilitated by the presence of chromophore groups in the ligand. In the present work, spectral deconvolution was used to improve the accuracy in the determination of formation constants of complexes between Arsenazo III and lanthanides. It turns out that all the 11 lanthanides studied here form the same type of complexes with this ligand. For each of them, the three formation constants correspond, respectively, to Ln to Ar ratios 1:1, 1:2 and 2:2. Their approximate values are in the ranges: K 11=10 5; K 12=10 10; K 22=10 15. Accurate values are given in the text. The results are compared with those obtained with the classical iteration method which makes use of the whole titration curve. The two techniques give satisfactorily convergent results. The new methodology might be useful in the treatment of the competition of two ligands for the complexation of the same lanthanide, where the first stages of the titration of only one of the complexants is experimentally available.

Thermodynamics of Cationic Lipid-DNA Complex Formation as Studied by Isothermal Titration Calorimetry

The detailed analysis of the cationic lipid-DNA complex formation by means of isothermal titration calorimetry is presented. Most experiments were done using 1,2-dioleyl-sn-glycero-3-ethylphosphocholine (EDOPC), but basic titrations were also done using DOTAP, DOTAP:DOPC, and DOTAP:DOPE mixtures. Complex formation was endothermic with less than 1kcal absorbed per mole of lipid or DNA charge. This enthalpy change was attributed to DNA-DNA mutual repulsion within the lamellar complex. The exception was DOTAP:DOPE-containing lipoplex for which the enthalpy of formation was exothermic, presumably because of DOPE amine group protonation. Experimental conditions, namely, direction and titration increment as well as concentration of titrant, which dictate the structure of resulting lipoplex (whether lamellar complex or DNA-coated vesicle), were found to affect the apparent thermodynamics of complex formation. The structure, in turn, influences the biological properties of the lipoplex. If the titration of lipid into DNA was carried out in large increments, the ΔH was larger than when the injection increments were smaller, a finding that is consistent with increased vesicle disruption under large increments and which is expected theoretically. Cationic lipid-DNA binding was weak in high ionic strength solutions, however, the effective binding constant is within micromolar range because of macromolecular nature of the interaction.