0000000000139585
AUTHOR
M.c.garcía Alvarez-coque
Acid—base properties of azo dyes in the presence of surfactants
Abstract The changes in the acid—base properties of an azo dye produced by the presence of surfactants, at surfactant concentrations below and above the CMC, are quantitatively described using a simple model and spectrophotometric and potentiometric measurements. The azo dye formed by aniline and N -(1-naphthyl)-ethylenediamine, and the surfactants sodium dodecyl sulphate, Triton X-100 and N -cetylpyridinium chloride (NCPC) are used. The protonation constants of the free and surfactant-bound dye species, the binding constant of the protonated and unprotonated forms of the dye, and the average aggregation number of the dye—NCPC aggregates are evaluated.
Determination of tyrosine and phenylalanine by derivatization with nitric acid and differential pulse polarography
Abstract The determination of tyrosine and phenylalanine by differential pulse polarography, after separation by thin-layer chromatography and derivatization with nitric acid to form the nitro compounds, is proposed. Experimental conditions for the derivatization treatment are established and the polarographic determination is optimized.
Quantitation of hydrophobicity in micellar liquid chromatography
Abstract Micellar liquid chromatography (MLC) is shown to be a promising technique for measuring the hydrophobicity of solutes. The presence of micelles has a profound effect on the chromatographic characteristics of reversed-phase columns. The linear relationships between the logarithm, log k , of the retention factor and such diverse properties as the number of carbon atoms in homologous series, octanol–water partition coefficients and solvatochromic parameters, which are observed in conventional reversed-phase liquid chromatography (RPLC), are not usually valid in MLC. For series of compounds exhibiting a wide range of hydrophobicity, k itself is linearly related to these properties. The…
The catalytic-thermometric determination of molybdenum in steels based on the hydrogen peroxide-iodide indicator reaction
Abstract The Mo VI -catalysed hydrogen peroxide-iodide reaction has been optimised for thermometry in an HCl medium using the initial slope method. The substitution of sodium perborate by H 2 O 2 and the conversion of the reaction to a process showing a Landolt effect are considered and discussed. The linear range extended from 0.2 μg Mo VI ml −1 to 1.2 μg Mo VI ml −1 with a detection limit of 0.06 μg Mo VI ml −1 . Interferences were also studied. The optimised procedure was applied to the evaluation of molybdenum in steels after extraction with α-benzoinoxime in chloroform.
Some observations on the automation by flow injection analysis of the spectrophotometric determination of amino acids and proteins witho-phthalaldehyde
Automation by flow injection analysis with Spectrophotometric detection of the determination of total amino acids and proteins witho-phthalaldehyde is not straightforward. The use of spectrophotometry, instead of spectrofluorimetry, and of N-acetyl-L-cysteine, instead of the conventional mercaptoethanol is advantageous because of the lower variability of absorptivities with respect to fluorescence yields, and the larger stability of the derivatives. Under adequate working conditions and with leucine as reference, the procedure can be used for the evaluation of total amino acids. A similar procedure is proposed for the analysis of proteins in a sample. Limits of detection are ≈ 1 × 10−5M for…
Modelling of the elution behaviour in hybrid micellar eluents with different organic modifiers
Abstract The elution behaviour in reversed-phase liquid chromatography (RPLC), with mobile phases containing micelles, can be described according to a distribution pattern among three phases: bulk water, micellar pseudophase and stationary phase. Several mechanistic equations based on this pattern, previously used to predict the retention in hybrid mobile phases of sodium dodecyl sulphate and propanol, were checked with several organic modifiers and a group of solutes of diverse polarity. The modifiers were the alcohols 1-propanol, 1-butanol and 1-pentanol, and other organic solvents widely used in conventional RPLC such as acetonitrile and tetrahydrofuran. New models were also proposed to …
Reduction of Convective Low-Frequency Noise in Thermal Lens Spectrometry
The construction and optimization of a thermal lens spectrometer with coaxial pump/probe configuration is reported. The low-frequency noise of convective origin is studied in detail, and it is demonstrated that a horizontal slit, instead of a pinhole, can be used as a simple way of reducing the noise to a negligible level without losing sensitivity.
Spectrophotometric determination of mercury(II) and silver(I) with copper(II) and diethyldithiocarbamate in the presence of triton X-100
Abstract Procedures for the determination of mercury and silver by displacement of diethyldithiocarbamate (DDTC) from its copper complex in the presence of 1% Triton X-100, and measurement of the decrease in the Cu(DDTC) 2 absorbance, are described. The use of the surfactant avoids the need for an extraction step. Reproducibility within 1% and detection limits of 0.25 ppm Hg(II) and 0.45 ppm Ag(I) have been obtained, and linear calibration ranges up to 13 ppm Hg(II) and 15 ppm Ag(I). In the presence of 0.1 M EDTA very good selectivity is achieved.
Examination of the least-squares method applied to the evaluation of physicochemical parameters with linearized equations
Physicochemical parameters are frequently evaluated by the least-squares method after linearization of the equation which relates the experimental variables to the parameters of interest. When a given set of data is treated to evaluate (n+1) parameters. (n+2) nominally identical linearized equations are possible; each of these leads to different sets of values of the parameters which are also affected by different variances. These differences are also observed when statistical weights are used. Two methods for establishing which one of the (n+2) equations is best for fitting the data, i.e., simulation of experiments and propagation of errors, are discussed, compared and applied to a potenti…
Thermal lens spectrometric determination of cerium with oxine
Abstract The spectrophotometric and photothermal (TLS) procedures for cerium determination using 8-hydroxyquinoline (oxine), after extraction into chloroform, are compared. Photothermal measurements are made using a coaxial pump/probe thermal lens spectrometer. The use of high-purity reagents at low concentrations permits a decrease in the TLS blank signal and noise, leading to a limit of detection of 9 × 10−9 M (cerium extract concentration), 40-fold lower than the spectrophotometric value. The dynamic range extended up to 6 × 10−7 M and the relative standard deviation for 5 × 10−7 M cerium was 3.9%.
Spectrophotometric Determination of Cystine with O-Phthalaldehyde in the Absence of Thiol
Abstract A spectrophotometric method for the determination of cystine with o-phthalaldehyde (OPA) in the absence of thiol is described. When cystine is heated at 60[ddot]C for 30 min in a low excess of OPA (pH 9.5), a very stable derivative with 1:2 stoichiometry (cystine:OPA) and an absorption maximum at 335 nm (E = 4600) is formed. At pH < 1 the derivative is protonated (protonation constants: log K1 = 5.88 and log K2 = 3.70 at I = 0.1 and 20[ddot]C) and another absorption band at 440 nm (E = 3800) appears, which allows the determination of cystine in the presence of other amino acids.
Use of the o-Phthalaldehyde and N-Acetyl-L-Cysteine the Evaluation of Milk Proteins
Abstract o -Phthalaldehyde and N-acetyl-Lcysteine are used in the determination of milk proteins. Three procedures are proposed and compared. One of them is based on reaction of o -phthalaldehyde and N-acetyl-Lcysteine with the intact proteins and the two others on reaction of the reagents with the released amino acids after total acid hydrolysis of the protein samples. When the protein sample is hydrolyzed, calibration is performed either with a hydrolyzed protein standard or with isoleucine. A procedure for the measurement of the degree of enzymatic hydrolysis of milk proteins without separation of the unhydrolyzed protein, which makes use of the same reagents, is also described. In all c…
Determination of Amino Acids by Micellar High-Performance Liquid Chromatography and Pre-column Derivatization withO-Phthalaldehyde and N-Acetyl-L-cysteine
Abstract Micellar liquid chromatography of proteic primary amino acids with pre-column derivatization with o-phthalaldehyde (OPA) and N-acetyl-L-cysteine was studied, using mobile phases containing a short-chain alcohol. The modification of pH gave a large variation of the retention as a result of the protonation of the carboxylate group of amino acids. Maximum resolution and adequate retentions were achieved with a 0.05 M sodium dodecyl sulphate/3% propanol mobile phase at pH 3. The reproducibility was lower than 1.0% at a 1 × 10−4 M concentration level and between 0.6 and 2.2% for 1 × 10−6 M. The determination of glycine, lysine, methionine and threonine in pharmaceutical formulations gav…
Some observations on the determination of sulfite by reduction to sulfide with sodium borohydride
Abstract Sodium borohydride is studied as an agent for reducing sulfite to sulfide. Rapid reductions are attained with greater than 97% recovery in the range 700–1200 μg of S/50 ml. Sulfate is not reduced.
Determination of aniline in vegetable oils by diazotization and coupling in a microemulsion medium
Abstract A microemulsion containing sodium dodecyl sulphate and n -pentanol in a mass ratio of 1 : 4, water and a vegetable oil was investigated using pseudo-tenary phase diagrams. The medium can co-solve important amounts of vegetable oils and aqueous solutions over a wide range of ionic strengths. A procedure for the determination of 1.4–140 μg ml −1 of aniline in vegetable oils using ionic diazotization and coupling reactions was developed. The absorbance was measured in an optically clear microemulsion containing 4% or 20% of oil. The procedure is much simpler and rapid than the official chromatographic methods and gives almost the same limits of detection (ca. 05 μg ml −1 ) using no mo…
Direct injection of physiological fluids in micellar liquid chromatography
Abstract Micellar liquid chromatography (MLC), which uses mobile phases of surfactants above the critical micellar concentration, provides a solution to the direct injection of physiological samples by solubilizing the protein components, and coating the analytical column with surfactant monomers to avoid clogging. A review showing the advantages and limitations of this technique over other chromatographic techniques used in drug analysis, working protocols, and examples of application is presented. The possibility of direct sample introduction simplifies and greatly expedites the treatments with reduced cost, improving the accuracy of the procedures. Surfactant monomers and micelles appear…
Spectrophotometric determination of cystine by formation of an o-phthalaldehyde/N-acetyl-l-cysteine derivative
Abstract Cystine reacts with o -phthalaldehyde (OPA) in the absence and presence of a thiol compound to yield different compounds. The use of N -acetyl- l -cysteine as thiol leads to the formation of two derivatives, likely simple and double isoindoles, where the disulfide bond remains unbroken. In contrast, mercaptoethanol gives rise to the reduction of the amino acid to form a cysteine derivative. Obtaining cystine isoindoles makes it possible to spectrophotometrically determine the amino acid after Chromatographic separation and is further evidence of the large stabilization effect produced by N -acetyl- l -cysteine in the formation of OPA-thiol derivatives.
Formation rates and protonation constants of azo dyes in a sodium dodecylsulphate micellar solution.
The effects of a sodium dodecylsulphate micellar solution on the coupling rates of several diazotizated arylamines with N-(1-naphthyl)-ethylenediamine and the shifts in the protonation constants of the corresponding azo dyes are quantitatively studied. Aniline, o-, m- and p-aminobenzoic acids, ethyl p-aminobenzoate and several sulphadrugs are used, and the relationships among the intensity of the effects and the molecular structure of the diazonium ions and the dyes are discussed. A single simplified procedure for the determination of all the substances at pH = 1.3 +/- 0.3, where coupling is quickly completed to directly produce the protonated form of the dyes, is established and applied to…
Graphical representation of polynuclear acid-base and complex equilibria.
Abstract A calculation procedure for diagrams of log CM = ƒ(pL) at various values of the ratio of analytical concentrations, CL/CM, in polynuclear binary systems is described. The diagrams are useful in the resolution of analytical problems of practical interest involving acid-base equilibria (such as the preparation of buffer solutions) and complex equilibria. The presence of a solid phase is also considered.
Determination of total free amino acids with o-phthalaldehyde and N-acetyl-l-cysteine
Abstract A spectrophotometric procedure is proposed for the determination of total free amino acids after reaction with o-phthalaldehyde and N-acetyl- l -cysteine, using isoleucine as the reference. The procedure was applied to the analysis of five samples of widely different composition. Recoveries were 98–105%.
Available Lysine in Protein, Assay Using o-Phthalaldehyde/ N-Acetyl-L-cysteine Spectrophotometric Method
An assay was based on reaction of free e-amino groups in proteins with the o-phthalaldehyde/N-acetyl-L-cysteine reagent to form isoindoles, which absorb at 335 nm. The procedure was suitable for proteins or mixtures of proteins with available lysine contents of more than 5 moles lysine/mole protein and required absence of free amino acids and peptides. This method was simpler and more convenient than other methods, since it did not require hydrolysis, amino acid analysis, long heating periods or solvent extraction.
Evaluation of the proteolysis degree with the o-phthalaldehyde/N-acetyl-L-cysteine reagent
The o-phthalaldehyde/N-acetyl-L-cysteine (OPA-NAC) reagent is applied to the spectrophotometric evaluation of the proteolytic activity of enzymes. The high stability of the OPA-NAC isoindoles makes a strict control of the time of reaction unnecessary. A mathematical expression is proposed to calculate proteolysis degrees, where the absorbance decrease of the OPA-NAC derivative of the protein itself during the hydrolysis process is taken into account. The method is applied to bovine serum albumine, caseine, lysozyme, lactoglobuline and protamine sulphate as substrates, and pronase, papaine, trypsin and chymotrypsin as enzymes.
A kinetic-catalytic method for iodide based on the simultaneous addition of inhibitor and substrate
Abstract A kinetic-catalytic method of analysis, based on the addition of a mixture of inhibitor and substrate, at a constant rate, over the catalyst in the presence of the other component of the indicator reaction is studied. The method is applied to the determination of iodide using the Ce(IV)As(III) catalytic reaction and Hg(II) as inhibitor. Under suitable experimental conditions the absorbance of the unconsumed Ce(IV) is inversely proportional to the initial iodide concentration.
Micellar Liquid Chromatography: A Worthy Technique for the Determination of β-Antagonists in Urine Samples
Several beta-antagonists (acebutolol, atenolol, celiprolol, labetalol, metoprolol, nadolol, propranolol) were determined in urine samples with fluorometric detection after direct injection, in less than 15 min, with a micellar mobile phase of 0.1 M sodium dodecyl sulfate (SDS), 15% propanol, and 1% triethylamine at pH 3. The limits of detection (38 criterion) were usually between 3 and 30 ng/mL. The addition of propanol and triethylamine and the reduction of the pH of the mobile phase improved the efficiency of the chromatographic peaks that was rather low in pure micellar eluents. The selection of the composition of the mobile phase was easily performed through the use of an interpretive p…
High-performance liquid chromatographic determination of diuretics in urine by micellar liquid chromatography.
The use of micellar liquid chromatography for the determination of diuretics in urine by direct injection of the sample into the chromatographic system is discussed. The retention of the urine matrix at the beginning of the chromatograms was observed for different sodium dodecyl sulphate (SDS) mobile phases. The eluent strengths of a hybrid SDS-methanol micellar mobile phase for several diuretics were compared and related to the stationary phase/water partition coefficient with a purely micellar mobile phase. The urine band was appreciably narrower with a mobile phase of 0.05 M SDS-5% methanol (v/v) at 50 degrees C (pH 6.9). With this mobile phase the determination of bendroflumethiazide an…
On the preparation of simple and universal buffers including polynuclear species
A simple generalized procedure for the calculation of electrolyte concentrations in pH-buffers is proposed. Mixtures of acid-base systems and formation of polynuclear species at high ionic strengths are considered, and a diagram useful for the study and preparation of the buffers is shown.