Thermodynamic characteristics of adsorption of some 1,3,4-oxadiazoles and 1,2,4,5-tetrazines from water-acetonitrile solutions on phenyl-bonded silica gel

Standard differential molar changes of the Gibbs energy, enthalpy, and entropy during adsorption in the Henry region of 17 derivatives of 1,3,4-oxadiazole and 1,2,4,5-tetrazine from water-acetonitrile solutions with acetonitrile concentrations of 20 to 80 vol % and a variation step of 5 vol % on silica gel with grafted phenyl groups at column temperatures of 313.15 to 333.15 K with at a step of 5 K are determined under dynamic conditions by high-performance liquid chromatography. The dependence of the thermodynamic characteristics of adsorption on the molecular structure of adsorbates is discussed. It is established that the absolute values of the changes in the Gibbs energy and enthalpy during adsorption as a whole increase with increasing Van der Waals surface area and volume of the adsorbate molecules, while the adsorption of most 1,3,4-oxadiazoles is characterized by a dependence close to isentropic. It is assumed that the determining role in the change of entropy during adsorption from solutions is played by the polarity of the adsorbate molecule and the conformational flexibility of its substituents.     

Preparative high-performance liquid chromatography with reversed phase packed glass columns

An efficient system for preparative reversed-phase separations with packed glass columns is described. The advantage of this system is the use of relatively simple and inexpensive equipment. Column performance, load capacity, effect of the feed volume and the feed concentration on peak broadening are shown. The influence of the selectivity and the capacity factors on column load have been measured. The effect of the column dimensions is demonstrated by means of practical applications. The loading capacity of a column depends on the thermodynamic proporties of the separation system used. It is therefore not expedient in preparative chromatography to correlate the loading capacity of a column by means of grams dissolved per grams of adsorbent.     

The evaluation of “reversed phase” high-performance liquid chromatography packing materials

Summary For the evaluation of reversed phase packing materials a mixture of acetylacetone, I-nitronaphthalene and naphthalene is proposed. This will reveal the usual optimum kinetic chromatographic parameters (the naphthalene peak), the degree of activity or endcapping status of the column (the ratio of the I-nitronaphthalene and naphthalene retention times) and trace metal activity (the shape and intensity of the acetylacetone peak).     

Thermodynamics of the sorption of 1,3,4-oxadiazole and 1,2,4,5-tetrazine derivatives from solutions on hypercrosslinked polystyrene

The sorption of a series of aromatic heterocycles on neutral hypercrosslinked polystyrene from maximally dilute acetonitrile and water.acetonitrile solutions of them is investigated by high-performance liquid chromatography in a range of column temperatures from 308 to 348 K at 5 K intervals. It is found that for all heterocycles, the logarithm of the retention factor versus the reciprocal of the temperature dependences can be approximated by linear functions with r ² ≥ 0.999. It is established that the true compensation effect (which does not result from the correlation between enthalpy and entropy determination errors) occurs during the sorption of heterocycles on hypercrosslinked polystyrene from both acetonitrile and water-acetonitrile solutions. It is shown that in the compensation diagram, the points corresponding to sorbates are grouped together according to the type of sorbate-sorbent interactions. It is found that changes in the Gibbs energy, enthalpy, and entropy of sorption from acetonitrile solutions and for sorption from water-acetonitrile solutions are related to one another by linear dependences.     

A reversed phase high-performance liquid chromatography method for determination of cephalexin in human plasma

A reversed-phase HPLC method for the analysis of cephalexin (7-[(aminophenylacetyl)amino]-3-methyl-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2-carboxylic acid) by isocratic separation is described. A comparison is made between the ultrafiltration land the extraction procedure developed in our laboratory. The extraction procedure, based on the deproteinization of plasma with perchloric acid followed by the extraction with dichloroethane and separation of cephalexin on a reversed phase column, gave better recovery. Quantitative validation of our method has been performed by an external standard technique. The relative standard deviations were between 1.1 and 1.9% in the within-day assay and between 1.2 and 2.2% in the inter-day assay. The limit of detection was 0.14ug/ml and the limit of quantification was 0.28ug/ml. This method is rapid, sensitive, simple, and reproducible. The LC MS/MS technique was successfully used for identification and quantification of cephalexin in human plasma samples.     

Analysis of terpenes fromGinkgo biloba L. extracts by reversed phase high-performance liquid chromatography

Summary The main terpenes ofGinkgo biloba L. extracts (bilobalide, ginkgolide A, ginkgolide B and ginkgolide C) have been separated by isocratic elution on a 3 μm C₁₈ Spherical column using 2-propanol:water (10∶90) as eluent.     

The chemistry of hexahydro-1,2,4,5-tetrazines—II: Mass spectra of 1,4-dialkylhexahydro-1,2,4,5-tetrazines and 1,4-dialkyl-2,5-diacylhexahydro-1,2,4,5-tetrazines

The mass spectra of twenty-eight 1,4-dialkylhexahydro-1,2,4,5-tetrazines and 1,4-dialkyl-2,5-diacylhexahydro-1,2,4,5-tetrazines (1) have been recorded and interpreted with the aid of deu-terium labelling, high resolution measurements, and the metastable defocusing technique. The principal fragmentations involve initial scission of ring bonds, particularly C-N bonds, leading to characteristic ions of mass M/2 and M/2± 1, and to 2,3-diaza-1,3-butadienyl ions (II). In the spectra of acylated compounds, acyl migration occurs within the molecular ion. The elimination of formaldehyde from the principal fragment ion is discussed.     

Negentropy and retention in reversed phase liquid chromatography

Summary In the proposed concept an eluate is considered to be an information source and the chromatographic column as a non specific receptor capable of understanding the information content. Molecular negentropy, estimated using Shannon's equation has been used as a measure of the information content. Retention of some experimental solutes has been shown to vary with the respective negentropy.     

Role of the organic solvent modifier in the reversed phase high-performance liquid chromatography of polypeptides

Summary The retention behaviour of several polypeptide hormones on Bondapak C₁₈ columns has been examined using mobile phases of different acetonitrile percentage compositions containing 15 mM triethylammonium phosphate or 15 mM orthophosphoric acid buffers. Under these elution conditions, the capacity factors and the selectivity parameters of these polypeptide hormones show pronounced dependencies on the volume fraction of the organic solvent modifier. In the range 0–40% acetonitrile, the capacity factors were monotonously attenuated with increasing modifier percentages with the elution order essentially in accord with that hat anticipated for a reversed phase separation mode. At higher concentrations of acetonitrile, retention of the polypeptides to the octadecylsilica support progressively increased with elution order reversals indicative of a normal- or polarphase separation mode. These observations are discussed in terms of the interplay of hydrophobic and silanophilic interactions which occur between the ionised polypeptides and the stationary phase under these changing mobile phase conditions.High Performance Liquid Chromatography of Amino Acids, Peptides and Proteins, XXXII. For previous publication see ref. [1].     

Effect of the nature of binary water-organic solvents on the thermodynamic characteristics of adsorption of certain 1,3,4-oxadiazoles and 1,2,4,5-tetrazines on phenyl-bonded silica gel

The thermodynamics of adsorption in the Henry region for certain 1,3,4-oxadiazoles and 1,2,4,5-tetrazines from water-acetonitrile, water-methanol, and water-isopropyl alcohol solutions on silica gel with grafted phenyl groups is studied according to the dynamic method under conditions close to equilibrium. The effect of the nature of the organic component of a solvent on the values of standard differential molar changes in Gibbs energy, enthalpy, and entropy is shown upon adsorption of the investigated substances from solutions. The role of intermolecular interactions occurring in bulk solutions and at the interface during the adsorption of heterocycles from diluted water-organic solutions is discussed. The compensation effect and linear Gibbs energy relationship upon the adsorption of 1,3,4-oxadiazoles and 1,2,4,5-tetrazines from the investigated water-organic solutions on silica gel with grafted phenyl groups is determined.     

Effect of temperature in reversed phase liquid chromatography

The high temperature liquid chromatography (HTLC) reveals interesting chromatographic properties but even now, it misses some theoretical aspects concerning the influence of high temperature on thermodynamic and kinetic aspects of chromatography: such a knowledge is very essential for method development. In this work, the effect of temperature on solute behavior has been studied using various stationary phases which are representative of the available thermally stable materials present on the market. The thermodynamic properties were evaluated by using different mobile phases: acetonitrile-water, methanol-water and pure water. The obtained results were discussed on the basis of both type of mobile phases and type of stationary phases. Type of mobile phase was found to play an important role on the retention of solutes. The kinetic aspect was studied at various temperatures ranging from ambient temperature to high temperature (typically from about 30 to 200 degrees C) by fitting the experimental data with the Knox equation and it was shown that the efficiency is improved significantly when the temperature is increased. In this paper, we also discussed the problem of temperature control for thermostating columns which may represent a significant source of peak broadening: by taking into account the three main parameters such as heat transfer, pressure drop and band broadening resulting from the preheating tube, suitable rules are set up for a judicious choice of the column internal diameter.     

Characterization of retentivity of reversed phase liquid chromatography columns

There are dozens of commercially available reversed phase columns, most marketed as C-8 or C-18 materials, but with no useful way of classifying their retentivity. A useful way of ranking these columns in terms of column "strength" or retentivity is presented. The method utilizes a value for ln k'(w), the estimated retention of a solute from a mobile phase of 100% water, and the slope of the plot of ln k' vsE(T)(30), the solvent polarity. The method is validated with 26 solutes varying in ln k'(w) from about 2 to over 20, on 14 different reversed phase columns. In agreement with previous work, it is found that the phase volume ratio of the column is the most important parameter in determining retentivity. It is strongly suggested that manufacturers adopt a uniform method of calculating this value and that it be made available in advertising, rather than the uninterpretable "% carbon".     

N-Methylimidazolium anion-exchange stationary phase for high-performance liquid chromatography

In this paper, the preparation of a new anion-exchange phase based on N-methylimidazolium immobilized on silica is described. HPLC separations of common inorganic anions, including an iodate, chloride, bromide, nitrate, iodide, and thiocyanate, were performed using a HPLC column (200 x 4.6mm I.D.) packed with the said phase, and phosphate buffer solution and acetate buffer solution as the mobile phases respectively, with UV detection at 200 nm. The effects of pH and the concentration of the eluent on the separation of anions were studied. With the efficiency and resolution of the column calculated, the results showed that this new phase can be used in the analysis of these in organic anions with great prospects. At the same time, successful separations of some organic anions, amines and nucleotides have also been obtained respectively using this new phase. The phase displayed a main strong anion-exchange mechanism and a coexistent reverse-phase interaction, etc.     

Polysiloxane-encapsulated stationary phase for reversed-phase high-performance liquid chromatography

Summary Silica beads of 6-μm average diameter were silanized with methylvinyldiethoxysilane and then subjected to encapsulation with poly(methylvinylsiloxane). The resulting product is a new stationary phase for reversed-phase high performance liquid chromatography (RP-HPLC) which has superior ability for the separation of polar, non-polar and basic compounds. The chromatographic peaks are symmetric. Its stability has been studied; after continuous use for three months the carbon content and chromatographic behaviour of the phase were unchanged. on to the silica surface to given an uniform organic film. Material prepared in this way has both good chromatographic behaviour and superior selectivity. Because contact of the silica matrix with the mobile phase is avoided, the alkali-resisting ability of the stationary phase is increased. The non-specific adsorption of alkaline solutes on to the silica surface is also avoided because of the complete coverage of surface silanol groups. Reports of stationary phases encapsulated with polystyrene [6], polybutadiene [I] and octadecylsiloxane polymers have recently appeared in the literature [3]. In this paper we report the encapsulation of poly-(methylvinylsiloxane) (analogous to the phase SE-31 often used in GC) on to a silica matrix previously modified with methylvinyldiethoxysilane. The resulting phase has superior performance in reversed-phase HPLC.     

Novel imidazolium stationary phase for high-performance liquid chromatography

A new imidazolium anion-exchange phase immobilized on silica is synthesized. HPLC separations of common inorganic anions (IO3-, Cl-, NO2-, Br-, NO3-, I-, SCN-) have been performed using a HPLC column (200 mm x 4.6 mm I.D.) packed with this stationary phase, with a phosphate buffer solution as the mobile phase and UV detection at 200 nm. The effects of pH and concentration of eluent on the separation of anions have been studied. Chromatographic parameters are calculated and the results show that the new stationary phase is of significant potential for the analysis of these anions. Successful separations of some ordinary organic anions have also been achieved with the said stationary phase. Meaningfully, organic and inorganic anions can be determined simultaneously and satisfactorily with several neutral compounds using the column. The separation of some organic compounds including hydroxybenzenes, bases and amines by this stationary phase with only water as the eluent has been investigated.     

Speciation of Cr(III) and Cr(VI) by reversed phase high-performance liquid chromatography using UV-detection

Summary A method for the simultaneous determination of Cr(III) and Cr(VI) in water samples is described. The different reaction products of Cr(III) and Cr(VI) species with ammonium pyrrolidinedithiocarbamate (APDC) are extracted with ethyl acetate and determined by reversed phase HPLC using UV-detection. The procedure is optimized and its detection limit accordingly improved as compared to literature data. The detection limits achieved are 2.4 g/l for Cr(III) and 2.1 g/l for Cr(VI) and the calibration curves are linear between 5 g/l and 5000 g/l. For the speciation of Cr, APDC was demonstrated to be more suitable as chelating agent than sodium diethyldithiocarbamate (NaDDC). The procedure was applied to the determination of both Cr species in galvanic waste waters and its accuracy was approved by comparing the results (at the 100 g/l level) with those of a photometric determination of Cr(VI) species.Dedicated to Professor Dr. Wilhelm Fresenius on the occasion of his 80th birthday