Evaluation of a Simple HPLC Correlation Method for the Estimation of the Octanol-Water Partition Coefficients of Organic Compounds

Abstract

A simple reverse-phase high performance liquid chromatographic method is evaluated for the estimation of octanol-water partition coefficients (log P) of organic compounds by correlation with their chromatographic capacity factors (k′). Using an unmodified commercial octadecylsilane column and a mobile phase consisting of methanol and an aqueous buffer, a linear relationship is established between the literature log P values of 68 compounds and the logarithms of their k′ values. For the determination of the partition coefficients of unknowns, one of two sets of standards is used to calibrate the system, the choice being dependent on the hydrogen-bonding character of the compounds being evaluated. The overall method is shown to be rapid and widely adaptable and to give log P data which are comparable to results obtained by classical or other correlation methods.     

Selenium research in mammals using nuclear analytical methods and related techniques in conjunction with biochemical procedures

Due to the essential functions of selenium-containing enzymes and the relationships between changes in the selenium status and diseases, the determination of the element and its compounds is of great interest. Radiotracer studies with ⁷⁵Se have been valuable tools in selenium research. NAA and ICP-MS allow both total element and stable isotope measurements. ICP-MS in conjunction with chromatographic separation techniques and gel electrophoretic procedures coupled with scanning methods such as XRF, PIXE and laser ablation ICP-MS have been used in the determination of the selenium compounds. In this survey the application of these methods in selenium research is discussed with the help of examples on the regulation of the selenium metabolism and the detection and investigation of novel selenium-containing proteins. This revised version was published online in August 2006 with corrections to the Cover Date.     

The effect of stationary phase on lipophilicity determination of beta-blockers using reverse-phase chromatographic systems

Evaluation of lipophilicity parameters for basic compounds using different chromatographic stationary phases is presented. An HPLC method for determination of lipophilic molecule-stationary phase interactions was based on gradient analysis. Differences in correlation between the lipophilicity of compounds and experimental chromatographic results obtained in pseudo-membrane systems showed a strong influence of stationary phase structure and physico-chemical properties. beta-Blocker drugs with varying lipophilicity and bio-activity were chosen as test compounds. The stationary phases used for the study were monolithic rod-structure C18 and silica gel octadecyl phase SG-C18 as reference material. The second group was silica gel-based polar-embedded alkylamide and cholesterolic phases. The mobile phase was composed of acetonitrile or methanol with ammonium acetate, and a linear gradient of methanol and acetonitrile in mobile phase was performed. A linear correlation of plots of log k(g) = f(log P) was observed, especially for polar-embedded phases, and this allowed log P(HPLC) to be calculated. The behavior of stationary phases in methanol and acetonitrile buffer showed differences between obtained log P(HPLC) values.     

Evaluation of Octanol-Water Partition Coefficients Using Capillary Gas Chromatography With Cold On-Column Injection

Abstract

A general procedure is described for the determination of the octanol-water partition coefficients (log P) of very lipophilic compounds by the shake flask method, employing the analytical technique of capillary gas chromatography with cold on-column injection. The solute and an internal reference with a known log P value are partitioned between the layers of an octanol-water mixture and their relative concentrations are determined by analysis of each phase, using highly standardized chromatographic conditions. The overall method is demonstrated to be convenient, reproducible, and useful for the determination of the partition coefficients of a wide variety of compounds with log P values as high as 5.7.     

Determination of Lipophilic Character of a Series of Dermorphin-Related Oligopeptioes by Means of Reversed-Phase HPLC

Abstract

In recent times there has been a growing interest in the determination of chromatographic parameters of lipophilicity with regard to their use in the study of quantitative structure-activity relationship (1, 2). Very good correlations had been shown between the chromatographic parameters and the log P or π values as a measure of the partition coefficient between octanol and water (2). The reversed phase TLC R_m values in two different chromatographic systems and the reversed-phase HPLC log k′ values of a series of dermorphin-related oligopeptides have been previously determined (3, 4). The purpose of the present work was to study the relationship between log k′ values on one hand and R_m or ^Σπ values on the other one in view of QSAR studies. In fact the discovery of enkephalin and endorphins with high affinities for opioid receptors added new dimensions to the study of structure-activity relationship of opioid agonists (5, 6, 7, 8).     

High-performance liquid chromatographic separation and determination of 2-(diethylamino)ethyl diphenylpropylacetate and its metabolites from biological samples

A high-performance liquid chromatographic method for the determination of 2-(diethylamino)ethyl diphenylpropylacetate (SKF 525-A; Proadifen), its two deethylated metabolites and a hydrolytic metabolite in biological samples has been developed. From many solvent systems investigated the compounds are best separated on an ODS/Sil-X column using an acetonitrile-phosphate buffer system containing 0.2% decylamine. The use of acetonitrile overcomes limitations associated with previously published methods for separating basic compounds where decylamine was used in a methanolic eluent. With the sampling procedures used, overall recoveries from biological tissues were around 80%. The chromatographic system also separates three tricyclic antidepressants which can be used as internal standards.     

Relationships between LC retention, octanol-water partition coefficient, and fungistatic properties of 2-(2,4-dihydroxyphenyl)benzothiazoles

The retention behavior of newly synthesized compounds with antimycotic activity from the 2-(2,4-dihydroxyphenyl)benzothiazole group by high-performance liquid chromatography has been investigated. RP-18 stationary phase and methanol-acetate buffer aqueous mobile phases at pH 4 and 7.4 have been used. In the case of the mobile phase at pH 7.4, higher concentrations of water can be applied than at pH 4. The studied compounds showed regular retention behavior, their log k values decreasing linearly with an increasing concentration of methanol in the mobile phase. On the basis of these relationships, the lipophilicity (log kw), specific hydrophobic surface area (S), and isocratic chromatographic hydrophobicity index (psi0) were determined. Similar log kw values and sensitivity to changes in the structure of compounds studied for both mobile phases have been found. Moderate correlations between the chromatographic parameters and the calculated octanol-water log P values were found. Finally, the lipophilicity parameters were compared with the fungistatic properties of compounds expressed by log MIC (minimum inhibitory concentration) values to find quantitative structure activity relationship equations.     

An improved method for the determination of uranium isotopes in environmental samples by alpha-spectrometry

In order to improve the selectivity of the uranium isotopes determination in environmental samples, further studies have been carried out, including (1) interference of ²¹⁰Po with uranium isotope determination, (2) distribution coefficients of polonium between 5% TOPO in toluene and aqueous hydrochloric and nitric acids, (3) decontamination factor of uranium from polonium of the recommended procedure, and (4) leaching effect comparison of two different leaching procedures in a lichen sample. Based on the new findings, a more accurate extraction chromatographic/ a-spectroscopy method has been developed. For the method's validation, four kinds of reference materials supplied by the IAEA have been tested. It is observed that nearly all the ²³⁸U, ²³⁴U and ²³⁵U concentrations obtained are in good agreement with the recommended or information values, showing that the method can give reliable results. A comparison with existing uranium determination methods has also been made. It is concluded that due to involving preconcentration and chemical separation, the extraction chromatographic/a-spectroscopy method is a more selective, very sensitive and accurate, and low cost method. This revised version was published online in July 2006 with corrections to the Cover Date.     

Application of micellar and microemulsion electrokinetic chromatography for characterization of gallium(III) complexes of pharmaceutical significance

CE with conventional UV detection has recently been shown as a highly effective means to assaying cytotoxic gallium(III)-based compounds with regard to desirable drug-like properties such as the stability and binding to serum proteins. In this extension of that work, different CE techniques are used to further characterize a given set of gallium coordination compounds with established antiproliferating efficacy. Using free-zone CE mode, the electrophoretic profiles of complexes are recorded in order to assess their actual charge state under physiological buffer conditions. Micellar and microemulsion electrokinetic chromatographic techniques are tested as tools for the rapid estimation of the n-octanol-water partition coefficient (log P) that provides a rationale estimate of a drug's ability to cross biological membranes. A range of electrolyte buffer systems with varying (both in the nature and concentration) organic modifiers are examined to evaluate their effect on the relationship between experimental or calculated log P and the retention factors of compounds (log k'). Both methods were found to be better applicable for neutral than for cationic Ga complexes, the microemulsion mode demonstrating superior lipophilicity estimations as well as statistically meaningful log P versus log k' correlations when all the complexes were included in one regression set.     

An approach to the prediction of retention times in liquid chromatography

The utility of Rekker's hydrophobic fragmental constant has been examined for optimization of reversed-phase mode liquid chromatographic separations. The chromatographic behavior of about 60 non-ionic compounds was measured in different acetonitrile/water mixtures and the logarithm of their capacity factors (log k) was correlated with their calculated hydrophobicities (log P). Linear relations were found in each case between log k and log P. The slope of the various lines was related to the percentage concentration of acetonitrile in the mobile phase. It was shown that, by using nine stand ard compounds and measuring their capacity factors in five eluents with different acetonitrile concentrations, the retention time could be predicted for 60 compounds. Calculation of the concentration of the organic modifier was also possible in a system of well coated octadecyl bonded packings with acetonitrile/water mixtures as eluent. Prediction of the capacity factor was accomplished to within 5% error.     

硅胶基质固定相离子对反相液相色谱法测定强离解化合物的正辛醇/水分配系数

反相液相色谱(RPLC)是测定正辛醇/水分配系数(log P)的有效方法,但由于缺少同类型模型化合物,RPLC在测定强离解化合物的log P时遇到挑战.该文在硅胶基质C18色谱柱上,采用离子抑制反相液相色谱(IS-RPLC)和离子对反相液相色谱(IP-RPLC)分别对中性化合物、酚酸、羧酸、磺酸及部分两性化合物的保留行...     

Speciation of arsenic in natural waters by HPLC-NAA

Neutron activation analysis (NAA) in combination with mainly high-performance liquid chromatography (HPLC) has been developed for the determination of low levels of five arsenic species, namely As(III), As(V), monomethylarsonic acid (MMA), dimethylarsinic acid (DMA), and arsenobetaine (AsB) in water samples. Organically bound arsenic (OBAs) and total arsenic have also been determined. In addition to anion-exchange HPLC, solid phase extraction and open-column cation-exchange chromatographic methods have also been used. The detection limits of the method have been found to be 0.005 ng·cm⁻³ for OBAs, 0.02 ng·cm⁻³ for AsB, DMA, MMA, As(III), and As(V) and 0.12 ng·cm⁻³ for total arsenic. This revised version was published online in July 2006 with corrections to the Cover Date.     

Chromatographic methods for tetracycline analysis in foods.

The tetracyclines have served for decades as an important class of antibiotics in food animal health and production. As such, they have also been a source of concern for residue monitoring authorities around the world. In response to this concern a number of microbial inhibition, immunoassay and bacterial receptor methods have evolved for the detection of this class of compounds in various foods of animal origin. However, these methods often lack specificity and are subject to false positive and false negative results. For these reasons a number of chromatographic methods for the separation and determination of the tetracyclines isolated from foods have been developed that are capable of identifying and quantifying individual tetracycline drugs. We present here an overview of tetracycline analytical methods, including microbial inhibition, immunoassay and receptor technologies for detection, techniques for isolation from food matrices, and thin-layer chromatographic, high-performance liquid chromatographic, gas chromatographic and mass spectrometric procedures for determination of this class of compounds. A discussion of the variables involved in such methodology and a review of method criteria are offered.     

Method for measuring the logarithm of the octanol-water partition coefficient by using short octadecyl-poly(vinyl alcohol) high-performance liquid chromatography columns

A simple, quick, versatile and inexpensive HPLC method to estimate the logarithm of the octanol-water partition coefficient (log Pow) employing a methanol-water gradient and a short octadecyl-poly(vinyl alcohol) (ODP) column is described. This method is different from published HPLC-based log Pow methods because it uses retention times from a rapid methanol-water gradient to directly generate log Pow estimates, rather than from a series of isocratic mixtures extrapolated to 100% water. These HPLC log Pow values have good precision and correlate well with traditional shake-flask log Pow values. If necessary, the log Pow determination (including replications) can easily be carried out using only a milligram of sample. By suppressing ionization of acids and bases by the use of a buffer in the aqueous phase, the method can measure the log Pow of neutral organic molecules at any pH between 2 and 13. The method can be used with impure material and is rapid, 7 min per run and 4 min equilibration; it lends itself to and has been utilized for high-throughput hydrophobicity determinations (we have now carried out thousands of HPLC log Pow measurements by this method).     

Sequential injection chromatographic determination of ambroxol hydrochloride and doxycycline in pharmaceutical preparations

A new separation method based on a novel reversed-phase sequential injection chromatography (SIC) technique was used for simultaneous determination of ambroxol hydrochloride and doxycycline in pharmaceutical preparations in this contribution.The coupling of short monolith with SIA system results in an implementation of separation step to until no-separation low-pressure method.A Chromolith^® Flash RP-18e, 25-4.6 mm column (Merck, Germany) and a FIAlab^® 3000 system (USA) with a six-port selection valve and 5 ml syringe were used for sequential injection chromatographic separations in our study. The mobile phase used was acetonitrile-water (20:90, v/v), pH 2.5 adjusted with 98% phosphoric acid, flow rate 0.48 ml min⁻¹, UV detection was at 213 nm.The validation parameters have shown good results: linearity of determination for both compounds including internal standard (ethylparaben) >0.999; repeatability of determination (R.S.D.) in the range 0.5-5.4% at three different concentration levels, detection limits in the range 0.5-2.0 μg ml⁻¹, and recovery from the pharmaceutical preparation in the range 99.3-99.9%. The chromatographic resolution between peak compounds was >5.0 and analysis time was <9 min under the optimal conditions. The method was found to be applicable for routine analysis of the active compounds ambroxol hydrochloride and doxycycline in various pharmaceutical preparations.     

Calculation of gas chromatographic retention indices of organic compounds from the boiling points of their structural analogs

The possibility of calculating gas chromatographic retention indices (1) is discussed. The indices are important parameters for chromatospectral identification of organic compounds from the boiling points of their structural analogs (T _b ^* ) using the linear logarithmic equation log I = a log T _b ^* + bA + c, where A are the structural parameters reflecting the one- to- one position of the compounds being compared in the corresponding taxonomic groups, including homologous series.     

Chromatographic determination of plant saponins--an update (2002-2005)

The developments during 2002-2005 in the methods used for saponin analyses in plant material are presented. There were number of papers published on isolation and identification of new saponins by chromatographic techniques. Some new developments can be found in separation techniques or solid and mobiles phases used. Separation of individual saponins is still complicated and time consuming. This is due to the fact that in most of the plant species saponins occur as a multi-component mixture of compounds of very similar polarities. Thus, to isolate single compound for structure elucidation or biological activity testing, a combination of different chromatographic techniques has to be used, e.g. first separation of the mixture to simpler sub-fractions on reversed phase C18 has to be followed by further purification on normal phase Silica gel column. Especially difficult is determination of saponins in plant material as these compounds do not possess chromophores and their profiles cannot be registered in UV. Most HPLC methods apply not only specific registration at 200-210 nm, but these methods are not applicable for determination of many saponins in plant material at levels lower than 200-300 mg/kg. Some new or improved techniques for quantification of saponins in plant material were published in reviewed period. These include further progress in the application of evaporative light scattering detection (ELSD) for saponin profiling and quantification, which is also not only specific but also more sensitive in comparison to 200-210 nm detection. Some progress in development of new applications for liquid chromatography-electrospray mass spectrometry (LC/ESI/MS) for saponin determination has also been done. This method gives highest sensitivity and on line identification of separated saponins and should be recommended for specialized analyses of extracts and pharmaceutical formulas like the validation of a new assay. From non-chromatographic techniques for saponin determination, a sensitive and compound specific ELISA tests for some saponins were developed.