A calculation method for the prediction of effective plate height in capillary gas chromatography

The effective plate height, h_eff, is considered to be a better measure of the efficiency of capillary column than the conventional plate height, h, in isothermal conditions. By using experimental data of 1-alcohols and n-alkanes, 2-ketones and 1-alkenes measured on capillary columns coated with non-polar stationary phases in isothermal and isobaric conditions, the peak width at half height is predicted with a function similar at that of adjusted retention time. The results obtained under different analytical conditions as the head pressure and the temperature of the column confirm the validity of the model, whose parameters are linear, and as a consequence a unique solution is obtained.     

超高效液相色谱塔板高度方程的推导

近年来,分析工作者采用超高效液相色谱(UPLC)完成了许多过去不能完成的分离分析工作。但是在阐述UPLC原理时不少人却采用了van Deemter方程。这是不对的。本文研究了UPLC色谱过程动力学,从热传导方程出发运用色谱动力学原理推导了包括考虑流动相摩擦生热影响的UPLC塔板高度方程H=2γD_m/u+((2λd_pu^1/3)/(u^1/3+ω(D_m/d_p)^1/3))+((2ku)/((1+k)²(1+κ₀)κ_d))+ ((θ(κ₀+κ₀k+k)²d_p²u)/(D_mκ₀(1+κ₀)²(1+k)²)) +(κ_i(κ₀+κ₀k+k)²d_p^5/2u^2/3)/(3κ₀Ω D_m^2/3(1+κ₀)²(1+k)²+(r₀²(κ₀+κκ₀k+k)u)/(4(1+k)D_r)·exp(-Kr₀²α)。上述方程右端最后一项代表了流动相摩擦生热对塔板高度的贡献。当流动相线速度较低时,流动相摩擦生热对塔板高度的贡献趋近于零,塔板高度方程还原成Horvath和Lin的方程;当流动相线速度较高时,由于流动相摩擦生热,柱轴心与边缘温差增加,流动相线速度径向分布差异导致柱效率降低,而柱轴心与边缘的温差与流动相线速度平方成正比。作者明确指出:UPLC的柱效率与柱内径密切相关,采用细内径柱有利于实现高效率;过高的流动相线速度将导致柱效率崩溃。     

The chromatographic processes in the S-chamber with the counter plate

For the first time the chromatographic processes in the new variant of a S(min)-chamber with a counter plate (a S(min)(CP)-chamber) positioned at a small distance above a separating plate have been studied, given the adsorption layers of the both plates (the separating plate and the counter plate) are turned to each other. It should be noted that the features of this method have not been completely studied, only two publications were devoted to the practical using of the S(CP)-chamber. Using of a dry counter plate in the S(min)-chamber having been proposed by us recently leads to the marked increase in the volume of the mobile phase that continuously migrates through the separating plate. Using of the S(min)-chamber with the dry counter plate for separation allows, first, substantially (up to 50%) increasing the value of R(f) (especially, in the range of the small meanings of R(f) (0-0.3)), second, increasing the efficiency of separation by 2.0-2.5 times, and, third, increasing the peak resolution of the method by ～25%. It should be noted that although when separating a mixture in the S(min)(CP)-chamber a slight increase in the experiment duration also occurs (by 20-50% depending on the sizes of the used plate), in all the cases chromatographic characteristics markedly improve (in comparison with separation in the same conditions, but without using of the counter plate).     

Influence of development distance on resolution in thin-layer chromatography

Theoretical equations are given to relate resolution with development distance. Comparative experiments undertaken with quaternary ammonium compounds as test substances and using a selection of basic drugs are presented. As expected, resolution decreases with shorter development distance. However, this effect is less pronounced for compounds with small R_f values and when the spot size at the starting point is kept small. It was also shown that, for correctly assessing resolution in relation to development distance, the amounts of material spotted have to be decreased for shorter development distances.     

Influence of various salts on the reversed-phase retention of some dansylated amino acids in TLC

Summary The retention of 14 dansylated amino acid derivatives was determined using aqueous LiCl, NaCl, KCl, RbCl and CsCl solutions as eluents in reversed-phase thinlayer chromatography. The salts exerted typical salting-out effects, the retention of each dansylated amino acid increased with increasing concentration of salt in the eluent. This effect has been tentatively explained by the suppression of the dissociation of the polar groups in the solute molecules resulting in increased apparent lipophilicity. The correlation between the increased retention of dansyl amino acids and the salt concentration was found to be linear. The hydrated radii and energy of hydration of cations as well as the hydrophobicity of free amino acids and the pK value of the -amino groups simultaneously influenced the retention.     

液相色谱塔板高度方程的统一形式

研究了色谱分离过程中物质的径向扩散和流动相发热对柱效能的影响。从热传导方程出发,运用色谱过程动力学原理推导了包括考虑流动相径向扩散、色谱柱发热影响的液相色谱塔板高度方程：

该方程概括了高效液相色谱（HPLC）、超高效液相色谱（UPLC）、毛细管电色谱（CEC）和消滞留层液相色谱（ESFLC）塔板高度与各种因素的关系。方程最后一项代表了径向扩散和柱发热对塔板高度的贡献。当流动相线速度较低且柱内径较细时,流动相摩擦生热和径向扩散对塔板高度的贡献趋近于零,塔板高度方程还原成Horvath和Lin的方程;当流动相线速度较高时,由于流动相摩擦生热,柱轴心与边缘温差增加,导致流动相线速度径向分布差异,使得柱效率降低。柱轴心与边缘的温差与流动相线速度平方成正比。该文指出,在流动相高线速度情况下,液相色谱的柱效率与柱内径密切相关,采用细内径柱有利于实现高速与高效率;过高的流动相线速度将导致色谱柱效率崩溃。     

The role of TLC in the screening of inherited metabolic diseases

Summary Thin-layer chromatography (TLC) is a rapid, reliable and inexpensive screening technique for diagnosis of inherited metabolic diseases (IMD). Our screening program encompasses five main situations where the use of TLC is considered to be vindicated: (i) analysis of amino acids; (ii) screening for sugar defects; (iii) detection of pathological oligosaccharidurias; (iv) screening for organic acid disorders; and (v) detection of abnormalities in tryptophan metabolism. Examples are presented of chromatograms obtained from pathological samples. Presented at the 21^st ISC held in Stuttgart, Germany, 15^th–20^th September, 1996     

Theory of reflectance measurements in quantitative evaluation in TLC and HPTLC. 1. Multilayer model of reflectance and transmittance

In the approach described in this paper, the layer of adsorbent is divided into n sublayers. The radiation density in each sublayer is given by the sum of the transmittance and the reflectance of the adjacent sublayers. Absorption of light by the layer or the substance will diminish this radiation density. Depending upon the factors of reflectance and transmittance some light will pass to the adjacent sublayers. Two polynomials are derived from this readily understood formulation to calculate the reflectance or the transmittance of a layer of finite thickness. The algorithms given describe the reflectance and transmittance encountered in practical measurements with large amounts of substance per spot, as well as with small amounts. There are no restrictions with respect to nonideal scattering or reflection by the adsorbent layer support.     

Influence of the peak height distribution on separation performances: Discrimination factor and effective peak capacity

Summary A new index of performance of the chromatographic separation between two adjacent peaks, the discrimination factor, d₀, is defined. It is normalized between 0 and 1 and is directly and easily determined from the chromatogram. It does not depend on any assumption regarding peak shape, except that the peak profiles of individual sample components have a single mode. Its value depends on the relative heights of the two peaks as well as on their separation. The separation power of a chromatographic system is classically measured by its peak capacity, defined on the basis of constant resolution between adjacent peaks. A previously developed statistical theory of the composition of mixtures makes it possible to extend the concept of peak capacity by taking into account the peak height distribution in typical average chromatograms. A new parameter, the effective peak capacity, is defined for this purpose on the basis of a constant discrimination factor between adjacent peaks. It allows to take into account the distribution of peak heights in statistical theories of the evaluation of complex chromatograms and in the measurement of the limit of determination in quantitative analysis. The characteristics of the two new parameters, the discrimination factor and effective peak capacity, are discussed and compared with those of their classical homologs, resolution and peak capacity, in the case of gaussian component peaks of equal widths.     

有机铁硫原子簇化合物薄层色谱展开剂优化快速选择

本文介绍了薄层色谱最佳展开剂组成的快速选择方法。首先利用对半优化法粗选最佳展开剂大概用量范围;其次绘制△Rt-ε0图预测展开剂系统:最后用数字分析法算出展开剂各组分最佳配比值。并用于有机铁硫原子簇化合物[1,2]的薄层分析。     

Two dimensional TLC determination of PAH in diluted automobile exhaust gases

A method is described for the determination of polycyclic aromatic hydrocarbons (PAH) in the diluted exhaust gas of diesel vehicles. Sampling is done by drawing off proportional streams from the dilution tunnel. The particulates deposited on filters are sublimed, and the sublimate is purified and prefractionated on silica gel. Further separation and quantitative determination of the PAH is performed by two dimensional TLC in conjunction with In situ fluorescence spectrometry. Results of theoretical considerations and experimental data on the distribution of the emitted PAH between particulate matter and the corresponding gas phase in diluted exhaust are presented. A modified Langmuir adsorption model is used to explain the effects of dilution ratio and sample temperature in the dilution tunnel. Comparison of the emission values for PAH obtained from diluted and undiluted exhaust shows good agreement. Furthermore, the PAH contents of the exhaust gas of several diesel-engined cars are also compared for various driving cycles.     

Determination of the lipophilicity of some anti-hypoxia drugs: Comparison of TLC and HPLC methods

Summary The lipophilicity of 14 anti-hypoxia drugs has been determined by reversed phase thin-layer (RPTLC) and reversed phase high performance liquid chromatography (RPHPLC) in eluent systems containing different concentrations of acetonitrile and potassium dihydrogen phosphate. There was significant correlation between lipophilicity and the specific hydrophobic surface area of the drugs in RPTLC, indicating that the drugs behave as an homologous series of compounds. In RPTLC the concentration of buffer has a negligible effect on the retention of the drugs whereas in RPHPLC the buffer concentration influenced the retention. This discrepancy can be explained by the lower sensitivity of RPTLC. There was strong correlation between lipophilicity values determined by both methods, proving that both are suitable for the determination of molecular lipophilicity.     

Retention behaviour of model solutes on mixed silica-magnesia adsorbents by TLC. Comparison with the adsorption properties of Florisil

Summary The retention behaviour of positional isomers (phenols, aniline derivatives, quinoline bases) and polyaromatic hydrocarbons on mixed silica-magnesia layers is examined using various binary eluents (ethyl acetate, dioxane or 2-propanol in n-heptane). The influence of magnesia content in mixed adsorbents and concentration of polar modifier in the eluent on retention were investigated. Results obtained in chosen systems were compared with retention of solutes on Florisil layers using the same eluents by theR _{M Florisil} vsR _{M mix} correlations. Analysis of the structural effects of solutes and the position of points onR _{M Florisil} vsR _{M mix} diagrams enables comparison of chemical character and distribution of active centers on Florisil and mixed adsorbent surfaces.     

Effect of halides on the TLC resolution of amino acids below their isoelectric points

TLC of a fifteen component mixture of amino acids has been carried out in two ways; firstly, the amino acids were treated with halides below their isoelectric points and chromatographed on plain silica plates, and secondly the amino acids in their cationic forms were chromatographed on silica plates impregnated with halides, keeping the same solvent system. The resolution is considered to be affected by hydrophobic interactions between silica gel and amino acid molecule and by the polarity and the flow of the mobile phase. The method provides resolution of 10–11 amino acids from the fifteen component mixture.     

Isocratic networks in supercritical fluid chromatography II: Selectivities and effective plate numbers

The dependence of the selectivity α and the effective plate number, N, on temperature and pressure is shown in three-dimensional diagrams. The specific system studied was pentane as the mobile phase, unmodified silica as the stationary phase, and a set of four polycyclic hydrocarbons as the test mixture. Temperature and pressure ranges were from 25 to 300°C and from 30 to 75 bar, respectively. The plots for α and N are found to resemble those for capacity ratio, k′, and resolution, R, studied earlier with the same system [1]. A distinct maximum is observed for N in the supercritical region, as found earlier for k′ and R. This applies also to α for the homologous substrate pair naphthalene/anthracene, while the resemblance for α is less pronounced for the pair anthracene/pyrene.     

Displacement and spacer displacement chromatography. Alteration of spot profile in displacement thin-layer chromatography

Summary Migration of spots in displacement development was investigated using a planar stationary phase. The effects of the presence or absence of the additive substances in the carrier were demonstrated when these compounds also played a role in the competition for the binding sites of the stationary phase; however, they were not members of the displacement train.In the case of displacement chromatography, compounds A+B are well separated but located very closely to each other. Using a spacer (C) in the sample, the peaks or spots of A and B are aparted by C and their virtual separation is increased.When compounds A+B were displaced by each other as well as by the displacer, the presence of the additive compound C in the carrier (carrier-spacer) can transfer an otherwise displaced component (A) into the so-called overloaded elution region of the chromatogram. At the same time the additive compound C does not unconditionally reduce the performance of displacement chromatography.Presented at the 17th International Symposium on Chromatography, September 25–30, 1988, Vienna, Austria.     

Thin-layer chromatography with high-boiling mobile phase. part 2: Effect of temperature on retention characteristics in TLC with high-boiling mobile phases

A study has been undertaken on the effect of temperature on retention characteristics in thin-layer chromatography (TLC) with low-volatility mobile phases (MP). It is shown that temperature variations in TLC in melts bring about variations in both the relative retention values and, in some cases, in the order of migration of the chromatographic zones across the layer. The variation in the capacity factor k' with temperature agrees well with Martin's equation. To explain the temperature dependence of R_f one must, in general, take into account the variations with temperature in both the partition coefficient and the phase ratio. To describe the variation in R_f with temperature in TLC with low-volatile MP one can use an approximate equation in which In R_f is a linear function of 1/T. The experiments indicate that temperature is a major factor in TLC in melts.     

TLC as a pilot technique for the optimization of gradient HPLC II. Use of data obtained from RP-18 plates for the prediction of gradient programs in reversed-phase liquid chromatography

Summary A method is described and experimentally verified permitting the determination of the optimal gradient programs in HPLC on the basis of isocratic TLC data. The proposed method has many advantages and can also be used in reversed-phase systems. A satisfactory agreement between the theoretical and experimental of V_R/V_O values has been confirmed. Some discrepancies for more strongly retained solutes are, probably due to the respective equilibrium and nonequilibrium state of the column in isocratic and gradient elution.Part I: Ref. [1].