Retention Behavior Of Solutes Research Articles

Linear solvent strength model on porous graphitic carbon stationary phase using high temperature liquid chromatographic method for allopurinol related substances analysis

A high-performance liquid chromatography (HPLC) method was developed for the analysis of Allopurinol and its Ph.Eur. impurities using a porous graphitic carbon (PGC) stationary phase. Retention behavior of solutes was studied across a wide temperature range (30–90 °C) and various gradient times (5–20 min). Analysis of the data revealed distinct retention mechanisms between reversed-phase and PGC phases. However, it was proved that the retention of Allopurinol and its Ph.Eur. impurities on PGC stationary phase can be effectively modeled using the linear solvent strength (LSS) theory. This allows for the utilization of LSS-based method development software to optimize methods under these conditions. By using commercial chromatographic modeling software, separation of Allopurinol and Ph.Eur. impurities was optimized within a large design space. At the optimized operating conditions (pH = 2.0, tG = 6 min, T = 60 °C), all solutes were separated within 6 min with baseline resolution. Comparison between predicted and experimentally measured chromatograms further confirmed the applicability of LSS theory in developing analytical methods for PGC-based HPLC systems. The presented approach offers a general framework for method development on PGC phases.

Studies on the retention mechanism of solutes in hydrophilic interaction chromatography using stoichiometric displacement theory II. HILIC/RPLC dual-retention mechanism of solutes in hydrophilic interaction chromatography over the entire range of water concentration in mobile phase

This paper is a continuation of research into the retention behavior and mechanism of solutes in hydrophilic interaction chromatography (HILIC) using stoichiometric displacement theory (SDT). A HILIC/reversed-phase liquid chromatography (RPLC) dual-retention mechanism was studied in detail using a β-CD HILIC column. The retention behaviors of three groups of solutes with varying polarities were studied over the entire range of water concentrations in the mobile phase on the β-CD column, resulting in the formation of “U-shape” curves when lgk’ was plotted against lg[H2O]. Additionally, the effect of hydrophobic distribution coefficient lgPO/W on the retention behaviors of solutes in HILIC and RPLC modes was also examined. A four-parameter equation derived from the SDT-R was found to accurately describe the “U-shaped” curves of solutes with RPLC/HILIC dual-retention mechanisms on β-CD column. The theoretical lgk’ values of solutes calculated using the equation were found to be in agreement with their experimental values, with correlation coefficients greater than 0.99. This indicates that the four-parameter equation derived from SDT-R can effectively describe the retention behaviors of solutes over the entire range of water concentrations in the mobile phase in HILIC. As such, SDT can be used as a theoretical guide for the development of HILIC, including the exploration of new dual-function stationary phases to enhance separation efficiency.

Retention modeling and adsorption mechanisms in reversed-phase liquid chromatography

To interpret the dependence of solute retention behavior on modifier content in reversed-phase liquid chromatography, a theoretical framework, based on the concentration dependence of solvophobic forces imposed on solutes and the competitive adsorptions of solutes and solvent modifiers, was proposed. The generality of the developed model was demonstrated by comparing the model with conventional retention models. The linear dependence of the Gibbs energy change of solute adsorption with respect to the modifier concentration was assumed, and the model was fitted to the experimental results, with good agreement demonstrated between the experimental data and the model. Retention behaviors were inferred to be determined by two key dimensionless groups that represented the reductions in the retention factors resulting from a weakened solvophobic interaction and modifier competitive adsorption. The retention behaviors were successfully deconvoluted for each contribution as a function of the modifier concentration by using the fitted parameters. The effects of both contributions on the retention behaviors were enhanced for the solutes with aromatic groups. The standard Gibbs energy change SLo of benzene adsorption was found to depend linearly on the number of modifier molecules present but independent of modifier identity. For the solutes associated with hydrogen-bonding groups, the degree of reduction in the solvophobic interactions was considerably reduced. Hence, the relative contributions of both mechanisms to solute retention depend greatly on the solute structure. Perturbation method was performed to investigate the modifier adsorption mechanisms. The results show that the standard Gibbs energy change SLo for the first-layer adsorption of modifiers changed linearly with the carbon number of modifier molecule. These results demonstrated that the proposed model can offer a physically consistent quantitative description of retention when solvent composition is varied.

Effects of buffer salt types and non-counter ions of ion-pair reagents on the retention behavior of strongly ionized acid compounds in ion-pair reversed-phase liquid chromatography

在离子对反相液相色谱(IP-RPLC)分析中,溶质保留受对离子(counter ion)的影响比较受人关注,但鲜有研究流动相中缓冲盐类型和离子对试剂中非对离子(non-counter ion)对溶质保留行为的影响。鉴于此,该文以14种磺酸化合物为研究对象,甲醇为有机调节剂,分别考察了3种缓冲盐体系(磷酸二氢铵、氯化铵和乙酸铵)和5种离子对试剂体系(四丁基溴化铵、四丁基磷酸二氢铵、四丁基硫酸氢铵、四丁基硝酸铵和四丁基乙酸铵)下强离解酸性化合物的IP-RPLC保留行为,通过比较不同流动相条件下得到的溶质log kw(100%水相作流动相时的保留因子)、S(线型溶剂强度模型线性回归得到的常数),以及CHI(色谱疏水指数,log kw/S),寻找保留行为规律。研究表明,流动相中的缓冲盐类型和离子对试剂非对离子均会影响化合物的log kw和S值,所有化合物在氯化铵缓冲盐体系下具有最大的log kw值。相对于无机阴离子,离子对试剂中弱离解性有机阴离子(乙酸根)的存在有利于增加磺酸化合物的S值。通过对比不同条件下的保留行为,推测磺酸化合物的IP-RPLC保留机理中同时存在着离子对模型和动态离子交换模型。与log kw和S值不同,化合物的CHI值受缓冲盐类型以及离子对试剂非对离子的影响较弱。此外,研究发现化合物的表观正辛醇/水分配系数(log D)与log kw、S、CHI之间均具有良好的线性相关性。不同缓冲溶液和不同离子对试剂非对离子条件下获得的log kw和S值存在着一定的差异,而CHI值相对稳定,因此,CHI更适用于IP-RPLC中定量结构-保留行为关系模型的建立。

Effect of 2-propanol content on solute retention mechanisms determined using amylose tris(3,5-dimethylphenylcarbamate) chiral stationary phase under normal- and reversed-phase conditions

The electrostatic interactions between chiral solutes and polysaccharide (PS)-based chiral selectors are the key to achieving chiral recognition; however, PS-based sorbents, derivatized of phenyl moieties, can exhibit considerably non-polar characteristics, and they are also useful for the separation of enantiomers in the reversed-phase mode. In this study, an immobilized amylose 3,5-dimethylphenylcarbamate-based sorbent was used to investigate the balance between electrostatic interactions and solvophobic interactions, with complementary effects on solute retention behavior when the isopropanol (IPA) concentration was altered. It was proposed that in both normal- and reversed-phase modes, information on the retention mechanisms could be obtained by observing the curvature of the logarithm of the retention factor versus the logarithm of the IPA concentration, and the slope values of the curves were related to the number of displaced IPA molecules upon solute adsorption. Using the proposed model and the two-site adsorption model, the retention behaviors of pantolactone (PL) enantiomers in both normal- and reversed-phase modes were investigated. The PL−sorbent interactions were classified into four types: electrostatic/enantioselective, electrostatic/nonselective, solvophobic/enantioselective, and solvophobic/nonselective. At IPA concentrations below 50 vol.% in n-hexane, the retention behaviors of PL were dominated by electrostatic/enantioselective sites, whereas at IPA concentrations beyond 50 vol.%, the solvophobic interactions of PL−sorbent were strengthened and mostly nonselective. By contrast, in the reversed-phase mode, a reverse in the enantiomeric elution order of PL was observed at 10 vol.% IPA, and considerably different enantioselectivity behaviors were found below and above 20 vol.%, indicating an abrupt change in the sorbent molecular environment. At IPA concentrations beyond 40 vol.%, the presence of PL-sorbent electrostatic interactions enhanced chiral recognition.

Extension of the linear solvent strength retention model including a parameter that describes the elution strength changes in liquid chromatography

Modelling the retention behaviour of solutes in liquid chromatography, based on the composition of the mobile phase is a common task in the chromatographic practice. Along the development of liquid chromatography (LC), several models have been proposed to help in understanding the retention mechanisms, and especially, allow the prediction of retention times with optimisation purposes. Particular models are used for different LC modes, such as normal phase (NPLC), reversed phase (RPLC), hydrophilic interaction (HILIC), and micellar (MLC). In this work, a general equation is proposed that includes a parameter (the elution degree, g), which characterises the way the elution strength varies with the modifier concentration. The elution degree adopts the value g = 1 when the system follows the linear solvent strength (LSS) model, where the elution strength is constant. When g > 1, the elution strength decreases, and for g < 1, it increases with the modifier concentration. The proposed equation was applied to experimental retention data obtained for several chromatographic systems in RPLC, HILIC, MLC, and microemulsion LC. It was found that values in the 1 < g < 2 range are most usual. The general behaviour of the proposed equation was studied for isocratic and gradient elution. A general expression to calculate the compression factor of chromatographic peaks in gradient elution was also developed. It is shown that an increasing g value makes retention factors close to zero more difficult, since the elution strength decreases as the modifier concentration increases. For this reason, the larger the g value, the harder it is to reach significant peak compression. In contrast, an elution mode with g < 1 would yield increased elution strength with the modifier concentration, giving rise to significant peak compression.

Solvent effects on the retention mechanisms of an amylose-based sorbent

The mobile phase, when used in combination with a polysaccharide-based sorbent, often contains hydrocarbons with polar modifiers. Studies have investigated the effect of solvent on the recognition mechanism of polysaccharide-based sorbents, but it remains unclear how these modifier molecules affect solute retention behavior. This study used an amylose 3,5-dichlorophenylcarbamate–based sorbent to systematically investigate the retention behavior of various solutes as a function of the concentration of four polar modifiers: ethanol, isopropanol, methyl tert-butyl ether, and acetone. The thermodynamic properties of adsorption for the four modifiers were thoroughly investigated using retention factor data, van’t Hoff plots, and adsorption isotherms of the modifier molecules. The adsorption data of acetone and isopropanol followed the Langmuir isotherm, whereas the bi-Langmuir isotherm more accurately fit the ethanol data. For methyl tert-butyl ether, the Brunauer–Emmett–Teller adsorption isotherm indicated multilayer adsorption with a low saturation capacity of the first adsorption layer. A multivalent retention model interpreted slopes of the plots of the logarithm of the retention factor versus the logarithm of the modifier concentration. Because the molecular polarity of acetone is stronger than that of methyl tert-butyl ether, the limiting absolute slope values of tetrahydrofuran at a very high acetone modifier concentration were smaller than that when methyl tert-butyl ether was used as the polar modifier. The higher absolute value of the slope for tert-butanol suggested the potential of CO groups of acetone molecules to form bifurcated H bonds on a tert-butanol molecule. When alcohol was employed as the polar modifier, the results suggested that the effect of isopropanol self-association on the retention factor was stronger than the effects of solute–IPA complexation and isopropanol adsorption. For alcohol modifiers, U-shaped retention curves were obtained for all aromatic solutes. When acetone or methyl tert-butyl ether was used, the absence of a U-shaped curve indicated the weak polarity of those modifiers.

Retention models and interaction mechanisms of benzene and other aromatic molecules with an amylose-based sorbent

Stoichiometric displacement models have been widely used for understanding the adsorption mechanisms of solutes in chromatography systems. Such models are used for interpreting plots of solute retention factor versus concentrations of polar modifier in an inert solvent. However, these models often assume that dispersion forces are negligible and they are unable to account for solutes with significant aromatic interactions. In this study, a systematic investigation of the relationship between retention behavior and aromatic groups was performed using five simple aromatic molecules—benzene, naphthalene, mesitylene, durene, and toluene—with a commercially available amylose tris(3,5-dimethylphenylcarbamate)-based sorbent. The enthalpy changes of adsorption, determined from van’t Hoff plots, were obtained separately in pure n-hexane and in pure isopropanol (IPA). In pure n-hexane, the solute adsorptions were driven by electrostatic interactions, favoring a T-shaped binding configuration (edge-to-face π-π interaction). The order of enthalpy change indicated the amount of effective T-shaped π-interactions. In pure IPA, solute adsorption was dominated by dispersion forces, favoring a sandwich binding configuration (face-to-face π-π interaction). The adsorption isotherms of toluene revealed that in pure IPA and in pure n-hexane, the isotherms were linear. The results suggested that the high solvent strength of IPA weakened the interactions between aromatic molecules. The retention behavior of the benzene, naphthalene, mesitylene, and durene as a function of IPA concentration was investigated. U-shaped retention curves were found for all aromatic solutes. A new retention model for monovalent aromatic solutes was developed for describing the U-shaped curves. Three key dimensionless groups were revealed to control the retention behavior. The models suggested that solvophobic interactions should be accounted for in the retention models used to investigate the retention behaviors of solutes associated with aromatic groups.

Application of solvation model to prediction of the solute retention in liquid chromatography over a wide range of mobile-phase compositions

Possibilities have been studied of using a solvation model to predict the retention behavior of solutes in liquid chromatography mobile phases of water-acetonitrile and water-methanol with the organic solvent content varying from 1 to 100 vol%. Twenty-one test solutes, both aliphatic and aromatic compounds, have been selected on the basis of two-level factorial designs. Using the multiple linear regression analysis, regression coefficients, which are characteristics of the stationary and mobile-phase system, were calculated for different mobile phases in the solvation model. Regression coefficients have been used for the prediction of the retention behavior. Unbiased results have been obtained by using two sets, one training and the other the testing set. The predicted retention has been compared with the experimental data. The methanol-water system provided good results at low and medium methanol concentrations; the retention prediction was unsatisfactory for mobile phases containing more than 90% of methanol. The acetonitrile-water system yielded similar results, but the retention prediction ceased to be at acetonitrile concentrations greater than 80%. The retention has primarily been determined by cohesive and acid-base interactions. The dependences of the regression coefficients on the mobile-phase composition were similar for the acetonitrile-water and methanol-water systems.

Chromatographic methods enabling the characterization of stationary phases and retention prediction in high-performance liquid chromatography and supercritical fluid chromatography.

In the scope of the present review, the current status of high-performance liquid chromatography/ultra-high performance liquid chromatography and supercritical fluid chromatography is briefly provided. These techniques and their retention mechanisms are compared. Various alternative approaches utilized for the determination and description of the retention processes in these two systems are mapped. Two frequently used concepts, linear-free energy relationships, and hydrophobic subtraction models, used for the characterization of the retention interactions, are discussed. Principles and selected applications of the both methods are also covered. Then the models applied for the prediction of retention behavior of solutes on stationary phases are outlined. The procedures utilized for the sorbent/column classification are also covered. Simple chromatographic tests frequently used for the basic characterization and mutual comparison of stationary phases are summarized and briefly commented on. The importance of a statistical evaluation of complex retention data obtained from the chromatographic measurements is outlined. Finally, computer simulations aiming at the facilitation of the quest to optimize separation conditions for a given mixture of analytes are touched upon.

Development and Evaluation of a New Fluorinated Double-Wall Carbon Nanotube HPLC Stationary Phase

A novel column based on silica-containing immobilized fluorinated double-wall carbon nanotubes (F-DWCNTs) was developed. This F-DWCNT stationary phase was synthesized to combine the analytical performance of carbon nanotubes and the fluorine-based unique selectivity for polar compounds. First, the chromatographic support was coated with DWCNTs in a noncovalent way to preserve the sp2 internal nanotube structure. Second, the DWCNT silica particles were functionalized with fluorine atoms via a solution of Br2 and BrF3 at room temperature. This F-DWCNT stationary phase was applied for a variety of separations. The solute retention behaviour was particularly studied under isocratic conditions with a high fraction of ACN in the ACN/water (v/v) mobile phase. The retention factors of the solute molecule do not depend linearly on the ACN fraction, but follow a quadratic relationship. This fluorinated stationary phase separated compounds based upon a combination of hydrophobic and polar selective stationary phase interactions. This F-DWCNT appeared to work best when fluorinated or halogenated compounds were encountered. They have longer retention time, better selectivity and work well with high fraction of organic modifiers. This novel stationary phase could thus be a good choice for LC–MS experiments.

APPLICATION OF MULTIVARIATE REGRESSION MODELS IN CHROMATOGRAPHY. NEW ADVANCES.

The aims of the review are the collection, concise description and evaluation of the multivariate mathematical-statistical methodologies used for the elucidation of the similarities and dissimilarities among retention parameters measured under various chromatographic conditions and the determination of the relationship between the physicochemical parameters and chromatographic retention behaviour of solutes.

Simultaneous Prediction of the Logarithmic Capacity Factor of Some Aliphatic and Aromatic Compounds on Five Different Stationary Phases in RP-LC Using Artificial Neural Network

The present work is a quantitative structure-retention relationship (QSRR) study based on multiple linear regressions (MLR) and artificial neural network (ANN) to predict the retention behavior of solutes in reverse-phase liquid chromatography based on their structures. To attain this goal, the capacity factors (log k′) of a collection of 65 aliphatic and aromatic solutes on five stationary phases (Zorbax SB-C18, Zorbax Rx-C18, Hypersil C18, Hypersil C8 and Zorbax C8) were selected as a data set. By using Dragon software, various descriptors were calculated for all molecules in the data set. The descriptors that appeared in this model were selected by stepwise multiple linear regression (MLR). These descriptors were used as inputs of the constructed ANN and its output was logarithm capacity factors of aliphatic and aromatic compounds on five different columns. Comparison between statistical results calculated for MLR and ANN model reveals that all statistics have improved considerably in case of the ANN model. The improved statistics by the ANN would suggest the existence of nonlinear relation between selected molecular descriptors and their retention in reverse phase liquid chromatography.

Multilinear gradient elution optimization in reversed-phase liquid chromatography based on logarithmic retention models: Application to separation of a set of purines, pyrimidines and nucleosides

The analytical solutions of the fundamental equation of the multilinear gradient elution are derived in two cases, when the dependence of the logarithm of the solute retention (ln k) upon the volume fraction of organic modifier ( φ) is a three-parameter logarithmic expression, and when a simple linear relationship between ln k and ln φ is adopted. The derived theoretical expressions for retention times under multilinear gradient conditions are embodied to simple algorithms for fitting gradient data and especially for resolution optimization. Their performance was examined by using a mixture of 16 model compounds chosen among purines, pyrimidine and nucleosides in eluting systems modified by acetonitrile. It was found that the accuracy of the predicted gradient retention times is very satisfactory even if the simple logarithmic expression for the retention behavior of solutes, i.e. the linear dependence of ln k upon ln φ, is used.

Solvent-gradient SMB to separate o-xylene and p-xylene

In batch chromatography, solvent-gradient operations (SG) produce significant improvement in terms of the enrichment of products and the separation time and the solvent consumption as compared with isocratic operations. This work studied solvent-gradient operation in reversed-phase simulated moving bed unit to separate ortho-xylene and para-xylene. In a solvent-gradient mode, different mobile phase compositions lead to a different retention behavior of solutes, i.e., different adsorption isotherms. Frontal analysis experiments for ortho-xylene and para-xylene were carried out with a reversed-phase column to measure adsorption parameters at several different mobile phase compositions, such as 45%, 50%, 60%, 75% and 90% acetonitrile. Therefore, the parameters in the retention model for solvent-gradient operation in the case of reversed-phase chromatography were estimated and applied to the design of an SMB system. A modified design method for solvent-gradient simulated moving bed chromatography (SG-SMB) was proposed. The robust operating conditions were obtained within the separation region on (φ R , φ E ) plane (φ R and φ E are the volumetric content of organic modifier in the raffinate and the extract streams, respectively). The performance results of isocratic and solvent-gradient SMB were compared. A partial-discard strategy and increasing of the solvent gradient level were also applied to improve the performance of the SG-SMB.

Prediction of the Gradient Retention Times of Purine Compounds in Reversed Phase High Performance Liquid Chromatography

Various retention models have been developed and adopted to predict the retention behaviors of solutes in high performance liquid chromatography. Although the retention times can be successfully predicted in a gradient elution, it is difficult to predict the fundamental parameters of the eluted peaks, such as the number of theoretical plates, the resolution between neighboring peaks, as well as asymmetry factors. Thus far, the Snyder retention model has widely been used to predict the retention behavior in both isocratic and gradient conditions. However, it has a critical defect in that it cannot closely follow the retention behaviors of solutes when a relatively low content of organic modifier is used in the mobile phase. This is a result of the nonlinear relationship between the logarithm value of the retention factor and the volumetric percentage of organic modifier in the mobile phase. To overcome this shortcoming, a modified retention model was adopted to predict the retention times in several linear gradient conditions. A numerical method that transforms any gradient condition into discrete step gradient conditions was also proposed to predict the retention times in a gradient elution. The model is suitable to apply to nonlinear and multilinear gradient conditions, including actual obtained gradient profiles. Two kinds of organic modifiers, methanol and acetonitrile, were employed, and four purine compounds were used as solutes. The predicted retention times obtained by the modified retention model are in good agreement with experimental data in terms of the achieved gradient conditions.

Peak crossover in high-performance liquid chromatography elution monitored using whole-column detection

A whole-column detection (WCD) system was fabricated using the parts from a typical A4 size optical scanner. Optimum layout of the fluorescence tube, beam condenser and photon transducers from the scanner made an effective devise to examine the dynamic chromatographic separation process. This system was applied in monitoring peak crossover in an ion-pair chromatographic elution. A stage-wise gradient was conducted in eluting a mixture of New Coccine and Carmoisine. Compound New Coccine lagging behind after sample injection moved passing Carmoisine at the column outlet. The complete on-column elution process could be observed directly using this detection system. This inexpensive and rugged WCD system provided a 0.3 mm spatial resolution and a 3.6 ms temporal resolution and was proven to be an adequate device to directly monitor solute retention behavior in the liquid chromatography column.

Retention prediction in ternary solvent reversed-phase liquid chromatography systems based on the variation of retention with binary mobile phase composition

An extension of the treatment adopted in a recent paper [P. Nikitas, A. Pappa-Louisi, P. Agrafiotou, J. Chromatogr. A 946 (2002) 33] was used to derive expressions describing the variation of solute retention k with composition in ternary reversed phase liquid chromatography, RP-LC, solvent systems. The equation of the partition model obtained in this way for a ternary mobile phase was identical to that previously derived using the solubility parameter concept. This equation as well as two new expressions of ln k versus organic modifiers content were tested in a variety of ternary solvent systems in order to examine the possibility of predicting retention behavior of solutes under ternary solvent mixture elution conditions from known retention characteristics in binary mobile phases. It was demonstrated the superiority of both new equations derived in this paper to that previously proposed and applied to date in ternary solvent mixtures.

Preliminary study of the retention behavior for different compounds using cryogenic chromatography at different initial temperatures

The Kováts indices ( I) of 24 volatile organic compounds were measured at the different initial temperatures by a laboratory-made cryogenic chromatography. The results show that the correlations of the Kováts indices and the temperature are nonlinear at low initial temperature. Seven QSRR models were built using the heuristic method at different initial temperatures by calculating the quantitative chemical parameters of 24 organic compounds to study the relationship between the retention behavior and the molecular parameters. By studying the relationship between carbon and retention behavior, the capacity factor, and resolution of selected compounds, the contribution of gas–liquid interfacial adsorption process, which influences the retention behavior of solutes, is becoming more and more important with the decrease of the initial temperature.

Quantitative structure–retention relationships for ionic and non‐ionic compounds in biopartitioning micellar chromatography

Quantitative structure-retention relationships, QSRRs, represent a powerful tool in chromatography. The objectives of QSRR studies are to predict the chromatographic retention behaviour of solutes based on their structural properties, to elucidate retention mechanisms, to optimize the separation of complex mixtures or to prepare experimental designs. In this paper, using the retention factors of 151 structurally unrelated solutes that cover a wide range of hydrophobicity, molecular size, hydrogen bonding properties and ionization degrees obtained in biopartitioning micellar chromatography (BMC) at different Brij35 micellar concentrations, several multivariate QSRR models are tested. It is demonstrated that the chromatographic retention of any molecule in BMC, independently of its family, can be adequately described by its hydrophobicity (expressed as log P) and its anionic and cationic total molar charge (expressed as alpha(A) and alpha(B)).