Direct determination of gentamicin components by capillary electrophoresis with potential gradient detection

A simple and fast method was developed to determine non-UV active compounds directly without derivatization. The usefulness of the method was demonstrated by detecting the major components in aminoglycoside antibiotic mixtures using capillary zone electrophoresis with potential gradient detection. Under optimized separation conditions (0.2 mM cetyltrimethylammonium bromide (CTAB), 1 mM ammonium citrate, pH 3.5), gentamicin was separated into three major peaks (C1, C1a, and C2+C2a) within 15 min. This method showed better sensitivity than other capillary electrophoresis (CE) methods for determining underivatized gentamicin. The linear range was from 10 to 500 ppm. Because of its good repeatability and simplicity, this new method could be a good alternative for the current assays given by US Pharmacopoeia and European Pharmacopoeia.     

Capacity factors and peak widths in the gradient elution reversed phase separation of high molar mass polystyrenes

Summary A simplex method for determining the constantsS andk _o from the equation lnk’=lnk _o−S ϕ (was developed and applied to the reversed phase separation of high molar mass polystyrenes using gradients of any curvature. Experimental retention times described using the equation had a standard deviation of 1.1%. The inclusion of a quadratic term in the equation was found to be unwarranted. BothS and lnk _o varied linearly with ln molar mass. The logarithm of peak width of an eluting polystyrene peak was found to be a linear function of the mobile phase composition at elution. The slope was equal toS.     

用TREF方法分离聚烯烃共混物

用ＴＲＥＦ方法分离聚烯烃共混物徐君庭，封麟先，杨士林（浙江大学高分子科学与工程学系，杭州，３１００２７）关键词升温淋洗分级，聚烯烃，共混物升温淋洗分级（ＴＲＥＦ）方法是８０年代发展起来的，它按结晶度将聚合物分级［１］，目前主要用于聚烯烃的分级［２，３...     

Optimization of stepwise gradient elution in reversed-phase chromatography

Summary Equations describing multi-step gradient elution with a mobile phase of constant composition in each step were derived. These equations useful for calculating the retention volumes in both gradient HPLC and TLC were derived on the basis of the relationship between the isocratic capacity factor and the volume fraction of the organic modifier. The validity of the equations was experimentally verified in a LiChrosorbRP-18-water/methanol system for 11 methyl- and chlorobenzenes and phenols. A satisfactory agreement between the theoretical and experimental k′ values was found.     

A practical approach to transferring linear gradient elution methods

Attempts to theoretically address the problems involved in transferring linear gradient elution methods have been somewhat ad hoc due to the simplifying assumptions usually made in conventional gradient elution theory. Until now, all equations based on the k* parameter of linear gradient elution theory used as the basis for predicting the separation selectivity have not explicitly included the effect of the dwell volume (VD). Using an exact equation for predicting k*, that is, one which fully accounts in an a priori fashion for VD, we find a set of simple yet exact equations which unequivocally must be satisfied to transfer an optimized linear gradient elution method from one system (column or instrument or both) to another. These relationships absolutely mandate that a change in the instrument dwell volume requires a proportional change in the column volume; in turn, a change in the column volume requires a proportional change in the flow rate and/or gradient time to maintain a constant gradient steepness. Although we are not the first to suggest these guidelines, this work provides a complete theoretical foundation for these exact guidelines for the maintenance of gradient selectivity for the case of transferring a linear gradient elution method between different columns packed with the same particles and/or between different instruments.     

Spherical coordinate representations of solvent composition for liquid chromatography method development using experimental design

One of the major techniques used for the method development of ternary and quaternary high performance liquid chromatography (HPLC) systems has been to use mixture designs, often referred to as "Glajch's Triangle". This technique does not allow for the systematic and simultaneous optimization of other factors such as gradient time, pH and temperature that affect the quality of separations. An alternative approach is to use experimental designs. The condition, however, that the composition of all components of the mobile phase must total 100% presents a problem when trying to mathematically represent ranges of each mobile phase constituent of a ternary or quaternary system. A method is described here, based on spherical coordinate representations, that adheres to the constraints of the mobile phase composition and allows experimental designs, such as central composite and factorial designs, to be applied to the simultaneous optimization of the mobile phase composition. Other factors, in particular temperature and gradient time, can then be included in the design. As a result of applying these designs to the HPLC separation of phenols and corticosteroids, it was found necessary to include three-way interactions between experimental factors in the model. The significance of these interactions shows that they need to be considered in HPLC method development.     

Retention modeling and simultaneous optimization of pH value and gradient steepness in RP‐HPLC using feed‐forward neural networks

A novel approach is proposed for the simultaneous optimization of mobile phase pH and gradient steepness in RP‐HPLC using artificial neural networks. By presetting the initial and final concentration of the organic solvent, a limited number of experiments with different gradient time and pH value of mobile phase are arranged in the two‐dimensional space of mobile phase parameters. The retention behavior of each solute is modeled using an individual artificial neural network. An “early stopping” strategy is adopted to ensure the predicting capability of neural networks. The trained neural networks can be used to predict the retention time of solutes under arbitrary mobile phase conditions in the optimization region. Finally, the optimal separation conditions can be found according to a global resolution function. The effectiveness of this method is validated by optimization of separation conditions for amino acids derivatised by a new fluorescent reagent.     

一种基于流动注射梯度技术识别异常峰及校正的方法

基于流动注射梯度信息提出了一种利用梯度比均值进行定量的校正方法。方法具有在线自动判别和修复异常峰并自行校正的功能，和通用的标准系列定量方法相比，两者测定精度相当。但本文提出的方法抗干扰能力明显优于后者，可适用于在线过程监测。     

Sequential optimisation of the separation of a complex mixture of plastics additives by HPLC with a quaternary gradient and a dual detection system

Summary The separation of two families of plastics additives (phenolic antioxidants and UV absorbers) has been achieved by high performance liquid chromatography with a quaternary gradient. A methodology of separation based on a sequential optimization is described. After a preliminary study of the effects of solvents on retention of compounds, the mobile phase is first chosen for each class of additives (Irganox and Tinuvin), then for the mixture of all the compounds and the separation is finally optimized. The importance of the column phase ratio is also reported. The use of two detectors, UV absorbance and light-scattering, enables all the compounds to be detected. The performance of the detectors has been compared and the effects of the nebulization temperature on the detection of low molecular mass compounds is reported.     

梯度淋洗离子色谱法同时分析有机酸与无机酸阴离子的应用研究

利用梯度离子色谱法和抑制电导检测器对有机酸与无机酸阴离子的色谱分析条件进行了研究,建立了最佳梯度程序。通过浓缩柱的富集,可使方法的检出限达1×10-3μg·L-1,各离子的相对标准偏差大部分在5%以下。此方法可用于火电厂水汽系统中痕量低分子有机酸和无机酸阴离子的快速、准确分析,结果满意。