How to prepare and stabilize very small nanoemulsions

Practical and theoretical considerations that apply when aiming to formulate by ultrasonication very small nanoemulsions (particle diameter up to 150 nm) with very high stability are presented and discussed. The droplet size evolution during sonication can be described by a monoexponential function of the sonication time, the characteristic time scale depending essentially on the applied power. A unique master curve is obtained when plotting the mean diameter size evolution as a function of sonication energy. We then show that Ostwald ripening remains the main destabilization mechanism whereas coalescence can be easily prevented due to the nanometric size of droplets. The incorporation of "trapped species" within the droplet interior is able to counteract Ostwald ripening, and this concept can be extended to the membrane compartment. We finally clarify that nanoemulsions are not thermodynamically stable systems, even in the case where their composition lies very close to the demixing line of a thermodynamically stable microemulsion domain. However, as exemplified in the present work, nanoemulsion systems can present very long-term kinetic stability.     

Size selectivity in field-flow fractionation: lift mode of retention with near-wall lift force

A simple theoretical model for the size selectivity, S(d), in the lift mode of retention in field-flow fractionation (FFF) is developed on the basis of the near-wall lift force expression. S(d) is made up of two contributions: the flow contribution, S(d,f), arising from the variation of the flow velocity at center of particle due to a change in particle position with particle size, and a slip contribution, S(d,s), arising from the concomitant change in the extent of retardation due to the presence of a nearby channel wall. The slip contribution is minor, but not negligible, and amounts to 10-20% of the overall size selectivity. It contributes to reduce S(d) in sedimentation FFF but to enhance it in flow FFF. S(d) would steadily increase with particle size if the flow profile was linear (Couette flow). Because of the curvature of the flow profile encountered in the classical Poiseuille flow, S(d) exhibits a maximum at some specific particle size. The model predicts a significant difference in S(d) between sedimentation FFF and flow FFF, arising from the different functional dependences of the field force with particle size between these two methods. The predictions are in good agreement with the various S(d) values reported in the literature in both sedimentation and flow FFF. On the basis of the model, guidelines are given for adjusting the operating parameters (carrier flow rate and field strength) to optimize the size selectivity. Finally, it is found that S(d) generally decreases with decreasing channel thickness.     

Practical implications of ionic strength effects on particle retention in thermal field-flow fractionation

Modification of ionic strength of an aqueous or non-aqueous carrier solution can have profound effects on the particle retention behavior in thermal field-flow fractionation (ThFFF). These effects can be considered as either advantageous or not depending on the performance criteria under consideration. Aside from the general increase in retention time of particulate material (latexes and silica particles), our experiments indicate improvement in resolution with increases in electrolyte concentration. Absence of an electrolyte in the carrier solution causes deviations from the theoretically expected linear behavior between the retention parameter lambda (a measure of the extent of interaction between the applied field and the particle) and the reciprocal temperature drop across the channel walls. A negative interaction parameter delta(w), of about -0.170 was determined for 0.105- and 0.220-microm polystyrene (PS) latex particles suspended in either a 0.25 or a 1.0 mM TBAP-containing acetonitrile carrier and for 0.220 microm PS in 0.50 and 1.0 mM NaCl-containing aqueous medium. This work also demonstrates that optimum electrolyte concentrations can be chosen to achieve reasonable experimental run-times, good resolution separations, and shifts in the steric inversion points at lower field strengths, and that too high electrolyte concentrations can have deleterious effects such as band broadening and sample loss through adsorption to the channel accumulation surface. The advantages of using ionic strength rather than field strength to effect desired changes are lowered power consumption and possible application of ThFFF to high temperature-labile samples.     

Temperature programmed retention indices: Calculation from isothermal data. Part 1: Theory

Direct conversion of isothermal to temperature programmed indices is not possible. In this work it is shown that linear temperature programmed retention indices can only be calculated from isothermal retention data if the temperature dependence of both the distribution coefficients and the column dead time are taken into account. Procedures are described which allow calculation of retention temperatures and from these, accurate programmed retention indices. Within certain limits the initial oven temperature and programming rate can be chosen freely. The prerequisite for this calculation is the availability of reliable isothermal retention data (retention times, retention factors, relative retention times, or retention indices) at two different temperatures for one column. The use of compiled isothermal retention indices at two different temperatures for the calculation of retention temperatures and thus temperature programmed indices is demonstrated. For the column for which programmed retention indices have to be determined, the isothermal retention times of the n-alkanes and the column dead time as a function of temperature have to be known in addition to the compiled data for a given stationary phase. Once the programmed retention indices have been calculated for a given column the concept allows the calculation of temperature programmed indices for columns with different specifications. The characteristics which can be varied are: column length, column inner diameter, phase-ratio, initial oven temperature, and programming rate.     

Prediction of protein retention at gradient elution conditions in ion-exchange chromatography.

This work presents a prediction procedure for protein retention in ion-exchange chromatography, where two linear gradient experiments of different length give the protein retention time at other linear gradients. The procedure predicts the retention time of early and late eluting proteins with similar precision and predictions by extrapolation deviate approximately 3% or less from the experimental retention times. By using the ionic strength, this procedure predicts protein retention times obtained with divalent ions in the eluent more accurately than a well-established procedure that uses the protein co-ion concentration.     

Fast quantitative characterisation of differential mobility responses

A chromatography-based method for producing mass flux-response surfaces for differential mobility spectrometers is described as a replacement for exponential dilution and mixing approaches. An exponential dilution or mixing experiment typically takes 150 min; while the exponential function in the Gaussian elution profile obtained from linear chromatography may be exploited in approximately 10 s. The approach was demonstrated with a gas chromatograph-mass spectrometer and the correlation of the calibration results to nominal on-column masses was within experimental error for 19 separate analyses. The method was then applied to a gas chromatographic (10.6 eV UV) differential mobility spectrometer. Mass fluxes in the range 10 pg s(-1) to 250 ng s(-1) were generated over the 5 s to 10 s associated with the elution of a chromatographic peak. The characterisations were repeated for a range of electrical field strengths from 10 kV cm(-1) to 30 kV cm(-1). Triplicate runs indicated that the approach was reproducible and that response surfaces could be generated rapidly from chromatographic data. The effects of trace impurities associated with the chromatographic eluent on the relationship between compensation voltage and electrical field strength was observed. This emphasised the importance of managing this aspect of the operation if reliable estimates of alpha functions for the compounds under study were to be obtained. Application of this approach to other detection systems with an 85% reduction in the analytical operations required to produce a reliable calibration function was also noted.     

Applicability of linear and nonlinear retention‐time models for reversed‐phase liquid chromatography separations of small molecules,peptides, and intact proteins

The applicability and predictive properties of the linear solvent strength model and two nonlinear retention‐time models, i.e., the quadratic model and the Neue model, were assessed for the separation of small molecules (phenol derivatives), peptides, and intact proteins. Retention‐time measurements were conducted in isocratic mode and gradient mode applying different gradient times and elution‐strength combinations. The quadratic model provided the most accurate retention‐factor predictions for small molecules (average absolute prediction error of 1.5%) and peptides separations (with a prediction error of 2.3%). An advantage of the Neue model is that it can provide accurate predictions based on only three gradient scouting runs, making tedious isocratic retention‐time measurements obsolete. For peptides, the use of gradient scouting runs in combination with the Neue model resulted in better prediction errors (<2.2%) compared to the use of isocratic runs. The applicability of the quadratic model is limited due to a complex combination of error and exponential functions. For protein separations, only a small elution window could be applied, which is due to the strong effect of the content of organic modifier on retention. Hence, the linear retention‐time behavior of intact proteins is well described by the linear solvent strength model. Prediction errors using gradient scouting runs were significantly lower (2.2%) than when using isocratic scouting runs (3.2%).     

Determination of calibration function in thermal field flow fractionation under thermal field programming

A new procedure for determining the calibration function able to relate retention and operative parameters to molecular weight of the species in thermal field flow (ThFFF) under thermal field programming (TFP) conditions is presented. The procedure involves determining the average values of retention parameters under TFP and determining a numerical function related to the temperature variations that occur during TFP. The calibration parameters are obtained by a procedure fitting the retention and operative parameters that hold true at the beginning of the TFP. The procedure is closely related to the one previously developed to calibrate the retention time axis under TFP ThFFF and, together, they constitute a full calibration procedure. Experimental validation was performed with reference to polystyrene (PS)-decalin and PS-THF systems. The calibration functions here obtained were compared to those derived by the classical procedure at constant thermal field ThFFF to obtain the calibration function at variable cold wall temperatures. Excellent agreement was found in all cases proving "universality" of the ThFFF calibration concept, i.e. it is independent of the particular system on which it was determined and can thus be extended to ThFFF operating under TFP. The new procedure is simpler than the classical one since it requires less precision in setting the instrumentation and can be obtained with fewer experiments. The potential applications for the method are discussed.     

Faster GC analyses performed by flow programming in short capillary columns

The technique of programming the carrier gas flow rate in gas chromatography, especially in connection with the use of capillary columns shorter than 10 m can significantly accelerate GC analyses. Equations for calculation of the parameters of the exponential flow function and retention data are described. The effects of flow programming in a short capillary column are shown in a few chromatograms. Different programming rates are tested and compared with temperature programming.     

pH gradient simulator for electrophoretic techniques in a windows environment

A new program is presented for optimizing mixtures of buffers and titrants for creating pH gradients for isoelectric focusing in immobilized pH gradients and in general for chromatographic processes. The program is written on a windows platform and it includes several novel features compared with previous simulators. First, the estimation of a pH gradient (a non-linear problem) has been transformed into a linear programming problem, thus allowing the use of the simplex as an optimization algorithm. Second, several types of pH gradients can be simulated and optimized, including linear, exponential, logarithmic and sigmoidal. Finally, an equation has been implemented in the program that accounts for the variation of the activity coefficients of ions as a function of the prevailing ionic strength in solution. This simulator was checked experimentally by eluting solutions from a two-vessel gradient mixer and verifying the shape of the various pH gradients. An excellent correlation was found between simulated and experimental data. The program allows the calculation and optimization of pH profiles (including the accompanying ionic strength and buffering power values) for mixtures of up to 50 different monoprotic or oligoprotic buffers and titrants. Calculations and optimizations are performed in a fraction of the time required by previous programs (often in less than 1 min, even for highly complex mixtures).     

DC-dielectrophoretic separation of microparticles using an oil droplet obstacle

A new dielectrophoretic particle separation method is demonstrated and examined in the following experimental study. Current electrodeless dielectrophoretic (DEP) separation techniques utilize insulating solid obstacles in a DC or low-frequency AC field, while this novel method employs an oil droplet acting as an insulating hurdle between two electrodes. When particles move in a non-uniform DC field locally formed by the droplet, they are exposed to a negative DEP force linearly dependent on their volume, which allows the particle separation by size. Since the size of the droplet can be dynamically changed, the electric field gradient, and hence DEP force, becomes easily controllable and adjustable to various separation parameters. By adjusting the droplet size, particles of three different diameter sizes, 1 microm, 5.7 microm and 15.7 microm, were successfully separated in a PDMS microfluidic chip, under applied field strength in the range from 80 V cm-1 to 240 V cm-1. A very effective separation was realized at the low field strength, since the electric field gradient was proved to be a more significant parameter for particle discrimination than the applied voltage. By utilizing low strength fields and adaptable field gradient, this method can also be applied to the separation of biological samples that are generally very sensitive to high electric potential.     

准弹性光散射研究苯乙烯-二乙烯基苯共聚超微粒的溶液性质

本文用光子相关光谱技术研究了聚苯乙烯-二乙烯基苯(PSt-DVB)共聚超微粒的溶液性质.该微粒的浓度涨落时间相关函数在波矢为q时,有指数衰减形式,其衰减速率г=q~2D_t.在精确的实验误差范围内,衰减速率的变化率为零。用累积量方法解析散射场的一阶时间相关方程,得到了作为浓度和温度函数的微粒扩散常数.与流体办学方程结合,计算出了微粒的流体力学尺寸.通过实验也确定了微粒在良溶剂中的分子形态.     

超临界流体色谱密度线性程升时保留规律的研究室

推导了密度线性程升时超临界流体色谱中保留值的运动方程和相应的理论板高方程式, 并在超临界色谱条件下用毛细管柱和微填充柱上的数据对保留值方程作了验证, 理论计算值和实验测量值之间的偏差最大不超过5.00%。在推导理论方程时,设一定密度范围内1nk和密度p之间为线性函数, 这个假设也得到了实验验证。     

网格搜索法优化毛细管气相色谱线性程序升温

 用一种新的计算程序进行毛细管气相色谱线性程序升温的优化。在OV 10 1固定相上对 2 1种Kovats保留指数为 6 0 0～ 10 0 0的组分进行了自动寻优工作 ,并分别在起始温度为 2 5℃和 30℃的线性程序升温条件下取得了较好的结果。这种优化方法不但节省了计算时间 ,而且在对计算程序进行适当修改后 ,可以用于多阶程序升温的优化工作。     

Generic approach to the method development of intact protein separations using hydrophobic interaction chromatography

We describe a liquid chromatography method development approach for the separation of intact proteins using hydrophobic interaction chromatography. First, protein retention was determined as function of the salt concentration by isocratic measurements and modeled using linear regression. The error between measured and predicted retention factors was studied while varying gradient time (between 15 and 120 min) and gradient starting conditions, and ranged between 2 and 15%. To reduce the time needed to develop optimized gradient methods for hydrophobic interaction chromatography separations, retention‐time estimations were also assessed based on two gradient scouting runs, resulting in significantly improved retention‐time predictions (average error < 2.5%) when varying gradient time. When starting the scouting gradient at lower salt concentrations (stronger eluent), retention time prediction became inaccurate in contrast to predictions based on isocratic runs. Application of three scouting runs and a nonlinear model, incorporating the effects of gradient duration and mobile‐phase composition at the start of the gradient, provides accurate results (improved fitting compared to the linear solvent‐strength model) with an average error of 1.0% and maximum deviation of –8.3%. Finally, gradient scouting runs and retention‐time modeling have been applied for the optimization of a critical‐pair protein isoform separation encountered in a biotechnological sample.     

Electrorheological fluid characterization via a vertical oscillation rheometer

A custom designed vertical oscillation rheometer (VOR) is used for the rheological measurements of electrorheological (ER) fluids consisting of 15 and 20 vol.% semiconducting polyaniline particles suspended in silicone oil. The viscoelastic material functions, including complex viscosity and complex shear modulus, are measured via geometric parameters, measured force, and applied strain of the VOR. Viscoelastic properties of the ER fluids are also measured as a function of applied electric field strength and particle concentration. The VOR, equipped with a high voltage generator, can easily be constructed and used to measure ER properties. It is further found that polyaniline suspensions behave as viscoelastic materials in an electric field. In linear viscoelastic conditions, elasticity was promoted with the increment of electric field due to particle chain structure in the presence of the applied electric field. It is also found that the applied electric field rather than particle concentration enhanced the elasticity of ER fluids.     

梯度液相色谱中任意等度、线性和阶梯梯度组合下的保留时间公式

基于线性溶剂强度模型,应用特征线分析的方法求解梯度洗脱模式下的理想液相色谱模型。在考虑到梯度延迟时间会对溶质的保留时间造成影响的情况下,得到适合于梯度液相色谱中任意等度、线性和阶梯梯度组合条件下的保留时间推导公式。应用这些公式计算任意的梯度条件下的保留时间,并将得到的结果与数值计f算的结果进行比较,二者完全一致,从而验证了推导得到的保留时间公式的正确性。由于这些公式具有形式简单、适用范围广等优点,因此可方便地应用于实际应用中,具有较高的实用价值。     

Investigation into the mechanism of tack of rubbers

The tack of cis-polybutadiene was measured by means of a Skewis tackmeter, and the results, which showed considerable variability, could be represented by a double exponential cumulative distribution function of Gumbel. It was concluded that there exists a linear relationship between the applied forces (contact and break) and the time that these forces are applied in the tack test. The mechanism of tack is considered in terms of two distinct processes: (a) the development of a bond and (b) the strength of the bond thus formed. The latter is rationalized by applying Eyring's absolute rate theory. A simple equation is derived which predicts an inverse linear relation between the breaking time and breaking force, which is in agreement with the experiment. The theory also suggests that the area of actual interpenetration of polymer interfaces depends largely on the contact pressure, whereas the depth of penetration appears to depend to a similar degree on both contact pressure and contact time.