Micellar electrokinetic chromatography with bis(2-ethylhexyl)sodium sulfosuccinate vesicles determination of synthetic food antioxidants

Capillary electrokinetic chromatography is suitable for the separation of mixtures of uncharged and charged solutes. In the present work the behavior of six synthetic food antioxidants--2[3]-tert.-butyl-4-hydroxyanisole, 2,6-di-tert.-butyl-p-cresol, tercbutylhydroquinone, 3,4,5-trihydroxybenzoic acid propyl ester, 3,4,5-trihydroxybenzoic acid octyl ester and 3,4,5-trihydroxybenzoic acid dodecyl ester--was studied in a capillary electrophoresis system using capillary electrokinetic chromatography with vesicles of the surfactant bis(2-ethylhexyl)sodium sulfosuccinate (AOT). Several studies aimed at calculating the critical aggregation concentration of the surfactant were conducted to check that under the conditions used the AOT was in a state of aggregation. Having checked the association shown by the surfactant, we then explored the greater or lesser capacity of the antioxidants to interact with this compound. We followed the evolution of the molecular absorption spectra of each of the antioxidants in the presence of the surfactant at different concentrations and the retention factors were calculated at different pH values. Additionally, in order to determine which species--anionic or neutral--was present at the pH of the buffer used (boric/borate), the pKa values in acetonitrile-water (20:80) were obtained. Resolution and quantification of the antioxidants demand optimization of the variables involved in the system, such as the percentage of acetonitrile, the concentration of AOT and boric/borate buffer, pH, voltage, etc. When this part of the study had been completed, calibrations were obtained for each of the antioxidants, obtaining good linear correlation coefficients in all cases. Finally, we propose a method that allows the resolution of the six most employed antioxidants in a capillary electrophoretic system in 15 min, using electrokinetic chromatography with AOT as the pseudostationary phase.     

Chemometric comparison of recent chromatographic and electrophoretic methods in a quantitative structure-retention and retention-activity relationship context

The retention characteristics of 21 basic pharmaceutical substances with a considerable difference in hydrophobicity (octanol-water partition coefficients, log P, between -0.026 and 6.45) are considered on an immobilized artificial membrane column, with a micellar liquid chromatography and a micellar electrokinetic capillary chromatography method. Utilising principal component analysis (PCA), it is seen that although the main retention principle is the same, the above methods as well as more classical RP-HPLC methods vary in secondary retention mechanisms. Combining the results of different methods a differentiation of the substances into their pharmacological families can be seen with PCA. The high correlations of the retention characteristics with log P and a biological parameter seem little affected by the method used.     

PH effects on drug interactions with lipid bilayers by liposome electrokinetic chromatography

Liposome electrokinetic chromatography (LEKC) provides convenient and rapid methods for studying drug interactions with lipid bilayers using liposomes as a pseudostationary phase. LEKC was used to determine the effects of pH on the partitioning of basic drugs into liposomes composed of zwitterionic phosphatidylcholine (PC), anionic phosphatidylglycerol (PG), and cholesterol, which mimic the composition of natural cell membranes. An increase in pH results in a smaller degree of ionization of the basic drugs and consequently leads to a lower degree of interaction with the negatively charged membranes. From the LEKC retention data, the fractions of drugs distributed in the bulk aqueous and the liposome phase were determined at various pH values. Finally, lipid mediated shifts in the ionization constants of drugs were examined.     

Analysis of antiepileptic drugs in biological fluids by means of electrokinetic chromatography

An overview of the electrokinetic chromatographic methods for the analysis of antiepileptic drug levels in biological samples is presented. In particular, micellar electrokinetic capillary chromatography is a very suitable method for the determination of these drugs, because it allows a rapid, selective, and accurate analysis. In addition to the electrokinetic chromatographic studies on the determination of antiepileptic drugs, some information regarding sample pretreatment will also be reported: this is a critical step when the analysis of biological fluids is concerned. The electrokinetic chromatographic methods for the determination of recent antiepileptic drugs (e.g., lamotrigine, levetiracetam) and classical anticonvulsants (e.g., carbamazepine, phenytoin, ethosuximide, valproic acid) will be discussed in depth, and their pharmacological profiles will be briefly described as well.     

Rapid estimation of octanol-water partition coefficients using synthesized vesicles in electrokinetic chromatography

Vesicle electrokinetic chromatography (VEKC) using vesicles synthesized from the oppositely charged surfactants cetyltrimethylammonium bromide (CTAB) and sodium octyl sulfate (SOS) and from the double-chained anionic surfactant bis(2-ethylhexyl)sodium sulfosuccinate (AOT) was applied to the indirect measurement of octanol-water partition coefficients (log Po/w). A variety of small organic molecules with varying functional groups, pesticides, and organic acids were evaluated by correlating log Po/w and the logarithm of the retention factor (log k') and comparing the calibrations. A linear solvation energy relationship (LSER) analysis was conducted to describe the retention behavior of the vesicle systems and compared to that of octanol-water partitioning. The solute hydrogen bond donating behavior is slightly different with the vesicle interactions using CTAB-SOS vesicles as compared to the octanol-water partitioning model. The AOT vesicle and octanol-water partitioning systems showed similar partitioning characteristics. VEKC provides rapid separations for determinations of log Po/w in the range of 0.5 to 5 using CTAB-SOS vesicles and 0 to 5.5 using AOT vesicles.     

Quantitative determination of drug encapsulation in poly(lactic acid) nanoparticles by capillary electrophoresis

Capillary electrophoretic (CE) methods were used for the quantitative determination of model drugs [salbutamol sulphate (SS), sodium cromoglycate (SCG) and beclomethasone dipropionate (BDP)] in poly(D,L-lactic acid) (PLA) nanoparticles, which were prepared by the nanoprecipitation method. Zeta potential and size distribution of the nanoparticles were determined by electrophoretic mobility determinations and photon correlation spectroscopy, respectively. Interactions between the drugs, the PLA nanoparticles and the fused-silica capillary were investigated by electrokinetic capillary chromatography (EKC). A quantitative CE method was developed for salbutamol sulphate and sodium cromoglycate, and the linearity and repeatability of migration times, peak areas and peak heights were determined. Microemulsion electrokinetic chromatography was used for the quantitative determination of beclomethasone dipropionate. According to this study, the applied electromigration techniques were suitable for the interaction, drug entrapment and dissolution studies of pharmaceutical nanoparticles. The results suggest that even quantitation of the drug located inside the nanoparticles was possible. Encapsulation of the more hydrophilic model drugs (SS, SCG) in the PLA nanoparticles was less efficient than in the case of BDP.     

Polymer-drug conjugates as inhalable drug delivery systems: A review

Accelerating interest by the pharmaceutical industry in the identification and development of less invasive routes of nanomedicine administration, coupled with defined efforts to improve the treatment of respiratory diseases through inhaled drug administration has fuelled growing interests in inhalable polymer-drug conjugates. Polymer-drug conjugates can alter the pharmacokinetic profile of the loaded drug after inhaled administration and enable the controlled and sustained exposure of the lungs to drugs when compared to the inhaled or oral administration of the drug alone. However, the major concern with the use of inhalable polymer-drug conjugates is their biocompatibility and long-term safety in the lungs, which is closely linked to lung retention times. A detailed understanding about the pharmacokinetics, lung disposition, clearance and safety of inhaled polymer-drug conjugates with significant translational potential is therefore required. This review therefore provides a comprehensive summary of the latest developments for several types of polymer-drug conjugates that are currently being explored as inhalable drug delivery systems. Finally, the current status and future perspective of the polymer-drug conjugates is also discussed with a focus on current knowledge gaps.     

SDS-Based Biomembrane Mimetic Chromatography for Prediction of Human Drug Transport as an in Vitro Technique

The main oral drug absorption barriers are fluid cell membranes, and generally drugs are absorbed by a passive diffusion mechanism. On the other hand, the blood–brain barrier (BBB) is considered to be the main barrier to drug transport into the central nervous system (CNS). The BBB restricts the passive diffusion of many drugs from blood to brain. Biopartitioning micellar chromatography (BMC), a mode of micellar liquid chromatography that uses micellar mobile phases in adequate experimental conditions, can be useful as an in vitro system in mimicking the drug partitioning process into biological systems. In this study, relationships between the BMC retention data of a heterogeneous set of 12 drugs and their pharmacokinetics parameters (human oral drug absorption and BBB penetration ability) are studied and the predictive ability of the models is evaluated. Modeling of log k _BMC of these compounds was established by multiple linear regression in two different concentrations (0.07 and 0.09 M) of sodium dodecyl sulfate (SDS). The results showed a fair correlation between human oral drug absorption and BMC retention data in 0.09 M SDS (R ² = 0.864) and a good correlation between the blood–brain distribution coefficient and BMC retention data in 0.07 M of SDS (R ² = 0.887). Application of the developed models to a prediction set demonstrated that the model is also reliable with good predictive accuracy. The external and internal validation results showed that the predicted values are in good agreement with the experimental value.     

Impurity analysis of pharmaceuticals using capillary electromigration methods

This review deals with the determination of impurities in pharmaceuticals by electromigration methods in the capillary format. These separation methods are either based on the different effective mobility of the charged analytes (as in zone electrophoresis and isotachophoresis) or include hybrid methods such as micellar electrokinetic chromatography, microemulsion electrokinetic chromatography and electrochromatography. The pharmaceutically active compounds under consideration belong to chemotherapeutic agents, central nervous system drugs, histamine receptor drugs, cardiovascular drugs, anticancer drugs, anti-inflammatory drugs and some other drugs. The review discusses about 150 publications from the period between 1980 and 2007 with special emphasis on the recent trends and gives details about the experimental conditions applied for analyses and the obtained analytical performance parameters.     

SDS-Based Biomembrane Mimetic Chromatography for Prediction of Human Drug Transport as an in Vitro Technique

The main oral drug absorption barriers are fluid cell membranes, and generally drugs are absorbed by a passive diffusion mechanism. On the other hand, the blood–brain barrier (BBB) is considered to be the main barrier to drug transport into the central nervous system (CNS). The BBB restricts the passive diffusion of many drugs from blood to brain. Biopartitioning micellar chromatography (BMC), a mode of micellar liquid chromatography that uses micellar mobile phases in adequate experimental conditions, can be useful as an in vitro system in mimicking the drug partitioning process into biological systems. In this study, relationships between the BMC retention data of a heterogeneous set of 12 drugs and their pharmacokinetics parameters (human oral drug absorption and BBB penetration ability) are studied and the predictive ability of the models is evaluated. Modeling of log k _BMC of these compounds was established by multiple linear regression in two different concentrations (0.07 and 0.09 M) of sodium dodecyl sulfate (SDS). The results showed a fair correlation between human oral drug absorption and BMC retention data in 0.09 M SDS (R ² = 0.864) and a good correlation between the blood–brain distribution coefficient and BMC retention data in 0.07 M of SDS (R ² = 0.887). Application of the developed models to a prediction set demonstrated that the model is also reliable with good predictive accuracy. The external and internal validation results showed that the predicted values are in good agreement with the experimental value.

     

Submitochondrial distribution of basic drugs in the isolated perfused lung

To clarify the mechanism by which basic drugs accumulate in the lung mitochondria, the binding selectivity of drugs to different submitochondrial components of the perfused rat lung was examined. The accumulation of basic drugs was the highest in the mitochondrial outer membrane fraction. The drug accumulation in this fraction increased with lipid solubility and was dose-dependent. It appears then that selective binding sites for basic drugs are present in the mitochondrial outer membrane.     

Hydrophilic interaction liquid chromatography/electrospray mass spectrometry determination of acyclovir in pregnant rat plasma and tissues

Reversed-phase chromatography is the most common means of separation for small drug molecules. However, polar drugs may suffer from poor retention and peak shape in reversed-phase high-performance liquid chromatography (RP-HPLC). Hydrophilic interaction liquid chromatography (HILIC) provides a viable alternative to RP-HPLC and is an excellent way to separate polar compounds. This paper describes a HILIC/ESI-MS/MS assay for the determination of acyclovir from rat plasma, amniotic fluid, placental tissue, and fetal tissue. The isocratic separation utilizes an underivatized silica column with an acetonitrile/formate buffer mobile phase (80:20). The method is validated over a range of 50 ng/mL to 50 micro g/mL with % error and % relative standard deviation of <15% over 3 days. All samples are prepared by acetonitrile protein precipitation, which yields high recovery (>84% for acyclovir). This assay can be applied to the pharmacokinetic study of the placental transfer of acyclovir.     

Calorimetric study of the alpha-tocopherol solubility in reversed AOT micelles

The experimental data of heat of mixing (Q) for heterogeneous system alpha-tocopherol/AOT/n-heptane with and without water at 25 degrees C are presented. The Q dependence on AOT (sodium bis (2-ethylhexyl) sulfosuccinate) concentration, and R parameter defined as R=[H2O]/[AOT] with flow calorimetric method were investigated. Using the D'Aprano model (which is formally identical to that used earlier by Magid et al.) the binding constant (K), the distribution constant of alpha-tocopherol (K distr) between hydrocarbon and the micellar phase, and the standard enthalpy of transfer (DeltaH tr 0) of alpha-tocopherol from the hydrocarbon to AOT reversed micelles were calculated. The solubility of alpha-tocopherol in AOT reversed micelles explored with the calorimetric technique was compared to the literature data obtained respectively with UV spectrophotometry for reversed micelles and by other techniques for the phospholipid bilayer.     

Effect of liposome composition on β-blocker interactions studied by capillary electrokinetic chromatography

Liposome capillary electrokinetic chromatography was used to investigate the interactions between three β-blockers of different hydrophobicity and various liposome solutions. The studied β-blockers comprised alprenolol, propranolol, and carvedilol. The composition of the liposome solutions, containing 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine, 1,2-dioleoyl-sn-glycero-3-phos-phoethanolamine, 1-palmitoyl-2-oleoyl-sn-glycero-3-phospho-l -serine, and cholesterol in various molar ratios, was designed by a response surface methodology-central composite design approach. Subsequently, after conducting the liposome capillary electrokinetic chromatography experiments and determining the retention factors from the electrophoretic mobilities of the compounds, and further calculating the distribution coefficients, an analysis of variance was performed. After extracting the statistical models, optimal operational conditions were obtained based on the developed models. To further investigate the interactions between the β-blockers and the liposomes, nanoplasmonic sensing experiments were carried out on two different liposome systems. The overall results demonstrate the strong influence of cholesterol and 1-palmitoyl-2-oleoyl-sn-glycero-3-phospho-l -serine on the distribution coefficients.     

Prediction of Human Drug Absorption Using Liposome Electrokinetic Chromatography

With the tremendously increasing numbers of novel drug candidates, there remains a compelling need for rapid screening methods for drug-like physiochemical and pharmacokinetic properties. Different technologies have emerged that enable rapid screening in vitro for sorting out new chemical entity (NCE) classes. It is invaluable for these technologies being developed early in the drug discovery process to avoid the loss of cost and time in late development due to poor absorption and/or bioavailability. In this study, liposome electrokinetic chromatography (LEKC) serves as a convenient, rapid and cost-effective tool to determine lipophilicity and to predict human oral absorption. Twenty-seven organic neutral molecules were evaluated by octanol/water system (log P _ow) and LEKC (log k), and linear solvation energy relationship (LSER) analysis was conducted to compare the retention mechanism between LEKC and octanol/water system. LEKC can provide a rapid indirect measurement of log P _ow for small organic neutral molecules. A clearly sigmoidal relationship could be seen by correlating log k with the fraction of 25 drugs absorbed in humans (Fa), and the outliers suggested the involvement of non-transcellular passive diffusion, e.g. active transport, paracellular route; on the contrary, it is not the case with the octanol/water system. Therefore, LEKC, in combination with other permeability prediction model, can provide a primary screen for a large number of drug candidates at early stage of the drug discovery process with high-throughput and at low-cost.     

Evaluation of hydrophobic interaction between acidic drugs and bovine serum albumin by reversed-phase high-performance liquid chromatography

The contribution of hydrophobic interaction to the protein binding of acidic drugs has been evaluated in terms of a new hydrophobic index (r-value), defined as the slope of the log-log plots of capacity factor vs. reciprocal of methanol concentration in an aqueous binary mobile phase, measured by the reversed-phase high-performance liquid chromatography. The logarithms of the binding constants (log K1) of the selected acidic drugs and the related aromatic carboxylic acids indicated linear relationship with their r-values, suggesting that the effect of hydrophobicity on protein binding can be explained similarly to that on the retention onto the reversed-phase stationary ligand.     

Measurement of histamine in rat bronchoalveolar lavage fluid by high-performance cation-exchange chromatography coupled with post-column derivatization to o-phthaldialdehyde.

To better define involvement of mast cell derived mediators in the pulmonary response to fibrogenic dusts, a rapid and accurate method was required to analyze samples of bronchoalveolar lavage fluid for histamine. Samples of rat lung lavage were analyzed for histamine via high-performance cation-exchange chromatography coupled with post-column derivatization with o-phthaldialdehyde. The fluorescent derivative could be detected to ca. 1 ng/ml of lavage. Recoveries averaged 94.2% with an average relative standard deviation of +/- 5.3%. There were no correlations between amount or fibrogenicity of inhaled dust and subsequent release of histamine into lavage fluid.     

Application of retention factors in affinity electrokinetic chromatography and capillary electrophoresis.

Affinity capillary electrophoresis has become an established approach for performing interaction studies. In affinity electrokinetic chromatography the retention factor, as in liquid chromatography, is useful for describing the migration behavior of the analytes, and is instrumental for assessing the affinity of an analyte for the pseudo-stationary phase. Erroneous use of the retention factor concept in affinity capillary electrophoretic studies has appeared in a number of recent papers. The errors and their origin are pointed out, and the correct use of retention factors in affinity electrokinetic chromatography and capillary electrophoresis is summarized.     

Monitoring of cefepime in human serum and plasma by micellar electrokinetic capillary chromatography: Improvement of sample preparation and validation by liquid chromatography coupled to mass spectrometry

Cefepime monitoring in deproteinized human serum and plasma by micellar electrokinetic capillary chromatography and liquid chromatography coupled to mass spectrometry in presence of other drugs is reported. For micellar electrokinetic capillary chromatography, sample preparation comprised dodecylsulfate protein precipitation at pH 4.5 using an increased buffer concentration compared to that of a previous assay and removal of hydrophobic compounds with dichloromethane. This provided robust conditions for cefepime analysis in the presence of sulfamethoxazole and thus enabled its determination in samples of patients that receive cotrimoxazole. The liquid chromatography assay is based upon use of a column with a pentafluorophenyl‐propyl modified and multiendcapped stationary phase and the coupling to electrospray ionization with a single quadrupole detector. The performances of both assays with multilevel internal calibration were assessed with calibration and control samples and both assays were determined to be robust. Cefepime levels monitored by micellar electrokinetic capillary chromatography in samples from patients that were treated with cefepime only and with cefepime and cotrimoxazole were found to compare well with those obtained by liquid chromatography coupled to mass spectrometry. Cefepime drug levels determined by micellar electrokinetic capillary chromatography could thereby be validated.