A study of the binding between polymers and peptides, using affinity capillary electrophoresis, applied to polymeric drug delivery systems

We have investigated the potential of affinity capillary electrophoresis (ACE) to evaluate binding constants between an anionic polydispersed polymer and four peptides. Nonlinear regression and three current linearization methods, the y-reciprocal, the x-reciprocal and the double-reciprocal, were employed for the estimation of the binding constants. The x-reciprocal and the double-reciprocal plots indicated the presence of two portions of straight lines for angiopeptin, triptorelin and the thyrotropin releasing hormone (TRH), and therefore the probable existence of a second-order interaction which causes the deviation from the 1:1 model. Peptide 1 exhibited a unique binding constant of 2.4 x 10(6)M(-1). In contrast, angiopeptin, triptorelin and TRH exhibited a K(1) of 4.0 x 10(6), 5.3 x 10(6) and 20.2 x 10(6)M(-1), respectively, and a K(2) of 0.4 x 10(6), 0.5 x 10(6) and 1.4 x 10(6)M(-1), respectively. The origin of the high scattering of the data points was further investigated. Neither the viscosity, nor the adsorption of the peptides to the capillary wall appeared to be the determining factor of data scattering. Finally, a possible adsorption of the polymer leading to the electroosmotic flow instability was supposed.     

Determination of oligonucleotide ISIS 2922 in nanoparticulate delivery systems by capillary zone electrophoresis

ISIS 2922 is an antisense oligonucleotide with antiviral activity against cytomegalovirus. However, its rapid degradation in biological fluids and its low capacity for diffusion across cell membranes limit its therapeutical use. One possibility to overcome these drawbacks consists of using nanoparticles as drug carriers. The aim of this study was to develop an analytical method for determining the amount of ISIS 2922 loaded into albumin nanoparticles. For this purpose, capillary zone electrophoresis (CZE) was performed on a fused-silica capillary filled with borate buffer (12.5 mM, pH 9.5). Paracetamol was used as an internal standard and a diode-array detection system was set at 270 nm. Under these conditions, the limit of quantitation of ISIS 2922 was 1.27 microg and the precision and accuracy of the method did not exceed 7%. Moreover, the use of paracetamol as internal standard and the quantification by means of a 'corrected area' procedure enabled us to reduce the peak variability and accurately determine the amount of oligonucleotide loaded in the albumin nanoparticles. In summary, this assay is a selective and sensitive CZE method for the accurate quantitation of ISIS 2922 oligonucleotide in albumin nanoparticles.     

Microfabricated capillary array electrophoresis DNA analysis systems

The equation of motion for a peak in a gas chromatography experiment is shown to be exact whenever the column outlet pressure is approximately zero. Based on steady-state assumptions, this equation is solved for an isothermal analysis with a single-ramp inlet-pressure or flow-rate program. Theoretical calculations of retention times are confirmed experimentally for two n-alkanes using a capillary column connected to a mass-selective detector. A slight but consistent discrepancy between theoretical and measured results is interpreted as a failure of the steady-state assumption when the magnitude of the rate of change of inlet pressure is large.     

Enantiospecific analysis by capillary electrophoresis: applications in drug metabolism and pharmacokinetics

Enantiospecific analysis has an important role in drug metabolism and pharmacokinetic investigations and its now no longer acceptable to determine total drug, or metabolite, concentrations following the administration of a racemate. Inspite of the fact that capillary electrophoresis (CE) has become an essential technique in pharmaceutical and enantiospecific analysis, the chromatographic methodologies remain the most commonly used approach for the determination of the enantiomeric composition of drugs in biological fluids. The application of CE to bioanalysis has been slow, which is in part associated with the complexity of biological matrices together with the relatively poor concentration limits of detection achievable. However, as a result of its versatility, high separation efficiency, minimal sample requirements, speed of analysis and low consumable expense CE is likely to play an increasingly significant role in the area. This review present an overview of enantiospecific CE in bioanalysis in which the approaches to enantiomeric resolution and the problems associated with biological matrices are briefly discussed. The application of enantiospecific CE to samples of biological origin is illustrated using examples where the methodology has either solved an analytical problem, or provided a useful alternative to the currently available chromatographic methods. Such improvements in methodology are associated with either the high separation efficiency and/or microanalytical capabilities of the technique. Enantiospecific CE will not replace the chromatographic methodologies but does provide the bioanalyst with a useful addition to his armamentarium.     

Fourier transform Raman spectroscopy of polymeric biomaterials and drug delivery systems

The characterization of drugs within polymer supports is an important prerequisite to our understanding of the chemical and/or physical mechanisms which control the release of drugs into the body from a biomedical polymer.The spectra of biomolecules obtained using a visible source are often accompanied by a strong persistent fluorescence background which is difficult to control. This has so far prevented the routine use of Raman spectroscopy in the analysis of biological compounds.FT Raman spectroscopy in the near infrared has proved on many occasions to be highly efficient in avoiding such problems. We have here reviewed the potential of FT Raman spectroscopy as an analytical technique for polymeric delivery systems by describing spectra of several significant drugs in clinically valuable polymeric supports.     

Pesticide analysis by capillary electrophoresis

In this work, a critical and updated revision of the current situation of the analysis of pesticides by Capillary Electrophoresis (CE) is presented. The review has been written in two main sections. The first one presents a thorough revision of the various offline and on-line sample preconcentration procedures that have been used in conjunction with CE to analyze these compounds. The second part reviews the various detection strategies (i.e., UV, LIF, MS, and electrochemical) and CE modes that have been applied to the analysis of pesticides. Future trends that can be expected from this hot research area are also discussed.     

Biodegradable polymeric matrix for long-acting and zero-order release drug delivery systems

Three types of biodegradable triblock copolymers, ABA, ABC and ACB, where A is polycaprolactone (PCL), B is polylactide (PLA) and C is polyglycolide (PGA), were used as matrix to test their controlled release behavior of Levo-Norgestrel in the form of microsphere. It was verified that the overall release kinetics changes from in vitro first order to zero order when the percentage of PCL segments in all of the block copolymers decreases from 85 % to 60 −65 % with a long-acting release time of 5 months or more. It is ascribed to the two release mechanisms, the diffusion release, contributed by the PCL segment, and the erosion-caused release by PLA or PGA segment. At a certain ratio of segments the two release ways make a compromise to a constant rate of release, i. e. a zero-order kinetics in these multiphase block copolymers.     

Low-density lipoprotein analysis in microchip capillary electrophoresis systems

Due to the mounting evidence for altered lipoprotein and cholesterol-lipoprotein content in several disease states, there has been an increasing interest in analytical methods for lipoprotein profiling for diagnosis. The separation of low- and high-density lipoproteins (LDL and HDL, respectively) has been recently demonstrated using a microchip capillary electrophoresis (CE) system [1]. In contrast to this previous study, the present report demonstrates that LDL analysis can be performed in an uncoated glass microchannel. Moreover, by adding sodium dodecyl sulfate (SDS) to the sample at a concentration well below the critical micellar concentration prior to injection, the LDL peak undergoes a focusing effect and exhibits an apparent efficiency of 2.2 x 10(7) plates/m. Laser light scattering experiments demonstrate that the low concentration of SDS used does not significantly alter lipoprotein particle size distribution within the time course that the analysis is performed. It is thus hypothesized that SDS nondisruptively coats LDL particles. The peak sharpening effect, observed only when SDS is added solely to the sample, is probably due to a mobility gradient created between the sample and the running buffer. The chip-based method demonstrated here has the potential for rapid analysis and sensitive detection of different LDL forms of clinical relevance.     

Physicochemical characterization of a PEGylated liposomal drug formulation using capillary electrophoresis

In this work, the applicability of using CE to perform a physicochemical characterization of a PEGylated liposomal drug formulation of the anti-cancer agent oxaliplatin was examined. Characterization of the liposomal drug formulation using CE instrumentation encompassed: determination of the electrophoretic mobilities, size determination by Taylor dispersion analysis and interaction studies. Electrophoretic mobilities determined by CE were compared with the results obtained by laser Doppler electrophoresis, which were found to be subject to larger variation. Average hydrodynamic diameters of the liposome preparations, as determined by Taylor dispersion analysis, were in the range of 61-84 nm and were compared with the results obtained by dynamic light scattering. Interactions between oxaliplatin (and paracetamol) and the PEGylated liposome were non-detectable by CE frontal analysis as well as by liposome electrokinetic chromatography. In contrast, for the more lipophilic compound propranolol, apparent liposome-aqueous phase distribution coefficients (D(lip) ) were successfully determined by both electrokinetic chromatography (log?D(lip) =2.10) and by CE frontal analysis (log?D(lip) =2.14). It is envisioned that CE and capillary-based techniques, including Taylor dispersion analysis, will be useful tools for the characterization of nanoparticulate (e.g. liposomal) drug formulations.     

Energy drink analysis by capillary electrophoresis

Caffeine and vitamins C, PP, and B6 have been determined in energy drinks by capillary electrophoresis. Its advantages and disadvantages over high-performance liquid chromatography have been considered and the influence of analysis conditions on the error of analysis and error sources in capillary electrophoresis have been estimated.     

Element speciation analysis by capillary electrophoresis

An overview of recent developments in the application of capillary electrophoresis to simultaneous separation and determination of different chemical forms of an inorganic element is presented, with particular emphasis placed on metal speciation analysis. Examples of species analysis are addressed, covering metal ions in different oxidation states, metal complexes with inorganic and organic ligands, metalloid oxoanions, organometallic compounds, ionic non-metal species, etc. The speciation performance of capillary electrophoresis is illustrated by a number of practically relevant applications. The method's strengths and current limitations with regard to chemical speciation studies are critically discussed.     

Separation of drug stereoisomers by capillary electrophoresis with cyclodextrins

Using capillary electrophoresis, the enantiomers and isomers of several chiral drug molecules were resolved with cyclodextrins. Parameters affecting the resolution between (+)- and (−)-epinephrine, such as pH, cyclodextrin concentration, buffer concentration, and capillary dimensions were investigated. In addition to this, the effect of cyclodextrin type (β and several derivatized β-cyclodextrins) on resolution between stereoisomers of several chiral drug was also investigated. This study showed that the structural features of the molecule, the derivative groups on the cyclodextrin, the buffer composition and the capillary dimensions influence resolution. The chiral drugs used in this study were propranolol, atenolol, betaxolol, dipivefrin, AL03152 (an aldose reductase inhibitor), AL03363 (an oxidation product of AL03152) and the cis/trans isomers of pilocarpine.     

Analysis of the antiepileptic drug keppra by capillary electrophoresis

A simple and rapid method for determination of the new antiepileptic drug keppra (levetiracetam) by capillary electrophoresis in borate buffer containing sodium dodecyl sulfate is described. The serum was injected without any treatment. The method compared well to high performance liquid chromatography. The mean of keppra in the serum of 35 patients was 25 mg/l (range 7-77 mg/l).     

Determination of organic acid drug counter-ions by capillary electrophoresis

Summary A capillary electrophoresis method is described for the novel application of quantifying levels of the simple organic acid counter-ions of a variety of basic drugs. These counter-ions are organic acids such as succinic and maleic. The method uses indirect UV detection and an electroosmotic flow modifier. Acceptable precision and detector linearity were obtained using internal standards. Method validation was completed and acceptable data was generated. The method is now in routine use for this type of testing.