CE-LIF method for the separation of anthracyclines: application to protein binding analysis in plasma using ultrafiltration.

Anthracyclines are chemotherapeutic drugs that are widely used in the treatment of cancers such as lung and ovarian cancers. The simultaneous determination of the anthracyclines, daunorubicin, doxorubicin and epirubicin, was achieved using CE coupled to LIF, with an excitation and emission wavelength of 488 and 560 nm, respectively. Using a borate buffer (105 mM, pH 9.0) and 30% MeOH, a stable and reproducible separation of the three anthracyclines was obtained. The method developed was shown to be capable of monitoring the therapeutic concentrations (50-50 000 ng/mL) of anthracyclines. LODs of 10 ng/mL, calculated at an S/N = 3, were achieved. Using the CE method developed, the in vitro protein binding to plasma was measured by ultrafiltration, and from this investigation the estimated protein binding was determined to be in the range of 77-94%.

Recent developments in amperometric detection for microchip capillary electrophoresis.

The interest in microfluidic devices has increased considerably over the past decade due to the numerous advantages of working within a miniature, microfabricated format. This review focuses on recent advances in coupling amperometric detection with microchip capillary electrophoresis (CE). Advances in electrochemical cell design, isolation of the detector from the separation field, and integration of both pre- and postseparation reaction chambers are discussed. The use of microchip CE with amperometric detection for enzyme/immunoassays, clinical and environmental assays, and the determination of neurotransmitters is described.

A review of membrane sampling from biological tissues with applications in pharmacokinetics, metabolism and pharmacodynamics.

This review provides an overview of membrane sampling techniques, microdialysis and ultrafiltration, and cites illustrations of their applications in pharmacokinetics, metabolism and/or pharmacodynamics. The review organizes applications by target tissue and general type of information gleaned. It focuses on recently published microdialysis studies (1999 to this writing) and offers the first review of ultrafiltration sampling studies. The advantages and limitations of using microdialysis and ultrafiltration sampling as tools for obtaining pharmacokinetic and metabolism data are discussed. Numerous examples are described including studies in which several types of data are collected simultaneously. Reports that study local metabolism of drug delivered through the probe are also presented.

Detection of homocysteine by conventional and microchip capillary electrophoresis/electrochemistry.

A method based on capillary electrophoresis (CE) with electrochemical (EC) detection for the determination of both total homocysteine (tHcy) and protein-bound homocysteine (pbHcy) in plasma is described. Both end-column and off-column amperometric detection were investigated. Off-column detection resulted in a more sensitive assay for the determination of homocysteine (Hcy). The detection limit for homocysteine was 500 nM using off-column EC detection and the response was linear over the range 1-100 microM. Therefore, this assay is appropriate for the quantification of Hcy over the physiological concentration ranges found in all disease states. Methodologies for the determination of tHcy and pbHcy in human plasma were investigated and optimized and the concentrations of both pbHcy and tHcy in plasma obtained from a healthy individual were determined to be 2.79+/-0.31 nuM (n = 4) and 3.37+/-0.15 microM (n = 3), respectively. The methodology was then transferred to a microchip CE-EC format and Hcy and reduced glutathione (GSH) were detected. Future work will focus on the development of ancillary methodologies to identify the other forms of Hcy in vivo.