Biomedical subjects

E N Fung

Publications and source records attributed to E N Fung.

4 recordsLinked to original sources

PubMed2001

Simultaneous determination of Ziagen and its phosphorylated metabolites by ion-pairing high-performance liquid chromatography-tandem mass spectrometry.

An ion-paring HPLC-MS-MS method with positive ion mode electrospray ionization has been developed to simultaneously quantify Ziagen, carbovir monophosphate, carbovir diphosphate and carbovir triphosphate. N',N'-Dimethylhexylamine was used as the ion-pairing agent. The presence of this ion-pairing agent allowed the retention and separation of the four compounds on a reversed-phase HPLC column as well as the detection of the nucleotides with positive ion mode electrospray ionization. The limits of detection were found to be better than 25 nM for all the analytes. Calibration curves of the analytes showed excellent linearity over the range of 25 nM to 5 microM. The relative standard deviations and accuracies for replicate analyses of quality control samples were less than 15%. The method has been successfully applied to the analysis of these compounds in human liver cells treated with Ziagen.

Anti-HIV Agents↗

PubMed1999

High-resolution capillary isoelectric focusing of complex protein mixtures from lysates of microorganisms.

High-resolution capillary isoelectric focusing separations of complex protein mixtures have been obtained for cellular lysates of Saccharomyces cerevisiae, Eschericia coli, and Deinococcus radiodurans. High quality separations are shown to be achievable for total protein concentrations of < 0.1 mg/mL. The separation reproducibility was examined, and the influence of the capillary inner wall coating on resolution investigated using fusedsilica capillaries coated with various hydrophilic polymers including hydroxypropyl cellulose, poly(vinyl alcohol), and linear polyacrylamide. Proteins having an isoelectric point (pI) difference of 0.004 are shown to be separated using a linear carrier ampholyte (linear pH gradient between two electrodes) of 3-10. Approximately 45 discrete peaks in the pI range of 5-7 were obtained for S. cerevisiae, approximately 80 peaks in the pI range of 4.5-8.5 for E. coli, and approximately 210 peaks in the pI range of 3-8.8 for D. radiodurans.

Bacterial Proteins↗

PubMed1998

Direct analysis of single rat peritoneal mast cells with laser vaporization/ionization mass spectrometry.

A linear time-of-flight mass spectrometer was used as a detector for flow cytometry. These two techniques were coupled by a laser vaporization/ionization interface. The estimated mass detection limit of the combined system was 20 amol of serotonin standard with one laser pulse. An aqueous buffer at physiological pH was used to ensure compatibility with cells. Rat peritoneal mast cells (RPMCs) were dispensed into the mass spectrometer in a single file confined within a 20-micron-i.d. capillary. By using the mass spectrometer as a detector, no precolumn staining or derivatization is required. Determination of serotonin and histamine in individual cells was demonstrated. With this method, hundreds of cells can be analyzed within a few minutes. The average amounts of histamine and serotonin per RPMC were found to be 0.75 +/- 0.33 and 0.11 +/- 0.06 fmol, respectively. No correlation was found between the amounts of the two amines in each cell.

Animals↗

PubMed1998

Fast DNA separations using poly(ethylene oxide) in non-denaturing medium with temperature programming.

We demonstrated fast DNA separations in low viscosity entangled solutions with a temperature gradient in a non-denaturing separation medium. The separations were carried out in a solution of commercially available poly(ethylene oxide) (PEO) [1 x Tris(hydroxymethyl)aminomethane borate buffer, without urea] with a temperature gradient of 2 degrees C/min. The performance was compared with that of a solution of PEO with urea at ambient temperature. We found that the former condition gives sufficient resolution for accurate base calling and that in general, it gave better separation for fragments larger than 450 base pairs (bp). Most importantly, the separation speed approaches 30 bp/min. In addition, we describe a simple yet reliable gel preparation protocol for such separations.

Base Sequence↗