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Biomedical subjects

J F Banks

Publications and source records attributed to J F Banks.

10 recordsLinked to original sources
PubMed1998

Capillary electrochromatography of biomolecules with on-line electrospray ionization and time-of-flight mass spectrometry.

Capillary electrochromatography (CEC) is considered a hybrid of liquid chromatography and capillary electrophoresis. It is expected to combine the high peak efficiency of capillary zone electrophoresis with the versatility and loading capacity of HPLC to bring about another high-performance MS-compatible chromatographic system. This paper explores the potential of CEC coupled with the electrospray ionization and time-of-flight mass spectrometry in biochemical analysis. The packed columns used in this study were tapered at the outlet to retain the packing material, thereby obviating the need for an outlet frit. Electrosmotically driven solvent gradients were employed for the separation of phenylthiohydantoin (PTH)-amino acids by reversed-phase chromatography, and a time-of-flight (TOF) mass spectrometer was employed as the detector for the CEC column effluent. The effect of CEC operating parameters, such as gradient shape, column length, and electric field, on the analytical results from the separation and MS detection of a standard mixture of PTH-amino acids was investigated. Particular attention was paid to the effect of sheath flow-rate, sheath composition and mass spectra acquisition rate on the performance of the electrospray TOF-MS.

Amino Acids↗
PubMed1997

Protein analysis by packed capillary liquid chromatography with electrospray ionization and time-of-flight mass spectrometry detection.

Packed capillary liquid chromatography (LC) has been interfaced on-line to electrospray (ES) ionization with time-of-flight (TOF) mass spectrometry (MS) detection for the separation and identification of standard proteins. Using this system, as little as 2.5 fmol of cytochrome c could be detected with a 0.1 mm I.D. column. The effect of column size on sensitivity has been investigated using various column diameters and a sensitivity increase of 428-fold has been observed between the 0.10- and 2.1 mm I.D. columns. Although the TOF-MS can ultimately record data at up to 100 mass spectra/s, the peak widths for these particular separations require only 1 spectra/s. Finally, the sensitivity obtained is a function of both the reduced diameter LC column and the high ion duty cycle of the TOF-MS.

Chromatography, Liquid↗
PubMed1997

Rapid peptide mapping by reversed-phase liquid chromatography on nonporous silica with on-line electrospray time-of-flight mass spectrometry.

Reversed-phase liquid chromatography (LC) using a nonporous silica support has been combined with electrospray (ES) time-of-flight (TOF) mass spectrometry (MS) for the fast separation and mass detection of peptides. Using this LC method, the resolution of a peptide mixture can be completed is less than 35 s. The resulting chromatographic peak widths are less than 1 s wide. Because of the unique nature of a TOF mass analyzer, complete mass spectra can be acquired at a rate which is sufficient to sample these narrow peaks. When compared with conventional LC, the same separation takes nearly 20 min to complete, and the signal-to-noise ratio observed in the total ion chromatogram is dramatically lower due to the influence of increased background noise in the mass spectra. The limit of detection for a low molecular weight peptide, Val-Pro-Leu, was found to be 6 pmol with the total ion chromatogram and 500 fmol with the reconstructed ion chromatogram. A peptide map of horse heart myoglobin, completed in 3.5 min, is shown as an example of the results which can be obtained from combining this fast LC method with fast ES/TOF/MS detection capability.

Animals↗
PubMed1997

Capillary electrophoresis with electrospray mass spectrometry detection for low-molecular-mass compounds.

Mass spectrometry (MS) detection using electrospray ionization (ESI) has been explored for the separation by capillary electrophoresis (CE) of a number of sample mixtures containing low-molecular-mass species. Optimal sheath liquid composition has been determined using a peptide mixture in which femtomolar quantities of analyte were easily observed. Effects of CE buffer choice were studied in detail. Also, a separation of basic drugs in cough syrup has been successfully detected by ESI-MS. Using negative ionization, a mixture of alkyl sulfonates and a mixture of food dyes were analyzed. All components were easily resolved and identified by molecular weight.

Alkanesulfonates↗
PubMed1997

Recent advances in capillary electrophoresis/electrospray/mass spectrometry.

Successful on-line interfacing of capillary electrophoresis (CE) with electrospray (ES) mass spectrometry (MS) has progressed substantially in recent years. Of particular note also is the development which has occurred in combining the more advanced capillary-based electromigration separation techniques, such as capillary gel electrophoresis (CGE), capillary isoelectric focusing (CIEF), capillary isotachophoresis (CIT), micellar electrokinetic chromatography (MEKC) and capillary electrochromatography (CEC), with ES/MS. The union of these electromigration schemes with MS detection provides a useful and sensitive analytical tool for the separation, quantitation and identification of biological, therapeutic, environmental and other important classes of chemical analytes. By making optimal use of the characteristics inherent with these separation mechanisms, greatly enhanced MS performance may be obtained. The following review summarizes the significant issues and challenges involved with CE/ES/MS analysis as well as results which have recently been obtained.

Animals↗
PubMed1996

Detection of fast capillary electrophoresis peptide and protein separations using electrospray ionization with a time-of-flight mass spectrometer.

Fast capillary electrophoresis (CE) separations on bare and coated fused silica have been detected on-line using a time-of-flight mass spectrometer which is capable of acquiring spectra at a rate suitable for the narrow peaks generated by CE. Protein and peptide separations producing peaks 1-2 s in width have been detected with an integration time of 0.125 s/spectrum, each spectrum being the sum of 1024 complete m/z range scans. The effects of the liquid sheath flow rate, electric field strength, and integration time on sensitivity and peak shape have been examined. A sensitivity limit of 8 fmol has been established for leucine enkephalin.

Amines↗
PubMed1994

LC/ESI-MS determination of proteins using conventional liquid chromatography and ultrasonically assisted electrospray.

Conventional unassisted electrospray ionization mass spectrometry (ESI-MS) has severe limitations as a liquid chromatography (LC) interface due to the few solvent compositions which can be electrosprayed without some type of assistance. LC mobile phases which have high flow rates (over 5 microL/min), high conductivity, or high surface tension are normally unsuitable for use with ESI. For this reason, an ultrasonic nebulizer has been developed which mechanically creates the fine spray of droplets needed for ESI and thus functions independently of the solvent composition. This device also operates at considerably higher liquid flow rates, up to several hundred microliters per minute. To characterize the system's performance, the ultrasonic nebulizer frequency, source electrode potentials, and drying and focusing gas flow rates were studied and optimized. Also, droplet size measurements were taken using a phase-Doppler anemometer, which showed consistent nebulizer performance up to a liquid flow rate of several hundred microliters per minute. Finally, the ultrasonic nebulizer was used for an LC application involving the separation of proteins on 1.0 and 2.1 mm i.d. columns. LC flow rates of up to 200 microL/min of mobile phases containing 0.1% trifluoroacetic acid could be easily nebulized. With the 1.0 mm i.d. column, as little as 32 fmol of cytochrome c could be detected in selected ion monitoring mode.

Chromatography, Liquid↗
PubMed1994

Ultrasonically assisted electrospray ionization for LC/MS determination of nucleosides from a transfer RNA digest.

The use of an ultrasonic nebulizer to assist electrospray ionization mass spectrometry (ESI-MS) has been described and demonstrated with the analysis of a transfer RNA digest by microcolumn LC. The restricted range of mobile-phase compositions amenable to the electrospray process has traditionally placed a severe limitation on the types of LC applications that can be used with ESI-MS. For this reason, an ultrasonic nebulizer configured for LC has been developed that can generate the fine dispersion of liquid required for ESI-MS from any type of mobile phase. In the case presented here, a transfer RNA was enzymatically digested into its substituent nucleosides, which were then analyzed by microcolumn LC. The required mobile-phase gradient (beginning at 5% methanol) falls outside the solvent range that can be used with conventional electrospray. The ultrasonic nebulizer, however, resolves this problem. The fundamental behavior of the four most common nucleosides (cytidine, adenosine, guanosine, uridine) was studied, and conclusions concerning the effects of solution chemistry were drawn. Specifically, signal from the H+ adducts of these species seems to be strongly dependent on the pKa value. Also, effects from several source operating variables were examined. These included capillary exit voltage, drying and focusing gases, and nebulizer frequency. Performance was found to be consistent over a wide range (0-100% methanol) of mobile-phase compositions. The limit of detection for adenosine injected onto a microcolumn was found to be 100 amol. Finally, nucleosides from as little as 150 fmol of RNA (amount prior to digestion) could be detected.

Chromatography, Liquid↗
PubMed1989

Capillary zone electrophoresis of oligonucleotides. Factors affecting separation.

The influence of various parameters affecting separation of oligonucleotides by capillary zone electrophoresis has been examined. The effects of pH, ionic strength, and various additives, including highly charged cations such as spermine, were studied. Using polycytidines as model compounds, it was demonstrated that pH in the range of 5-8 and ionic strength in the range of 20-200 mmol/l do not influence the separation of oligonucleotides substantially. However, with the addition of spermine to the background electrolyte, migration order was inverted as the effective mobilities of the larger oligonucleotides were greatly decreased. With the addition of spermine and sodium dodecyl sulfate, the separation+ of these model oligonucleotides was also significantly affected. The best separation of a homologous series of polycytidines was obtained with a background electrolyte containing 60 mmol/l histidine, 30 mmol/l glutamic acid, 50 mmol/l sodium dodecyl sulfate and 3 mmol/l spermine.

Electrophoresis↗