Capillary Electrophoresis Electrospray Ionization Research Articles

Application of capillary electrophoresis-electrospray ionization mass spectometry in the determination of molecular diversity.

By means of capillary electrophoresis coupled online to electrospray ionization MS, a library of theoretically 171 disubstituted xanthene derivatives was analyzed. The method allowed the purity and makeup of the library to be determined: 160 of the expected compounds were found to be present, and 12 side-products were also detected in the mixture. Due to the ability of capillary electrophoresis to separate analytes on the basis of charge, most of the xanthene derivatives could be resolved by simple capillary electrophoresis-MS procedures even though 124 of the 171 theoretical compounds were isobaric with at least one other molecule in the mixture. Any remaining unresolved peaks were resolved by MS/MS experiments. The method shows promise for the analysis of small combinatorial libraries with fewer than 1000 components.

Analysis of single cells with capillary electrophoresis electrospray ionization Fourier transform ion cyclotron resonance mass spectrometry.

Recent results with on-line capillary electrophoresis (CE) electrospray ionization (ESI) Fourier transform ion cyclotron resonance (FTICR) mass spectrometry suggest that CE/ESI-FTICR can provide a powerful technique for micro-sample analyses owing to the inherent sensitivity of the technique and the enhanced information content derived from high-performance mass measurements. Using micro-sampling methods and ion accumulation techniques based on quadrupolar excitation, we demonstrate that adequate sensitivity exists to characterize the hemoglobin from a single human erythrocyte (approximately 450 amol). In these studies mass spectra with average mass resolution in excess of 45 000 (FWHM) were obtained for both the alpha- and beta chain of hemoglobin following in-column lysing of a single erythrocyte.

Application of Sequential Paired Covariance to Capillary Electrophoresis Electrospray Ionization Time-of-Flight Mass Spectrometry: Unraveling the Signal from the Noise in the Electropherogram

The combination of capillary electrophoresis (CE) with electrospray ionization (ESI) time-of-flight mass spectrometry (TOF-MS) has recently been demonstrated. When CE is combined with TOF-MS using an electrospray interface, the method provides fast, high-resolution separations with rapid mass analysis and the potential for very high sensitivity. In analyzing the data, one first reconstructs an electropherogram from the data set and then identifies peaks in the mass spectra. The mass spectra corresponding to the peaks in the electropherogram are then inspected to enable solute identification. However, background noise often prevents detection of peaks in the reconstructed electropherogram, thus interfering with the analysis. In this work we demonstrate alternative electropherogram reconstruction techniques that enhance its signal-to-noise ratio, allowing more immediate identification of the number of components in a mixture and their corresponding retention times. The techniques described here should also be adaptable for on-line analysis of CE-ESI-TOF data and the combination with other separation techniques coupled to mass spectrometry (e.g., LC/MS), as well as any multichannel detection scheme. 21 refs., 4 figs., 1 tab.

Direct Electrospray Ion Current Monitoring Detection and Its Use with Online Capillary Electrophoresis Mass Spectrometry

A novel detection scheme for use with the capillary electrophoresis-electrospray ionization interface is presented, based upon the direct measurement of electrospray ion current after expansion into vacuum, that is shown to provide the basis for a simple detector or for simultaneous dual detection with mass spectrometry. The utility of this novel dual-detection scheme is illustrated with capillary electrophoresis Fourier transform ion cyclotron resonance (FTICR) mass spectrometry in which spectra are acquired only when a solute zone elutes from the CE capillary. This acquisition scheme affords significantly improved sensitivity and permits the acquisition of longer time domain signals without sacrificing separation efficiency. Mass resolving power in excess of 10[sup 5] (fwhm) is demonstrated by FTICR for components from a peptide/protein mixture comprised of analytes with molecular masses ranging from 2 to 29 kDa. 24 refs., 3 figs.

Determination of amino acids by on-line capillary electrophoresis-electrospray ionization mass spectrometry.

On-line capillary electrophoresis-electrospray ionization mass spectrometry (CE-ESMS) has been used for the separation and detection of amino acid mixtures. Four natural amino acids, histidine, tryptophan, phenylalanine, aspartic acid, and a tripeptide, glutathione, were separated and determined. Protonated molecules were detected in the CE-ESMS mode with detection limits of about one pmol. Optimum CE-ESMS operating conditions for amino acid analysis were determined employing acetic acid solutions as CE electrolytes. The examined parameters included capillary diameter (50-100 microns internal diameter), applied separation voltage (20-30 kV), and concentration of electrolyte (10-60% acetic acid). Stable working conditions were maintained when the CE currents were less than 18 microA. The use of electrolyte solutions such as those described here instead of true buffer solutions may have advantages for CE-ESMS systems which employ a "sheathless" interface.

Fumonisin B1 analysis with capillary electrophoresis-electrospray ionization mass spectrometry.

Fumonisin B1, a mycotoxin produced by a common fungal contaminant of corn, Fusarium moniliforme, was analyzed by capillary electrophoresis-electrospray ionization mass spectrometry. System performance was maximal with uncoated columns. System efficiencies of approximately 44,000 plates/m and reproducible analysis times of about 13 min were obtained. System efficiency with methyl-coated columns was approximately 24,000 plates/m. Reproducible analysis times of about 3.5 min were obtained with these columns. With uncoated columns, the concentration limit of detection was 156 ppb with a s/n ratio of approximately 10. The estimated injected mass at 156 ppb was 1.1 pg. Repeated injections of extracts containing constant fumonisin B1 concentrations showed that peak areas were slightly inconsistent, although generally similar to variations encountered with liquid chromatography-electrospray ionization mass spectrometry. The source of this inconsistency was traced to sample solubility, errors inherent in electrophoresis injections, and electrospray instability. Minimizing these problem areas will produce a technique with peak area reproducibilities comparable to liquid chromatography-electrospray ionization mass spectrometry, but with potentially greater resolving power.

Capillary electrophoresis Fourier transform ion cyclotron resonance mass spectrometry with sustained off-resonance irradiation for the characterization of protein and peptide mixtures

A new approach to protein and peptide analysis that involves the coupling of on-line capillary electrophoresis-electrospray ionization Fourier transform ion cyclotron resonance mass spectrometry with a variation of sustained off-resonance irradiation is described. With this technique, multiple irradiation frequencies are broadcast simultaneously, which yields fragmentation of species at different mass-to-charge ratio values from the same waveform. In conjunction with capillary electrophoresis, this technique can provide sequence information from small amounts of proteins or peptides in complex mixtures.Initial results obtained from a mixture of gramicidin S (1141 u), bee venom melittin (2845 u), and equine apomyoglobin (16,951 u) are presented.

Investigation of the metabolic fate of the neuroleptic drug haloperidol by capillary electrophoresis-electrospray ionization mass spectrometry

Combined capillary electrophoresis—mass spectrometry (CE—MS) is a powerful analytical tool for the characterization of components of complex mixtures. Furthermore, when interfaced with an electrospray ionization (ESI) source, skimmer induced fragmentation can yield structural information that aids compound identification. We show that these techniques can be used to probe the metabolic fate of drugs by investigation of the phase I metabolism of the widely used neuroleptic drug haloperidol. In addition, we demonstrate differences of in vitro metabolism of haloperidol by mouse and guinea pig hepatic microsomes.

Sensitivity considerations for large molecule detection by capillary electrophoresis-electrospray ionization mass spectrometry

The use of the electrospray ionization (ESI) method for interfacing capillary electrophoresis with mass spectrometry (CE-MS) is particularly well suited for the analysis of large molecules due to the multiple charging phenomenon. While ionization efficiency is very high, the available ion current is dispersed over more peaks so that the maximum peak intensity obtainable declines significantly for large molecules. Sensitivity with ESI can be improved by operation at very low flow-rates, an ideal situation for CE-MS. These and other considerations related to sensitivity are illustrated using ESI-MS measurements for cytochrome c.

Capillary zone electrophoresis and isotachophoresis— mass spectrometry of polypeptides and proteins based upon an electrospray ionization interface

The speical capabilities of the capillary electrophoresis electrospray ionization— mass spectrometer interface for the analysis of peptides and proteins with molecular weights extending to in excess 100 000 are reviewed. The dynamic combinations of both capillary zone electrophoresis and capillary isotachophoresis with electrospray ionization are illustrated for mixtures of peptides and proteins. Myoglobin and cytochrome c detection limits were ca 100 fmol. The potential extension of these methods for determination of the primary structure (sequence) of polypeptides using tandem mass spectrometry is shown to be facilitated by the high charge state of ions produced by the electrospray interface. The relevance of these results for advances in analytical biochemistry are discussed.