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To alleviate signal suppression resulting from nonvolatile positive ion additives, a hydrodynamic flow assisted double junction capillary electrophoresis-mass spectrometry (CE-MS) interface was proposed. The double junction interface which could alleviate the ion suppression due to nonvolatile negative ion additives was modified so that a hydrodynamic flow could be introduced to the interface. Using this setup, the apparent velocity of the ions was determined based on its electrophoretic mobility, electroosmotic flow in the transfer capillary, and hydrodynamic flow introduced by the syringe pump. CE-MS analysis of positively charged triazines was performed to demonstrate the practical value of this approach by using cetyltrimethylammonium ion (CTA(+)) as an additive. Because the separation column was dynamically coated with CTA(+), the EOF was reversed and the separation was performed under counter EOF mode. Under an appropriate hydrodynamic flow, the analytes (triazines) could be propagated toward the MS, whereas the additive (CTA(+)) ion was retained in the interface. Consequently, the problem of signal suppression by CTA(+) was alleviated, and the signals were enhanced more than 20-fold., (Copyright © 2014 Elsevier B.V. All rights reserved.)

A preconcentration approach for CE-MS using counterflow-assisted electrokinetic injection was proposed. The proposed preconcentration method was based on a counterflow-compatible sheathless interface. The interface was fabricated using a capillary-assembled PDMS microdevice that allowed the application of a counterflow and provided liquid-film electrical conduction. During electrokinetic injection, a hydrodynamic counterflow was introduced into the separation capillary to retard the movement of the stacking boundary to the CE outlet. Accordingly, a long injection time was achieved without a loss of CE separation. With the use of reverse polarity mode and a dynamic polybrene-coated separation capillary (filled with a 1% formic acid solution), a counterflow-assisted electrokinetic injection of peptide samples (in a aqueous solution containing 1% ammonia and 50% methanol) for 10 min provided a sensitivity enhancement of 750-1480 and a detection limit of 20 pM for the five peptide standards. The proposed technique was applied to the analysis of low nanomolar concentrations myoglobin tryptic peptides., (© 2013 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.)

This study develops a sheathless CE-MS interface using a robust PDMS membrane emitter and liquid-film electric conduction. A 3D mold was constructed for casting the device by using a one-step casting procedure. The interface consisted of a 125 μm-thick triangular emitter with a 50 μm-diameter microchannel, a conducting reservoir, and a 375 μm-diameter channel for assembling the separation capillary. The separation capillary was inserted into the 375 μm channel and connected to the emitter through the conducting reservoir. The electric contact for the CE outlet was established through a conductive liquid film in the space between the capillary terminus and the 375 μm channel. The one-step casting procedure and using a membrane emitter instead of a tapered emitter produced an easily fabricated and robust interface. A stable electrospray was obtained from 30 to 350 nL/min. Analyzing a five-peptide mixture in low-EOF (60 nL/min) and high-EOF (210 nL/min) conditions demonstrated the utility of the interface., (© 2013 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.)

A simple two column sheathless CE/MS interface was constructed using polydimethylsiloxane to fabricate a microdevice allowing facile column alignment and electrical connection. One conducting reservoir, two holes and one 1mm length microchannel between the holes were fabricated on the microdevice. The two holes were used for connecting separation capillary and ESI sprayer. The hole for ESI sprayer was fabricated at the edge of the conducting reservoir. The ESI sprayer was inserted through the reservoir to the hole so allowing it to be aligned with the separation column. Because the size of the hole was fabricated slightly larger than the outer diameter of the ESI sprayer, the electrical conduction was established through the thin conductive liquid film formed in the space between the ESI sprayer and the hole. The interface design presented was both easy to fabricate and operate and demonstrated good performance. The dead volume did not significantly affect operation as indicated by a demonstrated preservation of separation resolution. The intra-day precision and inter-day precision for peak areas and migration times observed using this interface were found to be less than 12% and 5%, respectively., (Copyright © 2012 Elsevier B.V. All rights reserved.)

An approach based on staggered multistep elution solid-phase extraction (SPE) capillary electrophoresis/tandem mass spectrometry (CE/MS/MS) was developed in the analysis of digested protein mixtures. On-line coupling of SPE with CE/MS was achieved using a two-leveled two-cross polydimethylsiloxane (PDMS)-based interface. Multistep elution SPE was used prior to CE to provide an additional dimension of separation, thus extending the separation capacity for the peptide mixture analysis. By decreasing in the number of co-eluting peptides, problems stemming from ionization suppression and finite MS/MS duty cycle were reduced. As a result, sequence coverage increased significantly using multistep elution SPE-CE/MS/MS compared to one-step elution SPE-CE/MS/MS in the analysis of a single protein tryptic digest (49% vs. 18%) and a six protein tryptic digest (22-71% vs. 10-44%). A staggered CE method was incorporated to increase the throughput. The electropherograms of consecutive CE runs were partially overlapped by injecting the sample plug at a fixed time interval. With the use of a 5 min injection interval, slightly poor results were obtained in comparison with the sequential CE method while the total analysis time was reduced to 28%., (Copyright © 2011 John Wiley & Sons, Ltd.)

Methodology is presented which greatly improves the sensitivity of ganglioside analysis by CE/MS without compromising separation efficiency. In this method, non-volatile additives were removed, where possible, to reduce ion suppression. Specifically, alpha-CD, used to break up ganglioside micelle formation, was replaced with isopropyl alcohol. To reduce ion suppression from sodium ions, ammonium borate was substituted for sodium borate in the preparation of borate buffer. Because borate anions were found to be essential for CE separation, a liquid-junction/low-flow interface was employed to restrict borate anions from entering the ESI ion source. With proper control over the EOF in the connecting capillary, borate anions were controlled to migrate away from the ESI ion source. With these modifications, the sensitivity of CE/MS analysis of gangliosides was improved significantly.

A two-leveled, two cross PDMS-based interface for on-line coupling of SPE with CE-MS was proposed. In this interface, the SPE column and the CE separation column were positioned orthogonally and two crosses were fabricated on the interface. With the two cross design, the operation of SPE could be performed independently without unexpected flow through leakage into the separation column. The performance of the interface was optimized using a peptide mixture. The position of the SPE column related to the CE separation channel was found to be critical to the performance of the system. Under the optimal position, the separation efficiency was similar to a CE-MS experiment without SPE. The peptide signals were enhanced 50- to 100-fold and the repeatability was within 4% RSD for migration time and 10% RSD for peak area. A tryptic digest of cytochrome c was used to demonstrate the feasibility of the interface in protein identification at a level of 1 ng/microL.

To alleviate ion suppression from phosphate buffer and to preserve separation integrity, a new capillary electrophoresis mass spectrometry (CE-MS) interface was developed. The interface consisted of a low-flow interface and a liquid junction. In this design, both the inlet reservoir and the liquid-junction reservoir were filled with phosphate running buffer. Because the phosphate anions in the column migrated toward the inlet reservoir (away from the electrospray ionization (ESI) source) the problem of ion suppression in ESI was avoided. The liquid junction was incorporated to eliminate issues of degraded separation observed when sheath liquid interfaces use different buffers for separation and MS analysis attributed to differences in anion velocity. The utility of the interface was demonstrated by the analysis of antihistamines at pH 3.5 and the analysis of perfluorocarboxylic acid at pH 9.5.

A multiplexed CE-MS interface using four low-flow sheath liquid ESI sprayers has been developed. Because of the limited space between the low-flow sprayers and the entrance aperture of the ESI source, multichannel analysis is difficult using conventional rotating plate approaches. Instead, a multiplexed low-flow system was achieved by applying an ESI potential sequentially to the four low-flow sprayers, resulting in only one sprayer being sprayed at any given time. The synchronization of the scan event and the voltage relays was accomplished by using the data acquisition signal from the IT mass spectrometer. This synchronization resulted in the ESI voltage being sequentially applied to each of the four sprayers according to the corresponding scan event. With this design, a four-fold increase in analytical throughput was achieved. Because of the use of low-flow interfaces, this multiplexed system has superior sensitivity than a rotating plate design using conventional sheath liquid interfaces. The multiplexed design presented has the potential to be applied to other low-flow multiplexed systems, such as multiplexed capillary LC and multiplexed CEC.

Antihistamines were analyzed by CE-ESI-MS using phosphate buffer. The separation was performed in an acidic environment so that phosphate ions had a net velocity flowing toward the inlet reservoir instead of the ESI source. To further reduce the effect of ion suppression, the sodium ion in sodium phosphate was replaced with an ammonium ion. Furthermore, with the combination of reducing the concentration of acid added to the sheath liquid and the use of a low-flow interface, phosphoric acid could be added to the sheath liquid. Because of the use of the same counterion (phosphate ion) in running buffer and in sheath liquid, the separation integrity (resolution, elution order, and peak shape) was preserved. In addition, ion suppression was also greatly alleviated because a minimal amount of phosphate flowed into the ESI source.

A sheathless interface has been developed for coupling CE with electrospray IT mass spectrometer. This interface utilized a pulsed ESI source. The use of a pulsed electrospray source allows the use of a sprayer with larger orifice, and thus alleviates the problem of column clogging during conductive coating and CE analysis. A pulsed ESI source operated at 20 Hz and 20% duty cycle was found to produce the optimal signals. For better signals, the maximum ion injection time in the IT mass spectrometer has to be set to a value close to the actual spraying time (10 ms). Using a sprayer with 50 microm od, more stable and enhanced signals were obtained in comparison with continuous CE-ESI-MS under the same flow rate (150 nL/min). The utility of this design is demonstrated with the analysis of synthetic drugs by CE-MS.

A low-flow electrophoresis-mass spectrometry (CE-MS) interface has been developed for interfacing capillary zone electrophoresis (CZE) with electrospray- ionization-mass spectrometry (ESI-MS). The interface consists of two parallel capillary columns (a separation column and a makeup column), and an emitter with a dual beveled edge. While maintaining a relatively low optimum flow rate, the dual-beveled-edge ESI emitter allows the use of a tip with larger orifice. Therefore, this interface is less prone to column blocking in comparison with a flat tip. Primarily attributed to low sample dilution and smaller initial droplet, the interface showed better sensitivity than a conventional sheath liquid interface. Furthermore, the interface was found to be more resistant to the presence of nonvolatile salts. By using 40 mM borate and 20 mM alpha-cyclodextrin (alpha-CD) as the running buffer, four major forms of gangliosides were detected by CE-MS.

A simple and durable sheath liquid interface for capillary zone electrophoresis-electrospray ionization-mass spectrometry (CZE-ESI-MS) has been developed. This interface utilized a beveled tip emitter and was found to be more sensitive than the conventional sheath liquid interface. The use of a beveled tip reduces the optimal flow rate and therefore decreases sample dilution. The interface utilized a 380 microm inner diameter and 400 microm outer diameter beveled tapered tip. Because of the large inner diameter and outer diameter of the tip, the interface is robust and can be easily implemented. The performance of this interface for CZE-ESI-MS and micelle electrokinetic capillary electrophoresis-electrospray-mass spectrometry, as demonstrated by the analysis of synthetic drugs and triazine mixtures, was significantly better than results obtained using a conventional sheath liquid interface.

A simple and time-saving procedure is proposed for preparing a silver-coated fused-silica capillary tip. The tapered capillary tip was coated with silver using an acrylic-based silver conductive pen. The fabrication of a silver-coated fused-silica capillary tip takes less than 5 minutes. The silver-coated fused-silica capillary tip is rugged and durable for sheathless capillary electrophoresis/electrospray mass spectrometry and also for microspray applications., (Copyright 2003 John Wiley & Sons, Ltd.)