ESI Spray Research Articles

Simple and Rapid Method for Determination of Abemaciclib in Human Serum using Supported Liquid Extraction Pretreatment and LC-MS/MS Analysis

We developed a simple and rapid method for the determination of abemaciclib in human serum using supported liquid extraction (SLE) method for pretreatment and LC-MS/MS. Abemaciclib was extracted using SLE method with methyltert-butyl ether (MTBE) as elution buffer, and analyzed by LC-QTOF MS system, LCMS9030 (Shimadzu). Abemaciclib and fluconazole (internal standard) were detected with ESI spray in positive ionization mode, and the transition were set at 507.3/393.1629 for abemaciclib and 307.1/220.0677 for fluconazole. The retention times of abemaciclib and fluconazole were 2.76 and 2.98 min, respectively, and good linearity was obtained from 20–1,000 ng/mL for abemaciclib. The regression equation (weight = 1/x2) describing the calibration curve in human serum was y = 0.0196 x – 0.056 (R2 = 0.999), where y is the peak area ratio of abemaciclib against the IS and x is the nominal concentration of abemaciclib. The intra- and inter-assay accuracy varied between -4.3–1.7%, and the precision varied between 0.90–6.19%. The mean recovery rate of abemaciclib was 87.7 ± 4.3%, and the mean matrix factor was 1.00 ± 0.083. Our method offers speed and simplicity of sample preparation, which is one of great advantages in the analysis of clinical specimens. We believe that the present method will contribute to establishing a methodology for determining the optimal dose of abemaciclib for individual patients. Key words: abemaciclib, SLE, supported liquid extraction, quantification

Electrohydrodynamics of Gas-Assisted Electrospray Ionization Mass Spectrometry.

Gas-flow assistance is commonly used in ESI-MS for improved transport and desolvation, and fundamental understanding of the underlying phenomena is essential for improvement of aerodynamic interfaces that couple ESI sources and MS. For this purpose, an electrohydrodynamic model is developed for simulation of charged droplet dynamics under the combined effects of gas flow and electric fields with consideration of space charge interactions within the charged aerosol plume. The model is implemented in COMSOL by exploiting a formalism for establishing the droplet trajectories as a sequence of successive droplets ejected at a frequency defined by the electrospray current. The model is used to assess the effect of two distinct flow configurations and compared to the baseline care of electrospray without assist gas. The simulated flows are jet flows oriented coaxially with the ESI spray, with and without imposed vorticity (swirling). Droplet trajectory simulations of a bimodal droplet population consisting of large primary droplets and small progeny droplets reveal a unique capability for vortical assist jet flow to selectively transmit smaller droplets into the MS due to inertial separation. ESI-MS analysis of fluorinated phosphazines subjected to the different gas flow conditions supports the model predictions. The electrohydrodynamic model developed in this work provides a versatile tool to analyze and design aerodynamic ESI interfaces with rigorous incorporation of drag, inertia, and space-charge repulsion and can be used as a powerful simulation methodology for optimizing charged droplet transmission and ultimately improved analytical performance of gas-assisted ESI-MS workflows.

Advanced molecular-fingerprinting analysis of dissolved organic sulfur by electrospray ionization-Fourier transform ion cyclotron resonance mass spectrometry using optimal spray solvent

Molecular level characterization of dissolved organic sulfur (DOS) by electrospray ionization-Fourier transform ion cyclotron resonance mass spectrometry (ESI-FTICR MS) is necessary for further understanding of the role of DOS in the environment. Here, ESI spray solvent, a key parameter for ion production during ESI process, was investigated for its effect on the molecular characterization of DOS by ESI-FTICR MS. 100% MeOH as spray solvent was found for the first time to remarkably enhance the ionization efficiency of the majority of CHOS-molecules in NOM, which facilitated a total of 1473 CHOS-molecular formulas with one sulfur atom to be detected. The number of CHOS-molecular formulas obtained using 100%MeOH as spray solvent increased notably over 740 in comparison with those using 50% MeOH aqueous solution (731) or 50% ACN aqueous solution (653). Moreover, due to the enhancement of ionization efficiency of DOS during ESI processes, the tandem mass spectra of the NOM CHOS-molecules could be easily obtained using 100% MeOH as spray solvent, which were hardly obtained using 50% MeOH aqueous solution as spray solvent. The results of the tandem mass spectra suggested the first discovery of organosulfates or sulfonic acids in Suwannee River NOM sample. A simple method based on 100% MeOH as ESI spray solvent for advanced molecular characterization of DOS by ESI-FTICR MS was proposed and applied, and the results revealed more molecular information of DOS in sea DOM samples.

Ionic liquid-copolymerized monolith based porous layer open tubular column for CEC-MS analysis

Developing novel capillary electrochromatography (CEC) column with high separation efficiency and capacity is an essential work in CEC. In this work, porous layer open tubular (PLOT) column has been developed by co-polymerization of quaternary ammonium typed ionic liquid (IL), 1-allyl-methylimidazolium chloride (AlMeIm+Cl−) with styrene (IL-styrene) and ethylene dimethacrylate (EDMA) for CEC-MS analysis. The PLOT-CEC column possesses high separation efficiency and capacity while keeping the inherent advantages of open tubular (OT) column unaltered, which is suitable for CEC analysis. Due to the uniform branch-like structure with large specific surface area, the IL-styrene based PLOT-CEC column showed excellent separation efficiency towards multiple kinds of analytes with theoretical plates up to N = 322,277 N/m for phloroglucinol. And thanks to the high permeability and large EOF mobility generated by positively charged ionic liquid, the PLOT-CEC column showed good performance in CEC-MS analysis of parabens, amino acids and peptides with stable separation and ESI spray. Good repeatability with relative standard deviations for intra-day, inter-day runs and between tubes less than 4.94% was obtained.

Corona discharge electrospray ionization of formate-containing solutions enables in-source reduction of disulfide bonds.

Disulfide bonds are critical linkages for maintaining protein structure and enzyme activity. These linkages, however, can limit peptide sequencing efforts by mass spectrometry (MS) and often require chemical reduction and alkylation. Under such conditions, information regarding cysteine connectivity is lost. Online partial disulfide reduction within the electrospray (ESI) source has recently been established as a means to identify complex cysteine linkage patterns in a liquid chromatography-MS experiment without the need for sample pre-treatment. Corona discharge (CD) is invoked as the causative factor of this in-source reduction (ISR); however, evidence remains largely circumstantial. In this study, we demonstrate that instrumental factors-nebulizing gas, ESI capillary material, organic solvent content, ESI spray needle-to-MS distance-all modulate the degree of reduction observed for the single disulfide in oxytocin, further implicating CD in ISR. Rigorous analysis of solution conditions, however, reveals that corona discharge alone can induce only minor disulfide reduction. We establish that CD-ESI of peptide solutions containing formic acid or its conjugate base results in a dramatic increase in disulfide reduction. It is also determined that ISR is exacerbated at low pH for complex peptides containing multiple disulfide bonds and possessing higher-order structure, as well as for a small protein. Overall, our results demonstrate that ESI of formate/formic acid-containing solutions under corona discharge conditions facilitates disulfide ISR, likely by a similar reduction pathway measured in γ-radiolysis studies nearly three decades ago.

Online Simultaneous Hydrogen/Deuterium Exchange of Multitarget Gas-Phase Molecules by Electrospray Ionization Mass Spectrometry Coupled with Gas Chromatography.

In this study, a hydrogen/deuterium (H/D) exchange method using gas chromatography-electrospray ionization/mass spectrometry (GC-ESI/MS) was first investigated as a novel tool for online H/D exchange of multitarget analytes. The GC and ESI source were combined with a homemade heated column transfer line. GC-ESI/MS-based H/D exchange occurs in an atmospheric pressure ion source as a result of reacting the gas-phase analyte eluted from GC with charged droplets of deuterium oxide infused as the ESI spray solvent. The consumption of the deuterated solvent at a flow rate of 2 μL min-1 was more economical than that in online H/D exchange methods reported to date. In-ESI-source H/D exchange by GC-ESI/MS was applied to 11 stimulants with secondary amino or hydroxyl groups. After H/D exchange, the spectra of the stimulants showed unexchanged, partially exchanged, and fully exchanged ions showing various degrees of exchange. The relative abundances corrected for naturally occurring isotopes of the fully exchanged ions of stimulants, except for etamivan, were in the range 24.3-85.5%. Methylephedrine and cyclazodone showed low H/D exchange efficiency under acidic, neutral, and basic spray solvent conditions and nonexchange for etamivan with an acidic phenolic OH group. The in-ESI-source H/D exchange efficiency by GC-ESI/MS was sufficient to determine the number of hydrogen by elucidation of fragmentation from the spectrum. Therefore, this online H/D exchange technique using GC-ESI/MS has potential as an alternative method for simultaneous H/D exchange of multitarget analytes.

Coupling of gas chromatography and electrospray ionization high resolution mass spectrometry for the analysis of anabolic steroids as trimethylsilyl derivatives in human urine

In this study, gas chromatography (GC) was interfaced with high resolution mass spectrometry (HRMS) with electrospray ionization source (ESI) and the relevant parameters were investigated to enhance the ionization efficiency. In GC-ESI, the distances (x-, y- and z) and angle between the ESI needle, GC capillary column and MS orifice were set to 7 (x-distance), 4 (y-distance), and 1 mm (z-distance). The ESI spray solvent, acid modifier and nebulizer gas flow were methanol, 0.1% formic acid and 5 arbitrary units, respectively. Based on these results, analytical conditions for GC-ESI/HRMS were established. In particular, the results of spray solvent flow indicated a concentration-dependent mechanism (peak dilution effect), and other parameters also greatly influenced the ionization performance. The developed GC-ESI/HRMS was then applied to the analysis of anabolic steroids as trimethylsilyl (TMS) derivatives in human urine to demonstrate its application. The ionization profiles of TMS-derivatized steroids were investigated and compared with those of underivatized steroids obtained from gas chromatography-electrospray ionization/mass spectrometry (GC-ESI/MS) and liquid chromatography-electrospray ionization/mass spectrometry (LC-ESI/MS). The steroids exhibited ionization profiles based on their structural characteristics, regardless of the analyte phase or derivatization. Groups I and II with conjugated or unconjugated keto functional groups at C3 generated the [M+H]+ and [M+H-TMS]+ ions, respectively. On the other hand, Groups III and IV gave rise to the characteristic fragment ions [M+H-TMS-H2O]+ and [M+H-2TMS-H2O]+, corresponding to loss of a neutral TMS·H2O moiety from the protonated molecular ion by in-source dissociation. To the best of our knowledge, this is the first study to successfully ionize and analyze steroids as TMS derivatives using ESI coupled with GC. The present system has enabled the ionization of TMS derivatives under ESI conditions and this method has potential as a novel ionization tool. It is also useful for the simultaneous analysis of steroids as TMS derivatives.

Hyphenation of Production-Scale Free-Flow Electrophoresis to Electrospray Ionization Mass Spectrometry Using a Highly Conductive Background Electrolyte.

In this technical note, we demonstrate the hyphenation of production-scale free-flow electrophoresis (FFE) and sheathless electrospray ionization mass spectrometry (ESI-MS). In contrast to previous hyphenation approaches, we used a highly conductive background electrolyte (BGE) required for production-scale FFE. We found that this kind of BGE as well as a production-scale setup leads to significant electric interference between FFE and MS. This interference prevents steady-state FFE operation. We examine this interference in detail and discuss possible solutions to this issue. We demonstrate that the straightforward grounding of the transfer line removes the influence of ESI-MS on FFE, but creates a current leak from the ESI interface, which adversely affects the ESI spray. Furthermore, we show that only the electrical disconnection of the ESI probe from the FFE-MS transfer line suppresses this undesirable current. In order to facilitate the electrical disconnection we used a low conductivity, silica-based ESI probe with withdrawn inner capillary. This approach allowed the interference-free hyphenation of production-scale FFE (using a highly conductive BGE) with ESI-MS.

A simple sheathless CE-MS interface with a sub-micrometer electrical contact fracture for sensitive analysis of peptide and protein samples

Online coupling of capillary electrophoresis (CE) to electrospray ionization mass spectrometry (MS) has shown considerable potential, however, technical challenges have limited its use. In this study, we have developed a simple and sensitive sheathless CE-MS interface based on the novel concept of forming a sub-micrometer fracture directly in the capillary. The simple interface design allowed the generation of a stable ESI spray capable of ionization at low nanoliter flow-rates (45–90 nL/min) for high sensitivity MS analysis of challenging samples like those containing proteins and peptides. By analysis of a model peptide (leucine enkephalin), a limit of detection (LOD) of 0.045 pmol/μL (corresponding to 67 attomol in a sample volume of ∼15 nL) was obtained. The merit of the CE-MS approach was demonstrated by analysis of bovine serum albumin (BSA) tryptic peptides. A well-resolved separation profile was achieved and comparable sequence coverage was obtained by the CE-MS method (73%) compared to a representative UPLC-MS method (77%). The CE-MS interface was subsequently used to analyse a more complex sample of pharmaceutically relevant human proteins including insulin, tissue factor and α-synuclein. Efficient separation and protein ESI mass spectra of adequate quality could be achieved using only a small amount of sample (30 fmol). In addition, analysis of ubiquitin samples under both native and denatured conditions, indicate that the CE-MS setup can facilitate native MS applications to probe the conformational properties of proteins. Thus, the described CE-MS setup should be useful for a wide range of high-sensitivity applications in protein research.

Spatial effects on electrospray ionization response

The position of the spray-head, the solvent, and a variety of additional instrumental parameters were independently adjusted during electrospray ionization mass spectrometric (ESI-MS) analysis of an equimolar mixture of two different ions. These parameters were found to have drastic effects on the distribution of signal intensity from one ion to another, and therefore the resulting usefulness of acquired spectra. The analytes studied were bis(triphenylphosphine)iminium (PPN) chloride and tetramethylammonium (TMA) chloride, two chemically distinct ions. The use of these two ions in a test solution yielded information regarding ESI probe spatial effects for two very different analytes, while probing the issue of sampling efficiency. Each experimental parameter was individually adjusted prior to rastering the spray head across the operational plane in order to observe how adjustment to a particular parameter affects analyte signal in relation to the distance from the MS aperture. Following acquisition, the intensities of both ions were plotted as ion contour maps demonstrating the intensity change with respect to capillary position in relation to the mass spectrometer aperture. The sharp contrast in ion intensity, and even differential ion activity, with relatively minor instrument changes (such as temperature programming, gas flow rates and solvent choice) clearly demonstrates the importance of finding the “sweet spot” for the ESI spray head, especially when signal intensity and a quality analysis are key.

Discovery of Mouse Spleen Signaling Responses to Anthrax using Label-Free Quantitative Phosphoproteomics via Mass Spectrometry

Inhalational anthrax is caused by spores of the bacterium Bacillus anthracis (B. anthracis), and is an extremely dangerous disease that can kill unvaccinated victims within 2 weeks. Modern antibiotic-based therapy can increase the survival rate to ∼50%, but only if administered presymptomatically (within 24-48 h of exposure). To discover host signaling responses to presymptomatic anthrax, label-free quantitative phosphoproteomics via liquid chromatography coupled to mass spectrometry was used to compare spleens from uninfected and spore-challenged mice over a 72 h time-course. Spleen proteins were denatured using urea, reduced using dithiothreitol, alkylated using iodoacetamide, and digested into peptides using trypsin, and the resulting phosphopeptides were enriched using titanium dioxide solid-phase extraction and analyzed by nano-liquid chromatography-Linear Trap Quadrupole-Orbitrap-MS(/MS). The fragment ion spectra were processed using DeconMSn and searched using both Mascot and SEQUEST resulting in 252,626 confident identifications of 6248 phosphopeptides (corresponding to 5782 phosphorylation sites). The precursor ion spectra were deisotoped using Decon2LS and aligned using MultiAlign resulting in the confident quantitation of 3265 of the identified phosphopeptides. ANOVAs were used to produce a q-value ranked list of host signaling responses. Late-stage (48-72 h postchallenge) Sterne strain (lethal) infections resulted in global alterations to the spleen phosphoproteome. In contrast, ΔSterne strain (asymptomatic; missing the anthrax toxin) infections resulted in 188 (5.8%) significantly altered (q<0.05) phosphopeptides. Twenty-six highly tentative phosphorylation responses to early-stage (24 h postchallenge) anthrax were discovered (q<0.5), and ten of these originated from eight proteins that have known roles in the host immune response. These tentative early-anthrax host response signaling events within mouse spleens may translate into presymptomatic diagnostic biomarkers of human anthrax detectable within circulating immune cells, and could aid in the identification of pathogenic mechanisms and therapeutic targets.

Self‐assembly formation of the magic ion of (H2O)20O+: observation of nanoscale cages of oxygenated water clusters induced from iron nanoparticles

For the first time, we observed a stable and intense ion (m/z 376) of the oxygenated water cluster ion ((H(2)O)(20)O(+)) produced from simply spraying an aqueous solution of iron nanoparticles (Fe NPs) into an electrospray mass spectrometry (ESI-MS) system. Tandem mass spectrometric (MS/MS and MS/MS/MS) results were applied to identify the assignments of the fragment ions of m/z 376 in order to explore the possible structures of this cluster ion. The possible structures of the (H(2)O)(20)O(+) ions are proposed as pentagonal dodecahedron water clathrate cages from the results of tandem mass spectrometry since eliminations of five water molecules were frequently observed in the MS/MS results for many subsequent fragment ions of m/z 376. The formation of this oxygenated water cluster ion ((H(2)O)(20)O(+)) in ESI-MS is attributed to the high surface reactivity and surface energy of Fe NPs during ESI processes (under high temperature and high voltage (5 kV) of ESI spray environment). We believe that the observation of self-assembly formation of oxygenated water clusters is an important issue in nanoscience as well as in the fields of water clusters.

Characterization of coordination complexes by desorption electrospray mass spectrometry with a capillary target

Metal coordination complexes were formed directly from liquid surfaces using desorption electrospray ionization (DESI) mass spectrometry, in which the analyte solutions were furnished by a target capillary that protrudes into the ESI spray. The approach is attractive because it separates complexities of ESI spray droplet formation from delivery of the analyte solution, and thereby gets around difficulty resulting from alteration of the spray process by changes in solution chemistry. Cs +, Ba 2+, and La 3+ coordination complexes were formed using 18-crown-6 (18c6) and triethylphosphate (TEP) as ligands (L), that had the general formula [M n+ (NO 3 −) n−1 (L) m ] +. Formation of singly charged cation complexes was preferred, with charge reduction at the metal site accomplished by attachment of nitrate. Using TEP as a model phosphoryl ligand, alkali metals coordinate with up to three ligands, with Cs + preferring fewer than Na +. Ba 2+ and La 3+ are formed as ion pair complexes [Ba(NO 3)] + and [La(NO 3) 2] +, and both will coordinate with up to four TEP ligands. Using 18c6, Cs + forms a bis-ligand complex. In contrast, [Ba(NO 3)] + prefers a single 18c6 ligand, while La forms mainly [La(NO 3) 2(18c6)] +, for which DFT calculations suggested a structure in which the nitrate ligands occupy pseudo-axial positions on opposing sides of the crown. Lower abundances of bis-18c6 complexes were also formed together with doubly charged [La(NO 3)(18c6) n ] 2+ complexes ( n = 2–4). The results suggest an alternative strategy for probing metal speciation in solution that is less perturbed by the droplet formation and ionization mechanisms operating in conventional electrospray ionization mass spectrometry.

On the Mechanism of Extractive Electrospray Ionization

Extractive electrospray ionization (EESI) is a powerful ambient ionization technique that can provide comprehensive mass spectrometric (MS) information on aerosols, complex liquids, or suspensions without any sample pretreatment. An understanding of the EESI mechanism is critical for defining its range of application, the advantages, and limitations of EESI, and for improving its repeatability, sensitivity, and selectivity. However, no systematic study of EESI mechanisms has been conducted so far. In this work, fluorescence studies in the EESI plume using rhodamine 6G and H-acid sodium salt directly demonstrate that liquid-phase interactions occur between charged ESI droplets and neutral sample droplets. Moreover, the effect of the composition of the primary ESI spray and sample spray on signals of the analyte in EESI-MS was investigated systematically. The results show that the analyte signals strongly depend on its solubility in the solvents involved, indicating that selective extraction is the dominant mechanism involved in the EESI process. This mechanistic study provides valuable insights for optimizing the performance of EESI in future applications.

Rapid fingerprinting and classification of extra virgin olive oil by microjet sampling and extractive electrospray ionization mass spectrometry

Microjet sampling in combination with extractive electrospray ionization (EESI) mass spectrometry (MS) was applied to the rapid characterization and classification of extra virgin olive oil (EVOO) without any sample pretreatment. When modifying the composition of the primary ESI spray solvent, mass spectra of an identical EVOO sample showed differences. This demonstrates the capability of this technique to extract molecules with varying polarities, hence generating rich molecular information of the EVOO. Moreover, with the aid of microjet sampling, compounds of different volatilities (e.g.E-2-hexenal, trans-trans-2,4-heptadienal, tyrosol and caffeic acid) could be sampled simultaneously. EVOO data was also compared with that of other edible oils. Principal Component Analysis (PCA) was performed to discriminate EVOO and EVOO adulterated with edible oils. Microjet sampling EESI-MS was found to be a simple, rapid (less than 2 min analysis time per sample) and powerful method to obtain MS fingerprints of EVOO without requiring any complicated sample pretreatment steps.

Development of Isotope Dilution-Liquid Chromatography/Tandem Mass Spectrometry as a Candidate Reference Method for the Determination of Acrylamide in Potato Chips

An isotope dilution-liquid chromatography/tandem mass spectrometric method was developed as a candidate reference method for the accurate determination of acrylamide in potato chips, starch-rich foodstuff cooked at high temperature. Sample was spiked with 13 C 3 -acrylamide and then extracted with water. The extract was further cleaned up with an Oasis HLB solid-phase extraction (SPE) cartridge and an Oasis mixed-phase cation exchange (MCX) SPE cartridge. The extract was analyzed by using LC/ESI/Tandem MS in positive ion mode. LC with a medium reversed-phase (C4) column was optimized to obtain adequate chromatographic retention and separation of acrylamide. MS was operated to selectively monitor [M+H] + ions of the analyte and its isotope analogue at m/z 72 and m/z 75, respectively. Sample was also analyzed by the LC/MS with selectively monitoring the collisionally induced dissociation channels of m/z 72 → m/z 55 and m/z 75 → 58. Compared to the LC/MS chromatograms, the LC/MS/MS chromatograms showed substantially reduced background chemical noises coming from solvent clusters formed during ESI spray processes and interferences from sample matrix. Repeatability and reproducibility studies showed that the LC/MS/MS method is a reliable and reproducible method which can provide a typical method precision of 1.0% while the LC/MS results are influenced by chemical interferences.

A severe peak tailing of phosphate compounds caused by interaction with stainless steel used for liquid chromatography and electrospray mass spectrometry

A severe peak tailing was observed for adenosine 5'-monophosphate in flow injection analysis with stainless steel tubing and water/methanol mixture (1:1, v/v) as carrier. The cause of the peak tailing was investigated by focusing on the chemical structure of the analytes, the material used for the analytical systems and the composition of the carrier. We clarified that the peak tailing was caused by the interaction between phosphate residues in the analytes and stainless steel. The severe peak tailing did not occur with stainless steel tubing when the phosphate compounds were analyzed with carrier containing phosphoric acid or phosphate buffer. The findings indicate that such ill peak profiles are usually not considerable in conventional HPLC separation because phosphoric acid or phosphate buffer is quite commonly used in eluents. In LC-MS, however, the use of phosphoric acid and phosphate buffer is usually avoided because of their non-volatility; therefore this interaction between stainless steel and phosphate compound becomes predominant and results in severe peak tailings. We also found an effective method for avoiding the interaction. When stainless parts, such as LC tubing and ESI spray capillary, were treated with phosphoric acid prior to analysis, the peak profiles of the phosphate compounds were dramatically improved, even when non-phosphate buffer is used as carrier.

Development of a capillary zone electrophoresis–electrospray ionisation tandem mass spectrometry method for the analysis of fluoroquinolone antibiotics

The applicability of a capillary zone electrophoresis–electrospray ionisation tandem mass spectrometric (CZE–ESI-MS–MS) method for the separation of nine fluoroquinolones was investigated. Method optimisation involved systematic trouble-shooting starting with the type and duration of capillary pre-washing and conditioning, the choice of both the CE run buffer, MS sheath liquid, CE run potential, ESI spray voltage, sheath gas flow-rate, MS capillary voltage and CE capillary and MS capillary temperatures. Another extremely important factor was found to be the degree to which the CE capillary protrudes into the ESI chamber as well as whether or not sheath gas and spray voltage are employed during the CE injection or not. The importance of the latter has, to our knowledge, not been addressed elsewhere. Nine fluoroquinolones have been separated and detected in a single run by this technique.

Highly robust stainless steel tips as microelectrospray emitters

Tapered stainless steel spray tips for sheathless microelectrospray ionization (microESI) have been developed. The fabrication procedure for the tapered stainless steel tips was optimized using an electropolishing technique followed by removal of the burr. Using the tip as the microESI emitter, a stable ESI spray was obtained at a flow rate of 20 nL/min. The sensitivity of the microESI system was almost two orders greater than that of the conventional ion spray system. The tip was highly stable, and was successfully used for over 1000 h. Moreover, these stainless steel tips were suitable for use with sheathless capillary electrophoresis/mass spectrometry (CE/MS) and capillary liquid chromatography/mass spectrometry (LC/MS) for routine analysis in proteomic and pharmaceutical applications.