Ghost Peaks of Aromatic Metabolites Induced by Corona Discharge Artifacts in LC-ESI-MS/MS.

LC-ESI-MS/MS is a preferred method for detecting and identifying metabolites, including those that are unpredictable from the genome, especially in basal metazoans like Cnidaria, which diverged earlier than bilaterians and whose metabolism is poorly understood. However, the unexpected appearance of a "ghost peak" for dopamine, which exhibited the same m/z value and MS/MS product ion spectrum during an analysis of Nematostella vectensis, a model cnidarian, complicated its accurate identification. Understanding the mechanism by which "ghost peaks" appear is crucial to accurately identify the monoamine repertoire in early animals so as to avoid misassignments.

Effects of anions on the electrospray ionization of proteins in strong acids.

The effect of anions on the positive electrospray ionization (ESI) of proteins in different strong acids with varying pH values from 3 to 1 is studied using high-pressure ESI. Reducing the pH from ∼2 to 1 caused a drastic shift in charge state from a high-charge-state distribution (HCSD) to a narrow low-charge-state distribution (LCSD). The shift in charge state was consistent with the circular dichroism result that showed a conformational change due to the "acid-induced folding" of proteins from an unfolding state to a compact molten globule state.

Direct ESI-MS of Ionic Liquids Using High-Pressure Electrospray From Large-Bore Emitters Made of Micropipette Tips.

Electrospray ionization mass spectrometry of neat undiluted ionic liquid (IL) and the analysis of protein with the doping of IL were performed using high-pressure electrospray. The use of disposable micropipette tips as emitters eased the handling of viscous and easy-to-clog samples and improved the reproducibility of the measurement. A high-pressure operation enabled the stable electrospray of the highly conductive IL from these relatively large bore emitters.

Tuning the Internal Energy of Ions Produced by Atmospheric and High-Pressure Electrospray by Modulating the Gas Throughput into the First Vacuum Stage.

In a high-pressure environment, electrospray ionization (ESI) can be achieved without discharge between the emitter and the counter electrode, thus enabling the generation of gas-phase ions from liquid with high surface tension, such as pure water, which requires a high onset voltage for stable ESI. In this study, the ion dissociation during the transferring of ions/charged droplets from a superatmospheric pressure environment to vacuum has been systematically investigated using benzyl ammonium thermometer ions. The ion source pressure did not affect the internal energy distribution of ions, whereas the gas throughput into the first vacuum stage clearly influences the internal energy distribution of the ions.

Formation of Alternating Surfactant-Enriched and Surfactant-Depleted Phases in the Taylor Cone of a Nanoelectrospray.

The electrospray ionization of highly conductive solutions containing Triton X-100, a nonionic surfactant, is found to induce alternating periods of surfactant enrichment and depletion when the concentration of the surfactant is near the critical micelle concentration (CMC) and when the flow rate is on the order of 10 nL/min. Analyzing the surfactant-protein mixture shows that the protein is partially denatured during the surfactant enrichment. The measurement of the phospholipid and oligosaccharide mixture prepared in the surfactant solution shows that the ion signal of the lipid is in phase with, and the hydrophilic oligosaccharide is out of phase with the surfactant signal.

Automated sample preparation for electrospray ionization mass spectrometry based on CLOCK-controlled autonomous centrifugal microfluidics.

We report a centrifugal microfluidic device that automatically performs sample preparation under steady-state rotation for clinical applications using mass spectrometry. The autonomous microfluidic device was designed for the control of liquid operation on centrifugal hydrokinetics (CLOCK) paradigm. The reported device was highly stable, with less than 7% variation with respect to the time of each unit operation (sample extraction, mixing, and supernatant extraction) in the preparation process.

Probing Acid-Induced Compaction of Denatured Proteins by High-Pressure Electrospray Mass Spectrometry.

Further increase in the acidity in the most denaturing acidic solution is known to induce compaction of the fully unfolded protein into a compact molten globule. The phenomenon of "" takes place at pH ∼1 in strong acid aqueous solutions with high electrical conductivity and surface tension, a condition that is difficult to handle using conventional electrospray ionization methods for mass spectrometry. Here, high-pressure electrospray ionization (HP-ESI) is used to produce well-resolved mass spectra for proteins in strong acids with pH as low as 1.

Feedback Control of Electrospray with and without an External Liquid Pump Using the Spray Current and the Apex Angle of a Taylor Cone for ESI-MS.

An electrospray operated in the steady cone-jet mode is highly stable but the operating state can shift to pulsation or multijet modes owing to changes in flow rate, surface tension, and electrostatic variables. Here, a simple feedback control system was developed using the spray current and the apex angle of a Taylor cone to determine the error signal for correcting the emitter voltage. The system was applied to lock the cone-jet mode operation against external perturbations.

Tuning oxidative modification by a strong electric field using nanoESI of highly conductive solutions near the minimum flow rate.

Oxidative modification is usually used in mass spectrometry (MS) for labeling and structural analysis. Here we report a highly tunable oxidation that can be performed in line with the nanoESI-MS analysis at the same ESI emitter without the use of oxidative reagents such as ozone and HO, and UV activation. The method is based on the high-pressure nanoESI of a highly conductive (conductivity >3.

Bipolar Electrospray from Electrodeless Emitters for ESI without Electrochemical Reactions in the Sprayer.

A bipolar ESI source is developed to generate a simultaneous emission of charged liquid jets of opposite polarity from an electrodeless sprayer. The sprayer consists of two emitters, and the electrosprays are initiated by applying a high potential difference (HV) across the counter electrodes facing each emitter. The sprayer and the liquid delivery system are made of all insulators without metal components, thus enabling the total elimination of electrochemical reactions taking place at the liquid-electrode interface in the typical electrosprayer.

A Subtle Change in Nanoflow Rate Alters the Ionization Response As Revealed by Scanning Voltage ESI-MS.

The small charged droplet generated from the nanoelectrospray ionization (nanoESI) source at nL/min flow rate gives its unique feature of high-performance ionization. A continuous scan of the flow rate in this regime can trace the effect of droplet size in greater detail for a better understanding of the ionization process. To date, such practical implementation is hindered by the lack of a suitable liquid pump and the reproducibility of microcapillaries-based systems.

Generation of Ions from Aqueous Taylor Cones near the Minimum Flow Rate: "True Nanoelectrospray" without Narrow Capillary.

Generating ultrafine charged droplets using electrospray is crucial for attaining high ionization efficiency for mass spectrometry. The size of the precursor charged droplets depends on the spray flow rate, and conventional wisdom holds that an electrospray of a nL/min flow rate (nanoelectrospray) is only possible using narrow capillaries with an inner diameter of ∼1 μm or smaller. Here, the electrospray of aqueous solutions with high electric conductivities generated from a large off-line capillary of 0.

Electrospray Ionization Inside the Ion Inlet Tube: Multijet Mode Operation.

We investigated the electrospray ionization inside the narrow channel of the ion inlet tube. An insulating emitter capillary made of fused silica with a 0.2 mm outer diameter was inserted into the ion inlet tubes with a 0.

Miniaturized String Sampling Probe and Electrospray Extraction/Ionization within the Ion Inlet Tube for Mass Spectrometric Endoscopy.

A moving string sampling probe and a new ESI based ionization source that can be readily incorporated into the existing endoscopes are developed for performing mass spectrometry during the endoscopic procedure. The medical-grade silk suture driven by a stepping motor is used to perform the sampling on the region of interest when the probe head is brought gently into contact with the surface of the gastrointestinal tissue. The tissues and the compounds adhered to the sampling string are transported to an ionization region inside the ion inlet tube in which they are extracted and ionized by the charging droplets generated from an electrospray outside the ion inlet.

High-pressure ESI-MS made easy using a plug-and-play ion source and its application to highly conductive aqueous solutions.

The performance of a compact high-pressure electrospray ionization (HP-ESI) source that can be readily used for commercial atmospheric pressure ionization (API) mass spectrometers is reported. The ion source employs a converging-diverging outlet nozzle, and ions/droplets generated inside the high-pressure compartment are carried by the high-velocity air jet toward the mass spectrometry (MS) ion inlet placed under the atmospheric pressure. With the use of a shielding electrode, the HP-ESI can also be operated with its emitter held at ground potential.

A Plug-and-Play High-Pressure ESI Source with an Emitter at Ground Potential and Its Application to High-Temperature Capillary LC-MS.

A new high-pressure ESI source that can be readily used for commercial API mass spectrometers in a plug-and-play manner without any modification on the ion sampling interface is introduced. The emitter can be operated at ground potential, and the positive mode electrospray is generated by applying a negative high potential to the counter electrode. A shielding electrode effectively shields the opposing electric field and improves the ion transmission.

High-Temperature Liquid Chromatography and the Hyphenation with Mass Spectrometry Using High-Pressure Electrospray Ionization.

Increasing the operating temperature of the liquid chromatography (LC) column has the same effect as reducing the diameter of the packing particles on minimizing the contribution of -term in the van Deemter equation, flattening the curve of plate height linear velocity in the high-speed region, thus allowing a fast LC analysis without the loss of plate count. While the use of smaller particles requires a higher pumping pressure, operating the column at higher temperature reduces the pressure due to lower liquid viscosity. At present, the adoption of high-temperature LC lags behind the ultra-high-pressure LC.

Real-time analysis of living animals and rapid screening of human fluid samples using remote sampling electrospray ionization mass spectrometry.

Real-time and in-situ mass-spectrometry analyses of living animal and biological sample were performed using a novel remote sampling electrospray ionization (RS-ESI) probe. Unlike conventional ESI, in which injection or syringe loading is required for sample introduction, the RS-ESI probe ionizes the samples when the sampling capillary is in contact with the sample. As the sampling capillary is electrically held at ground potential, the safety of the animal and operator is assured.

Hyphenation of high-temperature liquid chromatography with high-pressure electrospray ionization for subcritical water LC-ESI-MS.

High-pressure electrospray ionization (HP-ESI) performed under super-atmospheric pressure allows a stable and efficient electrospray of pure aqueous and/or superheated solutions even under a μL min-1 flow rate regime. In this paper, we report the direct coupling of the HP-ESI source to high-temperature liquid chromatography (HT-LC) operated at ≤30 μL min-1 flow rates. In addition to ESI, the ion source functions as a back-pressure regulator to keep the mobile phase in the liquid phase when the column is heated to >100 °C.

Analytical characteristics of nano-electrospray operated under super-atmospheric pressure.

High-pressure nanoelectrospray ionization (nanoESI) source is a recently developed technique in which the electrospray ionization is generated inside an enclosed chamber with gas pressure higher than the atmospheric pressure. In this paper, the performance of nanoESI under different gas pressures, emitter position, ion inlet temperature, additive for desalination are presented. Under a pressure of 2 bars, the nanoESI is almost eased from the electrical discharge problem, and that offers a wider tuning window for the emitter potential to produces a higher and more stable ion signal.

Electrospray ionization source with a rear extractor.

A new electrospray source design is introduced by having an extractor electrode placed at 1 to 2 mm behind the emitter tip. The extractor was integrated into the sprayer body as a single device. An insulating tube was used to isolate the emitter from the extractor and to deliver the sheath gas for the electrospray.

Towards Practical Endoscopic Mass Spectrometry.

In this paper, we briefly review the remote mass spectrometric techniques that are viable to perform "endoscopic mass spectrometry," , and MS analysis inside the cavity of human or animal body. We also report our experience with a moving string sampling probe for the remote sample collection and the transportation of adhered sample to an ion source near the mass spectrometer. With a miniaturization of the probe, the method described here has the potential to be fit directly into a medical endoscope.

In vivo endoscopic mass spectrometry using a moving string sampling probe.

At present, endoscopy relies almost exclusively on optical microscopy and the accurate analysis such as MS interrogation is performed ex situ using biopsy. In this work, a novel probing system is developed to perform in situ and in vivo endoscopic mass spectrometry using a moving string for the sampling and transportation of material. A prototype of a mass spectrometric endoscope is constructed using an industrial endoscope and a commercial mass spectrometer.

Desorption in Mass Spectrometry.

In mass spectrometry, analytes must be released in the gas phase. There are two representative methods for the gasification of the condensed samples, , ablation and desorption. While ablation is based on the explosion induced by the energy accumulated in the condensed matrix, desorption is a single molecular process taking place on the surface.