Selected Ion Chromatograms Research Articles

Particulate organic emissions from incense-burning smoke: Chemical compositions and emission characteristics

Incense burning is a common practice in Asian cultures, releasing hazardous particulate organics. Inhaling incense smoke can result in adverse health effects, yet the molecular compositions of incense-burning organics have not been well investigated due to the lack of measurement of intermediate-volatility and semi-volatile organic compounds (I/SVOCs). To elucidate the detailed emission profile of incense-burning particles, we conducted a non-target measurement of organics emitted from incense combustion. Quartz filters were utilized to trap particles, and organics were analyzed by a comprehensive two-dimensional gas chromatography-mass spectrometer (GC × GC–MS) coupled with a thermal desorption system (TDS). To deal with the complex data obtained by GC × GC–MS, homologs are identified mainly by the combination of selected ion chromatograms (SICs) and retention indexes. SICs of 58, 60, 74, 91, and 97 were utilized to identify 2-ketones, acids, fatty acid methyl esters, fatty acid phenylmethyl esters, and alcohols, respectively. Phenolic compounds contribute the most to emission factors (EFs) among all chemical classes, taking up 24.5 % ± 6.5 % of the total EF (96.1 ± 43.1 μg g−1). These compounds are largely derived from the thermal degradation of lignin. Biomarkers like sugars (mainly levoglucosan), hopanes, and sterols are extensively detected in incense combustion fumes. Incense materials play a more important role in shaping emission profiles than incense forms. Our study provides a detailed emission profile of particulate organics emitted from incense burning across the full-volatility range, which can be used in the health risk assessments. The data processing procedure in this work could also benefit those with less experience in non-target analysis, especially GC × GC–MS data processing.

Non-target scanning of organics from cooking emissions using comprehensive two-dimensional gas chromatography-mass spectrometer (GC×GC-MS)

Cooking emission is a major source influencing human health and air quality. To elucidate the emission pattern of cooking fumes, gaseous and particulate organics were sampled by Tenax TA desorption tubes and quartz filters, respectively. Organics were analyzed by a comprehensive two-dimensional gas chromatography-quadrupole mass spectrometer (GC × GC-qMS) coupled with a thermal desorption system (TDS). A non-target analysis was conducted identifying 166 (gas-phase) and 349 (particle-phase) compounds emitted from cooking fumes. We propose a four-step identification procedure, containing comparison with standards, homologs identification by selected ion chromatograms (SICs), searching spectrums in the National Institute of Standard Technology (NIST) library, and recognitions of unresolved complex mixtures (UCMs). The identified organics are then quantified or semi-quantified by surrogates in the same chemical class with an uncertainty of 27%. Intermediate volatility organic compounds (IVOCs) are extensively detected accounting for 20.5% and 81.4% of the total mass concentration and emission rates (ERs) in gas- and particle-phase samples. Aldehydes (30.6%) and acids (82.7%) dominate the mass concentration of organics in the gas- and particle-phase, respectively. Our work provides a detailed identification procedure in the non-target analysis and emphasizes the importance of IVOC measurement in cooking emissions.

An isotope dilution liquid chromatography-tandem mass spectrometry candidate reference measurement procedure for aldosterone measurement in human plasma.

Accurate quantitation of aldosterone is clinically important in standardized testing for primary aldosteronism. The results are often variable when performed by clinical immunoassays. To standardize and ensure the accuracy of clinical systems, reference measurement procedures (RMPs) with higher metrological order are required. A simple and reliable isotope dilution LC-IDMS/MS-based measurement procedure for human plasma aldosterone has been developed. This method involved plasma spiked with a deuterium-labelled internal standard, equilibrated for 0.5h, and extracted by liquid-liquid extraction (LLE) without derivatization. Aldosterone and its structural analogues were baseline separated with a C18-packed UHPLC column with gradient elution within 7min. The signal intensity variability and measurement imprecision were reduced by bracketing calibration during plasma aldosterone value assignment. The limit of detection (LoD) was 19.4pmol/L with a signal-to-noise ratio (S/N) > 3. The lowest limit of quantification (LLoQ) was 27.7pmol/L (S/N > 10 and CV < 10.0%). LLE was performed with 1mL of n-hexane/ethyl acetate (3:2, v/v), and the extraction recovery was determined to be 92.15 ± 3.54%. The imprecisions were ≤ 3.18% for samples at 124.8, 867.0, and 2628.5pmol/L. The recoveries were 98.11-101.61%. The relative bias between this candidate RMP and the established RMP was 2.76-1.89%. The linearity response ranged from 27.7 to 2774.4pmol/L with R2 = 0.999. The method performance met the requirements of RMPs (≤ 5% total CV and ≤ 3% bias). Furthermore, the developed method was applied to evaluate immunoassays through 41 patient sample comparisons. The calibration and measurement capability (CMC) of this method were also evaluated by measuring these samples. The candidate RMP can serve as an accurate reference baseline for routine methods and can be used for value assignment for reference materials. Selected ion chromatograms by LC-MS/MS using a C18 column for aldosterone and its structural analogues.

Generic multicriteria approach to determine the best precipitation agent for removal of biomacromolecules prior to non-targeted metabolic analysis

The removal of biomacromolecules from biofluids decreases the sample complexity and lower electrospray suppression effects. Furthermore, it can increase the analysis sensitivity, precision, and selectivity. Often removal approaches evaluate the model based on a single criterion, like protein removed or response of one of few specific metabolites. In this study, we used a multicriteria approach to test the effect of using the solvents methanol and acetonitrile (organic solvent precipitation), trichloroacetic acid (acidic precipitation) and ammonium sulphate (salting out) to remove biomacromolecules from a downstream recovery process from a bacillus fermentation. The downstream recovery process intermediates were analysed using reversed-phase ultra-high-pressure liquid chromatography with electrospray ionisation and high-resolution time-of-flight mass spectrometry detection. To evaluate the pre-treatment agents the following multicriteria was applied i) practical considerations, ii) total amino acid in the precipitated pellet, iii) putative identification of the molecules removed or created by the different treatments, iv) coherence between high quality extracted ion chromatograms (repeatability of DW-CODA) and v) replicate consistency from principal component analysis score values obtained by using the CHEMometric analysis of sections of Selected Ion Chromatograms (CHEMSIC) method. This study presents a generic workflow to find the best pre-treatment for removing bio-macromolecules from biofluids with a multicriteria approach. In our case, the best protein removal strategy for downstream recovery intermediates was acetonitrile precipitation. This method showed high precision, created few artefact peaks compared to simple sample dilution, and mainly removed small peptides.

Productivity and oil fingerprinting: Application of analytical chemistry in the assessment of reservoir quality

The ability to understand and possibly even predict the productivity of wells across a field is an important accomplishment in a production setting. In the tight Lower Cretaceous chalk field Valdemar, 5 wells have been observed to produce significantly larger oil volumes compared to nearby wells. This study was conducted to map the inter-well differences in the oil composition to better understand the productivity differences. This was done by analyzing any chemical differences of 21 samples from 16 different wells by GC-MS and principal component analysis of summed extracted ion chromatograms (SICs) using the chemometric analysis of selected ion chromatograms (CHEMSIC) method. The sterane (m/z 217 and m/z 218) and C4 (m/z 234) biomarker SICs were found to have chemically meaningful features described by principal component 1 (PC1). The association between these markers and the relative production was modeled to provide a better understanding of the productivity of the different wells. A correlation between oil saturation and productivity was established in the sense that the more mature and thus less viscous oil seem to have charged certain favorably placed reservoir sections first; these locations coincide with the location of the 5 wells in question.

Evaluation of a bracketing calibration-based isotope dilution liquid chromatography-tandem mass spectrometry candidate reference measurement procedure for 17α-hydroxyprogesterone in human plasma.

A candidate reference measurement procedure (RMP) based on isotope dilution coupled with liquid chromatography-tandem mass spectrometry (ID-LC-MS/MS) was developed and validated for the quantification of 17α-hydroxyprogesterone (17-OHP) in human plasma. 17-OHP spiked with a deuterium-labeled internal standard was extracted from plasma by liquid-liquid extraction with 1 mL n-hexane/ethyl acetate (3:2, v/v). Reversed-phase chromatography and positive electrospray ionization were used in the ID-LC-MS/MS. Gradient elution coupled with use of a C18-packed ultrahigh-performance liquid chromatography column allowed complete baseline resolution of 17-OHP from its structural analogue desoxycorticosterone in 6 min. To determine the 17-OHP level in human plasma, a bracketing calibration method was used to give higher accuracy and precision. The limit of detection and the lower limit of the measuring interval for the candidate RMP were 2.1 pg/mL (6.4 pmol/L) and 4.6 pg/mL (13.9 pmol/L), respectively. Extraction recovery was determined to be (96.08 ± 3.03)% (n = 3). Imprecision (intra-assay and interassay) was 4.03% or less at 0.83, 15.19, 64.22, and 313.46 ng/mL (2.51, 45.97, 194.34, and 948.56 nmol/L, respectively). Recoveries ranged from 98.05% to 102.24%. When comparing our RMP results with those obtained with an established RMP via International Federation of Clinical Chemistry and Laboratory Medicine external quality assessment scheme for reference laboratories in laboratory medicine (RELA) samples, we found that the biases ranged from -1.99% to 3.08% against the targets. No interference was observed, and the linear response ranged from 0.47 to 958.63 ng/mL (1.42 to 2900.90 nmol/L). Moreover, the candidate RMP was used to measure the concentration of 17-OHP in human plasma and was compared with an immunoassay using 40 plasma samples. The performance of the method meets the needs of an RMP (total coefficient of variation of 5% or less and bias of 3.08% or less). This method can be used for reference material value assignment of 17-OHP in human plasma matrix. It could also serve as an accurate reference baseline for routine methods to increase the accuracy and precision of certain clinical laboratory measurements. Graphical abstract Selected ion chromatograms obtained by liquid chromatography-tandem mass spectrometry with a C18 column for 17α-hydroxyprogesterone (17-OHP) from a plasma sample.

Removal of volatile gasoline compounds by indoor potted plants studied by pixel-based fingerprinting analysis

Indoor potted plants are able to remove volatile organic compounds (VOC) from air, but only few studies have investigated the removal of compounds in mixtures. Here, we present a non-targeted pixel-based fingerprinting analysis documenting the removal of a complex mixture of gasoline VOCs by Hedera helix under dynamic chamber conditions allowing for air exchange and continuous gasoline exposure. For 15 days, the entire potted plant was exposed to gasoline; subsequently, the epigeous plant parts were removed and the soil microcosm (i.e. soil, plant roots and microorganisms) was exposed to gasoline for another eight days. Quantitative analysis was performed for heptane, 3-methylhexane, toluene, ethylbenzene and m,p-xylenes, and the CHEMSIC method (CHEMometric analysis of Selected Ion Chromatograms) was used for non-targeted pixel-based fingerprinting analysis. The quantitative analysis demonstrated that the presence of potted plants or pots without epigeous plant parts led to a reduction of selected VOCs by 16.7–22.6%. The CHEMSIC method confirmed this and revealed that all gasoline VOCs were reduced in concentration when H. helix was present. The estimate for the total VOC removal was in the range of 11–32%. The removal was highest for samples where the epigeous plant parts were absent and compounds known to be hard to degrade by microorganisms such as dimethylcyclopentanes were removed the least compared to compounds more easily degraded by microorganisms such as heptane when epigeous plant parts were removed. All findings support the conclusion that the soil microcosm was the main responsible for the removal of VOCs.

Forensic Investigations of Diesel Oil Spills in the Environment Using Comprehensive Two-Dimensional Gas Chromatography-High Resolution Mass Spectrometry and Chemometrics: New Perspectives in the Absence of Recalcitrant Biomarkers.

Forensic investigations of oil spills aim to find the responsible source(s) of the spill. Oil weathering processes change the chemical composition of the spilled oil and make the matching of oil spill samples to potential sources difficult. Diesel oil spill cases are more challenging, because biomarkers recalcitrant to long-term weathering are absent. We developed and tested a new method for the analysis and matching of diesel oil spills using two-dimensional gas chromatography-high resolution mass spectrometry (GC × GC - HRMS) and 2D-CHEMSIC (2-Dimensional CHEMometric analysis of Selected Ion Chromatograms), an extension of the CHEMSIC method to GC × GC data. The 2D-CHEMSIC performs pixel-based analysis using chemometrics on concatenated sections of 2D extracted ion chromatograms to assess the overall chemical variability of the samples, with potential applications for matching spill-source pairs in forensic investigations. The method was tested on samples from a number of diesel oil spill cases, (i) distinguishing chemically similar source diesels, (ii) investigating weathering effects on spill samples to determine type and degree of weathering, and (iii) improving the matching of diesel oil spills affected by weathering. Positive matches for spill-source pairs were identified after excluding the signals from the hydrocarbons most susceptible to evaporation, and photo-oxidized spills were also matched due to the presence of unaffected hydrocarbons. Forensic diagnostics obtained by the 2D-CHEMSIC were validated by the conventional CEN-Tr method.

Analyte‐substrate interactions at functionalized tip electrospray ionization mass spectrometry: Molecular mechanisms and applications

Conventional electrospray ionization mass spectrometry (ESI-MS) commonly uses capillary tip for sample introduction and ionization. In recent years, ESI-MS using noncapillary substrate tips has attracted growing interest as it allows separation and enrichment of analytes from complex samples due to analytes-substrate interactions. In this work, model mixtures and functionalized tips were employed to investigate the molecular mechanism of the analyte-substrate interactions. The mixtures were directly loaded on substrate tips, and then temporal responses of analytes were investigated by monitoring selected ion chromatogram (SIC) responses of each analyte. It is found that all analytes are sprayed out together when bulk solution loaded substrate surface and then sequential ionization of analytes were observed. Sequential ionization of analytes was affected by the analytes-substrate interactions which caused analytes of weaker-interaction to be faster moved and the analytes of stronger-interactions to be retained on the substrate. The main molecular mechanisms of analyte-substrate interactions were revealed to be hydrophobic interactions and electrostatic interactions. Furthermore, based on the mechanistic insights, functionalized tips were further applied for rapid extractive sampling of target analytes from complex samples with good analytical performances. Overall, this study on the mechanism and applications of analyte-substrate interactions is useful for understanding the fundamental principles and further developments of functionalized tip electrospray ionization (TESI).

Study of characteristic ions in frequently-used traditional Chinese medicinal materials derived from animal bile and its application

A LC-MS method was established for study of characteristic ions in frequently-used traditional Chinese medicinal materials derived from animal bile. UPLC-Q-TOF was used in the data acquisition work, then, the software of MarkerLynxTM v.4.1 was performed in the chemometric analysis of data. Besides, selected ion chromatograms of these bile acid ions were comparative studied. Better results were gained in the specificity identification of pig bile, bear bile, cultivated cow-bezoar in vitro, artificial cow-bezoar and some compound preparations(Hugan tablet and Rengongniuhuang Jiaxiaozuo capsule) by this method. The method is suitable for the specificity identification of pig bile, bear bile, cultivated cow-bezoar in vitro, artificial cow-bezoar and compound preparations containing these medicinal materials.

Characterization of Plant Glycoproteins: Analysis of Plant Glycopeptide Mass Spectrometry Data with plantGlycoMS, a Package in the R Statistical Computing Environment.

plantGlycoMS is a set of tools, implemented in R, which is used to assess and validate glycopeptide spectrum matches (gPSMs). Validity of gPSMs is based on characteristic fragmentation patterns of glycopeptides (gPSMvalidator), adherence of the glycan moiety to the known N-glycan biosynthesis pathway in plants (pGlycoFilter), and elution of the glycopeptide within the observed retention time window of other glycopeptides sharing the same peptide backbone (rt.Restrict). plantGlycoMS also contains a tool for relative quantitation of glycoforms based on selected ion chromatograms of glycopeptide ion precursors in the mass spectrometry level 1 data (glycoRQ). This protocol walks the user through this workflow with example mass spectrometry data obtained for a plant glycoprotein.

New Approaches to the Application of DART Mass Spectrometry Coupled with Planar Chromatography for the Analysis of Mixtures of Organic Compounds

A desorption cell for TLC/DART mass spectrometry is developed. An analysis of mixtures of pharmaceutical substances has shown that the proposed approach can improve the sensitivity of the method and achieve the reproducibility of the recorded mass spectra and selected ion chromatograms.

Highly reproducible improved label-free quantitative analysis of cellular phosphoproteome by optimization of LC-MS/MS gradient and analytical column construction

Expanding the sequencing depth of the peptides with a statistically significant quantitative change derived from a biological stimulation is critical. Here we demonstrate that optimization of LC gradient and analytical column construction can reveal over 30,000 unique peptides and 23,000 phosphopeptides at high confidence. The quantitative reproducibility of different analytical workflows was evaluated by comparing the phosphoproteome of CD3/4 stimulated and unstimulated T-cells as a model system. A fritless, 50cm-long column packed with 1.9μm particles operated with a standard pressure HPLC significantly improved the sequencing depth 51% and decreased the selected ion chromatogram peak spreading. Most importantly, under the optimal workflow we observed an improvement of over 300% in detection of significantly changed phosphopeptides in the stimulated cells compared with the other workflows. The discovery power of the optimized column configuration was illustrated by identification of significantly altered phosphopeptides harboring novel sites from proteins previously established as important in T cell signaling including A-Raf, B-Raf, c-Myc, CARMA1, Fyn, ITK, LAT, NFAT1/2/3, PKCα, PLCγ1/2, RAF1, and SOS1. Taken together, our results reveal the analytical power of optimized chromatography using sub 2μm particles for the analysis of the T cell phosphoproteome to reveal a vast landscape of significantly altered phosphorylation changes in response to T cell receptor stimulation.

Source identification of beached oil at Al Zubarah, Northwestern Qatar

The Gulf was contaminated with a large quantity of crude oil during the Gulf War in 1991. Some of the oil beached on the shorelines of the Northwest coast of Qatar. It was discovered, during a sampling campaign in 2015, that the shorelines were contaminated with both fresh oil and heavily weathered oil that could be originating from the Gulf War. A total of 38 oil spill samples were collected during two sampling campaigns and analysed with gas chromatography - mass spectrometry (GC–MS). The chemical fingerprints of petroleum biomarkers and C0-C4 alkyl-substituted polycyclic aromatic compounds for these spill samples were compared to those of more than 250 crude oils from around the world applying the CHEMSIC method (chemometric analysis of selected ion chromatograms). The weathering degree varied significantly between samples, and relatively unweathered oil from the Gulf War oil spill is still found in the Al Zubarah area together with oil from more recent spills. The spill samples were found to originate from at least two sources with most resemblance to oils from Kuwait and Saudi Arabia.

Highly sensitive derivatization reagents possessing positively charged structures for the determination of oligosaccharides in glycoproteins by high-performance liquid chromatography electrospray ionization tandem mass spectrometry

We have developed three kinds of novel derivatization reagents (4-CEBTPP, 4-CBBTPP, 5-COTPP) with triphenylphosphine (TPP) as a basic structure carrying a permanent positive charge for resolution of the oligosaccharides in glycoprotein using high-performance liquid chromatography electrospray ionization tandem mass spectrometry (LC-ESI–MS/MS). The synthesized reagents reacted with the sialylglycosylamine of the sialylglycopeptide after treatment by PNGase F. The final derivatives were analyzed by ESI–MS and sensitively detected in the selected reaction monitoring (SRM) mode. Furthermore, the limits of detection (S/N=3) on the SRM chromatograms were at the fmol level (30fmol). Therefore, we used the limit of detection of the reagent products detected by the SRM and evaluated the utility of each reagent. Among the reagents, the positively charged 4-CEBTPP derivative’s peak area was the highest; 4-CEBTPP with a positively charged structure showed about a 20 times greater sensitivity for the glycosylamine of the SGP product compared to the conventional fluorescence reagent, Fmoc-Cl. In addition, various fragment ions based on the carbohydrate units also appeared in the MS/MS spectra. Among the fragment ions, m/z 627.37 (CE=40eV) corresponding to 4-CEBTPP-GlcNAc and m/z 120.09 (CE=100eV) corresponding to 4-CEBTPP are the most important ones for identifying the oligosaccharide. 4-CEBTPP-SGA was easily identified by the selected-ion chromatogram in the product ion scan (m/z 120.09) and in the precursor ion scan (m/z 627.37) by MS/MS detection. The derivatized analytes have a high ionization efficiency and they are detected with a high sensitivity in the electrospray ionization. The novel derivatization reagent with a multi-function provided a higher sensitivity for the oligosaccharide analysis, as well as a better specificity and feasibility. Furthermore, several oligosaccharides in fetuin and ribonuclease B were successfully identified by the proposed procedure.

Multi-mode acquisition (MMA): An MS/MS acquisition strategy for maximizing selectivity, specificity and sensitivity of DIA product ion spectra.

In proteomics studies, it is generally accepted that depth of coverage and dynamic range is limited in data-directed acquisitions. The serial nature of the method limits both sensitivity and the number of precursor ions that can be sampled. To that end, a number of data-independent acquisition (DIA) strategies have been introduced with these methods, for the most part, immune to the sampling issue; nevertheless, some do have other limitations with respect to sensitivity. The major limitation with DIA approaches is interference, i.e., MS/MS spectra are highly chimeric and often incapable of being identified using conventional database search engines. Utilizing each available dimension of separation prior to ion detection, we present a new multi-mode acquisition (MMA) strategy multiplexing both narrowband and wideband DIA acquisitions in a single analytical workflow. The iterative nature of the MMA workflow limits the adverse effects of interference with minimal loss in sensitivity. Qualitative identification can be performed by selected ion chromatograms or conventional database search strategies.

Source apportionment of polycyclic aromatic hydrocarbons (PAHs) in sediments from Khuzestan province, Iran

Khuzestan, Iran is heavily industrialised with petrochemical and refinery companies. Herein, sediment and soil samples were collected from Hendijan coast, Khore Mosa and Arvandroud River. The CHEMSIC (CHEmometric analysis of Selected Ion Chromatograms) method was used to assign the main sources of polycyclic aromatic hydrocarbon (PAH) pollution. A four-component principal component analysis (PCA) model was obtained. While principal component 1 (PC1) was related to the total concentration of PAHs, the remaining PCs described three distinct sources: PC2 and PC3 collectively differentiate between weathered petrogenic and pyrogenic, and PC4 is indicative for a diagenetic input. The sources of PAHs in the Arvandroud River were mainly relatively fresh oil with some samples corresponding to a weathered oil input. Further, perylene (indicator for diagenetic source) was identified. Samples from Khore Mosa revealed a mixture with high proportions of high-molecular-weight PAHs, indicating a pyrogenic/weathered petrogenic source. Samples from Hendijan coast contained low relative concentrations of PAHs, thus only little information on pollution sources.

PAH related effects on fish in sedimentation ponds for road runoff and potential transfer of PAHs from sediment to biota

Road runoff is an important source of pollution to the aquatic environment, and sedimentation ponds have been installed to mitigate effects on the aquatic environment. The purpose of this study was to investigate if a) fish from sedimentation ponds were affected by road pollution and; b) the transfer of PAHs from road runoff material to aquatic organisms was substantial.Minnow from a sedimentation pond (Skullerud) near Oslo (Norway) had higher levels of CYP1A enzyme and DNA stand breaks than minnow from the nearby river, but high concentrations of PAH-metabolites in bile revealed that both populations were highly exposed. Principal component analysis revealed that CYP1A and age of fish were correlated, while levels of PAH-metabolites were not correlated to CYP1A or DNA damage. Minnow from a lake un-affected by traffic had much lower levels of PAH-metabolites than the exposed fish, and also an improved condition. The latter results indicate that fish health was affected by road runoff.A closer investigation of PAH levels of the ecosystems of two sedimentation ponds (Skullerud and Vassum) and nearby environments were conducted. The concentration of the 16 EPA PAHs in sediments of the sedimentation ponds were high (1900–4200ngg−1), and even higher levels were observed in plants. Principal component analysis of selected ion chromatograms of PAHs showed a clear separation of plants vs. sediments. The plants preferentially accumulated the high molecular PAHs, both from sedimentation ponds with a petrogenic PAH isomer ratio in sediments; and from a lake with pyrogenic PAH isomer ratio in sediments.

Mobility of Proteins in Porous Substrates under Electrospray Ionization Conditions

Proteins are important substances in living organisms and characterization of proteins is an indispensible part for protein study. Analysis of proteins using electrospray ionization-mass spectrometry (ESI-MS) with porous substrates was investigated in this study. The results revealed that the ionization process had two stages. At the first stage, mobility and resulting spectra of proteins were similar to those obtained with conventional capillary-based ESI-MS. At the second stage, hydrophobic-hydrophobic interactions between proteins and the tip surfaces played an important role in mobility and detectability of protein ions, which were size and shape dependent, and a linear relationship could be found between the peak area of selected ion chromatogram and the cross section of protein ions. Preparative separation of proteins could be achieved by collecting the proteins remained on the porous substrates. These results led us to propose that electrospray ionization from porous substrates offer a potential approach for analysis of proteins and investigation of protein structures and conformations.

On an aerodynamic mechanism to enhance ion transmission and sensitivity of FAIMS for nano-electrospray ionization-mass spectrometry.

Simulations show that significant ion losses occur within the commercial electrospray ionization-field asymmetric waveform ion mobility spectrometer (ESI-FAIMS) interface owing to an angular desolvation gas flow and because of the impact of the FAIMS carrier gas onto the inner rf (radio frequency) electrode. The angular desolvation gas flow diverts ions away from the entrance plate orifice while the carrier gas annihilates ions onto the inner rf electrode. A novel ESI-FAIMS interface is described that optimizes FAIMS gas flows resulting in large improvements in transmission. Simulations with the bromochloroacetate anion showed an improvement of ~9-fold to give ~70% overall transmission). Comparable transmission improvements were attained experimentally for six peptides (2+) in the range of m/z 404.2 to 653.4 at a chromatographic flow rate of 300 nL/min. Selected ion chromatograms (SIC) from nano-LC-FAIMS-MS analyses showed 71% (HLVDEPQNLIK, m/z 653.4, 2+) to 95% (LVNELTEFAK, m/z 582.3, 2+) of ion signal compared with ion signal in the SIC from LC-MS analysis. IGSEVYHNLK (580.3, 2+) showed 24% more ion signal compared with LC-MS and is explained by enhanced desolvation in FAIMS. A 3-10 times lower limits of quantitation (LOQ) (<15% RSD) was achieved for chemical noise limited peaks with FAIMS. Peaks limited by ion statistics showed subtle improvement in RSD and yielded comparable LOQ to that attained with nano-LC-MS (without FAIMS). These improvements were obtained using a reduced FAIMS separation gap (from 2.5 to 1.5 mm) that results in a shorter residence time (13.2 ms ± 3.9 ms) and enables the use of a helium free transport gas (100% nitrogen).