Confined direct analysis in real time ion source and its applications in analysis of volatile organic compounds of Citrus limon (lemon) and Allium cepa (onion)

The DART (direct analysis in real time) ion source is a novel atmospheric pressure ionization technique that enables efficient ionization of gases, liquids and solids with high throughput. A major limit to its wider application in the analysis of gases is its poor detection sensitivity caused by open‐air sampling. In this study, a confined interface between the DART ion source outlet and mass spectrometer sampling orifice was developed, where the plasma generated by the atmospheric pressure glow discharge collides and ionizes gas‐phase molecules in a Tee‐shaped flow tube instead of in open air. It leads to significant increase of collision reaction probability between high energy metastable molecules and analytes. The experimental results show that the ionization efficiency was increased at least by two orders of magnitude. This technique was then applied in the real time analysis of volatile organic compounds (VOCs) of Citrus Limon (lemon) and wounded Allium Cepa (onion). The confined DART ion source was proved to be a powerful tool for the studies of plant metabolomics. Copyright © 2012 John Wiley & Sons, Ltd.     

Surface swabbing technique for the rapid screening for pesticides using ambient pressure desorption ionization with high-resolution mass spectrometry

A rapid screening method for pesticides has been developed to promote more efficient processing of produce entering the United States. Foam swabs were used to recover a multiclass mixture of 132 pesticides from the surfaces of grapes, apples, and oranges. The swabs were analyzed using direct analysis in real time (DART) ionization coupled with a high‐resolution Exactive Orbitrap? mass spectrometer. By using a DART helium temperature gradient from 100–350°C over 3 min, a minimal separation of analytes based on volatility differences was achieved. This, combined with the Exactive's mass resolution of 100 000, allowed the chromatographic step, along with the typical compositing and extraction steps associated with gas chromatography/mass spectrometry (GC/MS) or liquid chromatography/mass spectrometry (LC/MS) approaches, to be eliminated. Detection of 86% of the analytes present was consistently achieved at levels of 2 ng/g (per each apple or orange) and 10 ng/g (per grape). A resolution study was conducted with four pairs of isobaric compounds analyzed at a mass resolution of 100 000. Baseline separation was achieved with analyte ions differing in mass by 25 ppm and analyte ions with a mass difference of 10 ppm were partially resolved. In addition, field samples that had undergone traditional sample preparation using QuEChERS (quick, easy, cheap, rugged, and safe) were analyzed using both LC/MS and DART‐MS and the results from the two techniques were found to be comparable in terms of identification of the pesticides present. The use of swabs greatly increased sample throughput by reducing sample preparation and analysis time. Published in 2010 by John Wiley & Sons, Ltd.     

Ambient analysis by thermal desorption atmospheric pressure photoionization

Ambient mass spectrometry has attracted substantial attention in recent years. Among ambient ionization methods, thermal desorption ionization stands out because of two attributes: (1) simplicity, rendering the technique suitable for in-field applications, and (2) ability to couple with a variety of gas-phase ionization methods thereby broadening the range of molecules that can be analyzed with this method. Here, we report on improving the performance of a direct analysis in real time (DART) source by implementing atmospheric pressure photoionization (APPI) downstream of the desorption region. At identical desorption and ion sampling conditions, APPI leads to detection of radical molecular ions from non-polar compounds that are absent from the spectra generated by DART alone. Moreover, a factor of 3–5 improvement in sensitivity is observed using APPI for positive ions commonly detected by DART and DART-APPI. Using helium and nitrogen as desorption gases, APPI shows identical performance regardless of desorption gas type. In contrast, a dramatic decrease in sensitivity is observed for DART operated with nitrogen compared to DART with helium. Comparable performance for DART and DART-APPI are observed in negative ion mode, although both show a drastic improvement in the absence of the Vapur interface. This interface creates a differentially pumped chamber prior to inlet of the mass spectrometer and reduces the mass spectrometer gas load when helium is used as desorption gas.     

Analysis of multiple mycotoxins in cereals under ambient conditions using direct analysis in real time (DART) ionization coupled to high resolution mass spectrometry

Direct analysis in real time (DART) ionization coupled to an (ultra)high resolution mass spectrometer based on orbitrap technology (orbitrapMS) was used for rapid quantitative analysis of multiple mycotoxins isolated from wheat and maize by modified QuEChERS procedure. After initial evaluation of ionization efficiencies for major groups of mycotoxins achievable with DART technology, sample preparation procedure and instrument parameter settings were optimized to obtain sensitive and accurate determination of most intensively ionizing toxins (deoxynivalenol, nivalenol, zearalenon, actyldeoxynivalenol, deepoxy-deoxynivalenol, fusarenon-X, altenuene, alternariol, alternariolmethylether, diacetoxyscirpenol, sterigmatocystin). The lowest calibration levels (LCLs) estimated for the respective analytes ranged from 50 to 150 μg kg⁻¹. Quantitative analysis was performed either with the use of matrix-matched standards or by employing commercially available ¹³C-labeled internal standards (available for deoxynivalenol, nivalenol and zearalenon). Good recoveries (100-108%) and repeatabilities (RSD 5.4-6.9%) were obtained at spiking level 500 μg kg⁻¹ with isotope dilution technique. Based on matrix-matched calibration, recoveries and repeatabilities were in the range 84-118% and 7.9-12.0% (RSD), respectively. The trueness of data obtained for deoxynivalenol and zearalenon in wheat/maize by DART-orbitrapMS was demonstrated by analysis of certified reference materials (CRMs). Good agreement of these results with data generated by validated ultra-high pressure liquid chromatography-time-of-flight mass spectrometry method was documented.     

Analysis of isoflavones in soybeans employing direct analysis in real-time ionization-high-resolution mass spectrometry

A direct analysis in real‐time (DART) ion source coupled to a high‐resolution orbitrap mass spectrometer was used for the quantitative analysis of isoflavones isolated from soybeans. For the isolation of genistein, daidzein, glycitein, and their respective acetyl, malonyl, and glucoside forms, an extraction employing 80% aqueous MeOH enhanced by sonication was used. As far as the total isoflavones (expressed as aglycones) were to be determined, an acid hydrolysis with 80% aqueous EtOH and refluxing had to be employed, while in the latter case a good agreement of the results with the data generated by the UHPLC‐orbitrap MS method was achieved, in the case of the analysis of non‐hydrolyzed extracts, some overestimation of the results as compared with those generated by UHPLC‐orbitrap MS was observed. A careful investigation of this phenomenon showed that the free aglycones originated from the conjugated forms of isoflavones in the DART ion source, thus contributing significantly to the “free” genistein/daidzein/glycitein signals during the DART analysis. Good recoveries (95–102%) and repeatabilities (RSD: 7–15%) were obtained at the spiking levels of 0.5, 1, and 0.05 g/kg, for daidzein, genistein, and glycitein, respectively. The limits of detection estimated for the respective analytes were 5 mg/kg.     

热解析低温等离子体电离源用于糯高粱中农药残留的快速筛查

本研究设计并搭建了一套热解析低温等离子体电离源(TD-LTP),与质谱联用实现了糯高粱中农药残留的快速和高灵敏检测.TD-LTP由热解析装置和低温等离子体放电源两部分组成,农药残留样品首先在热解析进样器内汽化,再由载气载带进入等离子体区域被电离.热解析进样器使LTP产生的气相等离子体与样品之间的气-固或气-液相互作用转变为气-气相互作用,大大提高了难挥发样品(如农药)的电离效率;电离源与质谱进样口之间采用同轴连接,提高了离子的利用率和传输效率.与传统的LTP电离源相比,TD-LTP电离源的灵敏度提高了8倍以上,稳定性提高了4倍.本研究对热解析低温等离子体电离源的各参数进行了优化,并与自制的矩形离子阱质谱相结合,研究了12种农药在该电离源下的特征离子.最后,将此电离源与商品化的三重四极杆质谱仪联用,对糯高粱样品中的12种农药残留进行了快速筛查,结果表明,本方法灵敏度高,可以满足食品安全国家标准规定的谷物中农药残留最大限量检测要求.     

Cluster calibration in mass spectrometry: laser desorption/ionization studies of atomic clusters and an application in precision mass spectrometry

For accurate mass measurements and identification of atomic and molecular species precise mass calibration is mandatory. Recent studies with laser desorption/ionization and time-of-flight analysis of cluster ion production by use of fullerene and gold targets demonstrate the generation of atomic clusters for calibration purposes. Atomic ion results from the Penning trap mass spectrometer ISOLTRAP, in which a carbon cluster ion source has recently been installed, are presented as an application in the field of precision mass spectrometry.     

Atmospheric pressure laser desorption/chemical ionization mass spectrometry: a new ionization method based on existing themes

We have designed and constructed an atmospheric pressure laser desorption/chemical ionization (AP-LD/CI) source that utilizes a laser pulse to desorb intact neutral molecules, followed by chemical ionization via reagent ions produced by a corona discharge. This source employs a heated capillary atmospheric pressure inlet coupled to a quadrupole ion trap mass spectrometer and allows sampling under normal ambient air conditions. Preliminary results demonstrate that this technique provides approximately 150-fold increase in analyte ions compared to the ion population generated by atmospheric pressure infrared matrix-assisted laser desorption/ionization (AP-IR-MALDI).     

Optimization of direct analysis in real time (DART) linear ion trap parameters for the detection and quantitation of glucose

Presented here are findings for the development and optimization of a simple, high-throughput, and rapid method for the analysis of glucose. Because the applications of glucose and other six-carbon sugars is a growing field of interest especially in the production of biofuels, an efficient and rapid method for their quantitation from lignocelluloses is necessary. Glucose was analyzed using direct analysis in real time (DART) ionization and formed adducts (along with fragmentation) were observed with a linear ion trap (LIT) mass spectrometer. Since DART can be considered a complex thermal desorption ionization process, an optimization study of the helium gas temperature and introduction into the ionization region was performed. It was observed these parameters have a significant effect on the overall signal intensity as well as the signal-to-noise ratios in DART mass spectra. Using these optimized parameters, a set of different glucose concentrations (ranging from 10 to 3000 μM) were analyzed and used to determine a linear dynamic range (with the use of an internal standard). The analysis of the samples was done with minimal sample preparation and found to be reproducible on different days.     

Thin‐layer chromatography and mass spectrometry coupled using proximal probe thermal desorption with electrospray or atmospheric pressure chemical ionization

An atmospheric pressure proximal probe thermal desorption sampling method coupled with secondary ionization by electrospray or atmospheric pressure chemical ionization was demonstrated for the mass spectrometric analysis of a diverse set of compounds (dyestuffs, pharmaceuticals, explosives and pesticides) separated on various high‐performance thin‐layer chromatography plates. Line scans along or through development lanes on the plates were carried out by moving the plate relative to a stationary heated probe positioned close to or just touching the stationary phase surface. Vapors of the compounds thermally desorbed from the surface were drawn into the ionization region of a combined electrospray ionization/atmospheric pressure chemical ionization source where they merged with reagent ions and/or charged droplets from a corona discharge or an electrospray emitter and were ionized. The ionized components were then drawn through the atmospheric pressure sampling orifice into the vacuum region of a triple quadrupole mass spectrometer and detected using full scan, single ion monitoring, or selected reaction monitoring mode. Studies of variable parameters and performance metrics including the proximal probe temperature, gas flow rate into the ionization region, surface scan speed, read‐out resolution, detection limits, and surface type are discussed. Published in 2010 by John Wiley & Sons, Ltd.     

Detection of explosives on solid surfaces by thermal desorption and ambient ion/molecule reactions

A simple, fast and direct method is presented for detecting traces of solid explosives on cotton swabs or in particulate samples: ions are transferred into a mass spectrometer after thermal desorption and corona discharge chemical ionization in ambient air; specificity is enhanced using ambient ion/molecule reactions or by conventional tandem mass spectrometry.     

Analysis of cell cultures of Taxus wallichiana using direct analysis in real-time mass spectrometric technique

The applicability of a new mass spectrometric technique, DART (direct analysis in real time) has been studied in the analysis of the calli of Taxus wallichiana. The intact callus samples were directly analyzed by holding them in the gap between the DART source and mass spectrometer for measurements. Five C-14 oxygenated taxoids were characterized from the analysis of the calli of the Taxus wallichiana almost instantaneously. The confirmation of the structures of the identified taxoids was made through their accurate molecular formula determinations.     

Corona discharge secondary ionization of laser desorbed neutral molecules from a liquid matrix at atmospheric pressure

Matrix assisted laser desorption/ionization (MALDI) is studied at atmospheric pressure using liquid sampling methods. A time-of-flight mass spectrometer couples to an open sample stage accessed by a UV laser for desorption and ionization. Also coupled to the sampling state is a corona discharge for auxiliary ionization of desorbed neutral molecules. The interaction of the laser desorption and corona ionization is studied for a range of desorption conditions, showing enhanced analyte ionization, but the effect is analytically advantageous only at low desorption rates. The effect of corona discharge voltage was also explored. The decoupling of neutral molecule formation and subsequent ionization provides an opportunity to study each process separately.     

Coupling of planar chromatography with Direct Analysis in Real Time mass spectrometry

Direct Analysis in Real Time mass spectrometry (DART-MS) is an emerging and rapidly developing area of ambient desorption ionization mass spectrometric techniques. Its coupling with planar chromatography is especially promising, as compared to other ambient desorption ionization techniques, because it does not require the use of liquids that may distort the shape of a spot by diffusion effects. In the first publications on TLC/HPTLC-DART-MS, due to the fixed, horizontally aligned supply of the gas flow from the DART ionization source to the MS inlet, the introduction of HPTLC/TLC plates as cut strips was inconvenient for quantitation, and the repeatability was very low due to the manual positioning. Recently a new version of the DART ion source was suggested, which allows adjusting the angle of the DART gas stream and the use of a motorized rail, thereby, improving highly the capabilities of TLC/HPTLC-DART-MS. This comprehensive review describes the development and analytical capabilities of TLC/HPTLC-DART-MS, and the general DART-MS perspectives for surface analysis or imaging MS.      

Increasing the rate of sample vaporization in an open air desorption ionization source by using a heated metal screen as a sample holder

Rapid vaporization of sample into the ionizing gas exiting a direct analysis in real time (DART®) source has been enabled by directing a high electrical current through a metal wire screen to which sample has been applied. This direct heating of the screen enables rapid vaporization of sample as the wire temperature rises from room temperature to greater than 400°C in less than 20 s. Positioning the screen between the DART source and atmospheric pressure inlet of the mass spectrometer ensures that the ionizing gas is in close proximity to the sample molecules, resulting in efficient ionization while significantly reducing the time required for mass spectrometric analysis. The capability to modulate the electrical current flow through the wires facilitates either rapid desorption for the determination of single component samples or slower desorption where analysis of mixtures might be desired. The technology also enables deployment of strategies for the determination of chemicals present as powders that might otherwise require dissolution prior to analysis. Results from the use of this thermally assisted DART (‘TA‐DART’) system for the analysis of pure compounds, simple mixtures, solids and low vapor pressure samples are presented. Copyright © 2011 John Wiley & Sons, Ltd.     

Comparison of the internal energy deposition of direct analysis in real time and electrospray ionization time-of-flight mass spectrometry

The internal energy (E_int) distributions of a series of p-substituted benzylpyridinium ions generated by both direct analysis in real time (DART) and electrospray ionization (ESI) were compared using the “survival yield” method. DART mean E_int values at gas flow rates of 2, 4, and 6 L min⁻¹, and at set temperatures of 175, 250, and 325 °C were in the 1.92–2.21 eV range. ESI mean E_int at identical temperatures in aqueous and 50% methanol solutions ranged between 1.71 and 1.96 eV, and 1.53 and 1.63 eV, respectively. Although the results indicated that ESI is a “softer” ionization technique than DART, there was overlap between the two techniques for the particular time-of-flight mass spectrometer used. As a whole, there was an increase in E_int with increasing reactive and drying gas temperatures for DART and ESI, respectively, indicating thermal ion activation. Three dimensional computational fluid dynamic simulations in combination with direct temperature measurements within the DART ionization region revealed complex inversely coupled fluid-thermal phenomena affecting ion E_int values during atmospheric transport. Primarily, that DART gas temperature in the ionization region was appreciably less than the set gas temperature of DART due to the set gas flow rates. There was no evidence of E_int deposition pathways from metastable-stimulated desorption, but fragmentation induced by high-energy helium metastables was observed at the highest gas flow rates and temperatures.     

Novel approaches to analysis of 3-chloropropane-1,2-diol esters in vegetable oils

A sensitive and accurate method utilizing ultra-high performance liquid chromatography (U-HPLC) coupled to high resolution mass spectrometry based on orbitrap technology (orbitrapMS) for the analysis of nine 3-chloropropane-1,2-diol (3-MCPD) diesters in vegetable oils was developed. To remove the interfering triacylglycerols that induce strong matrix effects, a clean-up step on silica gel column was used. The quantitative analysis was performed with the use of deuterium-labeled internal standards. The lowest calibration levels estimated for the respective analytes ranged from 2 to 5 μg kg⁻¹. Good recovery values (89–120%) and repeatability (RSD 5–9%) was obtained at spiking levels of 2 and 10 mg kg⁻¹. As an alternative, a novel ambient desorption ionization technique, direct analysis in real time (DART), hyphenated with orbitrapMS, was employed for no separation, high-throughput, semi-quantitative screening of 3-MCPD diesters in samples obtained by chromatographic fractionation. Additionally, the levels of 3-MCPD diesters measured in real-life vegetable oil samples (palm oil, sunflower oil, rapeseed oil) using both methods are reported. Relatively good agreement of the data generated by U-HPLC-orbitrapMS and DART-orbitrapMS were observed. With regard to a low ionization yield achieved for 3-MCPD monoesters, the methods presented in this paper were not yet applicable for the analysis of these contaminants at the naturally occurring levels.     

On-line detection of human skin vapors

Vapors released by the skin in the hand of one human subject are detected in real time by sampling them directly from the ambient gas surrounding the hand, ionizing them by secondary electrospray ionization (SESI, via contact with the charged cloud from an electrospray source), and analyzing them in a mass spectrometer with an atmospheric pressure source (API-MS). This gas-phase approach is complementary to alternative on-line surface ionization methods such as DESI and DART. A dominating peak of lactic acid and a complete series of saturated and singly unsaturated fatty acids (C₁₂ to C₁₈) are observed, in accordance with previous off-line studies by gas chromatography-mass spectrometry. Several other metabolites have been identified, including ketomonocarboxylic and hydroxymonocarboxylic acids.     

Analysis of hairy root culture of Rauvolfia serpentina using direct analysis in real time mass spectrometric technique

The applicability of a new mass spectrometric technique, DART (direct analysis in real time) has been studied in the analysis of the hairy root culture of Rauvolfia serpentina. The intact hairy roots were analyzed by holding them in the gap between the DART source and the mass spectrometer for measurements. Two nitrogen-containing compounds, vomilenine and reserpine, were characterized from the analysis of the hairy roots almost instantaneously. The confirmation of the structures of the identified compounds was made through their accurate molecular formula determinations. This is the first report of the application of DART technique for the characterization of compounds that are expressed in the hairy root cultures of Rauvolfia serpentina. Moreover, this also constitutes the first report of expression of reserpine in the hairy root culture of Rauvolfia serpentina.     

Comparison of dielectric barrier discharge, atmospheric pressure radiofrequency-driven glow discharge and direct analysis in real time sources for ambient mass spectrometry of acetaminophen

Three plasma-based ambient pressure ion sources were investigated; laboratory constructed dielectric barrier and rf glow discharges, as well as a commercial corona discharge (DART source). All were used to desorb and ionize a model analyte, providing sampling techniques for ambient mass spectrometry (MS). Experimental parameters were optimized to achive highest signal for acetaminophen as the analyte. Insight into the mechanisms of analyte desorption and ionization was obtained by means of emission spectrometry and ion current measurements. Desorption and ionization mechanisms for this analyte appear to be identical for all three plasma sources. Emission spectra differ only in the intensities of various lines and bands. Desorption of solid analyte requires transfer of thermal energy from the plasma source to sample surface, in the absence of which complete loss of MS response occurs. For acetaminophen, helium was the best plasma gas, providing 100- to 1000-fold higher analyte response than with argon or nitrogen. The same trend was also evident with background ions (protonated water clusters). MS analyte signal intensity correlates with the ion density (expressed as ion current) in the plasma plume and with emission intensity from excited state species in the plasma. These observations support an ionization process which occurs via proton transfer from protonated water clusters to analyte molecules.