Identification of inorganic improvised explosive devices by analysis of postblast residues using portable capillary electrophoresis instrumentation and indirect photometric detection with a light-emitting diode

A commercial portable capillary electrophoresis (CE) instrument has been used to separate inorganic anions and cations found in postblast residues from improvised explosive devices (IEDs) of the type used frequently in terrorism attacks. The purpose of this analysis was to identify the type of explosive used. The CE instrument was modified for use with an in-house miniaturized light-emitting diode (LED) detector to enable sensitive indirect photometric detection to be employed for the detection of 15 anions (acetate, benzoate, carbonate, chlorate, chloride, chlorite, cyanate, fluoride, nitrate, nitrite, perchlorate, phosphate, sulfate, thiocyanate, thiosulfate) and 12 cations (ammonium, monomethylammonium, ethylammonium, potassium, sodium, barium, strontium, magnesium, manganese, calcium, zinc, lead) as the target analytes. These ions are known to be present in postblast residues from inorganic IEDs constructed from ammonium nitrate/fuel oil mixtures, black powder, and chlorate/perchlorate/sugar mixtures. For the analysis of cations, a blue LED (470 nm) was used in conjunction with the highly absorbing cationic dye, chrysoidine (absorption maximum at 453 nm). A nonaqueous background electrolyte comprising 10 mM chrysoidine in methanol was found to give greatly improved baseline stability in comparison to aqueous electrolytes due to the increased solubility of chrysoidine and its decreased adsorption onto the capillary wall. Glacial acetic acid (0.7% v/v) was added to ensure chrysoidine was protonated and to enhance separation selectivity by means of complexation with transition metal ions. The 12 target cations were separated in less than 9.5 min with detection limits of 0.11-2.30 mg/L (calculated at a signal-to-noise ratio of 3). The anions separation system utilized a UV LED (370 nm) in conjunction with an aqueous chromate electrolyte (absorption maximum at 371 nm) consisting of 10 mM chromium(VI) oxide and 10 mM sodium chromate, buffered with 40 mM tris(hydroxymethyl)aminomethane at pH 8.05. All 15 target anions were baseline separated in less than 9 min with limits of detection ranging from 0.24 to 1.15 mg/L (calculated at a signal-to-noise ratio of 3). Use of the portable instrumentation in the field was demonstrated by analyzing postblast residues in a mobile laboratory immediately after detonation of the explosive devices. Profiling the ionic composition of the inorganic IEDs allowed identification of the chemicals used in their construction.     

100,000-fold concentration of anions in capillary zone electrophoresis using electroosmotic flow controlled counterflow isotachophoretic stacking under field amplified conditions

An electroosmotic flow (EOF) controlled counterflow isotachophoretic stacking boundary (cf-ITPSB) system under field amplified conditions has been examined as a way to improve the sensitivity of anions separated by capillary zone electrophoresis. The system comprised a high concentration of a high-mobility leading ion (100 mM chloride) and a low concentration of low-mobility terminating ion (1-3 mM MES or CHES) added to the sample in an unmodified fused-silica capillary at pH 8.05, buffered with Tris. Computer simulation studies using the software GENTRANS showed an increase in sensitivity of at least 10-fold over the previous cf-ITPSB system for simple inorganic ions, nitrite and nitrate. The simulations also suggested that the cf-ITPSB became stationary within the capillary and that its stationary position was not adversely affected by the concentration of MES. This was in contrast to experimental results that showed a slow and continual movement of the cf-ITPSB. This was more pronounced at lower concentrations of terminator (i.e., <3 mM) and resulted in a loss of resolution due to the cf-ITPSB being closer to the detector upon separation. This discrepancy was attributed to the change in pH across the capillary due to electrolysis and low buffering capacity in the sample, a phenomenon that cannot be simulated by the GENTRANS software. Replacement of MES with CHES as a lower mobility ion with increased buffer capacity failed to reduce the movement of the cf-ITPSB but did provide a further 3-fold improvement in sensitivity. The potential of this approach for sensitivity enhancement was demonstrated for the co-EOF separation of a mixture of six inorganic and small organic ions, with detection limits at the single-figure nanogram per liter level. These detection limits are 100,000 times better than can be achieved by normal hydrodynamic injection (ions prepared in water) and 250 times better than has been achieved by other online preconcentration approaches. The application of the EOF-controlled cf-ITPSB with counter-EOF separation of two pharmaceutical pollutants, naproxen and diflunisal, was also demonstrated with an improvement in sensitivity of 1000 giving detection limits of 350 ng/L in sewage treatment wastewater without any offline pretreatment.     

Latex-coated polymeric monolithic ion-exchange stationary phases. 1. Anion-exchange capillary electrochromatography and in-line sample preconcentration in capillary electrophoresis

A sulfonated methacrylate monolithic polymer has been synthesized inside fused-silica capillaries of diameters 50-533-microm i.d. and coated with 65-nm-diameter fully functionalized quaternary ammonium latex particles (AS18, Dionex Corp.) to form an anion-exchange stationary phase. This stationary phase was used for ion-exchange capillary electrochromatography of inorganic anions in a 75-microm-i.d. capillary with Tris/perchlorate electrolyte and direct UV detection at 195 nm. Seven inorganic anions (bromide, nitrate, iodide, iodate, bromate, thiocyanate, chromate) could be separated over a period of 90 s, and the elution order indicated that both ion exchange and electrophoresis contributed to the separation mechanism. Separation efficiencies of up to 1.66 x 10(5) plates m(-1) were achieved, and the monoliths were stable under pressures of up to 62 MPa. Another latex-coated monolith in a 250-microm-i.d. capillary was used for in-line preconcentration by coupling it to a separation capillary in which the EOF had been reversed using a coating of either a cationic polymer or cationic latex particles. Several capillary volumes of sample were loaded onto the preconcentration monolith, and the analytes (inorganic anions) were then eluted from the monolith with a transient isotachophoretic gradient before being separated by electrophoresis in the separation capillary. Linear calibration curves were obtained for aqueous mixtures of bromide, nitrite, nitrate, and iodide. Recoveries of all analytes except iodide were reduced significantly when the sample matrix contained high levels of chloride. The preconcentration method was applied to the determination of iodide in open ocean water and provided a limit of detection of 75 pM (9.5 ng/L) calculated at a signal-to-noise ratio of 3. The relative standard deviation for migration time and peak area for iodide were 1.1 and 2.7%, respectively (n = 6). Iodide was eluted as an efficient peak, yielding a separation efficiency of 5.13 x 10(7) plates m(-1). This focusing was reproducible for repeated analyses of seawater.     

Separation and determination of polycyclic aromatic hydrocarbons by solid phase microextraction/cyclodextrin-modified capillary electrophoresis

A sensitive method for the determination of polycyclic aromatic hydrocarbons (PAHs) by solid phase microextraction coupled with cyclodextrin (CD)-modified capillary electrophoresis (CE) using UV detection has been developed. A glass fiber was prepared and used for absorbing 16 EPA priority PAHs from diluted samples until equilibrium was reached. After the glass fiber was connected to a separation capillary via an adapter, the absorbed analytes were directly released into the CE buffer stream, and electrophoretic separation was effected using a 50 mM borate, pH 9.2, buffer containing 35 mM sulfobutyloxy-β-CD, 10 mM methyl-β-CD, and 4 mM α-CD. Separation was effected since neutral PAHs differentially partitioned between the neutral and charged CD phases. Under 30 kV applied potential, separation was achieved in less than 15 min with high resolution and number of theoretical plates. Pyrene as low as 8 ppb was detected, while the highest limit of detection was 75 ppb for acenaphthene. Very satisfactory reproducibility with respect to migration time and peak area was obtained for repetitions using the same separation capillary and adapter, where only the extraction fiber was discarded after each analysis.     

Analysis of inorganic species by capillary electrophoresis-mass spectrometry and ion exchange chromatography-mass spectrometry using an ion spray source

Mixtures of inorganic ions separated by capillary electrophoresis (CE) and ion exchange chromatography (IC) are detected by mass spectrometry (MS) using an ion spray atmospheric pressure ionization source. The selectable degree of ion-adduct declustering and molecular fragmentation in the MS interface region allows the system to be operated as an elemental analyzer or as a molecular detector suitable for oxidation state determinations. Both inorganic anions and cations (including alkalis, alkaline earths, transition metals, and lanthanides) are analyzed by CE-MS. A variety of CE separation buffers are evaluated for the cation analyses (e.g., creatinine, ammonium acetate, and tris[hydroxymethyl]aminomethane). Only one of the buffers (i.e., creatinine) can be used for CE-indirect UV detection. A CE capillary permanently coated with strong anion exchange sites and a pyromellitic acid buffer (suitable for indirect UV detection) is used for the inorganic anion separations. The coated column eliminates the need for buffer modifiers to reverse the flow in the capillary, which then reduces background noise and mass spectral complexity. The separation and detection of 13 inorganic anions are also accomplished by IC using an anion exchange column with a carbonate-bicarbonate mobile phase, on-line suppressed conductivity detection, and mass spectrometric detection.     

High throughput profiling of charge heterogeneity in antibodies by microchip electrophoresis

A high throughput microchip capillary zone electrophoresis (CZE) method was developed for the analysis of charge heterogeneity in antibodies. The method utilizes high speed microchip electrophoresis separation and is well-suited for high throughput charge profiling of antibodies during process and formulation development. The method involves derivatization of protein molecules with Cy5 N-hydroxysuccinimide ester (NHS-ester), which does not change the protein charge profile and enables fluorescence detection on a commercial microchip instrument. The sample preparation can be performed in 96-well microtiter plates within 1 h, and each sample analysis takes only 80 s. Protein charge variants with a pI difference of 0.1 can be readily resolved in the 12.5 mm microfluidic channel. Charge profiles similar to those obtained using conventional CZE technology were found for all antibodies tested (pIs in the range of 7.5-9.2). The separation efficiency corresponds to 1.2 × 10(4) theoretical plates (1.0 μm plate height). Assay performance is assessed by demonstrating specificity, carryover, linearity, limit of detection, and precision.     

Focusing and Mobilization of Bacteria in Capillary Electrophoresis

An isotachophoretic method has been developed for mobilizing and focusing bacteria. This allows quantification and detection of bacteria in a narrow zone. Very good linearity was obtained for Micrococcus lysodeikticus (also called Micrococcus luteus, studied as a model of Gram+ bacteria) in the range of 0.4 × 10(8) cells/mL to 2.9 × 10(8) cells/mL, with correlation coefficients for peak height and peak area as a function of cell concentration of 0.999 and 0.998, respectively. This method is usable on both bare and hydroxypropyl cellulose-coated fused silica capillaries. The best results were obtained using 13.6 mM Tris, 150 mM boric acid as terminating electrolyte, and 4.5 mM Tris, 50 mM boric acid, and 3.31 mM HCl as leading electrolyte. With a 33.5 cm ×100 μm i.d. capillary, short migration times were obtained while maintaining very low electrical current in order to minimize any Joule heating and lysis of the bacteria. A UV area imaging detector (ActiPix D100, Paraytec) was used with a 109 cm × 100 μm i.d. capillary having three loops and four detection windows to monitor the migration behavior of M. luteus and to show the stability of the zone of the focused bacteria along the capillary. Similar results were obtained for Erwinia carotovora (a model of Gram- bacteria), and for Enterobacter cloacae and Vibrio splendidus.     

Electrochemical detectors prepared by electroless deposition for microfabricated electrophoresis chips

Microfabricated capillary electrophoresis (CE) chips with integrated electrochemical detection have been developed on glass substrates. An electroless deposition procedure was used to deposit a gold film directly onto the capillary outlet to provide high-sensitivity electrochemical detection for catechol and several nitroaromatic explosives. Scanning electron microscopy revealed that the electroless gold film contains nanoscopic gold aggregates (100-150 nm) with an average thickness of 79 nm. The electroless deposition procedure can be easily and routinely performed in any wet-chemistry laboratory, and electroless gold can be deposited onto complex and internal surfaces. Intimate coupling of electrochemical detection and CE chips obviates the need for a coupling mechanism or tedious alignment procedures. With nitroaromatic compounds as a working model, microchip capillary electrophoresis equipped with electroless gold has proven to provide high sensitivity and fast response times for sensor applications. The CE microchip system was capable of separation and determination of explosive compounds including TNT in less than 130 s with detection limits ranging from 24 to 36 microg/L, i.e., 4-fold enhancements in detection efficiency in comparison to thick-film technology.     

Multiresidue method for the determination of quinolone antibiotics in bovine raw milk by capillary electrophoresis-tandem mass spectrometry

A new analytical method based on capillary zone electrophoresis-tandem mass spectrometry (CZE-MS/MS) is proposed and validated for the identification and simultaneous quantification of eight quinolones for veterinary use in bovine raw milk. The studied quinolones include danofloxacin, sarafloxacin, ciprofloxacin, marbofloxacin, enrofloxacin, difloxacin, oxolinic acid, and flumequine, whose contents are regulated by the EU Council Regulation no. 2377/90 in animal edible tissues. Different parameters (i.e., separation buffer composition and electrospray conditions) were optimized in order to obtain both an adequate CE separation and a high sensitivity, using experimental design methodology to consider the interactions among the studied variables. MS/MS experiments using an ion trap as analyzer operating in the multiple reaction monitoring mode were carried out to achieve the minimum number of identification points according to the 2002/657/EC European Decision. For the quantification in bovine raw milk samples, a two-step solid-phase extraction procedure was developed using Oasis MAX and HLB cartridges without protein precipitation. Satisfactory results were obtained in terms of linearity (r2 between 0.989 and 0.992) and precision (RSD below 18%). The limits of detection and quantification (below 6 and 24 ppb, respectively) were in all cases lower than the maximum residues limits tolerated for these compounds in milk, the recoveries ranging from 81 to 110%, indicating the potential of the CZE-MS/MS for the analysis of regulated quinolone antibiotics in the food quality and safety control areas.     

Capillary electrophoresis with an integrated on-capillary tubular detector based on a carbon sol-gel-derived platform

An integrated on-capillary tubular electrochemical detector for capillary electrophoresis systems has been fabricated based on sol-gel technique. It consists of a sol-gel carbon composite tubular electrode attached permanently onto the outlet of the separation capillary. The device greatly eases the setting up of capillary electrophoresis with electrochemical detection (CEEC) as it makes possible electrode/capillary alignment without the aid of a micromanipulator since this integrated unit can be simply immersed in the CE separation buffer in an ordinary three-electrode stationary cell. To improve analytical performance of the integrated unit, the external wall of the exit capillary was etched with HF after the polyimide coating of the capillary had been removed. Influences of the working electrode length and the wall thickness at the outlet of capillary on the separation efficiency and amperometric sensitivity were assessed and optimized. The practical applicability of this configuration is demonstrated with the detection of both catecholamines and carbohydrates. The advantages, namely, versatility, convenience, ease of operation, and low-cost, of the new design combined with an excellent performance lead to high stability and low detection limits.     

Analysis of bile acids and their conjugates by capillary electrochromatography/electrospray ion trap mass spectrometry

Different macroporous, monolithic capillary columns were prepared to separate various bile acid mixtures through capillary electrochromatography (CEC) at high efficiency. These columns are shown to be ideally suitable for coupling to an electrospray ionization/ion trap mass spectrometer. Detection and structural identification of different bile acid derivatives in either the positive- or negative-ion mode necessitated column technologies with different polarities and the capabilities of a reversed electroosmotic flow. High column efficiencies (610,000 theoretical plates/meter for glycocholic acid in normal-phase separation) were preserved in the coupling to mass spectrometry (MS), with the detection limits of approximately 40 femtomole (for cholic acid) and identification through CEC/MS/MS.     

Application of diamond microelectrodes for end-column electrochemical detection in capillary electrophoresis

Highly boron-doped diamond microelectrodes were employed in an end-column electrochemical detector for capillary electrophoresis (CE). The diamond microline electrodes were fabricated from conducting diamond thin films (exposed surface area, 300 x 50 microm), and their analytical performance as CE detectors was evaluated in a laboratory-made CE installation. The CE-ED system exhibited high separation efficiency for the detection of several catecholamines, including dopamine (DA), norepinephrine (NE), and epinephrine (E), with excellent analytical performance, for example, 155,000 theoretical plates for DA. The diamond-based electrochemical detection system also displayed low detection limits (approximately 20 nM for E at S/N = 3) and a highly reproducible current response with 10 repetitive injections of mixed analytes containing DA, NE, and E (each 50 microM), with relative standard deviations (RSD) of approximately 5%. The performance of the diamond detector in CE was also evaluated in the detection of chlorinated phenols (CP). When compared to the carbon fiber microelectrode, the diamond electrode exhibited lower detection limits in an end-column CE detection resulting from very low noise levels and highly reproducible analyses without electrode polishing due to analyte fouling, which makes it possible to perform easier and more stable CE analysis.     

Isoelectric buffers for capillary electrophoresis. 2. Bismorpholine derivative of a carboxylic acid as a low molecular weight isoelectric buffer

A new compound class of synthetic isoelectric buffers is introduced, designed as a small molecule with one fully or prevailingly dissociated acidic group (such as sulfonic or carboxylic) and two partly pronated (buffering) basic amino groups attached onto a hydrophilic UV-transparent backbone. As an example, a new isoelectric compound 2,2-bis(4-morpholinylmethyl)propanoic acid (BMMPA) was synthesized by attaching two morpholine groups onto a molecule of pivalic acid. It was characterized as having an isoelectric point pI = 6.5 and exhibiting satisfactory buffering capacity at the pI. Solutions of BMMPA are transparent down to the low-UV spectral region, thus making it a potentially suitable buffer for a number of separation methods. Its use in capillary electrophoresis was demonstrated in a separation system for indirect photometric detection of anions based on an electrolyte with the anionic dye Orange G as the indirect detection probe and using BMMPA as a buffer. The use of an isoelectric buffering compound brings the advantages of a buffered electrolyte without the concomitant introduction of co-ions that would be detrimental to the indirect detection process. Submicromole per liter limits of detection for a number of inorganic and small organic ions were achieved. Optimal structural properties of the isoelectric buffer with respect to its buffering properties are discussed.     

Enhanced mixture analysis of poly(ethylene glycol) using high-field asymmetric waveform ion mobility spectrometry combined with fourier transform ion cyclotron resonance mass spectrometry

The combination of high-field asymmetric waveform ion mobility spectrometry (FAIMS) with Fourier transform ion cyclotron resonance mass spectrometry (FTICR-MS) makes possible lower detection limits, increased sensitivity, and increased dynamic range in the analysis of poly(ethylene glycol) (PEG) samples of low molecular weight. The signal gain obtained using FAIMS depends on ion identity, with a range between 1.8x and 14x obtained for various molecular ions of PEG 600. A 1.7-fold reduction in noise is obtained using FAIMS due to the elimination of chemical noise. The improved detection performance is predominantly due to a reduction in adverse Coulomb effects as a result of ions being selectively introduced into the mass spectrometer. The high ion transmission obtained using FAIMS combined with the high sensitivity of FTICR-MS detection make possible separation of multiple gas-phase conformers of PEG molecular cations that have low abundance (less than 0.2% relative abundance) and that have not been detected previously. Mixed dications of PEG that have the same nominal mass but differ by the number polymer subunits (m/Delta m up to 25,000) can be separately introduced into the mass spectrometer using FAIMS. Interactions of the carrier gas with the metal ions that are attached to the PEG molecules appear to be the most significant factor in these FAIMS separations.     

Sensitized enantioselective laser-induced phosphorescence detection in chiral capillary electrophoresis

The sensitivity of enantioselective cyclodextrin-induced room-temperature phosphorescence detection of camphorquinone (CQ) is enhanced using sensitization via a donor with a high extinction coefficient. The enantiomeric distinction is based on the different phosphorescence lifetimes of (+)-CQ and (-)-CQ after their complexation with α-cyclodextrin (α-CD). The collisional Dexter energy transfer from the selected donor 2,6-naphthalenedisulfonic acid (2,6-NS) to the acceptor CQ is still very efficient despite the inclusion of the acceptor into CD. For coupling to the chiral separation of (±)-CQ in cyclodextrin-based electrokinetic chromatography, the donor was added to the deoxygenated background electrolyte that consisted of 20 mM α-CD, 10 mM carboxymethyl-β-CD, and 25 mM borate buffer at pH 9.0. Time-resolved batch studies on sensitized phosphorescence show a significant enantioselectivity for (+)- and (-)-CQ in the presence of both α-CD and CM-β-CD although the lifetime difference is somewhat reduced with respect to direct excitation. The enantiomers were distinguished after their separation using an online time-resolved detection system. Excitation was performed at 266 nm with a pulsed, small-sized, quadrupled Nd:YAG laser. With 1 × 10(-5) M 2,6-NS, limits of detection of 4.1 × 10(-8) M and 5.2 × 10(-8) M were found for (+)-CQ and (-)-CQ, respectively. The online measured lifetimes were 238 ± 8 μs for (+)-CQ and 126 ± 10 μs for (-)-CQ. The method was used to determine the concentration of (±)-CQ leaching from a cured dental resin into water. The extracts contained 4.7 ± 0.1 × 10(-7) M of both (+)-CQ and (-)-CQ.     

Simultaneous enantioseparation and tandem UV-MS detection of eight beta-blockers in micellar electrokinetic chromatography using a chiral molecular micelle

The feasibility of using a new and more versatile polymeric chiral surfactant, i.e., poly(sodium N-undecenoxy carbonyl-L-leucinate (poly-L-SUCL) is investigated for simultaneous enantioseparation and detection of eight structurally similar beta-blockers with tandem UV and MS detection. Three optimization approaches, i.e., direct infusion-MS, capillary zone electrophoresis-MS, and chiral micellar electrokinetic chromatography-mass spectrometry (CMEKC-MS), were investigated to optimize sheath liquid parameters, spray chamber parameters, and CMEKC separation parameters for maximum sensitivity and chiral resolution. Compared to unpolymerized micelle of L-SUCL, the use of micelle polymer (i.e., poly-L-SUCL) provided significantly higher separation efficiency, lower separation current, and higher detection sensitivity for CMEKC-ESI-MS of beta-blockers. It was also observed that, unlike monomeric L-SUCL, polymeric L-SUCL provided enantioseparation of all beta-blockers even at the lowest surfactant concentration (i.e., 5 mM poly-L-SUCL). Under optimum CMEKC and ESI-MS conditions (15 mM poly-L-SUCL, 25 mM each of NH4OAc and TEA (pH 8.0); 80% (v/v) methanol sheath liquid containing 40 mM NH4OAc (pH 8.0); sheath liquid flow rate, 5.0 microL/min; drying gas flow rate, 5 L/min; drying gas temperature, 200 degrees C; nebulizing pressure, 6 psi (0.414 bar); capillary voltage, +2.5 kV; fragmentor voltage, 85 V), baseline enantioseparation of eight beta-blockers was achieved by tandem UV (in approximately 30 min) and MS (in approximately 60 min) detection. Calibration curves for all beta-blockers were linear in the range of 0.01-0.6 mM for both CMEKC-UV and CMEKC-MS methods, but the later method provided better concentration limit of detection with similar RSD for migration time and peak areas. The CMEKC-ESI-MS method appears suitable for use as a routine procedure for high-throughput separation of beta-blockers with high sensitivity.     

Fabrication of micro-nano structure nanofibers by solvent etching

Micro-Nano structure nanofibrous affinity membranes of poly(ether sulfones) (PES) blended with a functional polymer poly(ethyleneimine) (PEI) were fabricated by electrospinning technique followed by solvent etching in crosslinking solution. The surface SEM image of the water washed PES/PEI nanofibrous membrane confirmed that PEI was concentrated on the fiber surface. The nanofibrous PES/PEI membranes were crosslinked in a mixture of acetone and water with glutaraldehyde (crosslinking agent, GA), and the micro-nano structural surface of the nanofibrous membranes was created by solvent etching due to the solvation between PEI and the solvent water in the crosslinking solution during the crosslinking process. The influence of the component of the crosslinking bath on the mophology of the resulting PES/PEI nanofibers was investigated. It was found that the relatively uniform micro-nano spherules grew on the surface of the nanofibers when the content of water in crosslinking solution was more than 20 wt%, and the diameters of the spherules were in the range of 50-250 nm. The advantage of the micro-nano structrue for the heavy metal ions removal in wastwater has been demonstrated by taking a series of static adsorption experiments. It was found that the micro-nano structrue of PES/PEI nanofibrous membranes could bring high performance of adsorption capacity for heavy metal ions, indicating that the unique morphology could bring much more large surface area per unit mass and high effectivity for heavy metal ions removal from aqueous solutions.     

CE/electrospray ionization-MS analysis of underivatized d/l-amino acids and several small neurotransmitters at attomole levels through the use of 18-crown-6-tetracarboxylic acid as a complexation reagent/background electrolyte

A new capillary electrophoresis/mass spectrometry technique is introduced for attomole detection of primary amines (including several neurotransmitters), amino acids, and their d/l enantiomers in one run through the use of a complexation reagent while using only approximately 1 nL of sample. The technique uses underivatized amino acids in conjunction with an underivatized capillary, which significantly reduces cost and analysis time. It was found that when (+)-(18-crown-6)-2,3,11,12-tetracarboxylic acid (18-C-6-TCA, MW 440) was used as the background electrolyte/complexation reagent during the capillary electrophoresis/electrospray ionization-mass spectrometry (CE/ESI-MS) analysis of underivatized amino acids, stable complexes were formed between the amino acids and the 18-C-6-TCA molecules. These complexes, which exhibited high ionization efficiencies, were detectable at attomole levels for most amino acids. The detection limits of the AA/18-C-6-TCA complexes were on the average more than 2 orders of magnitude lower than that of the free amino acids in solution. In addition to lower detection limits under CE/ESI-MS, a solution of 18-C-6-TCA in the concentration range of 5-30 mM provided high separation efficiency for mixtures of l-amino acids as well as mixtures of d/l-amino acids. By using a solution of 18-C-6-TCA as the background electrolyte in conjunction with an underivatized, 130-cm-long, 20-microm-i.d., 150-microm-o.d. fused-silica capillary and by monitoring the m/z range of the amino acid/18-C-6-TCA complexes (m/z 515-700), most of the standard amino acids and many of their enantiomers were separated and detected with high separation efficiency and high sensitivity (nanomolar concentration detection limits) in one run. The solutions of 18-C-6-TCA also worked well as the CE/ESI-MS BGE for low-level detection of several neurotransmitters and some of their d/l enantiomers as well as for the analysis of amino acids at endogenous levels in lysed red blood cells.     

Single-channel microchip for fast screening and detailed identification of nitroaromatic explosives or organophosphate nerve agents

A single-channel chip-based analytical microsystem that allows rapid flow injection measurements of the total content of organic explosive or nerve agent compounds, as well as detailed micellar chromatographic identification of the individual ones, is described. The protocol involves repetitive rapid flow injection (screening) assays--to provide a timely warning and alarm--and switching to the separation (fingerprint identification) mode only when harmful compounds are detected. While micellar electrokinetic chromatography, in the presence of sodium dodecyl sulfate (SDS), is used for separating the neutral nitroaromatic explosive and nerve agent compounds, an operation without SDS leads to high-speed measurements of the "total" explosives or nerve agent content. Switching between the "flow injection" and "separation" modes is accomplished by rapidly exchanging the SDS-free and SDS-containing buffers in the separation channel. Amperometric detection was used for monitoring the separation. Key factors influencing the sample throughput, resolution, and sensitivity have been assessed and optimized. Assays rates of about 360 and 30/h can thus be realized for the "total" screening and "individual" measurements, respectively. Ultimately, such development will lead to the creation of a field-deployable microanalyzer and will enable transporting the forensic laboratory to the sample source.     

Indirect laser-induced fluorescence detection for capillary electrophoresis using a violet diode laser

The violet (415 nm) diode laser is used for indirect laser-induced fluorescence detection in capillary electrophoretic separations of inorganic anions and chemical warfare agent degradation products. Inorganic anions were detected using 8-hydroxypyrene-1,3,6-trisulfonic acid as the indirect probe and achieved submicromolar (40-80 ppb) detection limits in a 2-min separation. The chemical warfare agent degradation products methylphosphonic acid, ethyl methylphosphonate, isopropyl methylphosphonate, and pinacolyl methylphosphonate were detected using the porphyrin tetrakis(4-sulfophenyl)porphine as the indirect probe and achieved detection limits of 0.1 microM (9 ppb), which are 1 order of magnitude better than that achieved using indirect UV detection. Baseline stability achieved with the violet diode laser was excellent, with dynamic reserve (DR) values of > 1000, which are 15 times better than that achieved using an unstabilized HeCd laser.