Sensitive and selective gas chromatography-tandem mass spectrometry method for the detection of nitrobenzene in tobacco smoke.

Nitrobenzene, a potentially harmful compound found in tobacco smoke, has been largely excluded from prior analysis due to difficulties with quantification. Quantifying harmful compounds in cigarette smoke is useful to compare products, to examine the impact of design parameters on delivery, and to help estimate exposures. A sensitive high-throughput method has been developed for quantifying nitrobenzene in machine-generated mainstream cigarette smoke using isotope dilution gas chromatography-tandem mass spectrometry (ID-GC-MS/MS). This method has sufficient sensitivity to measure vapor phase nitrobenzene concentrations in the low nanogram range, with a 418 pg/cig method limit of detection. Precision estimates from two quality control cigarette products resulted in percent relative standard deviations of 11.5% and 14.9%; product variability estimates from 13 cigarette products resulted in percent relative standard deviations ranging from 2.8% to 16.9%. Nitrobenzene in the machine-generated, mainstream smoke from 15 cigarette products are reported and range from 18 to 38 ng/cig under the Health Canada Intense smoking regimen.

Rapid analysis of Lewisite metabolites in urine by high-performance liquid chromatography-inductively coupled plasma-mass spectrometry.

A high-throughput method has been developed for determining Lewisite [dichloro(2-chlorovinyl)arsine] exposure by measuring the urine metabolite 2-chlorovinylarsonous acid (CVAA) and the oxidized metabolite 2-chlorovinylarsonic acid (CVAOA). The rapid sample preparation included a simple dilution of 400 microL of urine with 40 microL of water and 1 mL of buffer containing an internal standard and brief centrifugation prior to analysis by high-performance liquid chromatography-inductively coupled plasma-mass spectrometry (ICP-MS). CVAOA and CVAA were eluted isocratically with retention factors of approximately 3.0 and 4.2, respectively, from a reversed-phase polar embedded column with a cycle time of 5 min per sample. The dynamic reaction cell, typically used to remove polyatomic isobaric interferences, was not required for ICP-MS analysis because of the resolution of chloride from arsenical peaks of interest. This method was used to detect CVAA and CVAOA in the urine of a rat administered Lewisite up to 24 h after exposure. The method demonstrated linearity over at least three orders of magnitude and had a method detection limit of 1.3 microg/L as CVAA (1.4 microg/L CVAOA). The relative standard deviations for quality control samples ranged from 3 to 6%. The method was sensitive and selective with no false positives in 100 different urine samples collected from individuals with no known exposure to Lewisite. Ninety-six samples could be analyzed in an 8-h day.

Nylon-6 capillary-channeled polymer (C-CP) fibers as a hydrophobic interaction chromatography stationary phase for the separation of proteins.

Nylon-6 capillary-channeled polymer (C-CP) fibers are used as the stationary phase for the hydrophobic interaction chromatography (HIC) separation of a synthetic protein mixture composed of ribonuclease A, lysozyme, and holotransferrin. Nylon is a useful polymer phase for HIC as it has an alkyl backbone, while the amide functionality is hydrophilic (in fact ionic) in nature. The combination of a nonporous polymer surface of the fiber phases and high column permeability yields very efficient mass transfer characteristics, as exhibited by narrowing of peak widths with increases in mobile phase linear velocity. Retention factors and resolution were evaluated at flow rates ranging from 0.5 to 9 mL/min (linear velocities of ca. 2 to 15 mm/s) and at gradient slopes between 3.3 and 20 %B/min. Optimum resolution was achieved by employing fast flow rates (9 mL/min) and slow gradients (3 %B/min), also resulting in the highest peak capacities.

Nylon-6 capillary-channeled polymer fibers as a stationary phase for the mixed-mode ion exchange/reversed-phase chromatography separation of proteins.

Capillary-channeled polymer (C-CP) fibers extruded from nylon-6 are used as the stationary phase for the ion-exchange/reversed-phase mixed-mode chromatographic separation of a three protein mixture. The nylon-6 C-CP fibers are packed collinearly in a 250 x 1.5-mm i.d. column with an interstitial fraction of approximately 0.6. The effects of four displacing salts at three different pHs are studied with regards to protein retention time, peak width, selectivity, and resolution for a synthetic mixture consisting of myoglobin, ribonuclease A, and lysozyme to determine the optimum mobile phase conditions. The net charge model is found to be inadequate in fully explaining the retention behavior, as the proteins are retained by anion and cation-exchange interactions, as well as hydrophobic interactions with the stationary phase. It is found that pH and displacing salt strength had a significant influence on the retention properties and resolution of the proteins.

Enhancing the response of alkyl methylphosphonic acids in negative electrospray ionization liquid chromatography tandem mass spectrometry by post-column addition of organic solvents.

A method to enhance the signal intensity and signal-to-noise of several alkyl methylphosphonic acids in negative electrospray ionization liquid chromatography tandem mass spectrometry (ESI LC-MS/MS) is presented. This class of compound represents the initial metabolites and environmental degradants of the nerve agents: VX, rVX (Russian VX), GB (Sarin), GF (Cyclosarin), and GD (Soman). Compared with the post-column addition of the mobile phase, the post-column addition of aprotic solvents and longer chain alcohols enhance the signal intensity and signal-to-noise ratio (S/N) of the chromatographic peaks by factors of up to 60 and 19, respectively. The post-column addition of water, methanol, and ethanol resulted in little or no relative signal enhancement. It is proposed that the post-column addition of these solvents do not result in the same enhancements due to stabilization of analyte solvation through hydrogen bonding.

Capillary-channeled polymer (C-CP) fibers as a stationary phase in microbore high-performance liquid chromatography columns.

Microbore columns utilizing polypropylene capillary-channeled polymer (C-CP) fibers as the stationary phase in high-performance liquid chromatography (HPLC) have been investigated. The polypropylene C-CP fiber diameter is approximately 50 microm, with eight channels along the periphery of the fiber ranging in diameter from approximately 12 to 35 microm. The polypropylene C-CP fibers were packed into fluorinated ethylene propylene (FEP) tubing, 1.3 mm inner diameter, with lengths of 500, 750, and 1,000 mm, to examine the effects of increased column length with regards to plate height, resolution and analysis time. The low backpressures characteristic of the C-CP fiber stationary phases allow the length of the column to be increased without significantly decreasing the specific permeability. The high specific permeability (approximately 5x10(-8) cm2) of the C-CP packed microbore columns yields a relatively low backpressure of 2.35 MPa at the highest flow rate of 17 microL/s (54 mm/s) for a 1,000 mm column. Radial compression of the soft-walled FEP tubing is accomplished by pulling the 1.7 mm o.d. column through a 1.4 mm diameter orifice. Reducing the inner diameter of the column from 1.3 to 1.0 mm lowered the interstitial fraction from 47% to 42%, decreased the A-term contributions to band broadening, resulted in a significant decrease in average plate height (approximately 30%), and increased resolution (approximately 36%) at identical linear velocities. Although the lower void volume of the radially compressed column increased the backpressure from 0.57 to 2.11 MPa at a linear velocity of approximately 20 mm/s, the specific permeability only decreased from approximately 7x10(-8) to 4x10(-8) cm2.

Hydrodynamic flow in capillary-channel fiber columns for liquid chromatography.

The flow characteristics of capillary-channel polymer (C-CP) fiber liquid chromatographic (LC) columns have been investigated. The C-CP fibers are manufactured with eight longitudinal grooves (capillary channels) extending the length of the fibers. Three C-CP fiber examples were studied, with fiber dimensions ranging from approximately 35 microm to 65 microm, and capillary-channel dimensions ranging from approximately 6 microm to 35 microm. The influence of fiber packing density and column inner diameter on peak asymmetry, peak width, and run-to-run reproducibility have been studied for stainless steel LC columns packed with polyester (PET) and polypropylene (PP) C-CP fibers. The van Deemter A-term was evaluated as a function of fiber packing density (approximately 0.3 g/cm(3)-0.75 g/cm(3)) for columns of 4.6 mm inner diameter (i.d.) and at constant packing densities for 1.5 mm, 3.2 mm, 4.6 mm, and 7.7 mm i.d. columns. Although column diameter had little influence on the eluting peak widths, peak asymmetry increased with increasing column diameter. The A-terms for the C-CP fiber packed columns are somewhat larger than current commercial, microparticulate-packed columns, and means for improvement are discussed. Applications in the area of protein (macromolecule) separations appear the most promising at this stage of the system development.