W. Clay Davis

An atmospheric pressure glow discharge optical emission source for the direct sampling of liquid media.

A glow discharge optical emission spectroscopy (GD-OES) source that operates at atmospheric pressure is described. This device utilizes an electrolytic solution containing the analyte specimen as one of the discharge electrodes. The passage of electrical current (either electrons or positive ions) across the solution/gas phase interface causes local heating and the volatilization of the analyte species. Collisions in the discharge region immediately above the solution surface result in optical emission that is characteristic of the analyte elements. Operation of this device with the analyte solution acting as either the cathode or anode is demonstrated. Current-voltage (i-V) plots reveal abnormal glow discharge characteristics, with operating parameters being dependent on the electrolyte concentration (i.e., solution conductivity) and the gap between the solution surface and the counterelectrode. Typical conditions include discharge currents of 30-60 mA, and potentials of 500-900 V. Electrolyte solutions having pH, pNa, or pLi values of 0.5-2 and interelectrode gaps of 0.5-3 mm produce stable plasmas in which the analyte solutions flow at rates of up to 3.0 mL/min. Preliminary limits of detection are determined for the elements Na, Fe, and Pb to be in the range of 11-14 ppm (approximately 60 ng) for 5-microL sample volumes.

Capillary-channeled polymer fibers as stationary phases in liquid chromatography separations

A method utilizing capillary-channeled polymer (C-CP) fibers as stationary phases in high-performance liquid chromatographic separations has been investigated. Polymeric fibers of differing backbones (polypropylene and polyester) having nominal diameters of approximately 50 and approximately 35 microm and a channeled structure on their periphery were packed into stainless steel tubing (305 x 4.6 mm I.D.) for use in reversed-phase separations of various mixtures. The fibers have eight channels running continuously along the axis which exhibit very high surface activity. As such, solvent transport is affected through the channels through wicking action. Bundles of 1000-3000 fibers are loaded co-linearly into the tubing, providing flow channels extending the entire length of the columns. As a result, backing pressures are significantly lowered (approximately 50% reduction) in comparison to packed-sphere columns. In addition, the capital costs of the fiber material (< US

Development of a Standard Reference Material for Metabolomics Research

0.25 per column) are very attractive. Flow-rates of up to 5 ml/min can be used to achieve near baseline separation of related compounds in reasonable run times, indicating very fast mobile phase mass transfer (C-terms). The polymer stationary phases demonstrate high selectivity for a wide variety of analytes with gradient elution employed successfully in many instances. Specifically, separations of three polyaromatic hydrocarbons (benzo[a]pyrene, chrysene, pyrene), mixtures of both organic and inorganic lead compounds [chlorotriethyllead, chlorotriphenyllead, lead nitrate, lead(II) phthalocyanine], and a lipid standard of triglycerides were accomplished on the polymeric stationary phases. Other species of biological interest, including groups of aliphatic and aromatic amino acids have also been effectively separated. The reversed-phase nature of the fiber surfaces is supported through atomic force microscopy measurements using hydrophilic and hydrophobic functionalized polystyrene beads as the probe tips. Separations of the various analytes demonstrate the feasibility of utilizing C-CP fibers as stationary phases in reversed-phase LC. It is envisioned that columns of this nature would be particularly useful in prep-scale separations as well as for immobilization matrices for organic constituents in aqueous environments.

Role of powering geometries and sheath gas composition on operation characteristics and the optical emission in the liquid sampling-atmospheric pressure glow discharge

The National Institute of Standards and Technology (NIST), in collaboration with the National Institutes of Health (NIH), has developed a Standard Reference Material (SRM) to support technology development in metabolomics research. SRM 1950 Metabolites in Human Plasma is intended to have metabolite concentrations that are representative of those found in adult human plasma. The plasma used in the preparation of SRM 1950 was collected from both male and female donors, and donor ethnicity targets were selected based upon the ethnic makeup of the U.S. population. Metabolomics research is diverse in terms of both instrumentation and scientific goals. This SRM was designed to apply broadly to the field, not toward specific applications. Therefore, concentrations of approximately 100 analytes, including amino acids, fatty acids, trace elements, vitamins, hormones, selenoproteins, clinical markers, and perfluorinated compounds (PFCs), were determined. Value assignment measurements were performed by NIST and the Centers for Disease Control and Prevention (CDC). SRM 1950 is the first reference material developed specifically for metabolomics research.

Detection, Identification, and Quantification of Selenoproteins in a Candidate Human Plasma Standard Reference Material

Characterization of the liquid sampling-atmospheric pressure glow discharge optical emission spectroscopy (LSAPGD-OES) source is described with regards to applications in low-flow separations such as capillary liquid chromatography and electrophoresis. Four powering modes are investigated, including the effects of the individual modes on current–voltage characteristics, analyte emission response, and temporal broadening of flow injection profiles. A concentric sheath gas is employed to stabilize the solution delivery at low liquid flow rates. Sheath gas composition ( No r He) effects analyte emission responses as well as gas phase rotational and excitation temperatures. 2 The respective powering modes both measures of temperature, with the OH rotationalgas temperatures ranging from ;2100 to 3000 K and the Fe (I) excitation temperatures ranging from ;2400 to 3600 K. Rotationaltemperature values increase slightly when helium is employed as a sheath gas as opposed to nitrogen, with the corresponding excitation temperatures increasing somewhat as well. Analytical response curves for Na and Hg in the various powering modes demonstrate good linearity, with the limits of detection for the analytes found to be on the order of ;4–10 ppm for 5 ml injections; equating to absolute detection limits of between 20 and 45 ng. It is believed that the approach demonstrated here suggests further improvements that will permit applications in a wide variety of aqueous solution analyses where low-flow rates and limited volumes are encountered. 2002 Elsevier Science B.V. All rights reserved.

Development of a new liquid chromatography method for the separation and speciation of organic and inorganic selenium compounds via particle beam-hollow cathode glow discharge-optical emission spectroscopy

To understand the effect of Se supplementation on health, it is critical to accurately assess the Se status in the human body by measuring reliable biomarkers. The preferred biomarkers of the Se status are selenoprotein P (SelP) and glutathione peroxidase 3 (GPx3) along with selenoalbumin (SeAlb), but there is still a real need for reference methods and reference materials to validate their measurements. Therefore, this work presents a systematic approach to provide quality control data in selenoprotein measurements. This approach combines online isotope dilution affinity liquid chromatography (LC) coupled to inductively coupled plasma mass spectrometry (ICPMS), laser ablation ICPMS, and tandem mass spectrometry (MS/MS) to identify and quantify SelP, GPx3, and SeAlb in a human plasma reference material SRM 1950. Quantitative determinations of SelP, GPx3, and SeAlb were 50.2 ± 4.3, 23.6 ± 1.3, and 28.2 ± 2.6 ng g(-1) as Se, respectively. The subsequent identification of the selenoproteins included nine SelP peptides, including two selenopeptides and nine GPx3 peptides, while albumin was identified with a protein coverage factor >95%. The structural elucidation of selenoproteins in the target Se affinity fractions in SRM 1950 provides information needed for method validation and quality control measurements of selenoproteins and therefore the selenium status in human plasma.

Development of a Common Procedure for the Determination of Methylmercury, Ethylmercury, and Inorganic Mercury in Human Whole Blood, Hair, and Urine by Triple Spike Species-Specific Isotope Dilution Mass Spectrometry

A system for the separation and detection of inorganic and organic selenium compounds utilizing particle beam-hollow cathode glow discharge-optical emission spectroscopy (PB-HC-OES) as a selenium-specific detector has been investigated. The PB interface includes a thermoconcentric nebulizer to generate a finely dispersed aerosol, a heated metal spray chamber for desolvation, and a two-stage momentum separator, which removes solvent vapor. The resulting beam of dry analyte particles are introduced into a heated (∼250 °C) hollow cathode, where they are vaporized, atomized and excited within the plasma. Using LC-PB-HC-OES the following compounds were examined: selenocystine, selenomethionine, selenoethionine, sodium selenate, and sodium selenite. A reverse phase ion-pairing chromatography method was developed to separate these five compounds (both organic and inorganic), with UV absorbance monitored at 210 nm. The Se I 204.0 nm atomic emission intensity was then monitored by coupling the LC column with the PB-HC-OES system. Emission responses for the selenium of both the organic and inorganic compounds in flow injection mode using 200 µL injection volumes indicate detection limits of ∼200 ppb (100 ng) with less than 10% RSD variability for triplicate injections over a range of 200–1960 ng. The retention times of the five analyte peaks are similar to those detected by UV absorbance, demonstrating the ability of the PB interface to preserve the chromatographic integrity of the separation. Optical emission detection of liquid chromatographic separations of the selenium-containing compounds demonstrates the feasibility of the PB-HC-OES system as a simple selenium-specific detector for liquid chromatography.

Atmospheric pressure, glow discharge, optical emission source for the direct sampling of liquid media

We report the first common methodology for the simultaneous determination of methylmercury (MeHg), ethylmercury (EtHg), and inorganic mercury (Hg(II)) in human blood hair and urine. With the exception of the initial sample mass (0.15 g for blood, 0.5 g for urine, and 0.1 g for hair), the same sample preparation and gas chromatography-inductively coupled plasma mass spectrometry (GC-ICPMS) measurement conditions are employed for the three matrixes providing experimental values in agreement with the certified values in the analysis of NIST SRM 955c (Caprine Blood) Level 3 and the certified human hairs IAEA 085 and IAEA 086. Also, the method provides quantitative recoveries for the three Hg species in the analysis of fortified human urine samples at 1, 2, and 5 ng Hg g-1. Mercury species concentrations for levels 2 and 4 of SRM 955c are reported here for the first time. A systematic interconversion of EtHg into Hg(II) was obtained for all matrixes reaching values up to 95% in blood, 29% in hair, and 11% in urine. MeHg dealkylation was also observed in a lesser extent in blood and hair analyses, but it was not observed when analyzing urine samples. Hg methylation was not observed in any matrix. The amount of NaBPr4 added for derivatization has been found to be the main factor responsible for Hg species interconversion. This work demonstrates for the first time that experimental conditions optimized for SRM 955c (caprine blood) are not valid for human blood samples as the optimum initial sample amount for a real sample is more than 3 times lower than that for SRM 955c.