Mycocerosic acid biomarkers for the diagnosis of tuberculosis in the Coimbra Skeletal Collection.

Tuberculosis has been a scourge of humans over many millennia, but questions remain regarding its evolution and epidemiology. Fossil biomarkers, such as DNA and long-chain mycolic acids, can be detected in ancient skeletal and other materials. The phthiocerol dimycocerosate waxes are also robust biomarkers for tuberculosis and sensitive methods are available for the detection of their mycocerosic acid components. The presence of mycocerosic acids was investigated in 49 individuals from the 1837-1936 Coimbra Identified Skeletal Collection (Portugal), half with documentary data indicating tuberculosis as a cause of death. Samples were hydrolysed, acidic components converted to pentafluorobenzyl esters, the non-hydroxylated long-chain esters isolated, and this fraction separated into multimethyl-branched and other esters by normal phase high performance liquid chromatography. Negative ion chemical ionisation gas chromatography mass spectrometry was used to detect diagnostic C29, C30 and C32 mycocerosic acids. Mycocerosic acids were detected in archaeological material for the first time, illustrating that they are valuable biomarkers for the diagnosis of ancient tuberculosis. A 72% correlation with the Coimbra burial record supported TB as the major cause of death. In addition, 30% of the skeletons, positive for mycocerosates, showed the presence of related long-chain mycolipenic acids.

The determination of trace metal pollutants in environmental matrices using ion chromatography.

A review is presented detailing the development of ion chromatography (IC) as a selective analytical tool for the determination of toxic metals and their organic species in many environmental sample matrices. A brief outline of ion chromatographic principles, together with an overview of the stationary phases used to separate metals, namely ion exchangers, modified ion pair sorbents and chelating ion exchangers, and the methods for detecting metal ions including hyphenation with spectroscopy and sample preparation schemes are also given, prior to a critical examination of developed methods for various metals including arsenic, chromium, cadmium, lead, mercury, beryllium, aluminium and uranium since 1990.

Dithizone derivatives as sensitive water soluble chromogenic reagents for the ion chromatographic determination of inorganic and organo-mercury in aqueous matrices.

Water-soluble sulfonate and the novel carboxylate analogues of dithizone, combined with ion interaction chromatography on a Dionex Acclaim 120 C18 silica column (250 x 4.6 mm id) with an eluent consisting of 10 mM tetrabutylammonium bromide and 60:40 methanol:water, have been developed as highly sensitive chromogenic ligands for the quantitative isocratic determination of inorganic and organo-mercury compounds in aqueous matrices in under 12 min. Using an optimised post column reagent system containing 0.65 mM dye, 0.5% Triton X-100 and 50 mM sodium hydroxide, good linearity (0-7.5 mg L(-1) R2 > 0.999), reproducibility using peak area measurements (RSD 0.69-1.38%, n = 8), and limits of detection (4-12 microg L(-1)) were achieved for methyl mercury, inorganic mercury and phenyl mercury.

Selectivity behaviour of a bonded phosphonate--carboxylate polymeric ion exchanger for metal cations with varying eluent compositions.

The chromatographic behaviour of a commercially available ion-exchange stationary phase (the Dionex IonPac CS12A column) is described for a wide range of transition and heavy metal ions with nitric acid eluents containing chloride and nitrate potassium salts. The separation selectivity was found to arise from simultaneous ion-exchange interactions and chelation with the attached carboxylic and phosphonic acid groups. These interactions were investigated by altering the ionic strength and pH of the eluent and also the column temperature. Strong affinity of the stationary phase towards heavy metal ions, in particular bismuth and the uranyl ion was observed at low pH under chelating ion-exchange conditions, with high efficiency separations of other ions including cadmium and lead being possible with short analysis times (approximately 5-15 min). Examples are given of separations obtained using 4-(2-pyridylazo)resorcinol or Arsenazo III as the post-column chromogenic reagents, demonstrating the potential versatility and utility of this stationary phase for heavy metal ion analysis.

Developments in suppressor technology for inorganic ion analysis by ion chromatography using conductivity detection.

A review is presented detailing the development and use of suppression devices for the conductimetric detection of inorganic ions by ion chromatography (IC). An overview of the general response equation for conductivity detection is also given. Topics of discussion include the role and function of suppressors, the development of early suppressors including packed column and membrane devices from 1975 to 1990 and the subsequent progression towards present day commercially available suppressors and recent innovations. Post-suppression devices for signal enhancement are also discussed.

Sensitive indirect photometric detection of inorganic and small organic anions by capillary electrophoresis using Orange G as a probe ion.

This study addresses the two major problems in the use of dyes as highly absorbing probes for indirect photometric detection in capillary electrophoresis (CE). First, effective electroosmotic flow (EOF) modification or suppression to allow separation and detection of a wide mobility range of analytes is not straightforward when electrolytes containing increased dye concentrations are used. The suppression of EOF to less than + 5x10(-9) m(2)V(-1)s(-1) was achieved with a combination of a poly(ethylenimine) (PEI)-coated capillary and the addition of the neutral polymer hydroxypropylmethylcellulose (HPMC) to the background electrolyte. Second, the deterioration of baselines due to adsorption of the dye probe to the capillary wall is generally a problem. In this work, baseline quality at higher probe concentrations was significantly improved by a rather unusual but highly effective combination of a simultaneous application of a slight overpressure (25 mbar) at the injection end during the separation, and the use of a relatively narrow capillary of 50 microm inner diameter. Both measures would appear to be counterproductive. Optimisation of the probe concentration with regard to signal-to-noise ratio resulted in an electrolyte of 4 mM Orange G, 0.05% HPMC buffered at pH 7.7 by the addition of 10.0 mM histidine isoelectric buffer. Very high separation efficiencies of 128 000-297 000 plates were made possible by the relatively high probe concentration. Combined with excellent detection sensitivity, even with the introduction of hydrodynamic flow and a reduced optical path length, these measures resulted in limits of detection ranging from 0.216 to 0.912 microM with a deuterium lamp light source (248 nm) and from 0.147 to 0.834 microM with a 476 nm blue light-emitting diode (LED) light source. Reproducibility over 30 consecutive runs without changing the electrolyte was excellent, with relative standard deviation (RSD) values of 0.14-0.80% for migration time, 1.27-3.36% for peak area and 0.88-5.12% for peak heights. The optimised electrolyte was used for the analysis of inorganic anions in air filter samples, providing good agreement with results obtained by ion chromatography.

Simulation and optimization of retention in ion chromatography using virtual column 2 software.

A new software package, Virtual Column 2, is described for the simulation and optimization of the separation of inorganic anions by ion chromatography (IC). The software uses a limited amount of experimental retention data acquired according to a correct experimental design to predict retention times for analytes over a designated search area of eluent compositions. The experimental retention data are used to solve a new retention model, called the linear solvent strength model, empirical approach (LSSM-EA), which then enables prediction of retention times for all eluent compositions in the search area. The theoretical development of LSSM-EA and the processes used for solving the equations are discussed. Virtual Column 2 can be used for eluents containing one or two competing ions, and the software contains retention databases for up to 33 analytes on the Dionex AS9A-HC, AS4A-SC, and AS14A analytical columns with carbonate-bicarbonate eluents and the Dionex AS10, AS15, and AS16 analytical columns with hydroxide eluents (results for the AS10 and AS15 columns are not discussed in the present study). Virtual Column 2 has been evaluated extensively and is shown to give predicted retention times that in most cases agree with experimentally determined data to within 5%. The software has uses in practical IC method development, education and training in IC, and refinement of existing IC methodology. A free version of this program is available by download at www.virtualcolumn.com.

Separation of polythionates and the gold thiosulfate complex in gold thiosulfate leach solutions by ion-interaction chromatography.

A method for the separation of the polythionates (SxO6(2-), x = 3-5) in gold thiosulfate leach solutions using ion-interaction chromatography with conductivity and ultraviolet (UV) detection is described. Polythionates were eluted within 18 min using an eluent comprising an acetonitrile step gradient at 0.0 min from 15% v/v to 28% v/v, 3 mM TBAOH, and 2.5 mM sodium carbonate, operated using a Dionex NS1-5 micron column with guard. The developed method was capable of separating the gold thiosulfate complex ion in standard solutions, but quantification of this species in realistic leach solutions proved impractical due to a self-elution effect that caused the gold peak to be eluted as a broad band. Detection limits for polythionates using a 10 microL injection volume ranged between 1-6 mg L(-1) (5-23 microM) for conductivity and 0.8-13 mg L(-1) (4-68 microM) for UV detection, based on a signal-to-noise ratio of 2. Calibration was linear over the ranges 5-2000, 10-2000 and 25-2500 mg L(-1) for trithionate, tetrathionate and pentathionate, respectively. The technique was applied successfully to leach liquors containing 0.5 M ammonium thiosulfate, 2 M ammonia, 0.05 M copper sulfate and 20 % m/v gold ore.

New conductivity detection response equation for anions eluted with fully and partially ionised eluents in non-suppressed ion chromatography.

A response equation for conductivity detection in ion chromatography has been derived. This equation is applicable to non-suppressed ion chromatography using both fully ionised and partially ionised eluents. A prime assumption of this equation is that when partially ionised eluents are used (such as benzoic acid), both the ionised and neutral components of the eluent species contribute to the detector response of anionic analytes. Experimental evidence is provided to support this assumption in that pH changes accompanying the elution of an analyte have been measured. These pH changes are proportional to the concentration of analyte injected onto the column, in accordance with predictions from the response equation. Furthermore, it is shown that protonated eluents (such as benzoic acid) give more sensitive detection than equivalent ionised eluents (such as potassium benzoate) and the signal enhancement achieved using a protonated eluent species is in accordance with theory.

Dynamic chelation ion chromatography of transition and heavy metal ions using a mobile phase containing 4-chlorodipicolinic acid.

The chromatographic behaviour of selected transition and heavy metal ions, the lanthanides, uranium and aluminium, on a neutral polystyrene-divinylbenzene (PS-DVB) stationary phase (7 microm Hamilton PRP-1) dynamically modified with 4-chlorodipicolinic acid, was investigated to evaluate retention characteristics. Complicated retention factor against pH plots were found for these metals demonstrating changes in retention order. It was concluded that complexation between the metal ions and the ligand adsorbed on the resin was strongly influenced by the decrease in dynamic loading with increase in pH, coinciding with changes in the metal-to-ligand ratio in the mobile phase. Possible reversed-phase interactions between metal-chlorodipicolinic acid complexes and the hydrophobic PS-DVB stationary phase also could not be ruled out. An eluent of 0.25 mM chlorodipicolinic acid, I M potassium nitrate at pH 2.2 was suitable for the separation of seven transition and heavy metal ions in under 20 min on a 250 x 4.6 mm column (with 50-mm guard column), determined in a certified water sample with good accuracy (R2 > or = 0.994) and reproducibility (RSD 1-4.2%). Pb(II), Cd(II) and Cu(II) were additionally analysed in <10 min in a more complicated certified rice flour matrix, using the same eluent but adjusted to pH 1.5, again with good accuracy (R2 > or = 0.998) and reproducibility (RSD 0.48-1.38%).