Liquid chromatography-mass spectrometry: reaching maturity rapidly.
Explore the source record for details and available documents.
Editorial↗
Biomedical subjects
Uwe Karst
Publications and source records attributed to Uwe Karst.
Reaction monitoring of enzyme-catalyzed ester cleavage by time-resolved fluorescence and electrospray mass spectrometry: method development and comparison.
Two complementary methods for reaction monitoring of the esterase-catalyzed cleavage of bis(2-pyridylmethyl)(2-acetoxyphenyl)amine are developed and compared. While enzyme-amplified lanthanide luminescence (EALL) allows for the time-resolved fluorescence determination of the intrinsically non-fluorescent product, both substrate and product of the enzymatic reaction may be determined simultaneously by electrospray mass spectrometry (ESI-MS). Excitation wavelength for the Tb(III) complex of the reaction product is 297 nm and emission was detected at 545 nm, which is the characteristic emission wavelength of the terbium(III) ion. In contrast to other EALL techniques, the presented method allows for the direct monitoring of an enzymatic conversion without any further sample preparation (e.g., rebuffering). For the mass spectrometric measurements the mass traces were set to m/z=306, 328, 348, and 370 for the protonated ester, the resulting phenol and their sodium adducts, respectively.
Catalysis↗
Analysis of phenothiazine and its derivatives using LC/electrochemistry/MS and LC/electrochemistry/fluorescence.
The on-line electrochemical conversion of phenothiazine and its derivatives after liquid chromatographic separation has been studied by mass spectrometry and fluorescence spectroscopy. In an electrochemical cell consisting of porous glassy carbon, the phenothiazines are readily converted to oxidized products, which can be detected by on-line fluorescence spectroscopy and mass spectrometry. The method allows rapid investigations on the electrochemical oxidation pathways, as demonstrated for phenothiazine itself. The phenothiazine derivatives are transferred into their strongly fluorescent sulfoxides. Based on this reaction, an LC/electrochemistry/fluorescence method was developed that allows for limits of detection between 5 x 10(-9) and 4 x 10(-8) mol/L and limits of quantification between 2 x 10(-8) and 1 x 10(-7) mol/L for the individual phenothiazines. The linear ranges comprised three decades starting at the limit of quantification.
Chromatography, Liquid↗
Liquid chromatography-electrochemistry-mass spectrometry of polycyclic aromatic hydrocarbons.
An efficient method for fast elucidation of the electrochemical reactions of polycyclic aromatic hydrocarbons (PAH) has been set up by applying post-column electrochemistry in liquid chromatography-mass spectrometry (LC-MS). With this set-up strong improvement of sensitivity in the LC-MS analysis of PAH is observed. Due to their low redox potentials, the non-polar PAH are converted into the respective radical cations, which may further react with constituents of the mobile phase and in additional electrochemical oxidation steps. Among other products, mono-, di-, and trioxygenated species are observed in aqueous solutions, alkoxylated compounds in alcohols, and solvent adducts in the presence of acetonitrile. While more different products are observed by using atmospheric pressure chemical ionization in the positive-ion mode (APCI(+)), the deprotonation of hydroxylated species results in very clear spectra in the negative-ion mode (APCI(-)). Deuterated PAH and deuterated solvents were used to gain additional information on the formation of the reaction products.
Journal Article↗
Strategies for the liquid chromatographic-mass spectrometric analysis of non-polar compounds.
Electrospray ionization and atmospheric pressure chemical ionization (APCI) have evolved recently as very useful tools for the liquid chromatographic-mass spectrometric (LC-MS) analysis of polar substances. Non-polar compounds, however, are difficult to analyze with these atmospheric pressure ionization techniques due to their soft ionization mechanism. Recently, new approaches have been introduced which are likely to overcome this obstacle, at least partly. On-line electrochemical conversion of the analytes to more polar reaction products, atmospheric pressure photoionization, atmospheric pressure electron capture negativeion-MS and coordination ionspray-MS are four techniques which are presented in detail compared and discussed critically with respect to their current status and future perspectives. Particular focus is directed from a chemical viewpoint on the substance groups which are accessible by each of the new approaches.
Atmospheric Pressure↗
Trace analysis of peroxide-based explosives.
The first method for quantitative trace analysis of peroxide-based explosives is described. A reversed-phase high-performance liquid chromatography method with post-column UV irradiation and fluorescence detection for the analysis of triacetone triperoxide (TATP) and hexamethylene triperoxide diamine (HMTD) has been developed. After separation, the analytes are degraded photochemically to hydrogen peroxide, which is subsequently determined on the basis of the peroxidase-catalyzed oxidation of p-hydroxyphenylacetic acid to the fluorescent dimer. This two-step reaction scheme in combination with the respective blanks (photochemical reactor switched off) provides for high selectivity. The limits of detection were 2 x 10(-6) mol/L for both TATP and HMTD, respectively. The method has been applied to the analysis of real samples.
Bridged Bicyclo Compounds, Heterocyclic↗
Liquid chromatography with on-line electrochemical derivatization and fluorescence detection for the determination of phenols.
A new methodological approach for the determination of monosubstituted phenols is described. After liquid chromatographic separation of the analytes, an on-line electrochemical derivatization is carried out and the reaction products are detected fluorometrically. Phenols are oxidized in the electrochemical cell to form fluorescent dimers and higher oligomers, which were identified by on-line electrochemistry/mass spectrometry. Major advantages of the proposed method include enhanced selectivity and sensitivity. Without prior enrichment of the analytes, limits of detection down to 2 x 10(-9) M (20 fmol) may be reached for selected phenols, e.g., for 4-octylphenol, 4-ethylphenol, and 4-(1-indanyl)phenol. Only readily available instrumentation is required for these measurements.
Journal Article↗
Diffusive sampling of methyl isocyanate using 4-nitro-7-piperazinobenzo-2-oxa-1,3-diazole (NBDPZ) as derivatizing agent.
A diffusive sampling method for the determination of methyl isocyanate (MIC) in air is introduced. MIC is collected using a glass fiber filter impregnated with 4-nitro-7-piperazinobenzo-2-oxa-1,3-diazole (NBDPZ). The urea derivative formed is desorbed from the filter with acetonitrile and analyzed by means of high-performance liquid chromatography (HPLC) using fluorescence detection (FLD) with lambdaex = 471 nm and lambdaex = 540 nm. Additionally, a method was developed using tandem mass spectrometric (MS-MS) detection, which was performed as selected reaction monitoring (SRM) on the transition [MIC-NBDPZ + H]+ (m/z 307) to [NBDPZ + H]+ (m/z 250). The diffusive sampler was tested with MIC concentrations between 1 and 35 microg m(-3). The sampling periods varied from 15 min to 8 h, and the relative humidity (RH) was set from 20% up to 80%. The sampling rate for all 15 min experiments was determined to be 15.0 mL min(-1) (using HPLC-FLD) with a relative standard deviation of 9.9% for 56 experiments. At 80% RH, only 15 min sampling gave acceptable results. Further experiments revealed that humidity did not affect the MIC derivative but the reagent on the filter prior to and during sampling. The sampling rate for all experiments (including long term sampling) performed at 20% RH was found to be 15.0 mL min(-1) with a relative standard deviation of 6.3% (N = 42). The limit of quantification was 3 microg m(-3) (LC-MS-MS: 1.3 microg m(-3)) for 15 min sampling periods and 0.2 microg m(-3) (LC-MS-MS: 0.15 microg m(-3)) for 8 h sampling runs applying fluorescence detection.
Absorption↗
A tripod ligand as new sensitiser for the enzyme amplified lanthanide luminescence determination of esterase.
Screening of a small library of tripod ligands resulted in the discovery of bis(2-pyridylmethyl)-(2-hydroxybenzyl)amine (HL1) as a new sensitiser, which is able to transfer its excitation energy to terbium(III). After synthesis of the acetic acid ester of HL1, a highly selective method for the determination of porcine liver esterase by means of enzyme amplified lanthanide luminescence (EALL) was developed. Enzyme-catalysed cleavage of the ester results in the formation of HL1. After excitation at 297 nm, the characteristic emission of Tb(III) at 545 nm is observed and used to determine the esterase concentration. In contrast to existing EALL methods, this method may be carried out at neutral pH and without further additives. Limit of detection for porcine liver esterase is 10(-9) mol l(-1) and limit of quantification is 3 x 10(-9) mol l(-1). A linear calibration range of two decades starting at the limit of quantification is observed.
Animals↗
LC-electron capture-APCI(-)-MS determination of nitrobenzoxadiazole derivatives.
Nitrobenzoxadiazole (NBD) derivatives are determined with limits of detection ranging down to 20 nmol l(-1) using liquid chromatography-mass spectrometry (LC-MS) with electron capture (EC) ionisation. An atmospheric pressure chemical ionisation (APCI) interface operated in the negative ion mode is used as ionisation source. Amine derivatives of 4-chloro-7-nitro-2,1,3-benzoxadiazole (NBDCl) as well as the isocyanate derivatives of 4-nitro-7-piperazino-2,1,3-nitrobenzoxadiazole (NBDPZ) have been analysed using this technique. The parameters favouring electron capture mechanisms have been investigated thoroughly under consideration of the competing mechanism of deprotonation to allow a better understanding of the electron capture process and to improve selectivity of the analysis.
Chromatography, Liquid↗
4-Nitro-7-piperazino-2,1,3-benzoxadiazole as a reagent for monitoring of airborne isocyanates by liquid chromatography.
4-Nitro-7-piperazino-2,1,3-benzoxadiazole (NBDPZ) is presented as a new reagent for the determination of mono- and diisocyanates in air samples. NBDPZ readily reacts with the airborne analytes, thus yielding the corresponding urea derivatives, which are subsequently separated by means of reversed-phase liquid chromatography. On a phenyl-modified stationary phase, excellent baseline separation for numerous mono- and diisocyanate derivatives is obtained. Both diode array and fluorescence detection are performed with limits of detection of 11-35 and 5-9 nmol/L for the individual derivatives, respectively. In contrast to established derivatizing agents for the analysis of isocyanates, NBDPZ provides for increased selectivity due to the favorable detection wavelengths in the visible range (UV/visible, absorption maximums approximately 480 nm; fluorescence, excitation maximums approximately 470 nm, emission maximums approximately 535 nm). In addition, the high molar absorptivities of the reagent and the derivatives provide excellent sensitivity that is superior to most literature-known methods. Finally, air sampling methods comprising both the use of impingers and test tubes are developed and successfully applied to the determination of isocyanates in gaseous samples. Excellent recovery reaching values of >90% is observed for each of the two techniques investigated.
Air Pollutants↗
Fast liquid chromatography-electrochemistry-mass spectrometry of ferrocenecarboxylic acid esters.
Rapid liquid chromatographic separations of ferrocenecarboxylic esters of various alcohols and phenols have been achieved on reversed-phase columns of 20 mm length. After separation, the ferrocene derivatives are oxidized electrochemically under formation of the charged ferrocinium species, which are easily detected by mass spectrometry using an atmospheric pressure chemical ionization source operated in the heated nebulizer mode. While a series of nine phenol derivatives was separated within less than 1.5 min, six alcohol derivatives eluted within 1 min. Limits of detection using a single quadrupole mass analyzer ranged from 60 to 190 nmol/l. Additional work was directed on the use of a graphite in-line filter instead of a silica-based reversed-phase column to achieve the separation.
Chromatography, Liquid↗
On-line electrochemistry--MS and related techniques.
This review summarizes publications on the on-line coupling of electrochemistry with mass spectrometry. After a brief historic introduction it is divided into three parts, organized in order of increasing complexity of the experimental arrangement. The first section deals with the use of the electrospray ion source as an electrochemical reactor for oxidation or reduction reactions. It is followed by the second part which covers the hyphenation of different kinds of electrochemical flow cell with a variety of ionization interfaces. The last section focuses on the on-line coupling of chromatographic techniques with electrochemical flow cells and mass spectrometry.
Journal Article↗
Aldehydes 2001--an unusual conference.
Explore the source record for details and available documents.
Editorial↗
LC-electron capture APCI-MS for the determination of nitroaromatic compounds.
The determination of selected nitroaromatic compounds in liquid chromatography-mass spectrometry with electron capture (EC) ionisation using a commercial atmospheric pressure chemical ionisation (APCI) interface in the negative mode is described. The electron capture effect is observed for nitroaromatics which do not easily undergo deprotonation under these conditions. Depending on the structure of the analytes, either dissociative or, for the first time in LC-MS, non-dissociative electron capture is observed. Limits of detection and linear range for the determination of the analytes match those obtained for nitroaromatics which undergo deprotonation. The investigated substances comprise numerous substituted nitrobenzenes and nitrobenzooxadiazoles.
Journal Article↗
A field test for the detection of peroxide-based explosives.
A rapid and simple field test for the detection of triacetone-triperoxide (TATP) and hexamethylenetriperoxidediamine (HMTD), two explosives which find significant illegal use, has been developed. Unknown samples are first treated with a catalase solution to remove hydrogen peroxide traces, in order to provide selectivity towards peroxide-based bleaching agents which are contained in commercial laundry detergents. Subsequently, the peroxide-based explosives are decomposed via UV irradiation, thus yielding hydrogen peroxide, which is determined by the horseradish peroxidase (POD) catalysed formation of the green radical cation of 2,2'-azino-bis(3-ethylbenzothiazoline)-6-sulfonate (ABTS). The limits of detection for this method are 8 x 10(-6) mol dm(-3) for TATP and 8 x 10(-7) mol dm(-3) for HMTD, respectively. As an option, p-hydroxyphenylacetic acid (pHPAA) may be used as peroxidase substrate, resulting in lower limits of detection (8 x 10(-7) mol dm(-3) for TATP and HMTD). The complete method uses a mobile setup to be applied under field conditions.
Explosions↗
Selective determination of hydrogen peroxide by adduct formation with a dinuclear iron(III) complex and flow injection analysis/tandem mass spectrometry.
A highly selective method for the determination of hydrogen peroxide is presented. In a flow injection analysis (FIA) instrument, the analyte is brought into contact with a dinuclear heptadentate iron(III) complex. The formation of the peroxide adduct is quantified using electrospray tandem mass spectrometry (ESI-MS/MS). Selected reaction monitoring (SRM) based on the transition from the triply charged peroxide adduct with m/z = 251.2 to the triply charged fragment ion of m/z = 240.5 is performed. The limit of detection for hydrogen peroxide is 10(-7) mol dm(-3), limit of quantification is 3 x 10(-7) mol dm(-3), and a linear range of 2.5 decades starting at the limit of quantification is observed.
Journal Article↗
Detection techniques for liquid chromatography.
Explore the source record for details and available documents.
Journal Article↗