Pterin chemistry. 46. Identification of substituted tetrahydropterins through gas chromatography and mass spectrometry

When treated with cyanide ion, 1,2:5,6-di-O-isopropylidene-α-_D-ribo-hexofuranos-3-ulose yields one or the other of the two corresponding epimeric cyanhydrines, depending on the experimental conditions. One of these epimers, reacted with ammonium cyanide, yields the corresponding amino-nitrile, the other being unreactive. Several derivatives of these compounds are briefly described.     

Derivatization procedures for the detection of estrogenic chemicals by gas chromatography/mass spectrometry

This work evaluated derivatization procedures for detecting both natural and synthetic estrogenic chemicals by gas chromatography/mass spectrometry (GC/MS). Different silylating agents, mainly trimethylsilylating (TMS) agents, were compared, and the roles of various content of trimethylchlorosilane (TMCS, as a stimulator) were investigated. The difference in the abundances of the derivatives was caused by the steric hindrance of multiple hydroxyl groups and ethynyl groups in the structures of estrogenic chemicals. The use of TMCS produces an increase in the derivatization yield, especially for the compounds with multiple hydroxyl groups (i.e., 17beta-estradiol (E(2)) and estriol (E(3))). Mass spectra of O-TMS derivatives and tentative fragmentation profiles are proposed. Molecular ions were the base peaks for all the derivatives, and were used as the quantitation ions to obtain maximum detection sensitivity and specificity. Sample enrichment was achieved by Oasis HLB solid-phase extraction cartridges. The quantitation limits of these compounds ranged from 5 to 10 ng/L in 1000-mL water samples. Recovery of the estrogenic chemicals in spiked various water samples ranged from 78 to 102% while relative standard deviation (RSD) ranged from 1 to 15%.     

The determination of steric purity of amines and amino acids by gas chromatography and mass spectrometry

Amines and amino acids have been condensed with a mixture of N-TFA-R-prolyl chloride and n-TFA-S-[1-²H]-prolyl chloride and the resulting four diastereoisomers have been separated by gas chromatography into two peaks. The amount of each diastereoisomer formed was then determined by chemical ionization mass spectrometry. The steric purity of the enantiomeric mixture can be calculated from these values. The method is not dependent on the diastereoisomer formation going to completion.     

Reaction gas chromatography/mass spectrometry. 4—structure elucidation of cyclic sulphides by gas chromatography/mass spectrometry with the use of on-line hydrodesulphurization

A method for structure determination of cyclic sulphides within mixtures is described. It involves the use of a vapour phase hydrodesulphurization micro-reactor (Raney nickel) located between the chromatographic column and the mass spectrometer (hydrogen and deuterium are used as carrier and reagent gases). A comparative examination of the mass spectra of the cyclic sulphides, as well as of the hydrocarbons and their deuterio analogues (deuterium atoms are bonded to carbon atoms which had been attached to sulphur) derived from the sulphides under the catalytic conditions employed, permits unambiguous determination of the size of the heterocycle and the position of the substituents.     

Reaction gas chromatography/mass spectrometry. 2—the use of catalytic on-line dehydrogenation for the investigation of alcohols by gas chromatography/mass spectrometry

A method for structure determination of aliphatic alcohols within mixtures is described. It involves the use of a vapour phase dehydrogenation micro-reactor (Cu, 300°C) located between the chromatographic column and the mass spectrometer or between the injection port and the column. Since primary and secondary alcohols are converted into corresponding carbonyl compounds, they can be readily differentiated from tertiary alcohols and dialkyl ethers. An examination of the mass spectra of alcohols and carbonyl compounds permits the determination of molecular mass, the location of hydroxyl group and the determination of branching at the β-carbon atom.     

Evaluation of time-of-flight mass spectrometric detection for fast gas chromatography

Separations below 1 s of a mixture of organic compounds ranging from C5 to C8 have been performed to investigate the performance of a time-of-flight mass spectrometer in fast gas chromatography. The gaseous samples were focussed on a cold trap, and then injected after thermal desorption to obtain the required narrow input band-widths. Also, to obtain a very fast separation, a short narrow bore column was used, operated at above-optimum inlet pressures. With this system, it was possible to identify ten compounds within 500 ms, showing peak-widths (2.354sigma) as narrow as 12 ms. The spectral acquisition rate used for these analyses was 500 Hz. The quality of the recorded spectra and the comparison with library spectra was very high. Deconvolution algorithms offer the possibility of identifying overlapping peaks. It is shown that the spectral scan speed of the time-of-flight mass spectrometer is high enough for very fast separations.     

Comparison of gas chromatography and liquid chromatography mass spectrometric measurements for high accuracy analysis of cholesterol in human serum by isotope dilution mass spectrometry

Cholesterol measurements are of vital clinical importance and reliable reference materials are essential for method validation. Gas chromatography with mass spectrometry (GC/MS) is usually used for the high accuracy analysis of cholesterol by isotope dilution. A certified reference material for cholesterol content in human serum was analysed by isotope dilution utilising GC/MS and liquid chromatography mass spectrometry (LC/MS). The use of LC/MS avoided the need for a derivatisation step. Both LC/MS and GC/MS produced results on the measurement of cholesterol that agreed within 0.5% of the certified value. Moreover, the precision obtained for ratio measurement using both techniques are comparable and lead to relative expanded standard uncertainties (with a coverage factor of 2) varying between 0.2 and 0.5%.     

Applied gas chromatography coupled to isotope ratio mass spectrometry.

Compound-specific isotope analysis (CSIA) by isotope ratio mass spectrometry (IRMS) following on-line combustion (C) of compounds separated by gas chromatography (GC) is a relatively young analytical method. Due to its ability to measure isotope distribution at natural abundance level with great accuracy and high precision, GC-C-IRMS has increasingly become the method of choice in authenticity control of foodstuffs and determination of origin in archaeology, geochemistry, and environmental chemistry. In combination with stable isotope labelled compounds, GC-C-IRMS is also used more and more in biochemical and biomedical application as it offers a reliable and risk-free alternative to the use of radioactive tracers. The literature on these topics is reviewed from the advent of commercial GC-C-IRMS systems in 1990 up to the beginning of 1998. Demands on sample preparation and quality of GC separation for GC-C-IRMS are discussed also.     

Identification of isopropyl substituted β-blocking agents in human urine by gas chromatography and tandem mass spectrometry

Summary A method for the sensitive detection of isopropyl substituted β-blocking agents in human urine is presented. The sample preparation step involves enzymatic hydrolysis, solid phase extraction and derivatisation withN-methyl-N-trimethylsilyl-trifluoroacetamide. The instrumental analysis was performed using gas chromatography-mass spectrometry with an ion trap mass spectromeler. The mass spectrometer was operated in the scan as well as in the MS-MS mode.     

The analysis of thiocarbamides by gas chromatography/negative-ion chemical-ionization mass spectrometry.

Thiocarbamides were converted to their di-N-pentafluorobenzyl (PFB) derivatives and analysed by gas chromatography/negative-ion chemical-ionization mass spectrometry with methane as reagent gas. The PFB derivatives of the 2-thiouracils gave mass spectra in which the ion current was carried largely by an ion arising from [M-PFB]-. The derivative was used in the determination of the uptake and metabolism of thiocarbamides by cultures of melanoma cells.     

Pyrolysis–gas chromatography mass spectrometry and field ionization mass spectrometry of polyquinones

Low- and high-molecular mass thermal decomposition products of five polyquinones with different linking aromatic structures have been analyzed by pyrolysis–gas chromatography and by direct (in-source) pyrolysis–field ionization mass spectrometry. The quantity of carboxyl groups present in the polymer is obtained by the amounts of carbon dioxide found by pyrolysis–gas chromatography. Assuming a radical thermal decomposition mechanism the distribution of ketoacidic and quinonoid segments along the macromolecular ladder could be estimated from the high-molecular mass products measured by pyrolysis–field ionization mass spectrometry. A random distribution of the two different segments was found for polyquinones with biphenylene and dibenzofuran subunits, while a structure built up of blocks of two or more identical segments was obtained for polyquinones with dibenzothiophene and diphenylmethane subunits. At the same time the anomalous structural moieties in the polyquinone ladders are also clarified with the help of the identification of the unexpected pyroysis products. Oxidated and bis-dibenzothiophene and bis-diphenylmethane subunits were found. The observed temperature dependence for the appearances of the thermal degradation products indicates that condensation and elimination reactions are taking place under the described pyrolysis conditions. Condensation in the ketoacidic segments forming new quinonoid segments proved to be important in the polymer which was a 100% poly(ketoacid), but negligible in the polyquinones containing ketoacidic segments up to 60%.     

Comprehensive two-dimensional gas chromatography in combination with rapid scanning quadrupole mass spectrometry in perfume analysis

Single column gas chromatography (GC) in combination with a flame ionization detector (FID) and/or a mass spectrometer is routinely employed in the determination of perfume profiles. The latter are to be considered medium to highly complex matrices and, as such, can only be partially separated even on long capillaries. Inevitably, several monodimensional peaks are the result of two or more overlapping components, often hindering reliable identification and quantitation. The present investigation is based on the use of a comprehensive GC (GC x GC) method, in vacuum outlet conditions, for the near to complete resolution of a complex perfume sample. A rapid scanning quadrupole mass spectrometry (qMS) system, employed for the assignment of GC x GC peaks, supplied high quality mass spectra. The validity of the three-dimensional (3D) GC x GC-qMS application was measured and compared to that of GC-qMS analysis on the same matrix. Peak identification, in all applications, was achieved through MS spectra library matching and the interactive use of linear retention indices (LRI).     

Medium- and long-chain 3-hydroxymonocarboxylic acids: analysis by gas chromatography combined with mass spectrometry

Medium- and long-chain 3-hydroxymonocarboxylic acids represent intermediates in the beta-oxidation of fatty acids: they accumulate in the plasma of patients with an inherited deficiency of long-chain 3-hydroxyacylcoenzyme A dehydrogenase. 3-Hydroxy acids with chain lengths varying from 6 to 16 were synthesized by a Reformatzky reaction. Capillary gas chromatography of the pertrimethylsilyl derivatives was performed on a CP-Sil 19 CB column, coupled to a quadrupole mass spectrometer in the electron impact mode. Calculation of the retention indices showed that the separation of the 3-hydroxy acids from the homologous straight-chain fatty acids may be troublesome, stressing the need for mass spectrometric identification.     

Combined use of gas chromatography and selected ion flow tube mass spectrometry for absolute trace gas quantification

The value of the gas chromatography (GC) and selected ion flow tube mass spectrometry (SIFT-MS) combination for the analysis of trace gases is demonstrated by the quantification of acetone in air samples using the three precursor ions available to SIFT-MS, viz. H3O+, NO+ and O2+, and by the separation of the isomers 1-propanol and 2-propanol, and their analysis using H3O+ precursor ions. It is shown that the GC/SIFT-MS combination allows for accurate trace gas quantification obviating the regular, time-consuming calibrations that are usually required for the more commonly used detectors of GC systems, and the positive identification of isomers in mixtures that is often challenging using SIFT-MS alone. Thus, the GC/SIFT-MS combination paves the way to more confident analyses of complex mixtures such as exhaled breath.     

Derivatization coupled to headspace programmed‐temperature vaporizer gas chromatography with mass spectrometry for the determination of amino acids: Application to urine samples

A new method based on headspace programmed‐temperature vaporizer gas chromatography with mass spectrometry has been developed and validated for the determination of amino acids (alanine, sarcosine, ethylglycine, valine, leucine, and proline) in human urine samples. Derivatization with ethyl chloroformate was employed successfully to determine the amino acids. The derivatization reaction conditions as well as the variables of the headspace sampling were optimized. The existence of a matrix effect was checked and the analytical characteristics of the method were determined. The limits of detection were 0.15–2.89 mg/L, and the limits of quantification were 0.46–8.67 mg/L. The instrumental repeatability was 1.6–11.5%. The quantification of the amino acids in six urine samples from healthy subjects was performed with the method developed with the one‐point standard additions protocol, with norleucine as the internal standard.     

Analysis of triacetone triperoxide by gas chromatography/mass spectrometry and gas chromatography/tandem mass spectrometry by electron and chemical ionization

The explosive triacetone triperoxide (TATP) has been analyzed by gas chromatography/mass spectrometry (GC/MS) and sub-nanogram detection limits are reported by ammonia positive ion chemical ionization (PICI), electron ionization (EI) and methane negative ion chemical ionization (NICI). Analysis by methane PICI and ammonia NICI gave detection limits in the low nanogram range. Analyses were carried out on (linear) quadrupole and ion trap instruments. Analysis of TATP by PICI using ammonia reagent gas is the preferred analytical method, producing low limits of detection as well as an abundant (greater than 60% of base peak) diagnostic adduct ion at m/z 240 corresponding to [TATP + NH4]+. Isolation of the [TATP + NH4]+ ion with subsequent collision-induced dissociation (CID) produces extremely low abundance product ions at m/z values greater than 60, and the m/z 223 ion corresponding to [TATP + H]+ was not observed. Density functional theory (DFT) calculations at the B88LYP/DVZP level indicate that dissociation of the complex to form NH4+ and TATP occurs at energies lower than peroxide bond dissociation, while protonation of TATP leads to cleavage of the ring structure. These results provide a method for pico-gram detection levels of TATP using commercial instrumentation commonly available in forensic laboratories. As a point of comparison, a detection limit of 15 ng was obtained by flame ionization detection.     

A single glucose derivative suitable for gas chromatography/mass spectrometry and gas chromatography/combustion/isotope ratio mass spectrometry

The incorporation of stable isotopes improves the assessment of glucose metabolism and, with some researchers using two tracers, (2)H-glucose assessed by gas chromatography/mass spectrometry (GC/MS) and (13)C-glucose by gas chromatography/combustion/isotope ratio mass spectrometry (GC/C/IRMS), a common derivative for both is advantageous. The most commonly used derivatives for GC/MS are inappropriate for GC/C/IRMS as additional functional groups dilute the label. We therefore considered the suitability of six derivatives for both GC/MS and GC/C/IRMS. Glucose alkylboronates were prepared by adding the appropriate alkylboronic acid (butyl- or methylboronic acid) in pyridine to desiccated glucose. The derivatisation was completed by reacting this with either (a) acetic anhydride or trifluoroacetic anhydride (acetate derivatives) or (b) bis(trimethylsilyl)trifluoroacetamide BSTFA (TMS derivatives). All six derivatives were assessed using GC/MS and (13)C GC/C/IRMS.Neither TMS derivative exhibited any signal intensity in the molecular ion, although a M-15 ion showed good agreement between experimental and theoretical data and, whilst still low in intensity, could be suitable for isotope work. Similarly, none of the acetate derivatives showed any intensity at the molecular ion although three key fragmentation series were identified. The most attractive sequence, initiated by the loss of 1,2 cyclic boronate, resulted in the main fragment ion of interest, m/z 240, corresponding to the fluorinated methylboronate derivate. Minimal carbon and hydrogen atoms are added to this derivative making it an excellent choice for stable isotope work, while proving suitable for analysis by both GC/MS and GC/C/IRMS.     

The location of the double bonds in substituted conjugated alkadienes by using the ethanethiol and gas chromatography/mass spectrometry

In order to locate the position of the double bonds in conjugated alkadienes substituted, several ethanethiol derivatives were studied. In all cases, on EI ionisation at 70 eV, the adducts obtained allowed easily the location of the double bonds in substituted conjugated alkadienes, contrary to the dimethyl disulfide (DMDS) derivatives.