Evaluation of different liquid chromatography-electrospray mass spectrometry systems for the analysis of heterocyclic amines

Three liquid chromatography-electrospray ionisation (LC-ESI) MS systems are evaluated for the analysis of heterocyclic amines (HAs). The electrospray sources and analysers (ion trap, single quadrupole and triple quadrupole) have been compared in terms of performance and quality parameters. In all cases, a C8 reversed-phase column and (acetic acid-ammonium acetate 30 mM pH 4.5)-acetonitrile (ACN) as mobile phase were used. Ionisation source parameters, post-column addition and working conditions for each acquisition mode (full scan, product ion scan, selected ion monitoring, and multiple reaction monitoring) were optimised for each instrument. The MS-MS spectra obtained with the ion trap and the triple quadrupole systems were very similar in both fragment ions and relative abundances, except for carbolines that showed adduct formation in the ion trap. Quality parameters were established and good precision (relative standard deviations (R.S.D.) < 12%) and very low limits of detection were obtained, mainly when using the triple quadrupole (< 9 pg injected). The content of HAs in a lyophilised beef extract was determined using the three instruments in order to compare their applicability for routine HAs analysis.     

Determination of heterocyclic aromatic amines by capillary electrophoresis coupled to mass spectrometry using in-line preconcentration

This paper shows the potentiality of capillary electrophoresis (CE) coupled to mass spectrometry (MS) for the analysis of heterocyclic aromatic amines obtaining good results in terms of sensitivity and precision. These compounds have a special interest since they can be carcinogenic for humans. The optimization of a CE-MS method was performed and the best conditions were obtained using a 16 mM formic acid/ammonium formate solution at pH 4.5 with 60% methanol as running electrolyte. For CE-MS coupling, a sheath liquid methanol/20 mM formic acid (75/25) solution at a flow rate of 3 microL/min and hydrodynamic injection of methanol mixtures for 10 s were used. Detection limits ranging from 18 ng/g to 360 ng/g and precisions up to 1.4% and 12% for migration time and concentration, respectively, were obtained. In order to improve sensitivity, field-amplified sample injection was applied as an in-line preconcentration method. Methanol/5 mM formic acid (50/50) as a sample solvent, 3 s hydrodynamic injection (0.5 psi) of a methanol plug, and 25 s of electrokinetic injection (10 kV) of the sample were found to be the optimum conditions. Detection limits up to 25 times lower and similar precisions than those reported for hydrodynamic injection were obtained.     

Synthesis of cis-bicyco¦13.3.0¦oct-3-ene-2,7-dione,a highly functionalized cyclopentanoid intermediate

cis-Bicyclo¦3.3.O¦oct-3-ene-2,7-dione (1) has been synthesized either from cis-bicyclo¦3.3.0¦octane-3,7-dione (2) or the ¦2+2¦-cycloadduct 10 of 5-trimethylsilylcyclopentadiene with dichloroketene. As anticipated, the conjugated double bond of acetal 9 acts as a Michael acceptor in front of nucleophiles such as PhSeCH₂Li to give the exo-1,4-addition derivative 16, from which intermediates with functionalized endo-side chains are easily worked up.     

Field amplified sample injection-capillary electrophoresis-tandem mass spectrometry for the analysis of acrylamide in foodstuffs

This paper shows the applicability of capillary electrophoresis (CE) coupled to mass spectrometry (MS) for the analysis of acrylamide (AA) in foodstuffs. In order to obtain an ionisable compound amenable to be analysed by CE, acrylamide was derivatised with 2-mercaptobenzoic acid. Spectra in positive and negative modes were studied in order to select the best ionisation mode and multistep tandem mass spectrometry was used to obtain structural information. Maximum signal was observed when negative mode was used and MS/MS and MS3 were selected for quantitation and confirmation, respectively. For the separation, a fused-silica capillary of 80 cm and 50 microm I.D. and 35 mM ammonium formate/ammonia solution at pH 10 as running electrolyte were used. The applicability of field amplified sample injection (FASI) in reversed polarity was evaluated in order to decrease detection limits. The developed FASI-CE-MS/MS method provided a detection limit of 8 ng g(-1) and good linearity (r=0.999) and precision (day-to-day lower than 15%). The method has been applied to the analysis of different representative food products and the results were compared with those obtained by LC-MS/MS.     

CEC separation of heterocyclic amines using methacrylate monolithic columns

Two methacrylate-based monolithic columns, one with a negatively charged group (sulfonic group) and another with a new monomer N,N-dimethylamino ethyl acrylate (DMAEA), were prepared and tested for the separation of basic compounds by CEC. This new monolithic stationary phase was prepared by the in situ polymerization of DMAEA with butyl methacrylate and ethylene dimethacrylate, using a ternary porogenic solvent consisting of water, 1-propanol and 1,4-butanediol. The performance of this column was evaluated by means of the analysis of a family of heterocyclic amines. Separation conditions such as pH, amount of organic modifier, ionic strength and elution mode (normal or counterdirectional flow) were studied. At the optimal running electrolyte composition, and using the counterdirectional mode, symmetrical electrochromatographic peaks were obtained, with the number of theoretical plates up to 30,000 and a good resolution between closely related peaks. The 2-acrylamido-2-methyl-1-propane-sulfonic acid column was used for CEC-MS, taking advantage of the compatibility of its elution mode (normal flow) with the MS coupling.     

Enantioselective organocatalytic asymmetric allylic alkylation. Bis(phenylsulfonyl)methane addition to MBH carbonates

The highly enantioselective asymmetric allylic alkylation of Morita-Baylis-Hillman carbonates with bis(phenylsulfonyl)methane is presented. The reaction is simply catalyzed by cinchona alkaloid derivatives affording the final alkylated products in good yields and enantioselectivities.     

Atmospheric pressure ionization–tandem mass spectrometry of the phenicol drug family

In this work, the mass spectrometry behaviour of the veterinary drug family of phenicols, including chloramphenicol (CAP) and its related compounds thiamphenicol (TAP), florfenicol (FF) and FF amine (FFA), was studied. Several atmospheric pressure ionization sources, electrospray (ESI), atmospheric pressure chemical ionization and atmospheric pressure photoionization were compared. In all atmospheric pressure ionization sources, CAP, TAP and FF were ionized in both positive and negative modes; while for the metabolite FFA, only positive ionization was possible. In general, in positive mode, [M + H]⁺ dominated the mass spectrum for FFA, while the other compounds, CAP, TAP and FF, with lower proton affinity showed intense adducts with species present in the mobile phase. In negative mode, ESI and atmospheric pressure photoionization showed the deprotonated molecule [M–H]^?, while atmospheric pressure chemical ionization provided the radical molecular ion by electron capture. All these ions were characterized by tandem mass spectrometry using the combined information obtained by multistage mass spectrometry and high‐resolution mass spectrometry in a quadrupole‐Orbitrap instrument. In general, the fragmentation occurred via cyclization and losses or fragmentation of the N‐(alkyl)acetamide group, and common fragmentation pathways were established for this family of compounds. A new chemical structure for the product ion at m/z 257 for CAP, on the basis of the MS³ and MS⁴ spectra is proposed. Thermally assisted ESI and selected reaction monitoring are proposed for the determination of these compounds by ultra high‐performance liquid chromatography coupled to tandem mass spectrometry, achieving instrumental detection limits down to 0.1 pg. Copyright © 2013 John Wiley & Sons, Ltd.     

Spontaneous Emergence of Chirality in the Limited Enantioselectivity Model: Autocatalytic Cycle Driven by an External Reagent

The model of limited enantioselectivity (LES) in closed systems, and under experimental conditions able to achieve chemical equilibrium, can give rise to neither spontaneous mirror symmetry breaking (SMSB) nor kinetic chiral amplifications. However, it has been recently shown that it is able to lead to SMSB, as a stationary final state, in thermodynamic scenarios involving nonuniform temperature distributions and for compartmentalized separation between the two autocatalytic reactions. Herein, it is demonstrated how SMSB may occur in LES in a cyclic network with uniform temperature distribution if the reverse reaction of the nonenantioselective autocatalysis, which gives limited inhibition on the racemic mixture, is driven by an external reagent, that is, in conditions that keep the system out of chemical equilibrium. The exact stability analysis of the racemic and chiral final outcomes and the study of the reaction parameters leading to SMSB are resolved analytically. Numerical simulations, using chemical kinetics equations, show that SMSB may occur for chemically reasonable parameters. Numerical simulations on SMSB are also presented for speculative, but reasonable, scenarios implying reactions common in amino acid chemistry.     

Preventing false negatives with high-resolution mass spectrometry: the benzophenone case

Benzophenone (BP) is one of the many contaminants reported as present in foodstuffs due to its migration from food packaging materials. Liquid chromatography/tandem mass spectrometry (LC/MS/MS) is acknowledged in the literature as the method of choice for this analysis. However, cases have been reported where the use of this methodology was insufficient to unambiguously confirm the presence of a contaminant. In previous work performed by the authors, the unequivocal identification of BP in packaged foods was not possible even when monitoring two m/z transitions (precursor ion – product ion), since ion ratio errors higher than 20% were obtained. In order to overcome this analytical problem a fast, sensitive and selective liquid chromatography/high‐resolution mass spectrometry (LC/HRMS) methodology has been developed and applied to the analysis of BP in packaged foods. A direct comparison between LC/HRMS and LC/MS/MS data indicated better selectivity when working with LC/HRMS at a resolving power of 50 000 FWHM (full width at half maximum) than when monitoring two m/z transitions by LC/MS/MS. The resolving power used enabled the detection and identification of Harman as the compound impeding the confirmation of BP by LC‐MS/MS. Similar quantitative results were obtained by an Orbitrap mass analyser (Exactive?) and a triple quadrupole mass analyser (TSQ Quantum Ultra AM?). Copyright © 2011 John Wiley & Sons, Ltd.