The Experts below are selected from a list of 29805 Experts worldwide ranked by ideXlab platform
Alexander Makarov - One of the best experts on this subject based on the ideXlab platform.
-
Resolving heterogeneous high-mass macromolecular machineries by Orbitrap-based single particle charge detection mass spectrometry
2019Co-Authors: Albert J. R. Heck, Tobias P. Woerner, Joost Snijder, Antonette Bennett, Mavis Agbandje-mckenna, Alexander MakarovAbstract:Abstract Here we show that single particle charge-detection mass spectrometry (CD-MS) can be performed on a ubiquitous Orbitrap mass analyser and applied to the analysis of high-mass (megadalton) heterogeneous biomolecular assemblies. We demonstrate that single particle high-mass ions can survive in the Orbitrap for seconds, whereby their measured signal amplitudes scale linearly with charge state over the entire m/z range. Orbitrap based single particle CD-MS can be used to resolve mixed ion populations, accurately predict charge states, and consequently also the mass of the ions. We successfully applied CD-MS to challenging natural and biotherapeutic protein assemblies, such as IgM oligomers, designed protein nano-cages, ribosome particles and intact, empty- and genome-loaded Adeno-associated virus particles. Single particle CD-MS combined with native MS on existing Orbitrap platforms will greatly expand its application, especially in the mass analysis of megadalton heterogeneous biomolecular assemblies.
-
Surface-Induced Dissociation of Noncovalent Protein Complexes in an Extended Mass Range Orbitrap Mass Spectrometer
Analytical chemistry, 2019Co-Authors: Zachary Vanaernum, Stevan Horning, Alexander Makarov, Joshua D. Gilbert, Mikhail E. Belov, Vicki H. WysockiAbstract:Native mass spectrometry continues to develop as a significant complement to traditional structural biology techniques. Within native mass spectrometry (MS), surface-induced dissociation (SID) has been shown to be a powerful activation method for the study of noncovalent complexes of biological significance. High-resolution mass spectrometers have become increasingly adapted to the analysis of high-mass ions and have demonstrated their importance in understanding how small mass changes can affect the overall structure of large biomolecular complexes. Herein we demonstrate the first adaptation of surface-induced dissociation in a modified high-mass-range, high-resolution Orbitrap mass spectrometer. The SID device was designed to be installed in the Q Exactive series of Orbitrap mass spectrometers with minimal disruption of standard functions. The performance of the SID-Orbitrap instrument has been demonstrated with several protein complex and ligand-bound protein complex systems ranging from 53 to 336 kDa....
-
Fundamentals of Orbitrap analyzer
Fundamentals and Applications of Fourier Transform Mass Spectrometry, 2019Co-Authors: Alexander Makarov, Dmitry Grinfeld, Konstantin AyzikovAbstract:Abstract This chapter discusses fundamentals of the Orbitrap™ mass spectrometry—the use of a purely electrostatic ion trap with harmonic properties to store ions and analyze their mass-to-charge distributions. The first section demonstrates principles of ion confinement in the quadro-logarithmic electrostatic field, where the ions perform hundreds of thousands of oscillations without losing their common phase. The discussion covers practical aspects of ion injection into the trap and the image current detection. The subsequent section introduces the reader to the Orbitrap aberration theory, gives a classification of electric field perturbations, and outlines the compensation methods. The next section concerns signal processing methods of the detected transients and covers both the traditional Fourier transform approaches and super-FT resolving methods. The chapter concludes with the overview of historical milestones of the Orbitrap technology and selected applications.
-
Coupling liquid chromatography to Orbitrap mass spectrometry
2017Co-Authors: Alexander Makarov, Michaela ScigelovaAbstract:The Orbitrap mass analyzer has become a mainstream mass spectrometry technique. In addition to providinga brief introduction to the Orbitrap technology and its continuing development, this article reviewsthe most recent publications quoting the use of the Orbitrap detection for a variety of chromatographicseparation techniques. Its coupling to reversed-phase liquid chromatography (LC) represents undoubtedlythe most ubiquitous approach to both small molecule and proteomic analyses. Multi-dimensional LCseparations have an important role to play in the proteomics applications while an ultra-high-pressureLC is more frequently encountered in the area of metabolomics and metabolite analysis. Recently, specialchromatographic techniques such as hydrophilic interaction chromatography and its variations have alsobeen also cited with the Orbitrap detection.
-
Evolution of Orbitrap Mass Spectrometry Instrumentation.
Annual review of analytical chemistry (Palo Alto Calif.), 2015Co-Authors: Shannon Eliuk, Alexander MakarovAbstract:We discuss the evolution of Orbitrap mass spectrometry (MS) from its birth in the late 1990s to its current role as one of the most prominent techniques for MS. The Orbitrap mass analyzer is the first high-performance mass analyzer that employs trapping of ions in electrostatic fields. Tight integration with the ion injection process enables the high-resolution, mass accuracy, and sensitivity that have become essential for addressing analytical needs in numerous areas of research, as well as in routine analysis. We examine three major families of instruments (related to the LTQ Orbitrap, Q Exactive, and Orbitrap Fusion mass spectrometers) in the context of their historical development over the past ten eventful years. We discuss as well future trends and perspectives of Orbitrap MS. We illustrate the compelling potential of Orbitrap-based mass spectrometers as (ultra) high-resolution platforms, not only for high-end proteomic applications, but also for routine targeted analysis.
Stevan Horning - One of the best experts on this subject based on the ideXlab platform.
-
Surface-Induced Dissociation of Noncovalent Protein Complexes in an Extended Mass Range Orbitrap Mass Spectrometer
Analytical chemistry, 2019Co-Authors: Zachary Vanaernum, Stevan Horning, Alexander Makarov, Joshua D. Gilbert, Mikhail E. Belov, Vicki H. WysockiAbstract:Native mass spectrometry continues to develop as a significant complement to traditional structural biology techniques. Within native mass spectrometry (MS), surface-induced dissociation (SID) has been shown to be a powerful activation method for the study of noncovalent complexes of biological significance. High-resolution mass spectrometers have become increasingly adapted to the analysis of high-mass ions and have demonstrated their importance in understanding how small mass changes can affect the overall structure of large biomolecular complexes. Herein we demonstrate the first adaptation of surface-induced dissociation in a modified high-mass-range, high-resolution Orbitrap mass spectrometer. The SID device was designed to be installed in the Q Exactive series of Orbitrap mass spectrometers with minimal disruption of standard functions. The performance of the SID-Orbitrap instrument has been demonstrated with several protein complex and ligand-bound protein complex systems ranging from 53 to 336 kDa....
-
Development of a GC/Quadrupole-Orbitrap mass spectrometer, part I: design and characterization.
Analytical chemistry, 2014Co-Authors: Amelia Peterson, Oliver Lange, Stevan Horning, Jan Peter Hauschild, Scott T. Quarmby, Dirk Krumwiede, Rachelle A. S. Lemke, Florian Grosse-coosmann, Timothy J. Donohue, Michael S. WestphallAbstract:Identification of unknown compounds is of critical importance in GC/MS applications (metabolomics, environmental toxin identification, sports doping, petroleomics, and biofuel analysis, among many others) and remains a technological challenge. Derivation of elemental composition is the first step to determining the identity of an unknown compound by MS, for which high accuracy mass and isotopomer distribution measurements are critical. Here, we report on the development of a dedicated, applications-grade GC/MS employing an Orbitrap mass analyzer, the GC/Quadrupole-Orbitrap. Built from the basis of the benchtop Orbitrap LC/MS, the GC/Quadrupole-Orbitrap maintains the performance characteristics of the Orbitrap, enables quadrupole-based isolation for sensitive analyte detection, and includes numerous analysis modalities to facilitate structural elucidation. We detail the design and construction of the instrument, discuss its key figures-of-merit, and demonstrate its performance for the characterization of u...
-
Mass Spectrometry-based Proteomics Using Q Exactive, a High-performance Benchtop Quadrupole Orbitrap Mass Spectrometer
Molecular & Cellular Proteomics, 2011Co-Authors: Annette Michalski, Oliver Lange, Andreas Wieghaus, Jan Peter Hauschild, Juergen Cox, Nagarjuna Nagaraj, Alexander Makarov, Eugen Damoc, Matthias Mann, Stevan HorningAbstract:Mass spectrometry-based proteomics has greatly benefitted from enormous advances in high resolution instrumentation in recent years. In particular, the combination of a linear ion trap with the Orbitrap analyzer has proven to be a popular instrument configuration. Complementing this hybrid trap-trap instrument, as well as the standalone Orbitrap analyzer termed Exactive, we here present coupling of a quadrupole mass filter to an Orbitrap analyzer. This "Q Exactive" instrument features high ion currents because of an S-lens, and fast high-energy collision-induced dissociation peptide fragmentation because of parallel filling and detection modes. The image current from the detector is processed by an "enhanced Fourier Transformation" algorithm, doubling mass spectrometric resolution. Together with almost instantaneous isolation and fragmentation, the instrument achieves overall cycle times of 1 s for a top10 higher energy collisional dissociation method. More than 2500 proteins can be identified in standard 90-min gradients of tryptic digests of mammalian cell lysate- a significant improvement over previous Orbitrap mass spectrometers. Furthermore, the quadrupole Orbitrap analyzer combination enables multiplexed operation at the MS and tandem MS levels. This is demonstrated in a multiplexed single ion monitoring mode, in which the quadrupole rapidly switches among different narrow mass ranges that are analyzed in a single composite MS spectrum. Similarly, the quadrupole allows fragmentation of different precursor masses in rapid succession, followed by joint analysis of the higher energy collisional dissociation fragment ions in the Orbitrap analyzer. High performance in a robust benchtop format together with the ability to perform complex multiplexed scan modes make the Q Exactive an exciting new instrument for the proteomics and general analytical communities.
-
a proteomics grade electron transfer dissociation enabled hybrid linear ion trap Orbitrap mass spectrometer
Journal of Proteome Research, 2008Co-Authors: Graeme C Mcalister, Stevan Horning, Alexander Makarov, Travis W Berggren, Jens Griepraming, Douglas H Phanstiel, George C Stafford, Danielle L Swaney, John E P Syka, Vlad ZabrouskovAbstract:Here we detail the modification of a quadrupole linear ion trap-Orbitrap hybrid (QLT-Orbitrap) mass spectrometer to accommodate a negative chemical ionization (NCI) source. The NCI source is used to produce fluoranthene radical anions for imparting electron transfer dissociation (ETD). The anion beam is stable, robust, and intense so that a sufficient amount of reagents can be injected into the QLT in only 4−8 ms. Following ion/ion reaction in the QLT, ETD product ions are mass-to-charge (m/z) analyzed in either the QLT (for speed and sensitivity) or the Orbitrap (for mass resolution and accuracy). Here we describe the physical layout of this device, parametric optimization of anion transport, an evaluation of relevant ETD figures of merit, and the application of this instrument to protein sequence analysis. Described proteomic applications include complex peptide mixture analysis, post-translational modification (PTM) site identification, isotope-encoded quantitation, large peptide characterization, and ...
-
higher energy c trap dissociation for peptide modification analysis
Nature Methods, 2007Co-Authors: Jesper V. Olsen, Oliver Lange, Stevan Horning, Alexander Makarov, Boris Macek, Matthias MannAbstract:Peptide sequencing is the basis of mass spectrometry–driven proteomics. Here we show that in the linear ion trap–Orbitrap mass spectrometer (LTQ Orbitrap) peptide ions can be efficiently fragmented by high-accuracy and full-mass-range tandem mass spectrometry (MS/MS) via higher-energy C-trap dissociation (HCD). Immonium ions generated via HCD pinpoint modifications such as phosphotyrosine with very high confidence. Additionally we show that an added octopole collision cell facilitates de novo sequencing.
Matthias Mann - One of the best experts on this subject based on the ideXlab platform.
-
Mass Spectrometry-based Proteomics Using Q Exactive, a High-performance Benchtop Quadrupole Orbitrap Mass Spectrometer
Molecular & Cellular Proteomics, 2011Co-Authors: Annette Michalski, Oliver Lange, Andreas Wieghaus, Jan Peter Hauschild, Juergen Cox, Nagarjuna Nagaraj, Alexander Makarov, Eugen Damoc, Matthias Mann, Stevan HorningAbstract:Mass spectrometry-based proteomics has greatly benefitted from enormous advances in high resolution instrumentation in recent years. In particular, the combination of a linear ion trap with the Orbitrap analyzer has proven to be a popular instrument configuration. Complementing this hybrid trap-trap instrument, as well as the standalone Orbitrap analyzer termed Exactive, we here present coupling of a quadrupole mass filter to an Orbitrap analyzer. This "Q Exactive" instrument features high ion currents because of an S-lens, and fast high-energy collision-induced dissociation peptide fragmentation because of parallel filling and detection modes. The image current from the detector is processed by an "enhanced Fourier Transformation" algorithm, doubling mass spectrometric resolution. Together with almost instantaneous isolation and fragmentation, the instrument achieves overall cycle times of 1 s for a top10 higher energy collisional dissociation method. More than 2500 proteins can be identified in standard 90-min gradients of tryptic digests of mammalian cell lysate- a significant improvement over previous Orbitrap mass spectrometers. Furthermore, the quadrupole Orbitrap analyzer combination enables multiplexed operation at the MS and tandem MS levels. This is demonstrated in a multiplexed single ion monitoring mode, in which the quadrupole rapidly switches among different narrow mass ranges that are analyzed in a single composite MS spectrum. Similarly, the quadrupole allows fragmentation of different precursor masses in rapid succession, followed by joint analysis of the higher energy collisional dissociation fragment ions in the Orbitrap analyzer. High performance in a robust benchtop format together with the ability to perform complex multiplexed scan modes make the Q Exactive an exciting new instrument for the proteomics and general analytical communities.
-
proteomics on an Orbitrap benchtop mass spectrometer using all ion fragmentation
Molecular & Cellular Proteomics, 2010Co-Authors: Tamar Geiger, Matthias MannAbstract:The Orbitrap mass analyzer combines high sensitivity, high resolution, and high mass accuracy in a compact format. In proteomics applications, it is used in a hybrid configuration with a linear ion trap (LTQ-Orbitrap) where the linear trap quadrupole (LTQ) accumulates, isolates, and fragments peptide ions. Alternatively, isolated ions can be fragmented by higher energy collisional dissociation. A recently introduced stand-alone Orbitrap analyzer (Exactive) also features a higher energy collisional dissociation cell but cannot isolate ions. Here we report that this instrument can efficiently characterize protein mixtures by alternating MS and “all-ion fragmentation” (AIF) MS/MS scans in a manner similar to that previously described for quadrupole time-of-flight instruments. We applied the peak recognition algorithms of the MaxQuant software at both the precursor and product ion levels. Assignment of fragment ions to co-eluting precursor ions was facilitated by high resolution (100,000 at m/z 200) and high mass accuracy. For efficient fragmentation of different mass precursors, we implemented a stepped collision energy procedure with cumulative MS readout. AIF on the Exactive identified 45 of 48 proteins in an equimolar protein standard mixture and all of them when using a small database. The technique also identified proteins with more than 100-fold abundance differences in a high dynamic range standard. When applied to protein identification in gel slices, AIF unambiguously characterized an immunoprecipitated protein that was barely visible by Coomassie staining and quantified it relative to contaminating proteins. AIF on a benchtop Orbitrap instrument is therefore an attractive technology for a wide range of proteomics analyses.
-
higher energy c trap dissociation for peptide modification analysis
Nature Methods, 2007Co-Authors: Jesper V. Olsen, Oliver Lange, Stevan Horning, Alexander Makarov, Boris Macek, Matthias MannAbstract:Peptide sequencing is the basis of mass spectrometry–driven proteomics. Here we show that in the linear ion trap–Orbitrap mass spectrometer (LTQ Orbitrap) peptide ions can be efficiently fragmented by high-accuracy and full-mass-range tandem mass spectrometry (MS/MS) via higher-energy C-trap dissociation (HCD). Immonium ions generated via HCD pinpoint modifications such as phosphotyrosine with very high confidence. Additionally we show that an added octopole collision cell facilitates de novo sequencing.
-
top down protein sequencing and ms3 on a hybrid linear quadrupole ion trap Orbitrap mass spectrometer
Molecular & Cellular Proteomics, 2006Co-Authors: Boris Macek, Jesper V. Olsen, Leonie F Waanders, Matthias MannAbstract:Top-down proteomics, the analysis of intact proteins (instead of first digesting them to peptides), has the potential to become a powerful tool for mass spectrometric protein characterization. Requirements for extremely high mass resolution, accuracy, and ability to efficiently fragment large ions have often limited top-down analyses to custom built FT-ICR mass analyzers. Here we explore the hybrid linear ion trap (LTQ)-Orbitrap, a novel, high performance, and compact mass spectrometric analyzer, for top-down proteomics. Protein standards from 10 to 25 kDa were electrosprayed into the instrument using a nanoelectrospray chip. Resolving power of 60,000 was ample for isotope resolution of all protein charge states. We achieved absolute mass accuracies for intact proteins between 0.92 and 2.8 ppm using the "lock mass" mode of operation. Fifty femtomole of cytochrome c applied to the chip resulted in spectra with excellent signal-to-noise ratio and only low attomole sample consumption. Different protein charge states were dissociated in the LTQ, and the sensitivity of the Orbitrap allowed routine, high resolution, and high mass accuracy fragment detection. This resulted in unambiguous charge state determination of fragment ions and identification of unmodified and modified proteins by database searching. Protein fragments were further isolated and fragmented in the LTQ followed by analysis of MS(3) fragments in the Orbitrap, localizing modifications to part of the sequence and helping to identify the protein with these small peptide-like fragments. Given the ready availability and ease of operation of the LTQ-Orbitrap, it may have significant impact on top-down proteomics.
Oliver Lange - One of the best experts on this subject based on the ideXlab platform.
-
Reprint of “Enhanced Fourier transform for Orbitrap mass spectrometry”
International Journal of Mass Spectrometry, 2015Co-Authors: Oliver Lange, Andreas Wieghaus, Eugen Damoc, Alexander MakarovAbstract:Abstract A novel method for processing of periodic signals, which combines absorption spectra presentation with magnitude spectra and finite-impulse-response filtering, is applied to image current transients acquired in Orbitrap mass spectrometry. Phasing of signal for absorption spectra is facilitated by the excitation-by-injection mechanism of forming coherent ion packets in the Orbitrap analyzer. In conjunction with extensive refinement of the trap and electronics design of the Orbitrap analyzer, this method allows completion of the excitation process and initiation of detection within a fraction of a millisecond after ejection of ions from an external storage device into the Orbitrap analyzer, thus avoiding baseline roll and large non-linear phase corrections. For most real-life analytes with limited signal decay over acquisition time, the method achieves a 2-fold increase of resolving power relative to the traditional Fourier transform processing method. For rapidly decaying signals of intact proteins, the relative increase in resolving power is reduced to about 1.4, which accords with theory. Peak shape and mass accuracy in LC/MS measurements obtained employing the novel method appear to be similar to those observed in traditional magnitude-mode FT spectra. Performance of the method was optimized using calibration mixtures and tested on different real-life samples, including complex peptide mixtures and proteins supplied to the mass spectrometer in infusion and LC/MS modes.
-
Development of a GC/Quadrupole-Orbitrap mass spectrometer, part I: design and characterization.
Analytical chemistry, 2014Co-Authors: Amelia Peterson, Oliver Lange, Stevan Horning, Jan Peter Hauschild, Scott T. Quarmby, Dirk Krumwiede, Rachelle A. S. Lemke, Florian Grosse-coosmann, Timothy J. Donohue, Michael S. WestphallAbstract:Identification of unknown compounds is of critical importance in GC/MS applications (metabolomics, environmental toxin identification, sports doping, petroleomics, and biofuel analysis, among many others) and remains a technological challenge. Derivation of elemental composition is the first step to determining the identity of an unknown compound by MS, for which high accuracy mass and isotopomer distribution measurements are critical. Here, we report on the development of a dedicated, applications-grade GC/MS employing an Orbitrap mass analyzer, the GC/Quadrupole-Orbitrap. Built from the basis of the benchtop Orbitrap LC/MS, the GC/Quadrupole-Orbitrap maintains the performance characteristics of the Orbitrap, enables quadrupole-based isolation for sensitive analyte detection, and includes numerous analysis modalities to facilitate structural elucidation. We detail the design and construction of the instrument, discuss its key figures-of-merit, and demonstrate its performance for the characterization of u...
-
Enhanced Fourier transform for Orbitrap mass spectrometry
International Journal of Mass Spectrometry, 2014Co-Authors: Oliver Lange, Andreas Wieghaus, Eugen Damoc, Alexander MakarovAbstract:Abstract A novel method for processing of periodic signals, which combines absorption spectra presentation with magnitude spectra and finite-impulse-response filtering, is applied to image current transients acquired in Orbitrap mass spectrometry. Phasing of signal for absorption spectra is facilitated by the excitation-by-injection mechanism of forming coherent ion packets in the Orbitrap analyzer. In conjunction with extensive refinement of the trap and electronics design of the Orbitrap analyzer, this method allows completion of the excitation process and initiation of detection within a fraction of a millisecond after ejection of ions from an external storage device into the Orbitrap analyzer, thus avoiding baseline roll and large non-linear phase corrections. For most real-life analytes with limited signal decay over acquisition time, the method achieves a 2-fold increase of resolving power relative to the traditional Fourier transform processing method. For rapidly decaying signals of intact proteins, the relative increase in resolving power is reduced to about 1.4, which accords with theory. Peak shape and mass accuracy in LC/MS measurements obtained employing the novel method appear to be similar to those observed in traditional magnitude-mode FT spectra. Performance of the method was optimized using calibration mixtures and tested on different real-life samples, including complex peptide mixtures and proteins supplied to the mass spectrometer in infusion and LC/MS modes.
-
Orbitrap mass spectrometry with resolving powers above 1,000,000
International Journal of Mass Spectrometry, 2012Co-Authors: Eduard Denisov, Oliver Lange, Eugen Damoc, Alexander MakarovAbstract:Experiments on a modified Orbitrap Elite™ instrument have shown that Orbitrap™ technology is capable of resolving power in excess of 1,000,000 when appropriate tolerance and tuning requirements are met. Such levels are achieved for m/z < 300–400 within a 3 s detection time which makes it compatible with several types of chromatographic separations. It was shown that resolving powers over 500,000–1,000,000 allow resolution of fine isotopic structure and direct identification of some important post-translational modifications of peptides.
-
ultra high resolution linear ion trap Orbitrap mass spectrometer Orbitrap elite facilitates top down lc ms ms and versatile peptide fragmentation modes
Molecular & Cellular Proteomics, 2012Co-Authors: Annette Michalski, Oliver Lange, Eugen Damoc, Jae C. Schwartz, Eduard Denisov, Philip M. Remes, Dirk Nolting, Mathias Muller, Rosa Viner, Michael W BelfordAbstract:Although only a few years old, the combination of a linear ion trap with an Orbitrap analyzer has become one of the standard mass spectrometers to characterize proteins and proteomes. Here we describe a novel version of this instrument family, the Orbitrap Elite, which is improved in three main areas. The ion transfer optics has an ion path that blocks the line of sight to achieve more robust operation. The tandem MS acquisition speed of the dual cell linear ion trap now exceeds 12 Hz. Most importantly, the resolving power of the Orbitrap analyzer has been increased twofold for the same transient length by employing a compact, high-field Orbitrap analyzer that almost doubles the observed frequencies. An enhanced Fourier Transform algorithm—incorporating phase information—further doubles the resolving power to 240,000 at m/z 400 for a 768 ms transient. For top-down experiments, we combine a survey scan with a selected ion monitoring scan of the charge state of the protein to be fragmented and with several HCD microscans. Despite the 120,000 resolving power for SIM and HCD scans, the total cycle time is within several seconds and therefore suitable for liquid chromatography tandem MS. For bottom-up proteomics, we combined survey scans at 240,000 resolving power with data-dependent collision-induced dissociation of the 20 most abundant precursors in a total cycle time of 2.5 s—increasing protein identifications in complex mixtures by about 30%. The speed of the Orbitrap Elite furthermore allows scan modes in which complementary dissociation mechanisms are routinely obtained of all fragmented peptides.
Vadims Bartkevics - One of the best experts on this subject based on the ideXlab platform.
-
Analytical capabilities of high performance liquid chromatography - Atmospheric pressure photoionization - Orbitrap mass spectrometry (HPLC-APPI-Orbitrap-MS) for the trace determination of novel and emerging flame retardants in fish.
Analytica chimica acta, 2015Co-Authors: Dzintars Zacs, Vadims BartkevicsAbstract:A new analytical method was established and validated for the analysis of 27 brominated flame retardants (BFRs), including so called "emerging" and "novel" BFRs (EBFRs and NBFRs) in fish samples. High performance liquid chromatography (HPLC) coupled to Orbitrap mass spectrometry (Orbitrap-MS) employing atmospheric pressure photoionization (APPI) interface operated in negative mode was used for the identification/quantitation of contaminants. HPLC-Orbitrap-MS analysis provided a fast separation of selected analytes within 14 min, thus demonstrating a high throughput processing of samples. The developed methodology was tested by intralaboratory validation in terms of recovery, repeatability, linear calibration ranges, instrumental and method limits of quantitation (i-LOQ and m-LOQ), and where possible, trueness was verified by analysis of certified reference materials (CRMs). Recoveries of analytes were between 80 and 119%, while the repeatability in terms of relative standard deviations (RSDs) was in the range from 1.2 to 15.5%. The measured values for both analyzed CRMs agreed with the provided consensus values, revealing the recovery of reference concentrations in 72-119% range. The elaborated method met the sensitivity criterion according to Commission Recommendation 2014/118/EU on monitoring of BFRs in food products for majority of the compounds. The concentrations of polybrominated diphenyl ethers (PBDEs) in real samples determined by HPLC-APPI-Orbitrap-MS method and validated gas chromatography-high-resolution mass spectrometry (GC-HRMS) method were found to be in a good agreement.
-
the development and validation of a rapid method for the determination of antimicrobial agent residues in milk and meat using ultra performance liquid chromatography coupled to quadrupole Orbitrap mass spectrometry
Journal of Pharmaceutical and Biomedical Analysis, 2015Co-Authors: Guntis Cepurnieks, Jekaterina Rjabova, Dzintars Zacs, Vadims BartkevicsAbstract:Abstract A new multi-class method has been developed for the identification and quantification of the residues of 26 antibiotics from different classes (sulfonamides, macrolides, tetracyclines, penicillins, and quinolones) in milk and meat by ultra performance liquid chromatography coupled to hybrid quadrupole – high resolution Orbitrap mass spectrometry (UPLC–qOrbitrap). The sample preparation included extraction of two analytical portions with acetonitrile and 5% trichloroacetic acid, respectively, followed by centrifugation and filtration. The method was validated over three days at 50% of MRL (maximum residue limit) set in the European Union. Experiments on spiked meat and milk samples showed that the average recovery of the antibiotics ranged from 83% to 112%, and the coefficients of variation were between 8.9% and 39%.