Dalton T. Snyder

Miniature and Fieldable Mass Spectrometers: Recent Advances

Dalton T. Snyder,† Christopher J. Pulliam,† Zheng Ouyang,‡ and R. Graham Cooks*,† †Department of Chemistry and Center for Analytical Instrumentation Development, Purdue University, West Lafayette, Indiana 47907, United States ‡Weldon School of Biomedical Engineering, Purdue University, West Lafayette, Indiana 47907, United States ■ CONTENTS Ambient Ionization and Sampling 2 Vacuum Systems 4 Analyzers and Configurations 6 MEMS and Other Alternative Fabrication Methods 8 Simulations 9 Electronics and Data Systems 12 Portable Systems 19 Mesoscale Systems 20 Planetary and Space Science Systems 21 Practical Applications 22 Expectations 23 Author Information 23 Corresponding Author 23 Notes 23 Biographies 23 Acknowledgments 24 References 24

Experimental Characterization of Secular Frequency Scanning in Ion Trap Mass Spectrometers

AbstractSecular frequency scanning is implemented and characterized using both a benchtop linear ion trap and a miniature rectilinear ion trap mass spectrometer. Separation of tetraalkylammonium ions and those from a mass calibration mixture and from a pesticide mixture is demonstrated with peak widths approaching unit resolution for optimized conditions using the benchtop ion trap. The effects on the spectra of ion trap operating parameters, including waveform amplitude, scan direction, scan rate, and pressure are explored, and peaks at black holes corresponding to nonlinear (higher-order field) resonance points are investigated. Reverse frequency sweeps (increasing mass) on the Mini 12 are shown to result in significantly higher ion ejection efficiency and superior resolution than forward frequency sweeps that decrement mass. This result is accounted for by the asymmetry in ion energy absorption profiles as a function of AC frequency and the shift in ion secular frequency at higher amplitudes in the trap due to higher order fields. We also found that use of higher AC amplitudes in forward frequency sweeps biases ions toward ejection at points of higher order parametric resonance, despite using only dipolar excitation. Higher AC amplitudes also increase peak width and decrease sensitivity in both forward and reverse frequency sweeps. Higher sensitivity and resolution were obtained at higher trap pressures in the secular frequency scan, in contrast to conventional resonance ejection scans, which showed the opposite trend in resolution on the Mini 12. Mass range is shown to be naturally extended in secular frequency scanning when ejecting ions by sweeping the AC waveform through low frequencies, a method which is similar, but arguably superior, to the more usual method of mass range extension using low q resonance ejection. Graphical Abstractᅟ

Calibration procedure for secular frequency scanning in ion trap mass spectrometers

RATIONALE Mass spectra can be recorded using ion traps by scanning the frequency of an alternating current (ac) signal that corresponds to the secular frequency of a trapped ion. There is a considerable simplification in the instrumentation needed to perform such a scan compared with conventional scans of the radiofrequency (rf) amplitude. However, mass calibration is difficult. An algorithm that can be used to achieve mass calibration is investigated and the factors that affect ion mass assignments are discussed. METHODS Time domain data, recorded using a commercial benchtop linear ion trap mass spectrometer, are converted to the m/z domain using ion Mathieu parameter qu values which are derived from the dimensionless frequency parameter βu expressed as a continuing fraction in terms of qu . The relationship between the operating parameters of an ideal ion trap and the ion m/z ratio is derived from the Mathieu equations and expressed as an algorithm which through successive approximations yields the Mathieu qu value and hence m/z values and peak widths. The predictions of the algorithm are tested against experiment by sweeping the frequency of a small supplementary ac signal so as to cause mass-selective ejection of trapped ions. RESULTS Calibration accuracy is always better than 0.1%, often much better. Peak widths correspond to a mass resolution of 250 to 500 in the m/z 100-1800 range in secular frequency scans. CONCLUSIONS A simple, effective method of calibration of mass spectra recorded using secular frequency scans is achieved. The effects of rf amplitude, scan rate, and ac amplitude on calibration parameters are shown using LTQ linear ion trap data. Corrections for differences in ion mass must be made for accurate calibration, and this is easily incorporated into the calibration procedure. Copyright

Single analyzer precursor scans using an ion trap

RATIONALE Precursor ion and neutral loss scans are general survey methods of tandem mass spectrometry (MS/MS) used for detecting structurally related compounds. Until now they have been performed in multiple analyzer instruments, e.g. triple quadrupoles and hybrid MS/MS instruments. Implementation of precursor ion scans in single mass analyzers would be advantageous in reducing instrument complexity. METHODS Adoption of secular frequency scanning as a method of mass-selective excitation is shown to enable precursor scans in a single ion trap in a miniature mass spectrometer. A small supplementary alternating current (ac) signal is swept in frequency so as to cause mass-selective excitation of trapped ions. Simultaneously, a higher fixed amplitude ac signal is applied at the fixed secular frequency of a product ion, ejecting the mass-selected product ion and providing temporal data corresponding to a precursor ion spectrum. RESULTS Precursor scanning in a single ion trap is demonstrated using a mixture of three illicit drugs: cocaine, 3,4-methylenedioxyamphetamine (MDA), and 3,4-methylenedioxymethamphetamine (MDMA). Acquisition of the spectra as a function of the frequency of the product ejection waveform demonstrates that the signals acquired represent precursor ion scans. CONCLUSIONS Secular frequency scanning is a nonconventional method of mass scanning that in combination with product ion ejection enables precursor scans in single ion traps. This phenomenon is demonstrated here for a miniature linear ion trap, but the concepts described also apply to quadrupole mass filters. Copyright

Multigenerational Broadband Collision-Induced Dissociation of Precursor Ions in a Linear Quadrupole Ion Trap

AbstractA method of fragmenting ions over a wide range of m/z values while balancing energy deposition into the precursor ion and available product ion mass range is demonstrated. In the method, which we refer to as “multigenerational collision-induced dissociation”, the radiofrequency (rf) amplitude is first increased to bring the lowest m/z of the precursor ion of interest to just below the boundary of the Mathieu stability diagram (q = 0.908). A supplementary AC signal at a fixed Mathieu q in the range 0.2–0.35 (chosen to balance precursor ion potential well depth with available product ion mass range) is then used for ion excitation as the rf amplitude is scanned downward, thus fragmenting the precursor ion population from high to low m/z. The method is shown to generate high intensities of product ions compared with other broadband CID methods while retaining low mass ions during the fragmentation step, resulting in extensive fragment ion coverage for various components of complex mixtures. Because ions are fragmented from high to low m/z, space charge effects are minimized and multiple discrete generations of product ions are produced, thereby giving rise to “multigenerational” product ion mass spectra. Graphical Abstractᅟ

Ion isolation and multigenerational collision-induced dissociation using the inverse Mathieu q scan: Isolation and fragmentation using inverse Mathieu q scan

RATIONALE In a bid to develop a mass spectrometer using ac frequency scanning for ion isolation, ion activation, and ion ejection, we have developed scan functions for each process using the inverse Mathieu q scan. METHODS Ion isolation is accomplished by frequency hopping, that is, by skipping past the ranges of frequencies corresponding to the ions to be isolated during the frequency sweep. Multigenerational collision-induced dissociation is demonstrated by scanning the frequency of excitation from low to high so that multiple generations of product ions can be observed in the product ion mass spectra. Because the excitation frequency is scanned quickly across a large range, fragmentation of some precursor ions can be too limited. However, by first fixing the excitation frequency on the precursor ion and then scanning the frequency using the inverse Mathieu q scan, a higher abundance of product ions can be obtained. RESULTS Isolation of a single mass-to-charge (m/z) as well as nonadjacent m/z ions is demonstrated with isolation efficiency greater than 70%. Fragmentation of caffeine and noroxycodone is demonstrated, the latter of which shows multiple generations of product ions. CONCLUSIONS The results demonstrated here provide strong evidence that an ion trap mass spectrometer can be operated without using an rf amplitude ramp for any operation, and instead ac frequency scanning can be used for all mass-selective operations. Copyright

Analysis of bacteria using zero volt paper spray

The application of zero volt paper spray to the discrimination between species of bacteria is demonstrated here. While absolute signal intensities of representative lipids from bacterial membranes were three orders of magnitude lower than for conventional paper spray performed at high potential (kilovolts), the significant reduction in noise offset this disadvantage, resulting in clear separation in principal component analysis space between Gram positive and Gram negative bacteria as well as excellent separation between bacteria species.

Simultaneous and Sequential MS/MS Scan Combinations and Permutations in a Linear Quadrupole Ion Trap

Methods of performing precursor ion scans as well as neutral loss scans in a single linear quadrupole ion trap have recently been described. In this paper we report methodology for performing permutations of MS/MS scan modes, that is, ordered combinations of precursor, product, and neutral loss scans following a single ion injection event. Only particular permutations are allowed; the sequences demonstrated here are (1) multiple precursor ion scans, (2) precursor ion scans followed by a single neutral loss scan, (3) precursor ion scans followed by product ion scans, and (4) segmented neutral loss scans. (5) The common product ion scan can be performed earlier in these sequences, under certain conditions. Simultaneous scans can also be performed. These include multiple precursor ion scans, precursor ion scans with an accompanying neutral loss scan, and multiple neutral loss scans. We argue that the new capability to perform complex simultaneous and sequential MSn operations on single ion populations represents a significant step in increasing the selectivity of mass spectrometry.

Single Analyzer Neutral Loss Scans in a Linear Quadrupole Ion Trap Using Orthogonal Double Resonance Excitation

In this follow-up paper to our previous work on single analyzer precursor ion scans in a linear quadrupole ion trap (Snyder, D. T.; Cooks, R. G. Single analyzer precursor ion scans in a linear quadrupole ion trap using orthogonal double resonance excitation. J. Am. Soc. Mass Spectrom. 2017, DOI: 10.1007/s13361-017-1707-y), we now report the development of single analyzer neutral loss scans in a linear quadrupole ion trap using orthogonal double resonance excitation. Methodologically, there are three key differences between single analyzer precursor ion scans and neutral loss scans under constant radiofrequency (rf) conditions: (1) in the latter experiment, both excitation and ejection frequencies must be scanned, whereas in the former the ejection frequency is fixed, (2) the need to maintain a constant neutral loss while incrementing both precursor and product ion masses, complicated by the complex relationship between secular frequency and mass, requires use of two simultaneous frequency scans, both linear in mass, and (3) because the ejection frequency is scanned, a third ac signal occurring between the ac excitation and ac ejection frequency scans must also be applied and scanned in order to reject artifact peaks caused by ejection of unfragmented precursor ions. Using this methodology, we demonstrate neutral loss scans on a commercial linear ion trap using mixtures of illicit drugs and acylcarnitines. We also demonstrate neutral loss scanning on a Populus deltoides leaf and on a lignin sample, both significantly more complex mixtures.

Successive Resonances for Ion Ejection at Arbitrary Frequencies in an Ion Trap

AbstractThe use of successive resonances for ion ejection is demonstrated here as a method of scanning quadrupole ion traps with improvement in both resolution and sensitivity compared with single frequency resonance ejection. The conventional single frequency resonance ejection waveform is replaced with a dual-frequency waveform. The two included frequencies are spaced very closely and their relative amplitudes are adjusted so that the first frequency that ions encounter excites them to higher amplitudes where space charge effects are less prominent, thereby giving faster and more efficient ejection when the ions come into resonance with the second frequency. The method is applicable at any arbitrary frequency, unlike double and triple resonance methods. However, like double and triple resonance ejection, ejection using successive resonances requires the rf and AC waveforms to be phase-locked in order to retain mass accuracy and mass precision. The improved performance is seen in mass spectra acquired by rf amplitude scans (resonance ejection) as well as by secular frequency scans. Graphical Abstractᅟ