Development of a high-Performance MALDI-TOF mass spectrometer utilizing a spiral ion trajectory

A novel MALDI-TOF mass spectrometer that utilizes a spiral ion trajectory was developed. In this mass spectrometer, the ions sequentially passed through four toroidal electrostatic sectors and revolved along a figure-eight-shaped orbit on a particular projection plane. Each toroidal electrostatic sector had eight stories, and during multiple revolutions, the ion trajectory shifted perpendicular to the projection plane in every cycle, thereby generating a spiral trajectory. The flight path length of one cycle was 2.1 m; therefore, when the ions completed eight cycles, the total flight path length was 17 m. By adopting an ion optical system that had a flight path length five times longer than that in the commonly used reflectron ion optical system, the mass dependence on the mass resolving power was reduced, while improving the mass accuracy of the mass measurements. The basic performance of the system was tested by using standard peptides or the tryptic digest of bovine serum albumin. A mass resolving power of 80,000 (full width at half maximum) was achieved at m/z = 2564 (ACTH18-39). An improved mass accuracy less than 2 ppm was realized over a wide m/z range of 500 to 3000 by correction using one or two internal standard substances.     

High-resolution time-of-flight spectra obtained using the MULTUM II multi-turn type time-of-flight mass spectrometer with an electron ionization ion source

This paper describes experiments demonstrating the high mass-resolving power of the MULTUM II multi-turn type time-of-flight (ToF) mass spectrometer with a 1.308-meter circuit controlled by four toroidal electric sector fields(1) and an electron ionization (EI) ion source. A mass resolution of 250,000 [full-width at half maximum: (FWHM)] was obtained for N(2)(+) after a flight time of 9.0 ms (flight cycles: 1,200, flight length: 1,500 M). A doublet of (12)C(5)H(5)(14)N and (13)C(12)C(5)H(6) (m/Deltam = 9,746; Deltam: mass difference of doublet, m: mass of lighter ion of doublet) was separated and a mass resolution of 91,000 (FWHM) was obtained. A doublet of CDCl(2) and CH(2)Cl(2) (m/Deltam = 54,162) was also separated. A mass resolution of 115,000 (FWHM) was then achieved. When one peak of these doublets was used as a calibrant, the mass of the other peak was determined within a few ppm by mass difference. The ToF depending on the square of m/z was significantly larger than the systematic errors in the ToF, so that good mass accuracy was obtained by one-point mass determination.     

Multi-turn time-of-flight mass spectrometers with electrostatic sectors

The mass resolution of a time-of-flight (TOF) mass spectrometer is directly proportional to its total flight pathlength. Multi-turn or multi-passage ion optical geometries are necessary to obtain fight distances of sufficient length within reasonable size limitations. We have investigated ion optics for a multi-turn TOF mass spectrometer with electrostatic sectors. The concept of 'perfect' focusing conditions is introduced. Furthermore, a new type of multi-turn TOF mass spectrometer, the MULTUM Linear plus, was developed. It consists of four cylindrical electric sectors and 28 electric quadrupole lenses. It has a vacuum chamber 60 x 70 x 20 cm in size. Mass resolution is demonstrated to increase according to the number of ion cycles. A mass resolution of 350 000 (m/z = 28, FWHM) was achieved after 501.5 cycles. The MULTUM Linear plus analyzer is not simple, however; 28 electric quadrupole lenses are used. In order to reduce the number of ion optical parts, an improved multi-turn TOF mass spectrometer, the MULTUM II, consisting of only four toroidal electric sectors, was also developed. The possibility of tandem mass spectrometric applications using multi-turn TOF mass spectrometers is also discussed.     

Application of a multi-turn time-of-flight mass spectrometer, MULTUM II, to organic compounds ionized by matrix-assisted laser desorption/ionization

The circuit shape of the ion path, or the multi-turn, provides a solution for achieving unrestricted mass resolution from time-of-flight mass analyzers. The potential of a multi-turn type mass spectrometer, the MULTUM II, with a 1.308 m circuit controlled by four toroidal electric sector fields in biological applications was examined. With matrix-assisted laser desorption/ionization, the ion flight of 18 cycles gave a mass resolution of 10,000 for MH+ of protophorphyrin IX. This resolution was correlated with the flight length, and a resolution of 61,000 was achieved for MH+ of angiotensin I after 75 cycles or a 98.75 m total flight. The results demonstrate that the multi-turn mass spectrometer allows not only high resolution but also very high separation of the ions of molecular species from organic compounds.     

Tandem Time-of-Flight Mass Spectrometer with High Precursor Ion Selectivity Employing Spiral Ion Trajectory and Improved Offset Parabolic Reflectron

In this study, we have developed a tandem time-of-flight mass spectrometry (TOF/TOF) technique involving the use of a matrix-assisted laser desorption/ionization ion source that exhibits high precursor ion selectivity. An ion optical system with a 17 m spiral ion trajectory was used in the first time-of-flight mass spectrometer. High precursor ion selectivity was achieved by realizing a 15 m flight path, which is considerably longer than that of the conventional MALDI-TOF/TOF before the precursor ion selection by an ion gate; monoisotopic ions could be selected properly up to m/z 2500. Furthermore, the first time-of-flight mass spectrometer was composed of electrostatic sectors and could eliminate post-source decay (PSD) ions. Precursor ions with 20 keV kinetic energy were selected and injected into a collision cell, leading to the generation of fragment ions by high-energy collision-induced dissociation (HE-CID). The optimized second time-of-flight mass spectrometer included a post-acceleration region and an offset parabolic reflectron to record product ion spectra in the entire mass range. Our system could generate a simple HE-CID product ion spectrum because each fragment pathway could be observed as a single peak by the selection of monoisotopic ions of all precursor ions and HE-CID fragment pathways could be predominantly observed by the PSD ion elimination.     

Design of a new multi‐turn ion optical system ‘IRIS’ for a time‐of‐flight mass spectrometer

A new multi‐turn ion optical system ‘IRIS’ has been designed for use with a high‐performance time‐of‐flight (TOF) mass spectrometer, which satisfies the new design concepts of time focusing and phase space stability. It has an elliptical flight path composed of four toroidal electric sectors, with a flight path length for one lap of 0.974 m. Dimensions and voltages of sector electrodes have been optimized to satisfy theoretical requirements by simulations using surface charge method. Generally, multi‐turn instruments require an injection and ejection system to inject and eject ions. On the basis of this ion optical study, we have designed an injection and ejection ion optical system, which achieves time focusing for the total system. Furthermore, we have designed novel field‐adjusting electrodes (FAEs) for the perforated sectors in the injection and ejection systems, which accurately correct the electric potential around the perforated sector's hole. We have also used simulations to evaluate mass resolving power and ion transmissions for various lap numbers or flight path lengths. Through these we have confirmed that mass resolving powers of over 100 000 can be achieved with reasonable ion transmissions for a given set of initial conditions. Usually a multi‐turn TOF mass spectrometer with a closed optic axis has mass range limitations from overtaking ions. To solve this problem, a TOF segmentation method is proposed that identifies all peaks in a TOF spectrum, including those from overtaking ions. Copyright © 2008 John Wiley & Sons, Ltd.     

半圆弧面线性离子阱性能优化的模拟研究

半圆弧面线性离子阱具有电极结构简单、便于加工和安装精度高等优点。为进一步提升半圆弧面线性离子阱的分析性能,本研究在实验室原有半圆弧面线性离子阱的基础上提出了一种四面开槽的半圆弧面线性离子阱,并对其电极半径与场半径之比r/r0以及离子出射方向上电极的“拉伸”距离进行了优化。模拟结果表明：当r/r0=5：5,离子出射方向上的电极向外“拉伸”0.8~1.2 mm时,离子阱的性能有较大提升,尤其是“拉伸”距离为0.9 mm时所得质量分辨率最高,当扫描速率为409 Da/s时, m/z=609 Da的离子质量分辨率可达到6264( M/△M, FWHM)。作为对比,本研究同时对双曲面线性离子阱的性能进行了仿真优化,结果表明,经过优化后的半圆弧面线性离子阱的性能可与双曲面线性离子阱相媲美。     

Optimizing signal and mass resolution for matrix-assisted laser desorption utilizing a linear time-of-flight mass spectrometer.

Results are presented for various instrumental configurations employed for matrix-assisted laser desorption mass spectrometry. Mass resolution is determined for a linear time-of-flight mass spectrometer for various lengths of the field-free region. A wire ion guide is utilized and is shown to improve ion transport efficiencies for longer field-free regions. It is also determined experimentally that a modest mass resolution increase is often obtained in configurations employing the wire ion guide when compared to the mass resolution obtained with the same geometry without the wire ion guide. Optimal applied potentials are determined for the wire ion guide. No mass dependence on the optimal applied potential (-100 V) for the wire ion guide is observed for samples of equine myoglobin (MW 16,951.5 Da) and a bacterial protease (MW 27,228.4 Da). The optimal applied voltage was also found to be identical (-100 V) for the singly through quadruply charged molecular ion species of rabbit gamma globulin (MW approximately 150,000 Da). It is shown that a 2 m flight tube with a wire ion guide provides better signal-to-noise mass spectra than a 1 m flight tube without the wire ion guide and can more than double the mass resolution obtainable. Utilization of a 4 m flight tube gives minimal mass resolution enhancement at the expense of signal-to-noise.     

Improved sensitivity and mass range in time-of-flight bioaerosol mass spectrometry using an electrostatic ion guide

Bioearosol mass spectrometry (BAMS) analyzes single particles in real time from ambient air, placing strict demands on instrument sensitivity. Modeling of the BAMS reflectron time of flight (TOF) with SIMION revealed design limitations associated with ion transmission and instrument sensitivity at higher masses. Design and implementation of a BAMS linear TOF with electrostatic ion guide and delayed extraction capabilities has greatly increased the sensitivity and mass range relative to the reflectron design. Initial experimental assessment of the new instrument design revealed improved sensitivity at high masses as illustrated when using standard particles of cytochrome C (m/z approximately 12,000), from which the compound's monomer, dimer (m/z approximately 24,000) and trimer (m/z approximately 36,000) were readily detected.     

新型三角形电极圆环离子阱的理论模拟研究

圆环离子阱由于其离子储存能力明显优于相同体积下的三维离子阱,近年来被认为是离子阱小型化发展的另一个重要方向。为进一步优化圆环形离子阱的质谱性能,特别是质量分辨能力,本研究提出了一种由三角形电极构建的新型圆环离子阱,它由两个完全等同的、截面为三角形的圆环电极及两个大小不等的圆筒型电极所组成,离子通过共振激发方式弹出。通过理论模拟和对电极结构的优化,获得了具有非对称性的三角形电极结构,通过改善圆环结构,优化电场分布,提高了离子引出效率和离子阱的质量分辨能力,其中一种最优化结构的圆环离子阱对m/z 609离子的质量分辨率达到1486。     

Rapid sequencing of a peptide containing a single disulfide bond using high-energy collision-induced dissociation

A peptide containing a single disulfide bond was sequenced using high-energy collision-induced dissociation (HE-CID) in conjunction with a high mass resolution time-of-flight tandem mass spectrometer equipped with a matrix-assisted laser desorption/ionization source. This mass spectrometer, which has spiral ion trajectory, allowed both high mass resolution and high precursor ion selectivity. It is difficult to obtain sufficient product ions from peptides containing disulfide bonds using HE-CID due to the single collision in the gas phase. To compensate for insufficient dissociation, the disulfide bond was cleaved via an in-source reduction process using 1,5-diaminonaphthalene, a reducing matrix. After applying the reduction in the ionization, subsequent sequencing using HE-CID provided the detailed structural information of the peptide containing the single disulfide bond.     

小型高分辨电子轰击离子源反射式飞行时间质谱仪的研制

常用的气体分析质谱仪使用四极杆质谱作为分析器,分辨率一般低于300,无法解决同质量数离子带来的干扰问题.本实验自行研制了一种小型高分辨气体分析质谱仪,它采用电子轰击离子源反射式飞行时间质量分析器.仪器腔体总长45 cm,在m/z 28的位置,质量分辨率达到3000(Full width at half maximum,FWHM),实现了CO和N2的半峰谷分离;在m/z 69的位置,仪器分辨率达到5000(FWHM).在直接大气压进样条件下,可以检测到空气中136Xe(含量7.8 μ g/m3)和80Kr(含量2.8 μg/m3).使用ADC采集时,仪器的动态范围为1 06.该仪器将作为高端气体质谱仪,应用于过程监测在线分析、环境有机挥发物研究、热分析质谱及催化反应监测等领域.     

High resolution on a quadrupole ion trap mass spectrometer

By using a modified ion trap mass spectrometer, resolution in excess of 30,000 (FWHM) at m I z 502 is demonstrated. The method of increasing resolution in the ion trap mass spectrometer operated in the mass-selective instability mode depends on decreasing the rate of scanning the primary radio frequency amplitude as well as using resonance ejection at the appropriate frequency and amplitude. A theoretical basis for the method is introduced.     

Quantitation of small molecules using high‐resolution accurate mass spectrometers – a different approach for analysis of biological samples

The quantitative capabilities of a linear ion trap high‐resolution mass spectrometer (LTQ‐Orbitrap™) were investigated using full scan mode bracketing the m/z range of the ions of interest and utilizing a mass resolution (mass/FWHM) of 15000. Extracted ion chromatograms using a mass window of ±5–10 mmu centering on the theoretical m/z of each analyte were generated and used for quantitation. The quantitative performance of the LTQ‐Orbitrap™ was compared with that of a triple quadrupole (API 4000) operating using selected reaction monitoring (SRM) detection. Comparable assay precision, accuracy, linearity and sensitivity were observed for both approaches. The concentrations of actual study samples from 15 Merck drug candidates reported by the two methods were statistically equivalent. Unlike SRM being a tandem mass spectrometric (MS/MS)‐based detection method, a high resolution mass spectrometer operated in full scan does not need MS/MS optimization. This approach not only provides quantitative results for compounds of interest, but also will afford data on other analytes present in the sample. An example of the identification of a major circulating metabolite for a preclinical development study is demonstrated. Copyright © 2009 John Wiley & Sons, Ltd.     

Persistence length of wormlike micelles composed of ionic surfactants: self-consistent-field predictions

The persistence length of a wormlike micelle composed of ionic surfactants C(n)E(m)X(k) in an aqueous solvent is predicted by means of the self-consistent-field theory where C(n)E(m) is the conventional nonionic surfactant and X(k) is an additional sequence of k weakly charged (pH-dependent) segments. By considering a toroidal micelle at infinitesimal curvature, we evaluate the bending modulus of the wormlike micelle that corresponds to the total persistence length, consisting of an elastic/intrinsic and an electrostatic contribution. The total persistence length increases with pH and decreases with increasing background salt concentration. We estimate that the electrostatic persistence length l(p,e)(0) scales with respect to the Debye length kappa(-1) as l(p,e)(0) approximately kappa(-p) where p approximately 1.98 for wormlike micelles consisting of C(20)E(10)X(1) surfactants and p approximately 1.54 for wormlike micelles consisting of C(20)E(10)X(2) surfactants. The total persistence length l(p,t)(0) is a weak function of the head group length m but scales with the tail length n as l(p,t)(0) approximately n(x) where x approximately 2-2.6, depending on the corresponding head group length. Interestingly, l(p,t)(0) varies nonmonotonically with the number of charged groups k due to the opposing trends in the electrostatic and elastic bending rigidities upon variation of k.     

Orthogonal extraction ion mobility spectrometry

Ion Mobility Spectrometry is a powerful tool for the study of molecular conformations, separation of mass isomers, and analysis of complex mixtures and suppression of chemical background. The factors that limit the capabilities of the technique include its relatively low resolving power and duty cycle. New principle of gas-phase ion separation, based on ion focusing under the influence of electrostatic field and stationary in time gas flow, is proposed. Both analytical calculations and a numerical simulation show that a diffusion-limited resolution of several hundred can be achieved. The new type of ion mobility analyzer is called orthogonal extraction IMS. The proposed ortho-IMS can be interfaced with commercial mass spectrometers and offers the theoretical resolution of several hundred and ion transmission close to 100%.     

Photodissociation investigation of doubly charged ethanol clusters induced by inner-shell electron ionization

Fragmentation of doubly charged ethanol clusters [(C(2)H(5)OH)(n)] following the O 1s ionization has been investigated by means of the photoelectron-photoion-photoion coincidence (PEPIPICO) method. The dominant fission channel of (C(2)H(5)OH)(n)(2+) was the formation of protonated cluster ion pairs [H(C(2)H(5)OH)(l)(+)/H(C(2)H(5)OH)(m)(+)]. The fragmentation mechanisms of these ion pairs were discussed based on the analysis of the PEPIPICO contour shape. It was clarified that the prominent fragmentation channel was a secondary decay mechanism, where neutral evaporation occurs after charge separation. On the other hand, the formation of small fragment ions was suppressed, excluding the formation of certain specific fragments (H(3)O(+), C(2)H(5)(+)/COH(+), and C(2)H(4)OH(+)). The formation of small fragment ions was suppressed due to the cooling effect caused by the neutral evaporation and the decrease in the electrostatic repulsive force caused by charge separation.     

Optimization of a quadrupole ion storage trap as a source for time‐of‐flight mass spectrometry

Designs of a quadrupole ion trap (QIT) as a source for time‐of‐flight (TOF) mass spectrometry are evaluated for mass resolution, ion trapping, and laser activation of trapped ions. Comparisons are made with the standard hyperbolic electrode ion trap geometry for TOF mass analysis in both linear and reflectron modes. A parallel‐plate design for the QIT is found to give significantly improved TOF mass spectrometer performance. Effects of ion temperature, trapped ion cloud size, mass, and extraction field on mass resolution are investigated in detail by simulation of the TOF peak profiles. Mass resolution (m/Δm) values of several thousand are predicted even at room temperature with moderate extraction fields for the optimized design. The optimized design also allows larger radial ion collection size compared with the hyperbolic ion trap, without compromising the mass resolution. The proposed design of the QIT also improves the ion–laser interaction volume and photon collection efficiency for fluorescence measurements on trapped ions. Copyright © 2011 John Wiley & Sons, Ltd.     

Ion trajectories in an electrostatic ion guide for external ion source fourier transform ion cyclotron resonance mass spectrometry

An electrostatic ion guide (EIG) that consists of concentric cylinder and central wire electrodes can transport ions efficiently from an external ion source to an ion cyclotron resonance (ICR) ion trap for mass analysis, with several advantages over current injection methods. Because the electrostatic force of the EIG captures ions in a stable orbit about the wire electrode, ions with initially divergent trajectories may be redirected toward the ICR ion trap for improved ion transmission efficiency. SIMION trajectory calculations (ion kinetic energy, 1–200 eV; elevation angle, 0.30 °; azimuthal angle, 0.360°) predict that ions of m/z 1000 may be transmitted through a strong (0.01 → 3.0-T) magnetic field gradient. Judicious choice of ion source position and EIG potential minimizes the spread in ion axial kinetic energy at the ICR ion trap. Advantages of the EIG include large acceptance angle, even for ions that have large initial kinetic energy and large radial displacement with respect to the central z-axis, low ion extraction voltage (5–20 V), and efficient trapping because ions need not be accelerated to high velocity to pass through the magnetic field gradient.     

Small gas-phase dianions of Zn3O(4)(2-), Zn4O(5)(2-), CuZn2O(4)(2-), Si2GeO(6)(2-), Ti2O(5)(2-) and Ti3O(7)(2-)

We have searched for new species of small oxygen-containing gas-phase dianions produced in a secondary ion mass spectrometer by Cs+ ion bombardment of solid samples with simultaneous exposure of their surfaces to O2 gas. The targets were a pure zinc metal foil, a copper-contaminated zinc-based coin, two silicon-germanium samples (Si(1-x)Ge(x)(with x= 6.5% or 27%)) and a piece of titanium metal. The novel dianions Zn3O(4)(2-), Zn4O(5)(2-), CuZn2O(4)(2-), Si2GeO(6)(2-), Ti2O(5)(2-) and Ti3O(7)(2-) have been observed at half-integer m/z values in the negative ion mass spectra. The heptamer dianions Zn3O(4)(2-) and Ti2O(5)(2-) have been unambiguously identified by their isotopic abundances. Their flight times through the mass spectrometer are approximately 20 micros and approximately 17 micros, respectively. The geometrical structures of the two heptamer dianions Ti2O(5)(2-), and Zn3O(4)(2-) are investigated using ab initio methods, and the identified isomers are compared to those of the novel Ge2O(5)(2-) and the known Si2O(5)(2-) and Be3O(4)(2-) dianions.