Mass spectrometer hyphenated techniques

Several types of LC and one nonchromatographic separation system for liquids have been interfaced with MS. HPLC is widely used to separate nonvolatile organic compounds of all polarities and molecular weights. Coupled to a mass spectrometer, the technique is called LC-MS. Supercritical fluid chromatography (SFC) and the nonchromatographic separation technique of capillary electrophoresis (CE) are also used with mass spectro-metric detection. The interfacing, ionization sources, operation, and applications of these hyphenated methods are covered in Chapter 13. 

Spectrometry Chemical Ionization in Mass Spectrometry Chemical Structure Information from Mass Spectrometry Forensic Science, Applications of Mass Spectrometry Hyphenated Techniques, Applications of in Mass Spectrometry Laser Microprobe Mass Spectrometers MS-MS and MS" Peptides and Proteins Studied Using Mass Spectrometry Quadru-poles. Use of in Mass Spectrometry. 

Multidimensional or hyphenated instmments employ two or more analytical instmmental techniques, either sequentially, or in parallel. Hence, one can have multidimensional separations, eg, hplc/gc, identifications, ms/ms, or separations/identifications, such as gc/ms (see CHROMATOGRAPHY Mass spectrometry). The purpose of interfacing two or more analytical instmments is to increase the analytical information while reducing data acquisition time. For example, in tandem-mass spectrometry (ms/ms) (17,18), the first mass spectrometer appHes soft ionization to separate the mixture of choice into molecular ions the second mass spectrometer obtains the mass spectmm of each ion. 

The combination of chromatography and mass spectrometry (MS) is a subject that has attracted much interest over the last forty years or so. The combination of gas chromatography (GC) with mass spectrometry (GC-MS) was first reported in 1958 and made available commercially in 1967. Since then, it has become increasingly utilized and is probably the most widely used hyphenated or tandem technique, as such combinations are often known. The acceptance of GC-MS as a routine technique has in no small part been due to the fact that interfaces have been available for both packed and capillary columns which allow the vast majority of compounds amenable to separation by gas chromatography to be transferred efficiently to the mass spectrometer. Compounds amenable to analysis by GC need to be both volatile, at the temperatures used to achieve separation, and thermally stable, i.e. the same requirements needed to produce mass spectra from an analyte using either electron (El) or chemical ionization (Cl) (see Chapter 3). In simple terms, therefore, virtually all compounds that pass through a GC column can be ionized and the full analytical capabilities of the mass spectrometer utilized. 

Principles and Characteristics Extraction or dissolution methods are usually followed by a separation technique prior to subsequent analysis or detection. While coupling of a sample preparation and a chromatographic separation technique is well established (Section 7.1), hyphenation to spectroscopic analysis is more novel and limited. By elimination of the chromatographic column from the sequence precol-umn-column-postcolumn, essentially a chemical sensor remains which ensures short total analysis times (1-2 min). Examples are headspace analysis via a sampling valve or direct injection of vapours into a mass spectrometer (TD-MS see also Section 6.4). In... 

Gas chromatography is a most favourable case for interfacing to a mass spectrometer, as the mobile phases commonly used do not generally influence the spectra observed, and the sample, being in the vapour phase, is compatible with the widest range of mass-spectral ionisation techniques. The primary incompatibility in the case of GC-MS is the difference in operating pressure for the two hyphenated instruments. The column outlet in GC is typically at atmospheric pressure, while source pressures in the mass spectrometer range from 2 to... 

FTIR in multiply hyphenated systems may be either off-line (with on-line collection of peaks) [666,667] or directly on-line [668,669]. Off-line techniques may be essential for minor components in a mixture, where long analysis times are required for FT-based techniques (NMR, IR), or where careful optimisation of the response is needed. In an early study a prototype configuration comprised SEC, a triple quadrupole mass spectrometer, off-line evaporative FTIR with splitting after UV detection see Scheme 7.12c [667]. Off-line IR spectroscopy (LC Transform ) provides good-quality spectra with no interferences from the mobile phase and the potential for very high sensitivity. Advanced approaches consist of an HPLC system incorporating a UV diode array, FTIR (using an ATR flow-cell to obtain on-flow IR spectra), NMR and ToF-MS. 

Principles and Characteristics The main reasons for hyphenating MS to CE are the almost universal nature of the detector, its sensitivity and the structural information obtainable, including assessment of peak purity and identity. As CE is a liquid-phase separation technique, coupling to the mass spectrometer can be achieved by means of (modified) LC-MS interfaces. Because of the low flow-rates applied in CE, i.e. typically below lOOnLmin-1, a special coupling device is required to couple CE and the LC-MS interface. Three such devices have been developed, namely a... 

MSn are of particular interest. MSn stands for the n-fold coupling of mass spectrometers, alternatively serving as separation and detection instrument. By hyphenated techniques the dimensionality of analytical information (see Sect. 3.4) and, therefore, also the information amount (see Sect. 9.3) is significantly increased (Eckschlager and Danzer [1994]). 

VDU screen via suitable electronic amplifying circuitry where the data are presented in the form of an elution profile. Although there are a dozen or more types of detector available for gas chromatography, only those based on thermal conductivity, flame ionization, electron-capture and perhaps flame emission and electrolytic conductivity are widely used. The interfacing of gas chromatographs with infrared and mass spectrometers, so-called hyphenated techniques, is described on p. 114 etseq. Some detector characteristics are summarized in Table 4.11. 

Figure 2.10. Schematic of a MALDI-TOF mass spectrometer with reflector and time-lag focusing. Reprinted from A. Westman-Brinkmalm and G. Brinkmalm (2002). In Mass Spectrometry and Hyphenated Techniques in Neuropeptide Research, J. Silberring and R. Ekman (eds.) New York John Wiley Sons, 47-105. With permission of John Wiley Sons, Inc.

In a typical analysis, one approach would be to carry out the analysis by first using Cl and quadrupole MS. The fragments from this first MS would then be directed to an El and a TOF mass spectrometer. Different fragments will be observed and this will yield additional information about the sample. In many cases, the MS-MS analysis is applied to samples eluting from either LC, HPLC, or GC chromatographic separation techniques. For additional information on this topic, see Triple Hyphenated Methods. ... 

Instrumental developments (e.g., of sector field instruments with multiple ion collection, introduced in 1992, or the insertion of collision and reaction cells in order to reduce disturbing isobaric interferences), the progress in applications for ultratrace analysis, also in combination with on line hyphenated separation techniques (HPLC, CE), especially routine capability as well as decreasing price and user friendly maintenance mean that sales are increasing by 10 % every year. To improve the analytical performance of ICP mass spectrometers for precise isotope ratio measurements (e.g., for geochronology or for the study of fine isotope variation in nature) powerful instrumentation with high mass dispersion and multiple ion collector systems instead of single ion collection are commercially available on the analytical market. 

Transient signals are typically obtained in atomic spectrometry when samples are introduced by flow injection techniques or when the spectrometer is used as an element-specific detector in hyphenated techniques. Inductively coupled plasma mass spectrometry has nowadays become the detection technique of choice for multielement-specific detection in speciation as it allows multielemental... 

The identification of a compound only by its retention time is somewhat arbitrary. A better method consists of using two different techniques that are complementary. Coupling a chromatograph with a second instrument on-line such as a mass spectrometer or an infrared spectrophotometer realises these objectives. These coupled methods, often called hyphenated techniques, allow us to obtain two different types of information that are independent. Therefore, it is possible to determine with more certainty the nature and concentration of the components in a mixture in nanograms or smaller units of measurement. 

Complex mixtures are often analysed by hyphenated techniques, in which a separation method is interfaced with a mass spectrometer. Coupling of a gas chromatograph to a mass spectrometer (GC/MS) is simple the chromatographic capillary column is directly inserted into the ionisation chamber. Compounds eluting from the column in a gaseous state are introduced into the ion source and analysed in the order in which they exit the CG column. The vacuum system of the mass... 

Figure A.3A.3 LC/MS analysis of a dietary supplement consisting of extract of Trifolium pratense (red clover). Reversed-phase C18 HPLC and negative ion electrospray ionization mass spectrometry were used with a quadrupole mass spectrometer analyzer (Agilent also see Table A.3A.1). The map illustrates the abundance of information provided by this hyphenated technique with HPLC mass chromatograms in one dimension and mass spectra in another dimension.

Numerous papers have relied on only UV-visible spectra for their identification of phenolics, but for positive identification purposes, HPLC-mass spectrometry (MS) is another detection mode that can provide detection of all phenolic compounds in foods. This technique involves a hyphenated instrument that uses a mass spectrometer as a detector for HPLC or uses HPLC as cleanup step for mass spectrometry. After preparative HPLC, the MS technique has frequently been employed for structural identification of phenolics in many foods and essential oils because of its sensitivity and selectivity and its ability to provide structural information. 

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