FAB

A connnon feature of all mass spectrometers is the need to generate ions. Over the years a variety of ion sources have been developed. The physical chemistry and chemical physics communities have generally worked on gaseous and/or relatively volatile samples and thus have relied extensively on the two traditional ionization methods, electron ionization (El) and photoionization (PI). Other ionization sources, developed principally for analytical work, have recently started to be used in physical chemistry research. These include fast-atom bombardment (FAB), matrix-assisted laser desorption ionization (MALDI) and electrospray ionization (ES). 

Finally, following Mead and Truhlar [10], it may be seen that an interchange of A and B is equivalent to a sign reversal of <() followed by a rotation perpendicular to the AB bond, under the latter of which Aab) is invariant and Fab) changes sign. The net effect is therefore to induce the tiansitions... 

If now there exists a representation in which A is zero, then in this representation Fab is also zero [by Eq. (99)]. Now, in a (/-transformed representation (which can be chosen to be completely general), one finds that... 

Fast-Atom Bombardment (FAB) and Liquid-Phase Secondary Ion Mass Spectrometry (LSIMS) Ionization... 

A big step forward came with the discovery that bombardment of a liquid target surface by abeam of fast atoms caused continuous desorption of ions that were characteristic of the liquid. Where this liquid consisted of a sample substance dissolved in a solvent of low volatility (a matrix), both positive and negative molecular or quasi-molecular ions characteristic of the sample were produced. The process quickly became known by the acronym FAB (fast-atom bombardment) and for its then-fabulous results on substances that had hitherto proved intractable. Later, it was found that a primary incident beam of fast ions could be used instead, and a more generally descriptive term, LSIMS (liquid secondary ion mass spectrometry) has come into use. However, note that purists still regard and refer to both FAB and LSIMS as simply facets of the original SIMS. In practice, any of the acronyms can be used, but FAB and LSIMS are more descriptive when referring to the primary atom or ion beam. 

When the liquid target is not a static pool but, rather, a continuous stream of liquid, the added description of dynamic is used. Thus, dynamic FAB and LSIMS refer to bombardment of a continuously renewed (flowing) liquid target. 

If the liquid that is being bombarded contains ions, then some of these will be ejected from the liquid and can be measured by the mass spectrometer. This is an important but not the only means by which ions appear in a FAB or LSIMS spectrum. Momentum transfer of preformed ions in solution can be used to enhance ion yield, as by addition of acid to an amine to give an ammonium species (Figure 4.3). 

An example of enhanced ion production. The chemical equilibrium exists in a solution of an amine (RNH2). With little or no acid present, the equilibrium lies well to the left, and there are few preformed protonated amine molecules (ions, RNH3+) the FAB mass spectrum (a) is typical. With more or stronger acid, the equilibrium shifts to the right, producing more protonated amine molecules. Thus, addition of acid to a solution of an amine subjected to FAB usually causes a large increase in the number of protonated amine species recorded (spectrum b). 

A typical FAB mass spectrum of glycerol alone, showing a protonated molecular ion at m/z 93 accompanied by decreasing numbers of protonated cluster ions (m/z, 1 + nx92 n = 2, 3, 4,. ..). 

In addition to low volatility, the chosen liquid should be a good all-around solvent. Since no one liquid is likely to have the required solvency characteristics, several are in use (Table 4.1). If a mass spectmm cannot be obtained in one solvent, it is useful to try one or more others before deciding that an FAB spectrum cannot be obtained. 

In general, FAB and LSIMS will give excellent molecular mass information in the range (approximately) of m/z 100-2000. Above this value, the abundance of molecular ions tends to diminish until, in the region of m/z 4000-5000, they become either nonexistent or very difficult to... 

The basic principles of fast-atom bombardment (FAB) and liquid-phase secondary ion mass spectrometry (LSIMS) are discussed only briefly here because a fuller description appears in Chapter 4. This chapter focuses on the use of FAB/LSIMS as part of an interface between a liquid chromatograph (LC) and a mass spectrometer (MS), although some theory is presented. 

The FAB source operates near room temperature, and ions of the substance of interest are lifted out from the matrix by a momentum-transfer process that deposits little excess of vibrational and rotational energy in the resulting quasi-molecular ion. Thus, a further advantage of FAB/LSIMS over many other methods of ionization lies in its gentle or mild treatment of thermally labile substances such as peptides, proteins, nucleosides, sugars, and so on, which can be ionized without degrading their. structures. 

Liquid chromatography is a separation method that is often applied to nonvolatile, thermally labile materials such as peptides, and, if their mass spectra are required after the separation step, then a mild method of ionization is needed. Since FAB/LSIMS is mild and works with a liquid matrix, it is not surprising that attempts were made to utilize this ionization source as both an inlet... 

In dynamic FAB, this solution is the eluant flowing from an LC column i.e., the target area is covered by a flowing liquid (dynamic) rather than a static one, as is usually the case where FAB is used to examine single substances. The fast atoms or ions from the gun carry considerable momentum, and when they crash into the surface of the liquid some of this momentum is transferred to molecules in the liquid, which splash back out, rather like the result of throwing a stone into a pond (Figure 13.2). This is a very simplistic view of a complex process that also turns the ejected particles into ions (see Chapter 4 for more information on FAB/LSIMS ionization). 

Liquids examined by FAB are introduced into the mass spectrometer on the end of a probe inserted through a vacuum lock in such a way that the liquid lies in the target area of the fast atom or ion beam. There is a high vacuum in this region, and there would be little point in attempting to examine a solution of a sample in one of the commoner volatile solvents such as water or dichloromethane because it would evaporate extremely quickly, probably as a burst of vapor when introduced into the vacuum. Therefore it is necessary to use a high-boiling solvent as the matrix material, such as one of those listed in Table 13.1. 

Having considered the various parts of a dynamic-FAB system (atom gun, ionization, and matrix), it is now necessary to see how these are put together in a working inlet/ion source interface. 

A dynamic-FAB probe having a simple copper target. The narrow fused-silica tube passes through the shaft, its end lying flush with the target surface. 

A typical TIC chromatogram from an analysis of peptides resulting from enzymatic digest of myoglobin. The peaks represent individual peptides eluting from an LC column and being mass measured by a spectrometer coupled to it through a dynamic-FAB inlet/ion source. 

A dynamic-FAB probe tip incorporating a screen and wick assembly at the target surface. 

Mostly, positive-ion FAB yields protonated quasi-molecular ions [M -i- H]+, and the negative-ion mode yields [M - H]. In the presence of metal salts (e.g., KCl) that are sometimes added to improve efficiency in the LC column, ions of the type [M -i- X]+are common, where X is the metal. Another type of ion that is observed is the so-called cluster, a complex of several molecules with one proton, [M -i- H]+ with n = 1, 2, 3,. .., etc. Few fragment ions are produced. 

The LC/TOF instmment was designed specifically for use with the effluent flowing from LC columns, but it can be used also with static solutions. The initial problem with either of these inlets revolves around how to remove the solvent without affecting the substrate (solute) dissolved in it. Without this step, upon ionization, the large excess of ionized solvent molecules would make it difficult if not impossible to observe ions due only to the substrate. Combined inlet/ionization systems are ideal for this purpose. For example, dynamic fast-atom bombardment (FAB), plas-maspray, thermospray, atmospheric-pressure chemical ionization (APCI), and electrospray (ES)... 

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