Atmospheric pressure chemical ionization applications

A number of analytical techniques such as FTIR spectroscopy,65-66 13C NMR,67,68 solid-state 13 C NMR,69 GPC or size exclusion chromatography (SEC),67-72 HPLC,73 mass spectrometric analysis,74 differential scanning calorimetry (DSC),67 75 76 and dynamic mechanical analysis (DMA)77 78 have been utilized to characterize resole syntheses and crosslinking reactions. Packed-column supercritical fluid chromatography with a negative-ion atmospheric pressure chemical ionization mass spectrometric detector has also been used to separate and characterize resoles resins.79 This section provides some examples of how these techniques are used in practical applications. 

In the following chapters, the basic principles of HPLC and MS, in as far as they relate to the LC-MS combination, will be discussed and seven of the most important types of interface which have been made available commercially will be considered. Particular attention will be paid to the electrospray and atmospheric-pressure chemical ionization interfaces as these are the ones most widely used today. The use of LC-MS for identification and quantitation will be described and appropriate applications will be discussed. 

Figure 4.21 Schematic of an atmospheric-pressure-chemical-ionization probe. From applications literature published by Micromass UK Ltd, Manchester, UK, and reproduced with permission.

Most reported triazine LC applications are reversed-phase utilizing C-8 and C-18 analytical columns, but there are also a few normal-phase (NH2,CN) and ion-exchange (SCX) applications. The columns used range from 5 to 25-cm length and from 2 to 4.6-mm i.d., depending on the specific application. In general, the mobile phases employed for reversed-phase applications consist of various methanol and/or acetonitrile combinations in water. The ionization efficiency of methanol and acetonitrile for atmospheric pressure chemical ionization (APcI) applications were compared, and based on methanol s lower proton affinity, the authors speculated that more compounds could be ionized in the positive ion mode when using methanol than acetonitrile in the mobile phase. 

As with GC/MS, LC/MS offers the possibility of unequivocal confirmation of analyte identity and accurate quantiation. Similarly, both quadrupole and ion-trap instruments are commercially available. However, the responses of different analytes are extremely dependent on the type of interface used to remove the mobile phase and to introduce the target analytes into the mass spectrometer. For pesticide residue analyses, the most popular interfaces are electrospray ionization (ESI) and atmospheric pressure chemical ionization (APCI). Both negative and positive ionization can be used as applicable to produce characteristically abundant ions. 

The most important techniques belonging to this class are electrospray ionization (ESI), atmospheric pressure chemical ionization (APCI) and, more recently, atmospheric pressure photoionization (APPI). At present the latter does not have applications in cultural heritage, so it will be not described here. 

Jones JJ, Kidwell H, Games DE. 2003. Application of atmospheric pressure chemical ionization mass spectrometry in the analysis of barbiturates by high-speed analytical countercurrent chromatography. Rapid Commun Mass Spec-trom 17 1565. 

Kagan M, Chlenov M, Kraml CM. 2004. Normal-phase high-performance liquid chromatographic separations using ethoxynonafluorobutane as hexane alternative. II. Liquid chromatography-atmospheric pressure chemical ionization-mass spectrometry applications with methanol gradients. J Chromatogr A 1033 321. 

Isoo, K., Otsuka, K., and Terabe, S. (2001). Application of sweeping to micellar electrokinetic chromatography-atmospheric pressure chemical ionization-mass spectrometric analysis of environmental pollutants. Electrophoresis 22, 3426-3432. 

X. Xu, A.M. van der Craats, E.M. Kok and P.C.A.M. de Bruyn, Trace analysis of peroxide explosives by high performance liquid chromatography — atmospheric pressure chemical ionization — tandem mass spectrometry (HPLC-APCl-MS/MS) for forensic applications ,... 

Gallagher, R. T Balogh, M. P. Davey, P. Jackson, M. R. Sinclair, L Southern, L. J. Combined electrospray ionization-atmospheric pressure chemical ionization source for use in high-throughput LC-MS applications. Anal. Chem. 2003, 75, 973-977. 

Atmospheric pressure chemical ionization (APCI) is the most common alternative to ESI in LC/MS. Whereas the application of ESI is generally recommended in the case of polar compounds, which may have a high molecular weight (up to 1 million Da), the problem-solving domain of APCI is limited to compounds with an MW smaller than 1000 Da. However, less polar compounds are accessible than with ESI. 

In parallel with these developments, other techniques have been introduced that were especially applicable to the combination of liquid chromatography with MS. The most interesting, from the point of view of structural studies of flavonoid glycosides, are thermospray (TSP) and atmospheric pressure ionization (API) methods, which include electrospray ionization (ESI) and atmospheric pressure chemical ionization (APCI). TSP was the first... 

Grayer, R.J. et al.. The application of atmospheric pressure chemical ionization liquid chromatography-mass spectrometry in the chemotaxonomic study of flavonoids characterization of flavonoids from Ocimum gratissimum var., Phytochem. Anal., 11, 257, 2000. 

Coupling of liquid chromatography with mass spectrometry allows unequivocal online spectrometric identification of all nitrofurans at the very low residue concentrations required by regulatory agencies for confirmatory analysis in animal-derived foods. Typical examples of mass spectrometry applications in confirming nitrofuran residues in edible animal products employ thermospray (174, 176), ionspray (166), or atmospheric pressure chemical ionization (157) interfaces. 

LC-MS, as a technique, is very much dependent upon ionization (and ion vaporization) techniques that are suited to LC conditions, i.e. techniques where a relatively large solvent flow can be accommodated, which restricts us to just two ionization methods electrospray ionization (ESI) and atmospheric pressure chemical ionization (APCl). Both techniques are very similar in their modes of operation (see Section 5.2.1), relying on the formation of a spray from a solvent flow at atmospheric pressure, and hence they are ideally suited to use in LC-MS applications. 

Recent advances in electrospray ionization (ESI), atmospheric-pressure chemical ionization (APCI), thermospray, and particle beam LC-MS have advanced the analyst toward the universal HPLC detector, but price and complexity are still the primary stumbling blocks. Thus, HPLC-MS remains expensive and the technology has only recently been described. Early commercial LC-MS uses particle beam and thermospray sources, but ESI and APCI interfaces now dominate. Liquid chromatography MS can represent a fast and reliable method for structural analyses of nonvolatile compounds such as phenolic compounds (36,37), especially for low-molecular-weight plant phenolics (38), but the limited resolving power of LC hinders the widespread use of its application for phenolics as compared to GC-MS. 

They are still the workhorses of coupled mass spectrometric applications, as they are relatively simple to run and service, relatively inexpensive (for a mass spectrometer), and provide unit mass resolution and scanning speeds up to approximately 10,000 amu/s. This even allows for simultaneous scan/ selected ion monitoring (SIM) operation, in which one part of the data acquisition time is used to scan an entire spectrum, whereas the other part is used to record the intensities of selected ions, thus providing both qualitative information and sensitive quantitation. They are thus suitable for many GC-MS and liquid chromatography-mass spectrometry (LC-MS) applications. In contrast to GC-MS with electron impact (El) ionization, however, LC-MS provides only limited structural information as a consequence of the soft ionization techniques commonly used with LC-MS instruments [electrospray ionization (ESI) and atmospheric pressure chemical ionization (APCI)]. Because of this limitation, other types of mass spectrometers are increasingly gaining in importance for LC-MS. 

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