Liquid Chromatography-Mass Spectrometry Coupling LC-MS

GC/MS with capillary columns has been the gold standard for more than 20 years, but LC/MS has become a complementary method due to the success in interface development with atmospheric pressure ionisation (API) for low molecular weight compounds and the appHcation to biopolymers. For many areas of analytical chemistry, LC/MS has become indispensible due to its advantages over GC/MS for polar and thermolabile analytes. A Hmiting factor for LC/MS has been the incompatibility between the hquid eluting from the LC and the mass spectrometer vacuum. This could be overcome in electrospray ionisation with the use of a nebuliser gas ( ion spray ) or additional heated drying gas ( turbo ion spray ) (70, 71]. Due to its high sensitivity and selectivity, APl-MS has become a standard tool for the stracture elucidation of analytes from complex mixtures. 

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Nowadays, ESI is the leading member of the group of atmospheric pressure ionization (API) methods and the method of choice for liquid chromatography-mass spectrometry coupling (LC-MS, Chap. 12). [10-13] Currently, ESI and MALDI (Chap. 10) are the most commonly employed ionization methods and they opened doors to the widespread biological and biomedical application of mass spectrometry. [5,10,11,13-17] Moreover, ESI serves well for the analysis of ionic metal complexes [18,19] and other inorganic analytes. [20-22]... 

The ion spray liquid chromatography/mass spectrometry (LC-MS) interface coupled via a postsuppressor split with an ion chromatography (IC) has been used in the analysis of alcohol sulfates. The IC-MS readily produces the molecular weight while the tandem mass spectrometric detection IC-MS-MS provides structural information [305]. 

The system relies upon preliminary fractionation of the microbial crude extract by dualmode countercurrent chromatography coupled with photodiode array detection (PDA). The ultraviolet-visible (UV-Vis) spectra and liquid chromatography-mass spectrometry (LC-MS) of biologically active peaks are used for identification. Confirmation of compound identity is accomplished by nuclear magnetic resonance (NMR). Use of an integrated system countercurrent chromatography (CCC) separation, PDA detection, and LC-MS rapidly provided profiles and structural information extremely useful for metabolite identification (dereplication, Figure 14.1). 

B. Ardrey, Liquid Chromatography-Mass Spectrometry An Introduction (Chichester, UK Wiley, 2003) W. M. A. Niessen. Liquid Chromatography-Mass Spectrometry, 2nd ed (New York Marcel Dekker, 1999) M. S. Lee, LC/MS Applications in Drug Development (New York Wiley, 2002) J. Abian, Historical Feature The Coupling of Gas and Liquid Chromatography with Mass Spectrometry, J. Mass Spectrom. 1999, 34, 157. 

The use of liquid chromatography-mass spectrometry (LC-MS) is becoming more popular because of the increasing number of LC-MS interfaces commercially available thermospray (TSP), particle beam (PB), and atmospheric pressure ionization (API). Coupled with mass spectroscopy, HPLC provides the analyst with a powerful tool for residue determination. 

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. 

As an example, consider a chromatogram in which a number of compounds are detected with different elution times, at the same time as a their spectra (such as UV or mass spectra) are recorded. Coupled chromatography, such as high-performance chromatography-diode array detection (HPLC-DAD) or liquid chromatography-mass spectrometry (LC-MS), is increasingly common in modern laboratories, and represents a rich source of multivariate data. These data can be represented as a matrix as in Figure 4.3. 

An extensive list that defines acronyms and abbreviations in the field of mass spectrometry was published in 2002 [6], A single analytical technique or a type of instrument is abbreviated without hyphens or slashes. However, it is customary to use hyphens for a description of an instrument whereas an abbreviation that describes the method uses slashes. For example, LC-MS is an instrument where a liquid chromatograph is coupled with a mass spectrometer, while LC/MS is the method of liquid chromatography/mass spectrometry. Thus, one uses an LC-MS instrument to obtain a LC/MS spectrum. 

Analytical techniques such as liquid chromatography/mass spectrometry (LC/ MS) to evaluate nonvolatile organics, gas chromatography/mass spectrometry (GC/ MS) to evaluate semivolatile organics, and inductively coupled plasma (ICP) spectroscopy to detect and quantitate inorganic elements should be a part of this study. 

GC-FID gas chromatography with flame ionization detector, HPLC-UV high performance liquid chromatography with UV detector, LC-MS liquid chromatography coupled with mass spectrometry, GC-FPD gas chromatography with flame photometric detector... 

The possibility for unambiguous identification of the analytes is offered by liquid chromatography-mass spectrometry (LC-MS). Mass spectrometry detection systems use the difference in mass-to-charge ratio (m/z) of ionized atoms or molecules to separate them from each other. Molecules have distinctive fragmentation patterns that provide structural information to identify structural components. The on-line coupling of LC with MS for the determination of drug residues in food has been under investigation for almost two decades. 

Atomic absorption spectrometry with flame (AA-F) or electrothermal atomization furnace (AA-ETA), inductively coupled plasma-emission spectroscopy (ICP-ES), inductively coupled plasma-mass spectrometry (ICP-MS), and high-performance liquid chromatography-mass spectrometry (LC-MS) are state-of-the-art analytical techniques used to measure metals in biological fluids. They are specific and sensitive and provide the cfinical laboratory with the capability to measure a broad array of metals at clinically significant concentrations. For example, ICP-MS is used to measure several metals simultaneously. Photometric assays are also available but require large volumes of sample and have limited analytical performance. Spot tests are also... 

Most of these measurements are still routinely perfonned with immunoassays because of their simplicity, rapidity and relatively low cost, and despite occasional concerns about their reliability and validity. Serum total T has also been quantitated by gas chromatography-mass spectrometry (GC-MS), liquid chromatography-mass spectrometry (LC-MS) and LC-MS-MS. MS-based methods are often used in research smdies or to confirm immunoassay results. The advantage of chromatography coupled with MS is the high specificity not available with immunoassays because they are susceptible to cross-reactions. An immunoassay might measure a host of structurally related compounds in addition to the target analyte. 

Detection by on-line coupled liquid chromatography-mass spectrometry (LC-MS) is at present at an experimental stage. The technique and instrumentation in LC-MS is improving rapidly at present, and recent reviews on the subject should be consulted for up-to-date information. 

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