Liquid chromatography-mass instrumentation

Liquid chromatography/mass spectrometry (LC/MS) is an analytical technique combining the advantages of an LC instrument with those of a mass spectrometer. 

The need to understand the fate of pesticides in the environment has necessitated the development of analytical methods for the determination of residues in environmental media. Adoption of methods utilizing instrumentation such as gas chro-matography/mass spectrometry (GC/MS), liquid chromatography/mass spectrometry (LC/MS), liquid chromatography/tandem mass spectrometry (LC/MS/MS), or enzyme-linked immunosorbent assay (ELISA) has allowed the detection of minute amounts of pesticides and their degradation products in environmental samples. Sample preparation techniques such as solid-phase extraction (SPE), accelerated solvent extraction (ASE), or solid-phase microextraction (SPME) have also been important in the development of more reliable and sensitive analytical methods. 

Niessen, W.M.A., Advances in instrumentation in liquid chromatography-mass spectrometry and related liquid-introduction techniques, ]. Chromatogr. A, 794, 407, 1998. 

Other combinations are available. For example, liquid chromatographs connected to mass spectrometers (known as liquid chromatography-mass spectrometry [LC-MS]) are fairly common. Almost any combination of two instruments that can be thought of has been built. In addition, two of the same instruments can be connected so that the output from one is fed directly into the other for further separation and analysis. Examples include two mass spectrometers in an MS-MS arrangement and two different gas chromatography columns connected in a series, known as GC-GC. To keep up with these advances, one needs to have a working knowledge of the fundamental principles involved in the techniques and of the abbreviations used for the various instrumentation methods. 

The different ways a particle beam liquid chromatography mass spectrometer can be configured reflect the versatility of the system in accommodating both the application and the availability of existing instrumentation. The system consists of these elements ... 

Many of the more established techniques have been validated through collaborative studies which becomes of greater importance as laboratories seek to become accredited via ISO, EN or related systems where the use of official or well validated methods is mandatory. New instrumental techniques are constantly being reported in the literature but it often requires many years before procedures are introduced, validated and then applied within the food industry. Recent techniques that can be included in this category are capillary electrophoresis and liquid chromatography-mass spectrometry (LC-MS). In time procedures based on these techniques will also become accepted as routine methods and are likely to be adopted by some of the official international bodies like the AOAC International, CEN, ISO, etc. 

Of particular note is liquid chromatography/mass spectrometry (LC/MS) detection. LC/M S technology is a critical technique for DMPK studies due to its ability to analyze samples with very high sensitivity and specificity particularly within complex mixtures. It is not uncommon to find LC/MS based sample analysis residing within its own functional department due to the specialized facility requirements and technical skills of the operators. Additionally with LC/MS instrumentation becoming lower cost and simpler to operate, they are also becoming a workhorse... 

W. M. A. Niessen and A. P. Tinke, Liquid chromatography-mass spectrometry. General principles and instrumentation , 7. Chromatogr. 703 37-57 (1995). 

Liquid chromatography/mass spectrometry (LC/MS)-based techniques provide unique capabilities for pharmaceutical analysis. LC/MS methods are applicable to a wide range of compounds of pharmaceutical interest, and they feature powerful analytical figures of merit (sensitivity, selectivity, speed of analysis, and cost-effectiveness). These analytical features have continually improved, resulting in easier-to-use and more reliable instruments. These developments coincided with the pharmaceutical industry s focus on describing the collective properties of novel compounds in a rapid, precise, and quantitative way. As a result, the predominant pharmaceutical sample type shifted from nontrace/pure samples to trace mixtures (i.e., protein digests, natural products, automated synthesis, bile, plasma, urine). The results of these developments have been sig-... 

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. 

Biological samples (plasma, serum, blood, and urine) are very complex. They contain a wide variety of matrix components such as proteins, lipids, and salts. To quantify trace amount of analytes (e.g., drug and its metabolites) in complex biological samples by liquid chromatography-mass spectrometry (LC-MS), the samples should be properly treated prior to being injected onto an LC-MS instrument,... 

As an instrumental approach to conventional electrophoresis, capillary electrophoresis offers the capability of on-line detection, micropreparative operation and automation (6,8,45-47). In addition, the in tandem connection of capillary electrophoresis to other spectroscopy techniques, such as mass spectrometry, provides high information content on many components of the simple or complex peptide under study. For example, it has been possible to separate and characterize various dynorphins by capillary electrophoresis-mass spectrometry (33). Therefore, the combination of CE-mass spectrometry (CE-MS) provides a valuable analytical tool useful for the fast identification and structural characterization of peptides. Recently, it has been demonstrated that the use of atmospheric pressure ionization using Ion Spray Liquid Chromatography/ Mass Spectrometry is well suited for CE/MS (48). This approach to CE/MS provides a very effective and straightforward method which allow the feasibility of obtaining CE/MS data for peptides from actual biological extracts, i.e., analysis of neuropeptides from equine cerebral spinal fluid (33). 

W.M.A. Niessen, Progress in liquid chromatography-mass spectrometry instrumentation and its impact on high-throughput screening, 7. Chromatogr., A, 1000, 413-436 (2003). 

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. 

The molecular mass of protein and peptide samples was determined by electrospray ionization mass spectrometry using a Perkin-Elmer Sciex API 100 mass spectrometer. The sample was introduced either by infusion or by on-line liquid chromatography/mass spectrometry (LC/MS) using a splitter. The data were obtained by scanning from 450 to 2000 Da with a scan time of 5 s and a step size of 0.25 Da with 1.0 ms dwell time per mass step. The molecular mass of the sample was obtained using the software provided by the instrument manufacturer. 

Over 30 years of liquid chromatography-mass spectrometry (LC-MS) research has resulted in a considerable number of different interfaces (Ch. 3.2). A variety of LC-MS interfaces have been proposed and built in the various research laboratories, and some of them have been adapted by instmment manufacturers and became commercially available. With the advent in the early 1990 s of interfaces based on atmospheric-pressure ionization (API), most of these interfaces have become obsolete. However, in order to appreciate LC-MS, one carmot simply ignore these earlier developments. This chapter is devoted to the older LC-MS interfaces, which is certainly important in understanding the histoiy and development of LC-MS. Attention is paid to principles, instrumentation, and application of the capillary inlet, pneumatic vacuum nebulizers, the moving-belt interface, direct liquid introduction, continuous-flow fast-atom bombardment interfaces, thermospray, and the particle-beam interface. More elaborate discussions on these interfaces can be found in previous editions of this book. 

Liquid chromatography-mass spectrometiy (LC-MS) based on atmospheric-pressure ionization (API) was demonstrated as early as 1974 (Ch. 3.2.1). However, it took until the late 1980 s before API was starting to be widely applied. Today, it can be considered by far the most important interfacing strategy in LC-MS. More than 99% of the LC-MS performed today is based on API interfacing. In this chapter, instrumentation for API interfacing is discussed. First, vacuum system for MS and LC-MS are briefly discussed. Subsequently, attention is paid to instrumental and practical aspects of electrospray ionization (ESI), atmospheric-pressure chemical ionization (APCI), and other interfacing approaches based on API. The emphasis in the discussion is on commercially available systems and modifications thereof. Ionization phenomena and mechanisms are dealt with in a separate chapter (Ch. 6). Laser-based ionization for LC-MS is briefly reviewed (Ch. 5.9). 

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