Detectors, HPLC mass spectrometer

However, by no means are microbore HPLC columns nonoptimal when sample volumes are limited. Because of the substantially reduced mobile-phase volumes necessary to carry out a given separation, microbore columns are more easily interfaced to many useful detection systems, for example, electron-capture detectors, nitrogen-specific thermionic detectors, and mass spectrometers. 

Spectroscopic detectors measure partial or complete energy absorption, energy emission, or mass spectra in real-time as analytes are separated on a chromatography column. Spectroscopic data provide the strongest evidence to support the identifications of analytes. However, depending on the spectroscopic technique, other method attributes such as sensitivity and peak area measurement accuracy may be reduced compared to some nonselective and selective detectors. The mass spectrometer and Fourier transform infrared spectrometer are examples of spectroscopic detectors used online with GC and HPLC. The diode array detector, which can measure the UV-VIS spectra of eluting analytes is a... 

A decade ago, a system such as the one described would require many PCs to control all the component parts. The HPLC, mass spectrometer, fraction collector and even the diode array would each require a separate PC and. separate software. Modem systems are controlled by more integrated software and the MS-prep system is now controlled by one computer and one software platform. The demands on this computer should not be underestimated. The computer controls the HPLC system including the diode array detector and gradient conditions, the mass spectrometer, the make-up pump and the fraction collector. The same computer is also required to acquire and store data from each of these component instruments. 

The next generation of HPLC detectors, namely mass spectrometers, are now available and being deployed for water-soluble vitamin determinations in dietary supplements with electrospray ionization interface (ESI) (Holler et al. 2006). There remain obstacles to overcome for multivitamin analyses in complex food matrices, as co-elution of vitamins or excipients can compromise... 

In addition, the appHcation of the mass spectrometer (ms) as a detector for gas—Hquid chromatography has made the positive identification of peaks possible. High performance Hquid chromatography (hplc), which involves various detectors, can be used to measure hydrophilic and hydrophobic organic compounds in water. 

High-pressure pumps operating at up to 6000 psi are required to force solvent through a tightly packed HPLC column, and electronic detectors are used to monitor the appearance of material eluting from the column. Alternatively, the column can be interfaced to a mass spectrometer to determine the mass spectrum of every substance as it elutes. Figure 12.18 shows the results of HPLC analysis of a mixture of 10 fat-soluble vitamins on 5 jam silica spheres with acetonitrile as solvent. 

A more definitive identification may be obtained by combining retention characteristics with more specific information from an appropriate detector. Arguably, the most information-rich HPLC detectors for the general identification problem are the diode-array UV detector, which allows a complete UV spectrum of an analyte to be obtained as it elutes from a column, and the mass spectrometer. The UV spectrum often allows the class of componnd to be determined but the... 

HPLC was a widely used analytical technique before the development of LC-MS and continues to be used in many laboratories with other forms of detector. What are the advantages of the mass spectrometer when used as a detector ... 

The mass spectrometer provides the most definitive identification of aU of the HPLC detectors. It allows the molecular weight of the analyte to be determined - this is the single most discriminating piece of information that may be obtained - which, together with the structural information that may be generated, often allows an unequivocal identification to be made. 

In this chapter, an HPLC system has been described in terms of its component parts and the effect of each of these on the use of a mass spectrometer as a detector... 

To appreciate the ways in which mass spectral data may be processed to utilize fully the selectivity and sensitivity of the mass spectrometer as a detector for HPLC. 

A number of different types of HPLC detector have been discussed in the previous chapter. In comparison to these, a mass spectrometer is a relatively expensive detector and there need to be considerable advantages associated with its use to make the significant financial investment worthwhile. What are these advantages In order to answer this question, we must first consider what it is we are trying to achieve when using chromatography ... 

The application areas for LC-MS, as will be illustrated later, are diverse, encompassing both qualitative and quantitative determinations of both high-and low-molecular-weight materials, including synthetic polymers, biopolymers, environmental pollutants, pharmaceutical compounds (drugs and their metabolites) and natural products. In essence, it is used for any compounds which are found in complex matrices for which HPLC is the separation method of choice and where the mass spectrometer provides the necessary selectivity and sensitivity to provide quantitative information and/or it provides structural information that cannot be obtained by using other detectors. 

GC = gas chromatography ECD = electron capture detector EIA = enzyme-immunoassay GPC = gel permeation chromatography HPLC = high-performance liquid chromatography ITMS = ion trap mass spectrometer LSE = liquid solid extraction MS = mass spectrometry RSD = relative standard deviation SPE = solid phase extraction... 

Specifications for modem detectors in HPLC are given by Hanai [538] and comprise spectroscopic detectors (UV, F, FUR, Raman, RID, ICP, AAS, AES), electrochemical detectors (polarography, coulometry, (pulsed) amperometry, conductivity), mass spectromet-ric and other devices (FID, ECD, ELSD, ESR, NMR). None of these detectors meets all the requirement criteria of Table 4.40. The four most commonly used HPLC detectors are UV (80%), electrochemical, fluorescence and refractive index detectors. As these detectors are several orders of magnitude less sensitive than their GC counterparts, sensor contamination is not so severe, and... 

The mass spectrometer is mainly used as a mass detector in chromatography (GC, SFC, HPLC, SEC, TLC). With the great variety of interfaces, ionisation modes and mass spectrometers, chromatography-mass spectrometry is highly diversified. High-resolution separations combined with accurate mass measurements and element-sensitive detection (MIP, ICP) have been reported. 

Consider one small molecule, phenylalanine. It is an essential amino acid in our diet and is important in protein synthesis (a component of protein), as well as a precursor to tyrosine and neurotransmitters. Phenylalanine is one of several amino acids that are measured in a variety of clinical methods, which include immunoassay, fluorometry, high performance liquid chromatography (HPLC see Section 4.1.2) and most recently MS/MS (see Chapter 3). Historically, screening labs utilized immunoassays or fluorimetric analysis. Diagnostic metabolic labs used the amino acid analyzer, which was a form of HPLC. Most recently, the tandem mass spectrometer has been used extensively in screening labs to analyze amino acids or in diagnostic labs as a universal detector for GC and LC techniques. Why did MS/MS replace older technological systems The answer to this question lies in the power of mass spectrometer. 

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