High performance liquid mass analyzers

Figure 2.1 Mass spectrometric approach. Dl, direct inlet GC, gas chromatography HPLC, high performance liquid chromatography CZE, capillary zone electrophoresis El, electron ionization Cl, chemical ionization ESI, electrospray ionization DESI, desorption electrospray ionization APCI, atmospheric pressure chemical ionization MALDI, matrix assisted laser desorption ionization B, magnetic analyzer E, electrostatic analyzer...

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. 

A more traditional but still successful method for the detection of a protein phosphorylation is by radioactive labeling with 35P. The labeled protein is digested, the peptides are separated by high-performance liquid chromatography, and the phosphorylated peptides are detected in specific fractions via their radioactivity. The fraction with the phosphorylated peptides can be further analyzed by mass spectrometry (Figeys et al., 1999). 

Many methods have been used to quantify steroidal compounds. These include RIA, gas chromatogra-phy-mass spectrometry (GC/MS), high-performance liquid chromatography (HPLC), and liquid chroma-tography-mass spectrometry (LC/MS). Although these techniques are successful in the analysis of steroids, it has been difficult to achieve quantitative analysis of small samples of neurosteroids because of their low concentrations in nervous tissues. Highly specific analytical methods are required to analyze small quantities of neurosteroids and their sulfates. Only with extremely sensitive methods of analysis is it possible to discover whether neurosteroids are synthesized in nervous tissues in quantities sufficient to affect neuronal activity, and whether these neurosteroids are distributed uniformly in brain. 

D Commercial COTS controlled by external computer Hybrid systems such as automated dissolution workstation with high-performance liquid chromatography (HPLC) or ultraviolet-visible (UV-Vis) interface Liquid chromatographs, gas chromatographs, UV/Vis spectrophotometers, Fourier transform infrared (FTIR) spectrophotometers, near-infrared (NIR) spectrophotometers, mass spectrometers, atomic absorption spectrometers, thermal gravimetric analyzers, COTS automation workstations... 

More recently, butterfat composition has been analyzed by capillary GC on polarizable liquid phases (122,123), reversed-phase high-performance liquid chromatography (124,125), and GC and HPLC with mass spectrometry (MS) (126), as well as by MS-MS (127). These studies have led to an extensive resolution of butterfat triacylglycerols, but the possible presence of small amounts of positional and reverse isomers as well as of species containing two or three short-chain acids per molecule has not been addressed. 

---