High pressure chromatography-coupled mass spectrometry

W.E. Neff, W.C. Byrdwell, Characterization of model triacylglycerol (triolein, trilinolein and trilinolenin) autoxidation products via high performance liquid chromatography coupled with atmospheric pressure chemical ionization mass spectrometry, Journal of Chromatography A, 818, 169 186 (1998). 

In recent years, several techniques have been developed for mass spectrometry, whereby samples are ionized and analysed from a condensed phase, without prior volatilization. These desorption techniques have permitted the extension of mass spectrometric analyses to sulfate and glutathione conjugates, as well as to underivatized and labile glucuronic acid conjugates. Primary among these techniques are field desorption 6, plasma desorption (7), laser desorption (8), fast atom bombardment (or secondary ion mass spectrometry with a liquid sample matrix) ( ) and thermospray ionization ( O). The latter can also serve to couple high pressure liquid chromatography and mass spectrometry for analysis of involatile and thermally labile samples. 

El-Hela, A.A., Al-Amier, H.A., and Ibrahim, T.A. 2010. Comparative study of the fla-vonoids of some Verbena species cultivated in Egypt by using high-performance liquid chromatography coupled with ultraviolet spectroscopy and atmospheric pressure chemical ionization mass spectrometry. J. Chromatogr. A 1217 6388-6393. 

GuiUarme, D., Schappler, J., Rudaz, S., and Veuthey, J.L. 2010. Coupling ultra-high-pressure liquid chromatography with mass spectrometry. Trends Arud. Chem. 29 15-27. 

The use of separation techniques, such as gel permeation and high pressure Hquid chromatography interfaced with sensitive, silicon-specific aas or ICP detectors, has been particularly advantageous for the analysis of siUcones in environmental extracts (469,483—486). Supercritical fluid chromatography coupled with various detection devices is effective for the separation of siUcone oligomers that have molecular weights less than 3000 Da. Time-of-flight secondary ion mass spectrometry (TOF-sims) is appHcable up to 10,000 Da (487). 

Whilst these methods are informative for the characterisation of synthetic mixtures, the information gained and the nature of these techniques precludes their use in routine quantitative analysis of environmental samples, which requires methods amenable to the direct introduction of aqueous samples and in particular selective and sensitive detection. Conventionally, online separation techniques coupled to mass spectrometric detection are used for this, namely gas (GC) and liquid chromatography (LC). As a technique for agrochemical and environmental analyses, high performance liquid chromatography (HPLC) coupled to atmospheric pressure ionisation-mass spectrometry (API-MS) is extremely attractive, with the ability to analyse relatively polar compounds and provide detection to very low levels. 

Henchiz Y. Guillarme D. Martel S. Rudaz S. Veuthey J-L. Carrupt P-A. Fast log P determination by ultra-high-pressure liquid chromatography coupled with UV and mass spectrometry detections. Analytical andBioanalytical Chemistry, 2009, 394, 1919-1630. 

Mass spectrometry (MS) is probably a famhiar tool to chemistry and biology students as a technique commonly used to measure the molecular mass of a sample. Often, MS is used in tandem with other techniques for chromatic separation of the sample before mass measurement. Some common hyphenated techniques include HPLC-MS, high-pressure liquid chromatography coupled to MS GC-MS, gas chromatography coupled to MS or CE-MS, capillary electrophoresis coupled to MS. 

In recent years the rapid development of high-sensitivity analytical techniques such as mass spectrometry (MS) and liquid chromatography (LC) supported the investigation of the endocannabinoids as part of a complex lipid network. The identification of lipid components of the endocannabinoid system can be achieved in a single analytical step by state-of-the-art platforms such as tandem mass spectrometry (MS/MS), which provides the detailed structural information necessary for characterization of lipids and increases specificity in complex biological matrices. Furthermore, the implementation of ionization techniques such as electrospray ionization (ESI) and atmospheric pressure chemical ionization (APCI) allow the coupling of LC to MS, and permits the separation and analysis of endocannabinoids with greater speed and accuracy. 

Figure 4 Schematic diagram of high-pressure liquid chromatography inductively coupled plasma mass spectrometry (HPLC-ICP-MS) system with dual-inlet system for isotope dilution analyses. (From Ref. 32.)...

Strege summarized the technique of high-performance liquid chromatography-electrospray ionization mass spectrometry (HPLC-ESI-MS) in dereplication of natural products. In contrast to earlier electron impact ionization (El), ESI technique is applicable to virtually any ion in solution with a soft ionization process. A comparison of ESI with fast atom bombardment (FAB), matrix assisted laser desorption ionization (MALDI), atmospheric pressure chemical ionization (APCI) and other techniques demonstrates its superior sensitivity, compatibility and reliability when coupled with HPLC [51]. 

MALDI and ESI represent the predominant ionization techniques in mass spectrometry-based proteomics, as recognized by the Nobel Prize in chemistry in 2002. MALDI is mainly used to volatize and ionize simple polypeptide samples for mass spectrometric (MS) analysis at high speed. The analysis of more complex peptide mixtures is usually conducted via ESI mass spectrometry (ESI MS) coupled online with a high-pressure liquid chromatography (HPLC) system to concentrate and separate peptides prior to MS analysis. 

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