LC-MS and LC-MS/MS: Principles and Applications

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Introduction

Mass spectrometry (MS) is a method used to analyse a sample and measure the mass-to-charge ratio thereof. It can be used to determine the mass of samples as well as the composition of the sample.

Liquid chromatography (LC) or more specifically known as high-performance liquid chromatography (HPLC) is a technique that makes use of chromatography to separate a mixture of complex compounds into its constituent molecules and can further be used to identify, quantify and purify these components.

The combination of MS and LC is a highly specialised analytical technique with high sensitivity and combines the separation capability of HPLC with the mass analysis of MS (COVEY, et. al., 1986). Liquid chromatography-mass spectrometry (LC-MS also known as HPLC-MS) can be used for the specific detection of a certain chemical substance in the presence of numerous other chemicals closely related to the required substance. Thus the identification of a certain substance in a complex mixture can be done (ARDREY, 2003).

Chromatography decreases analysis speed but offers two main benefits. Metabolites with identical masses, but different retention times, can be distinguished and the separation of metabolites from interfering substances allows for improved quantitative accuracy (LAST, et. al., 2007).

2. LC-MS

2.1 Principles of LC-MS and LC-MS/MS

The combination of the separation technique of LC allows pure compounds to enter the identification by mass spectrometry and therefore increases the chance of successful identification of an unknown substance. This is based on the principle that many compounds with identical retention characteristics have different mass spectra and in this way can be differentiated. The...

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 NANDAKUMAR, R., MADAYIPUTHIYA, N. & FOUAD, A.F. 2009. Proteomic analysis of endodontic infections by liquid chromatography-tandem mass spectrometry. Oral microbioligal immunology. 24:347-352

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