Esi-qtof-ms coupled to hplc and its application for food safety  (Page 40/3)

Introduction

High-performance liquid chromatography (HPLC) is a very powerful separation method widely used in environmental science, pharmaceutical industry, biological and chemical research and other fields. Generally, it can be used to purify, identify and/or quantify one or several components in a mixture simultaneously.

Mass spectrometry (MS) is a detection technique by measuring mass-to-charge ratio of ionic species. The procedure consists of different steps. First, a sample is injected in the instrument and then evaporated. Second, species in the sample are charged by certain ionized methods, such as electron ionization (EI), electrospray ionization (ESI), chemical ionization (CI), matrix-assisted laser desorption/ionization (MALDI). Finally, the ionic species wil be analyzed depending on their mass-to-charge ratio ( m / z ) in the analyzer, such as quadrupole, time-of-flight (TOF), ion trap and fourier transform ion cyclotron resonance.

The mass spectrometric identification is widely used together with chromatographic separation. The most common ones are gas chromatography-mass spectrometry (GC-MS) and liquid chromatography-mass spectrometry (LC-MS). Because of the high sensitivity, selectivity and relatively low price of GC-MS, it has very wide applications in drug detection, environmental analysis and so forth. For those organic chemistry research groups, it is also a daily-used and convenient equipment. However, GC-MS is ineffective if the molecules have high boiling point and/or will be decomposed at high temperature.

In this module, we will mainly talk about liquid chromatography and electrospray ionization quadrupole time-of-flight mass spectrometry (LC/ESI-QTOF-MS). As mentioned above, the LC has an efficient capacity of separation and MS has a high sensitivity and strong ability of structural characterization. Furthermore, TOF-MS, has several distinctive properties on top of regular MS, including fast acquisition rates, high accuracy in mass measurements and a large mass range. The combination of LC and ESI-TOF-MS allow us to obtain a powerful in the quantitative and qualitative analysis of molecules in complex matrices by reducing the matrix interferences. It may play an important role in the area of food safety.

How it works

Generally, LC-MS has four components, including an autosampler, HPLC, ionization source and mass spectrometer, as shown in [link] . Here we need to pay attention to the interface of HPLC and MS so that they can be suitable to each other and be connected. There are specified separation column for HPLC-MS, whose inner diameter (I.D.) is usually 2.0 mm. And the flow rate, which is 0.05 - 0.2 mL/min, is slower than typical HPLC. For the mobile phase, we use the combination of water and methanol and/acetonitrile. And because ions will inhibit the signals in MS, if we want to modify to mobile phase, the modifier should be volatile, such as HCO ₂ H, CH ₃ CO ₂ H, [NH ₄ ][HCO ₂ ] and [NH ₄ ][CH ₃ CO ₂ ].