High performance liquid chromatography incompatibilities with

This is not the case when high performance liquid chromatography (HPLC) and MS are considered where, due to the incompatibilities of the two techniques, they cannot be linked directly and an interface must be used, with its prime purpose being the removal of the chromatographic mobile phase. Unfortunately, no single... 

In the separation of biomolecules, sample preparation almost always involves the use of one or more pretreatment techniques. With high-performance liquid chromatography (HPLC), no one sample preparation technique can be appHed to all biological samples. Several techiques may be used to prepare the sample for injection. For example, complex samples require some form of preffactionation before analysis, samples that are too dilute for detection require concentration before analysis, samples in an inappropriate or incompatible solvent require buffer exchange before analysis, and samples that contain particulates require filtration before injection into the analytical instrument. 

One of the major advantages of SFC over HPLC (high-performance liquid chromatography) is its compatibility with the flame ionization detector (FID), a universal and sensitive detector for carbon compounds. Unfortunately, most modifiers used in SFC are incompatible with FID. Therefore, a search for polar modifiers that have less response to FID led to the use of water, formic acid, and formamide. These modifiers produce acceptably low background noise and enable the use of FID. Because both water and formic acid have poor solubilities in carbon dioxide, they have been used as modifiers at very low concentrations. However, the modifier effect is significant even at this low level. For example, when water or formic acid was used as modifiers, the resolution of free fatty acids was significantly improved. 

The determination of the different forms (e. g. compounds or complexes) in which an element occurs (often referred to as the speciation of an element and speciation analysis, respectively [28]) is in most cases performed by hyphenated techniques. These are the combination of a high-performance separation technique such as gas or liquid chromatography, or electrophoresis, and an element- or compound-specific detector [29]. While the former provides the separation of the different elemental species prevalent in the sample, the latter brings selective and sensitive detection. In the case of AAS, only the hyphenation with gas and Hquid chromatography, respectively, has gained importance. The combination of atomic absorption spectrometry and electrophoresis has never proven successfiil, obviously due to the incompatibility of the extremely low flow rates of electrophoretic separations with the aspiration volumes of flame atomisers and the difficulties of interfacing the two techniques. 

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