High performance liquid chromatography and its difference from gas chromatography

<p> Abstract: High performance liquid chromatography is an important branch of chromatography. It uses liquid as the mobile phase and uses a high-pressure infusion system to pump a single solvent with different polarities or mixed solvents and buffers with different proportions into a fixed phase. The phase chromatography column, after the components in the column are separated, enter the detector for detection, so as to realize the analysis of the sample.

High performance liquid chromatography:

Especially refers to a chromatographic separation and analysis method using liquid as the mobile phase. It uses a high-pressure pump, a chemically bonded stationary phase high-efficiency separation column, a highly sensitive special detector and other new experimental techniques to establish a liquid chromatography analysis method based on classic chromatographic theory.

High pressure: 150-350 * 105 Pa

High efficiency: more than 30,000 trays / meter

High sensitivity: 10-9g (UV detection), 10-11g (fluorescence detection)

The difference between liquid chromatograph and gas chromatograph:

1. Analysis of the difference between objects

GC: suitable for samples that can be vaporized, have good thermal stability, and low boiling point; but for high boiling point, poor volatility, poor thermal stability, ionic and polymer samples, especially for most biochemical samples. 20% of organic matter

HPLC: suitable for samples that can be made into a solution after being dissolved (including organic medium solutions), not limited by the volatility and thermal stability of the sample, for biochemical samples with large molecular weight, difficult to gasify, and poor thermal stability, polymers and ions Type samples can be tested for a wide range of uses, accounting for 80% of organic matter

2. Differences in mobile phase differences

GC: The mobile phase is inert, the gas components have no affinity with the mobile phase, and only interact with the stationary phase.

HPLC: The mobile phase is a liquid, and the mobile phase and the components have an affinity force, which can improve the selectivity of the column, improve the resolution, and play a positive role in the separation. And there are many types of mobile phases, and there is a wide range of choices. Changing the polarity and pH of the mobile phase also plays a role in regulating separation. When two or more liquids with different ratios are used as mobile phases, the separation selectivity can also be increased.

3. Operating conditions

GC: heating operation

HPLC: room temperature; high pressure (large liquid viscosity, small peak broadening)

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