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High performance liquid chromatography is based on classical chromatography, citing the theory of gas chromatography. Technically, the mobile phase is changed to high pressure delivery (maximum delivery pressure up to 4.9'107Pa); the column is used in a special way. The small particle size filler is filled, so that the column efficiency is much higher than the classical liquid chromatography (the number of plates per meter can reach tens of thousands or hundreds of thousands); at the same time, the column is connected with a highly sensitive detector, which can flow out The material is continuously tested.
1. High pressure: Liquid chromatography uses a liquid as the mobile phase (called a carrier liquid). The liquid flows through the column and is subject to high resistance. In order to pass the column quickly, a high pressure must be applied to the carrier liquid. Generally up to 150 ~ 350 × 105Pa.
2. High speed: The flow rate of the mobile phase in the column is much faster than that of the classical chromatography, generally up to 1~10ml/min. The analytical time required for high performance liquid chromatography is much less than that of classical liquid chromatography, typically less than 1 h.
3. Efficient: Recently, many new stationary phases have been developed to greatly improve the separation efficiency.
4. High sensitivity: High-performance liquid chromatography has widely used high-sensitivity detectors to further improve the sensitivity of the analysis. Such as fluorescence detector sensitivity up to 10-11g. In addition, the amount of sample used is small, generally a few microliters.
5. Wide adaptability: comparison between gas chromatography and high performance liquid chromatography: gas chromatography has the advantages of good separation ability, high sensitivity, fast analysis speed and convenient operation, but it is too high in boiling point due to technical conditions. Substances with poor material or thermal stability are difficult to analyze by gas chromatography. In high performance liquid chromatography, only the sample can be made into a solution without gasification, so it is not limited by the volatility of the sample. For high-boiling, poor thermal stability and relatively high molecular weight (more than 400) organic matter (these substances account for almost 75% to 80% of the total organic matter), high-performance liquid chromatography can be used for separation and analysis. According to statistics, among the known compounds, about 20% can be analyzed by gas chromatography, and about 70 to 80% can be analyzed by liquid chromatography.
This is the end of the introduction of Characteristics of high performance liquid chromatography(HPLC). I hope it can help you.