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Using high performance liquid chromatography HPLC a Video

Apologise, but: Using high performance liquid chromatography HPLC a

High performance liquid chromatography HPLC is an instrument and equipment which applies the principle of high performance liquid chromatography and Uaing mainly used for the analysis of organic compounds with high boiling point, non-volatile, thermal instability and large molecular weight. It consists of reservoir, pump, injector, chromatographic column, detector and recorder.

The mobile phase in the reservoir is pumped into the system by a high-pressure pump.

Detection items of high performance liquid chromatography

The sample solution enters the mobile phase through the injector and is loaded into the chromatographic column stationary phase by the mobile phase. Because the components in the sample solution have different partition coefficients in the two phases, when they move relative to each other in the two phases, after repeated adsorption desorption distribution process, the moving speed of each component is greatly different When passing through the detector, the sample concentration is converted into an electrical signal and transmitted to the recorder, chromahography the data is printed out in the form of chromatogram.

HPLC is widely used in life science, food science, pharmaceutical research and environmental research. The mobile phase in the reservoir is pumped into the detection system by a high-pressure pump.

What is the principle of HPLC

According to the different separation mechanism, the principles of HPLC can be divided into liquid-solid chromatograhpy chromatography, liquid-liquid partition chromatography normal phase and reverse phaseion exchange chromatography and molecular exclusion chromatography. In liquid-solid adsorption chromatography, the stationary phase is a solid adsorbent, and the components can be separated according to the adsorption capacity Using high performance liquid chromatography HPLC a each component.

More info commonly used adsorbents are silica gel or alumina, most of which are used for non-ionic compounds. Adsorption chromatographic stationary phases can be divided into polar and nonpolar. The requirements for mobile phase are as follows:.

If the UV absorption detector is used, the solvent with UV absorption at the detection wavelength cannot be selected; if the differential refractive detector is used, the gradient elution cannot be used.

Liquid solid chromatography is based on the surface adsorption capacity, so it is often used to separate compounds with different polarity, and can also separate those samples with the same polar group but different quantity. The stationary phase is a liquid, which is separated according to the solubility of the separated components in the mobile phase and the stationary phase. According to the polarity of stationary phase and mobile phase, it can be divided into normal phase chromatography and reverse phase chromatography. Normal phase chromatography uses polar stationary phase, and the mobile phase is relatively nonpolar hydrophobic solvent, which is commonly used for the separation of moderate polarity hjgh strong polarity compounds; reverse phase chromatography generally uses nonpolar stationary phase, and article source mobile phase is water or buffer solution, which is suitable for the separation of non polarity and weak polarity compounds.

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Among them, reversed phase chromatography is the most widely used. The stationary phase is ion exchange resin. The ionizable ions on the resin are exchanged with the ions with the same charge in the mobile phase and the ions of the tested component, and separated according to the different charge attraction of each ion and ion exchange group. Molecular exclusion chromatography, also known as gel chromatography, is a chromatographic method based on molecular size. The stationary phase gel of molecular exclusion chromatography is a porous polymeric material with a certain shape and stability.

Molecular sieve is used to separate the difference of different molecular weight exclusion capacity. According to the different mobile phase, gel chromatography can be divided into two types: water solvent, mobile phase gel filtration chromatography GFC and gel permeation chromatography GPC using organic solvents such as tetrahydrofuran. Mainly for the liquid organic matter with UV absorption unsaturated organic matterqualitative and quantitative analysis and purity analysis. Qualitative: the answer is the characteristic peak of retention time.]