Thin-layer chromatography silica gel plate production and operation method

by:Keyuan     2020-12-04
Thin-layer chromatography silica gel plate is what we usually call silicon sheet, to understand the thin-layer chromatography silica gel plate production and operation must know: thin-layer chromatography ( TLC) The principle of; Thin layer chromatography is a trace and rapid analysis method, easy operation, low equipment requirement, but for every kind of method of the separated material need to be independent, TLC is mainly used in amino acids, peptides, sugar, lipid, nucleotide and alkaloids separation and identification of a variety of material. Its main principle is according to the sample composition and adsorbent adsorption force and its distribution in the exhibition solvent layer coefficient of different and separate the mixture. Thin-layer chromatography silica gel plate operation method: a. Silica gel plate 30 min at 100 ℃ oven activation; After cooling, lines from the bottom 2 cm, with a pencil lightly marked sample point position, two points 1 - sample average distance 1. 5厘米; b。 General sample points, each point sample amount is zero. 8 μL( As the case may be) Sample points, points to small as far as possible, if you need to add quantity, stay dry sample points on the same point again after the second and third spot. Every sample point to point at the last point, after drying to prevent the spread of some sample excessive influence the results; c。 At room temperature after drying, quickly put some good silicon sheet in have an agent in advance chromatography cylinder, sealed chromatography cylinder, began to show layer; d。 To show layer of silica gel plate along the distance on the upper edge of 2 cm when take out the silica gel plate, room temperature is 50 ℃ after drying oven drying 5 min, take out, after waiting for silica gel plate cooling into the second exhibition layer chromatography cylinder; Recommended products: foaming silica gel plate e. Show layer after the completion of room temperature dry and choose the suitable method to color color and calculated the Rf value. Color rendering methods: TLC method of color has a lot of, some common methods of color is as follows: uv ZhaoSheFa, fluorescent reagent method and ultraviolet ZhaoSheFa, etc; But according to the different of the object under test to choose the right color method. Above is the thin-layer chromatography silica gel plate making and operating methods for reference only, the rubber to remind you: at the time of operation or experiment should pay attention to the points. 1. Sample: the dissolution of similar sample of the soluble in developing solvent polarity, high volatile organic solvents. 2. Point: sample weight control, baseline point sample from the bottom 2. 0 cm, diameter control sample ( 5 mm) And the distance between the point ( 1 - 1. 5 cm) , avoid because of diffusion and impact test ( The figure below) , point sample must pay attention to don't damage the thin layer surface. 3. Fair layer: layer containers must be sealed and in advance to join the solvent, the solvent vapour equilibrium. An agent cannot be higher than sample, otherwise, cannot obtain the Rf value crude product and accurate judgment on the thin layer plate of each component in the relative position. 4. Developing solvent selection criteria: have good solubility, separated for separation between components, the Rf value of the component under test in 0. 2 to 0. Between 8 and doesn't react with the component under test or adsorbent, boiling point, moderate viscosity smaller, unfolds component spots round and concentrated, mixed solvent had better use fresh preparation. 5. Edge effect, edge effect is a sample in chromatography, thin layer plate on both sides of the spots than middle spots moving fast, deflection and out to the sides. The reason for this is that when using mixed solvent, weak polarity and lower boiling point solvents in the thin layer plate two edge is volatile, made the agents on the thin layer plate have different proportion, and edge effect, the most effective way to solve the edge effect is cut off at the end of the thin layer plate into an agent two horns. More about the principle of thin layer chromatography silica gel plate production, please continue to pay attention: silica gel plate manufacturers relevant article recommended: foaming silica gel plate in everyday life really to remove silica gel products smell silicone mold manufacturing method, what | silicon mold making process
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