High-efficient extraction method applicable to ligneous plant endogenous hormone determination

Abstract

The invention belongs to the technical field of ligneous plants, and discloses a high-efficient extraction method applicable to ligneous plant endogenous hormone determination. The method comprises the steps of weighing lemon leaves, quick-freezing through liquid nitrogen, grinding into powder, adding a methanol solution, placing in a 4-DEG C refrigerator, and staying overnight; adding methanol for washing residues, repeatedly operating for two times, and combining supernatants; transferring the filtered supernatant into a rotary evaporation bottle, vacuum evaporating to obtain a water phase at 36 DEG C under weak light, getting the water phase, and using pure water for washing the rotary evaporation bottle for 1 time; refrigerating at minus 80 DEG C for 10min, and then freeze-drying; dissolving through 4ml of PBS with the pH being equal to 3.0; extracting through a 4ml centrifuge tube with 10ml of ethyl acetate for 3 times, remaining an ester phase, and removing the water phase; transferring an ethyl acetate phase into the rotary evaporation bottle, vacuum evaporating until drying at 36 DEG C under weak light, and using a moving phase of a pump A for dissolving an undersize column; refrigerating the water phase, freeze-drying, and directly storing; dissolving through a 5mL of moving phase, thus obtaining ZT; before carrying out HPLC (High Performance Liquid Chromatography) analysis, and filtering a sample through a microfiltration membrane with the hole diameters being 0.22 micron, wherein the filtered sample is used as a solution for test.

Description

technical field [0001] The invention belongs to the technical field of woody plants, and in particular relates to a high-efficiency extraction method suitable for measuring endogenous hormones of woody plants. Background technique [0002] High performance liquid chromatography (HPLC) has the characteristics of high efficiency and high sensitivity, and has become an ideal analysis method for plant endogenous hormones. The most commonly used extraction method at present is to extract with 80% icy methanol, extract overnight, collect the filtrate, spin evaporate to dryness, dissolve the raffinate with a small amount of water, freeze-thaw overnight, after melting, centrifuge to get the supernatant, adjust the pH, Add ethyl acetate for extraction, take the organic phase, and then rotate it to evaporate, and finally dissolve it with the mobile phase, and load it for determination. Woody plants have the characteristics of significantly higher content than herbaceous plants, compl...

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