Efficient extraction method applicable to measuring endogenous hormone of herbaceous plants

Abstract

The invention belongs to the technical field of herbaceous plants, and discloses an efficient extraction method applicable to measuring endogenous hormone of herbaceous plants. The efficient extraction method includes weighing 0.2 g of ludwigia repens, quickly freezing the ludwigia repens by the aid of liquid nitrogen, grinding the ludwigia repens to obtain powder, adding the powder into 10 ml ofcooled 80% methanol solution and placing the methanol solution in a refrigerator at the temperature of 4 DEG C overnight; washing residues by the aid of 5 mL of 80% methanol, repeatedly carrying out operation twice and combining supernatant; transferring filtered supernatant into rotary evaporating flasks, carrying out reduced-pressure evaporation in dim light at the temperatures of 36 DEG C untilwater phase is obtained, acquiring approximately totally 6 ml of water phase, and washing each rotary evaporating flask by the aid of pure water once; carrying out cold storage at the temperature of-80 DEG C for 10 min and then carrying out freeze-drying to obtain dry powder; dissolving the dry powder by the aid of mobile phase of a pump A to obtain samples and allowing the samples to flow through Pak-ODS-C18 small columns; allowing the samples to flow through microporous filter membranes with the sizes of 0.22 micrometer before HPLC (high-performance liquid chromatography) analysis is carried out. The samples which flow through the microporous filter membranes are used as liquid for test.

Description

technical field [0001] The invention belongs to the technical field of herbal plants, and in particular relates to a high-efficiency extraction method suitable for measuring endogenous hormones of herbal 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, com...

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