A method for determining the alkali content of steroidal alkaloids produced by tissue culture

Abstract

The invention relates to an alkali content determination method for steroid alkaloid produced by tissue culture method. The alkali content determination method comprises the following steps of (1) preparing culture media; (2) carrying out callus culture; (3) carrying out rooting culture; (4) extracting and preparing the alkaloid; (5) weighing a sample prepared by the step (4) precisely, concentrating and volatilizing by reduced pressure distillation to recover ethanol, preparing a solution by using 5 mL of glacial acetic acid and standing in a refrigerator for a certain time for use; (6) analyzing and determining a peak area value by using a high performance liquid chromatography; (7) establishing a standard curve; and (8) determining the steroid alkaloid content: calculating the steroid alkaloid content of the sample according to the peak area value determined by the step (6) and a regression equation of the step (7). The alkali content determination method takes use of an ultrasonic technique, is relatively small in solvent usage amount, reduces cost, takes use of a rotary evaporator to distill the solvent under the reduced pressure, reduces pressure in the system by virtue of a vacuum pump, reduces a boiling point of the liquid, and obtains an alkaloid concentrated solution at a relatively low temperature.

Description

Technical field [0001] The invention relates to a method for determining the alkali content of steroidal alkaloids produced by a tissue culture method. Background technique [0002] The traditional methods for extracting alkaloids include decoction, immersion, reflux and percolation. The operation is simple and easy, and more effective ingredients can be extracted. However, there are high energy consumption, high loss of effective ingredients and relatively high impurities in the extraction process. Many problems such as low efficiency. With the development of physical science, some new technologies have been applied to the extraction process of alkaloids in view of the problems existing in the traditional extraction process. The strengthening effect of the new technology or the extraction of fluids under supercritical conditions greatly improves the extraction efficiency and reduces Process energy consumption has become a research hotspot due to its significant advantages. Sun...

AI Technical Summary

Problems solved by technology

[0002] The traditional methods of extracting alkaloids include decoction, dipping, reflux and percolation, which are easy to operate and can extract more active ingredients, but there are many problems in the extraction process, such as large energy consumption, large loss of active ingredients, and relatively high impurities. Many, low efficiency and other issues

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