Method for carrying out extraction, inspection and content determination on 5-hydroxytryptamine in sea buckthorn
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A technology of serotonin and seabuckthorn, which is applied in the field of extraction, inspection and content determination of indole derivatives in seabuckthorn, can solve the problems of large amount of organic solvent, cumbersome operation steps, time-consuming and labor-consuming, and achieve fast development speed, simple equipment, The effect of easy operation
Active Publication Date: 2019-01-11
LULIANG UNIV
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There have been many reports at home and abroad on the methods of extracting and separating 5-HTP from Ghana seeds, but these methods generally have the disadvantages of cumbersome operation steps, large amount of organic solvents, not conducive to environmental protection, and residual solvents and resin monomers.
[0005] In order to solve the cumbersome extraction, inspection and content determination of 5-hydroxytryptamine in seabuckthorn, the process of manually scraping, extracting, recombining, and then injecting and analyzing on the mass spectrometer to obtain the mass spectrum is time-consuming and labor-intensive, and the content determination results are easily disturbed. Accurate problem, the present invention provides a time-saving, labor-saving, easy-to-operate method that can quickly extract and detect 5-hydroxytryptamine from seabuckthorn and determine its content
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[0042] Step a, crude extraction of 5-hydroxytryptamine:
[0043] Dry the seabuckthorn peels and branches in the shade, crush them, pass through a 40-mesh sieve, and store them in glass containers. Accurately weigh 10g of powder and add absolute ethanol respectively, the ratio of solid to liquid is 1:20 (w / v). The ultrasonic microwave extractor extracts in three stages, the time is 2min, 3min, 5min, the temperature is 60°C, 70°C, 80°C, the ultrasonic power is constant, the microwave power is 450W, and the ultrasonic power is 700W to obtain the extract. Centrifuge the extract at 5000r / min for 10min, filter, repeat 3 times and combine the filtrates. Set the temperature to 77°C, speed 30 to concentrate to 1 / 4, add 200mL methanol to dissolve, then add 200mL n-hexane to mix and extract, take the methanol phase at 66°C, speed 30 to concentrate to solid, add 100mL ethyl acetate to dissolve, wait until fully After dissolving, add 100mL ultrapure water, after separation, take ethyl ac...
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Abstract
The invention discloses a method for carrying out extraction, inspection and content determination on 5-hydroxytryptamine in sea buckthorn and relates to the field of extraction, inspection and content determination of indole derivatives in the sea buckthorn. Sea buckthorn fruit peels and fruit branches are taken as a raw material; the raw material is smashed and screened; absolute ethyl alcohol is added; ultrasonic microwave extraction is carried out; centrifugal filtration is carried out; the raw material is concentrated to 1/4; methanol is added for dissolution; normal hexane is added, andmixing, extraction and concentration are carried out; ethyl acetate is added for dissolution; after sufficient dissolution is carried out, ultrapure water is added; liquid separation is carried out; ethyl acetate phase is obtained and is concentrated to dry; little absolute ethyl alcohol is added for dissolution; volume is fixed at 50mL; content is determinated through TLC reflection scanning; molecular weight information of samples is determinated through utilization of a mass spectrometer; functional groups are detected through infrared; optical rotation and specific rotation are determinated through utilization of a polarimeter; evaporation and crystallization are carried out; and a melting point is determinated. According to the method provided by the invention, time and labor are reduced; a separation effect is relatively good; purity is high; a result is accurate; and the method for carrying out the extraction, inspection and content determination is simple in operation and highin efficiency.
Description
technical field [0001] The invention relates to the field of extraction, testing and content determination of indole derivatives in seabuckthorn, in particular to a method for extracting, purifying and testing 5-hydroxytryptamine in seabuckthorn. Background technique [0002] Seabuckthorn, belonging to the genus Hippophae rhamnoides, its chemical components mainly include vitamins, proteins and amino acids, oils and fatty acids, volatile oils, organic acids, sugars, 5-hydroxytryptamine, trace elements, flavonoids, terpenes And sterols, etc., there are more than 300 kinds of biologically active substances, among which 5-hydroxytryptamine exists in the root bark, stem bark and fruit of seabuckthorn in free form and compound form. [0003] 5-hydroxytryptamine, also known as serotonin, has the molecular formula C 10 h 12 N 2 O, referred to as 5-HT, widely exists in mammalian tissues, especially in the cerebral cortex and synapses. It is an inhibitory neurotransmitter and an i...
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