Extraction, purification method and application of alkaloids in soft-shelled turtle
An extraction method and a technology of the purification method are applied in the field of purification and extraction of alkaloids in the turtle, and can solve the problems of limiting the application of the turtle, the substance that cannot confirm the substantial function of the turtle, and the unclear effective components of the turtle.
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Embodiment 1
[0039] This embodiment is the extraction method of total alkaloids in soft-shelled turtle.
[0040] Fresh soft-shelled turtles were washed with distilled water, dried at 45°C, ground into powder, and stored at low temperature at 4°C for later use.
[0041] Get 20g ground turtle powder and use 95% ethanol 200ml (material ratio 1:10), under the condition of 70 ℃, Soxhlet extraction for 8 hours, filter; Filter the filter residue with 70% methanol 200ml again (material ratio 1:10), in Under the condition of 70°C, Soxhlet extraction was performed for 8 hours, filtered, and the two extracts were combined.
[0042] After the extract was dried under reduced pressure, it was dissolved with 50 ml of 0.5% dilute hydrochloric acid, then extracted with 150 ml of chloroform, and the extract was dried under reduced pressure to obtain 1.124 g of the total alkaloid components of the turtle with antibacterial and anticancer activities. The extraction yield was 5.62%.
Embodiment 2
[0044] In this example, 9-(2',2'-dimethylpropionylhydrazone)-3,6-dichloro-2,7-bis-[2-(diethylamine) was obtained after purification of the total alkaloids of the turtle - Specific examples of ethoxy]fluorene (GBA18).
[0045] (1), 5 g of total alkaloids of the turtle obtained in Example 1 were dissolved in 50 ml of 0.5% dilute hydrochloric acid, and the pH value was adjusted to 8.0 with dilute ammonia; then, extracted with 150 ml of chloroform, and the extract was dried under reduced pressure to obtain 89 mg of solid extract.
[0046] (2) Take 500g of 200-300 mesh silica gel, mix it with chloroform after activation, and put it into a chromatographic column (3cm*50cm) by wet method. 89 mg of the solid extract obtained in step (1) was dissolved in chloroform and then wet-applied, eluted with chloroform / methanol (9:1), and the eluent was dried under reduced pressure to obtain 50 mg of the solid extract.
[0047] (3) Take 500g of 200-300 mesh silica gel, after activation, mix it ...
Embodiment 3
[0052] This embodiment is a specific embodiment of GBA18 carbon spectrum and hydrogen spectrum analysis.
[0053] 3.1 1 H-NMR analysis
[0054] See further here Table 1 and figure 2 .
[0055] Table 1
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[0058] 3.2 13 C-NMR analysis
[0059] See further here Table 2 and image 3 .
[0060] Table 2
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