The use of total alkaloids in Jingdongshan orange
A technology of total alkaloids and drugs, applied in the field of medicine, can solve problems such as no patent literature reports, and achieve the effects of expanding value, simple process flow, and simple extraction process
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Embodiment 1
[0023] Embodiment 1: Preparation of total alkaloids of Jingdongshan orange
[0024] The aboveground part of Jingdongshan orange was collected from Bijie, Guizhou, dried naturally and powdered to obtain the Jingdongshan orange sample; 100g of Jingdongshan orange sample was weighed, and the ratio of solid to liquid was g:mL=1:10. % methanol aqueous solution for 30 minutes, filtered, and the filter residue was repeated ultrasonic extraction for 3 times, the filtrate was combined, evaporated and concentrated at 45°C to obtain the extract; the extract was dissolved in 0.3% hydrochloric acid aqueous solution, and the pH was adjusted to 2-3 , extracted with ethyl acetate, collected the water layer, then added ammonia water with a volume concentration of 5%, adjusted the pH to 9-10, then extracted with ethyl acetate, collected the ethyl acetate layer, evaporated and concentrated, and freeze-dried to obtain the scene Dongshan orange total alkaloids (TAMK).
Embodiment 2
[0025] Example 2: Evaluation of anti-inflammatory activity at the cell level of RAW 264.7
[0026] LPS-induced RAW 264.7 was used to establish a cell inflammation model, and the expression of the total alkaloids of Jingdongshan orange on the expression of NO and related inflammatory factors in RAW264.7 cells was determined to evaluate the anti-inflammatory activity of the total alkaloids of Jingdongshan orange;
[0027] 1. Cell culture and non-toxic dosage screening
[0028] (1) RAW 264.7 cells were inoculated in DMEM containing 10% fetal bovine serum (FBS), 1% penicillin (100 U / mL) and streptomycin (100 mg / mL) mixed antibiotics, at 37°C, 5 Culture in a humid environment with % CO2 and 95% air;
[0029] (2) The toxicity of the total alkaloids of Jingdongshan orange to RAW264.7 cells was determined by MTT method, and the cultured RAW264.7 cells were treated with 1×10 4 Each well was connected to a 96-well plate. After 24 hours of culture, the medium was sucked away. The sampl...
Embodiment 3
[0039] Example 3: Establishing a mouse ear swelling model to evaluate the anti-inflammatory activity of total alkaloids in Jingdongshan orange
[0040] The mouse ear swelling model was established by smearing xylene, and the anti-inflammatory activity of the total alkaloids of Jingdongshan orange was evaluated by measuring the weight change of the mouse ear before and after smearing;
[0041] 1. Establishment of mouse ear swelling model and activity evaluation
[0042] Male Kunming mice are 4-5 weeks old Kunming mice (18-22g), 8 mice in each group, and the experimental groups are as follows:
[0043] Group K: control group (distilled water gavage, K), Group M: model group (distilled water gavage, M), Group DEX: positive control group (dexamethasone 5mg / kg), Group TAMKL: total alkaloids of Jingdongshan orange Low-dose group 50mg / kg, GroupTAMKM: Jingdongshan orange total alkaloids middle-dose group 100mg / kg. Group TAMKH: Jingdongshan orange total alkaloid high dose group 200 m...
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