Extraction method and uses of traditional Chinese medicine cassia seed protein
An extraction method and technology of cassia seed, which is applied in the field of extraction and purification of active ingredients of traditional Chinese medicine, can solve the problems of cassia seed that have not been reported in literature, and the properties and functions of cassia seed protein that have not been seen
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Embodiment 1
[0044] according to figure 1 , 2 Process, the Semen Cassiae used in this example was purchased from a pharmacy and produced in Anhui. The extraction process was carried out at 4°C. Cassia seeds (1000g) were fully crushed and then soaked in petroleum ether for 24 hours to degrease, in 2 times. Recover the solvent, and soak the residue in 50mmol KH with a volume of 1g: 10ml 2 PO 4 -NaOH (pH8.0) for 10 h, repeated 3 times, and combined extracts. The extract was centrifuged at a high speed of 12,000r / min for 20min, and the supernatant was taken. Ammonium sulfate was added to the supernatant with stirring to 50% saturation. Centrifuge at high speed under the same conditions as above to retain the precipitate. The precipitate was desalted in a dialysis bag with a molecular weight of 3,000. The dialysate was double-distilled water, and the dialysate was changed every 6 hours. After the sample was fully desalted, it was first frozen in a -20°C refrigerator for 24 hours, and th...
Embodiment 2
[0056] Others are the same as in Example 1, except that Semen Cassiae (500g) is fully ground in step (1) and soaked in sherwood oil for 36h to degrease, which is carried out in 3 times. The residue was soaked in 50mmol KH at a volume of 1g:25ml 2 PO 4 -NaOH (pH8.0) for 12h, repeated 4 times. The extract was centrifuged at high speed for 25 minutes, and ammonium sulfate was added to the supernatant to reach a saturation of 45%. After high-speed centrifugation, the precipitate was desalted in a dialysis bag with a molecular weight of 5,000. After desalting, freeze in a -20°C refrigerator for 16 hours, and then transfer to a freeze dryer (-50°C) to freeze-dry for 72 hours; the crude protein in step (2) is dissolved in 15mM Tris-HCl (pH7.5) in the buffer. The molecular sieve column was equilibrated with 0.2mol NaCl, 15mmol Tris-HCl (pH7.5) buffer solution for 3 column volumes. The sample volume is about 240ul each time, and the flow rate is 0.3ml / min; in step (3), solution A: ...
Embodiment 3
[0058] Others are the same as in Example 1, except that Semen Cassiae (2000g) is fully ground in step (1) and then soaked in sherwood oil for 48h to degrease, which is carried out in 4 times. The residue was soaked in 50mmol KH at a volume of 1g:50ml 2 PO 4 -NaOH (pH8.0) for 8h, repeated 5 times. The extract was centrifuged at high speed for 30 min, the supernatant was added with ammonium sulfate to 55% saturation, centrifuged at high speed, and the precipitate was desalted in a dialysis bag with a molecular weight of 8,000. After desalting, freeze-dry in a freeze-dryer (-65° C.) for 72 hours; the crude protein in step (2) is dissolved in 25 mmol Tris-HCl (pH 8.0) buffer. The molecular sieve column was equilibrated with a buffer solution of 0.3mol NaCl and 25mmol Tris-HCl (pH8.0) for 5 column volumes. The sample volume is about 180ul each time, and the flow rate is 0.7ml / min; in step (3), solution A: 25mmol Tris-HCl (pH8.0); solution B: 1mol NaCl, 25mmol Tris-HCl (pH8.0). ...
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