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Method for efficiently separating active polysaccharides from fresh okra
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A technology of active polysaccharides and okra, applied in the field of efficient separation of active polysaccharides, three-phase system combined with ammonium sulfate precipitation, to achieve good anti-oxidation and hypoglycemic activities in vitro, low cost, and large economic benefits
Inactive Publication Date: 2020-03-17
JIANGSU UNIV
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[0005] After searching, there is no report on the use of a three-phase system composed of tert-butanol and inorganic salt ammonium sulfate combined with ammonium sulfate precipitation for efficient separation of active polysaccharides in fresh okra
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Embodiment 1
[0022] Step 1, fresh okra homogenization treatment: 500 g of commercially available fresh okra was cleaned, sliced, added 1500 g of distilled water for homogenization treatment, and filtered through gauze to obtain filtrate and filter residue;
[0023] Step 2, three-phase system separation: add 1000mL of okra filtrate obtained in step 1 to 200g of ammonium sulfate, stir to dissolve, adjust the pH value of the solution to 7.0, add 1500mL of tert-butanol, shake in a constant temperature water bath at 35°C for 30min, 4500rpm Centrifuge for 10 min, and take out the upper layer tert-butanol phase and the lower layer ammonium sulfate phase respectively;
[0024] Step 3, ammonium sulfate precipitation and separation of okra polysaccharides: add 100 g of ammonium sulfate to the 1000 mL of ammonium sulfate phase solution obtained in step 2, so that the mass fraction of ammonium sulfate in the solution is 30%, centrifuge, dissolve the precipitate in water, and dialyze for 48 hours. Conc...
Embodiment 2
[0027] The test process is the same as in Example 1, except that 300 g of ammonium sulfate is added in step 3 so that the mass fraction of ammonium sulfate in the solution is 50%. The total sugar content of okra polysaccharides measured is 93.5%, and the molecular weight distribution is a single peak, such as figure 2 Shown, the weight average molecular weight is 3.6×10 6 Da, IC for scavenging DPPH radicals 50 The value is 0.045mg / mL, the equivalent antioxidant capacity is 71.3μmol Trolox / g sample, and the ability to reduce ferrous ions is 169.3μmol FeSO 4 / g sample, IC of α-amylase inhibitory activity 50 The value is 0.10 mg / mL.
Embodiment 3
[0029] The test process is the same as in Example 1, except that 200 g of ammonium sulfate is added in step 3 so that the mass fraction of ammonium sulfate in the solution is 40%. The total sugar content of okra polysaccharides measured is 90.1%, and the molecular weight distribution is a single peak, such as image 3 Shown, the weight average molecular weight is 4.3×10 6 Da, IC for scavenging DPPH radicals 50 The value is 0.11mg / mL, the equivalent antioxidant capacity is 68.2μmol Trolox / g sample, and the ability to reduce ferrous ions is 161.8μmol FeSO 4 / g sample, IC of α-amylase inhibitory activity 50 The value is 0.15 mg / mL.
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Abstract
The invention discloses a method for efficiently separating active polysaccharides from fresh okra, and belongs to the technical field of bioactive polysaccharide separation. The method comprises thefollowing steps: cleaning and slicing okra, adding distilled water, homogenizing the okra and distilled water, and filtering the obtained mixture with a gauze; adding an inorganic salt ammonium sulfate and tert-butyl alcohol into the obtained okra filtrate, oscillating the obtained solution in a 35-DEG C constant-temperature water bath for 30 min, and carrying out centrifugal separation; carryingout rotary evaporation on an upper-layer organic phase, and recovering the tert-butyl alcohol; precipitating the lower ammonium sulfate phase solution with ammonium sulfate with the mass fraction of 30-50%, carrying out centrifugal separation, dissolving the obtained precipitate in water, dialyzing the obtained solution, and performing spray drying to obtain an okra polysaccharide solid; and recovering ammonium sulfate from the precipitated ammonium sulfate solution through evaporative crystallization. The method has the advantages of simplicity in operation, easiness in implementation, low cost, realization of green and efficient obtaining of high-purity active polysaccharides, bringing great economic benefits to enterprises, and suitableness for large-scale promotion. In addition, the method can also be universally applied to efficient and high-purity separation of active polysaccharides of other fresh fruits and vegetables such as garlic, onions, tomatoes and apples.
Description
technical field [0001] The invention relates to a method for efficiently separating active polysaccharides from fresh okra, in particular to a three-phase system composed of tert-butanol and inorganic salt ammonium sulfate combined with ammonium sulfate precipitation for efficient separation of fresh okra active polysaccharides, which belongs to biological activity Technical field of polysaccharide separation. Background technique [0002] Okra (Abelmoschus esculentus (L.) Moench), also known as okra, coffee sunflower, papaya, Vietnamese sesame, is an annual herb, rich in a variety of active substances such as protein, fat, carbohydrates and rich in vitamin A and B group and trace elements such as calcium, phosphorus, iron, zinc and selenium have high nutritional and health value, and have the functions of anti-fatigue, improving immunity, reducing lung damage and anti-cancer. Okra polysaccharide is a kind of high molecular acidic polysaccharide, mainly composed of rhamnose...
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