阅读说明：本技术 三叶木通果皮多酚的提取分离方法 (The extraction separation method of Akebia trifoliate koiaz Peels polyphenol ) 是由 张孟琴 田玉婷 罗彪 于 2019-07-19 设计创作，主要内容包括：本发明公开了一种三叶木通果皮多酚的提取分离方法,将三叶木通果皮洗净、干燥、粉碎、过滤,得到粉末；接着酶解,向粉末中加入所述粉末重量的纤维素酶、果胶酶、中性蛋白酶和水混合搅拌均匀,并用超声波处理,停止超声,于下水浴保温,得酶解液；向上述酶解液中加入所述酶解液重量的氯化铝,并将pH调整为5.5-6.5,静置12-24h后,于2000-2400r/min下离心10-15min,过滤分离出滤液,将混合沉淀用酸溶解成液体,并向液体中添加等量的乙酸乙酯萃取,乙酸乙酯的体积浓度为40-60％,得萃取液；用截留分子量为1-3万的PES膜对萃取液进行超滤,膜面流速为2-2.5m/s,操作温度为35-45℃,跨膜压力为0.1-0.2MPa,超滤后对滤液进行浓缩,干燥,得到三叶木通皮多酚粉末,提取成本低,适合大规模生产。(The invention discloses a kind of extraction separation methods of Akebia trifoliate koiaz Peels polyphenol, and Akebia trifoliate koiaz Peels are cleaned, is dry, are crushed, filtering, obtain powder；Then it digests, cellulase, pectase, neutral proteinase and the water that the powder weight is added into powder are mixed evenly, and with ultrasonication, stop ultrasound, keep the temperature in lower water-bath, obtain enzymolysis liquid；The aluminium chloride of the enzymolysis liquid weight is added into above-mentioned enzymolysis liquid, and pH is adjusted to 5.5-6.5, after standing 12-24h, 10-15min is centrifuged under 2000-2400r/min, filtrate is filtered to isolate, by mixed precipitation acid dissolution at liquid, and the ethyl acetate of addition equivalent extracts into liquid, the volumetric concentration of ethyl acetate is 40-60%, obtains extract liquor；Ultrafiltration, crossflow velocity 2-2.5m/s are carried out to extract liquor with the PES film that molecular cut off is 1-3 ten thousand, operation temperature is 35-45 DEG C, transmembrane pressure is 0.1-0.2MPa, and filtrate is concentrated after ultrafiltration, dry, threeleaf akebia skin polyphenol powder is obtained, extraction cost is low, is suitble to large-scale production.)

1. the extraction separation method of Akebia trifoliate koiaz Peels polyphenol, it is characterised in that: comprise the following steps that

1) Akebia trifoliate koiaz Peels are cleaned, are put into drying box, 10-12h is dried at 55-65 DEG C, crushed, cross 35-45 mesh, Powder is obtained, it is spare.

2) cellulase of the powder weight 0.6-0.8%, pectase, the 0.1-0.3% of 1.5-2.5% are added into powder Neutral proteinase and the water of 60-70% be mixed evenly, and with ultrasonication 6-9min, stop ultrasound, in 42-54 DEG C Lower water-bath keeps the temperature 3-5h, obtains enzymolysis liquid.

3) aluminium chloride of the enzymolysis liquid weight 2-5% is added in Xiang Shangshu enzymolysis liquid, and pH is adjusted to 5.5-6.5, stands After 12-24h, it is centrifuged 10-15min under 2000-2400r/min, filters to isolate filtrate, obtains primary sedimentation；Add into filtrate Enter the calcium chloride of the filtrate weight 3-8%, and pH be adjusted to 8-10, after standing 6-8h, under 2000-2400r/min from Heart 10-15min, filtering, obtains secondary precipitation；The primary sedimentation and secondary precipitation are mixed, mixed precipitation is obtained.

4) by above-mentioned mixed precipitation acid dissolution at liquid, and into the liquid, the ethyl acetate of addition equivalent is extracted, acetic acid The volumetric concentration of ethyl ester is 40-60%, obtains extract liquor.

5) ultrafiltration, crossflow velocity 2-2.5m/s, behaviour are carried out to above-mentioned extract liquor with the PES film that molecular cut off is 1-3 ten thousand Making temperature is 35-45 DEG C, and transmembrane pressure 0.1-0.2MPa is concentrated filtrate after ultrafiltration, dry, obtains threeleaf akebia skin Polyphenol powder.

2. the extraction separation method of Akebia trifoliate koiaz Peels polyphenol according to claim 1, it is characterised in that: the step (2) digest when, be added into powder the cellulase of the powder weight 0.6%, 0.3% pectase, 0.3% neutral egg White enzyme and 70% water are mixed evenly.

3. the extraction separation method of Akebia trifoliate koiaz Peels polyphenol according to claim 1, it is characterised in that: the step (2) power of ultrasonic wave is 150-250W, frequency 20-35KHz in.

4. the extraction separation method of Akebia trifoliate koiaz Peels polyphenol according to claim 1, it is characterised in that: the step (4) acid is food-grade nitric acid or food grade hydrochloric acid in.

5. the extraction separation method of Akebia trifoliate koiaz Peels polyphenol according to claim 1, it is characterised in that: the step (5) temperature being concentrated in is no more than 75 DEG C, and dry is vacuum drying.

Technical field

The present invention relates to extraction and separation technology fields, and in particular to the extraction separation method of Akebia trifoliate koiaz Peels polyphenol.

Background technique

Threeleaf akebia belongs to fallen leaves bejuco, is similar to banana, therefore claims " wild banana ", and fresh fruit is rich in Protein, soluble sugar, organic acid and amino acid needed by human, mineral element etc. are recorded, three Ye Mu according to " Japan hanako materia medica " It is logical that there is the effect of liver-smoothing, qi-regulating, promoting blood circulation and pain relieve, relieving restlessness diuresis.

Preliminary effect is had been achieved for the development and utilization of Trilobed Caulis Akebiae fruit at present, but its pericarp is thick and heavy, almost As many as 40%~69.1% of the total fruit weight of Zhan, is often dropped, and Akebia trifoliate koiaz Peels can extract pericarp polyphenol, can not only improve three The economic benefit of leaf caulis akebiae, but also can solve because pericarp abandons bring problem of environmental pollution.

Summary of the invention

(1) technical problems to be solved

In order to overcome the shortage of prior art, it is proposed that the extraction separation method of Akebia trifoliate koiaz Peels polyphenol passes through ultrasonic wave Assistance enzymolysis carries out broken wall to Akebia trifoliate koiaz Peels, precipitating extracts polyphenol, membrane filter technique carries out a series of technological means phases such as clean In conjunction with, keep product yield high, purity is high, and operating method is simple, equipment requirement is low, and extraction cost is low, is suitble to large-scale production.

(2) technical solution

The present invention is achieved through the following technical solutions: the invention proposes the extraction separation sides of Akebia trifoliate koiaz Peels polyphenol Method, which is characterized in that comprise the following steps that

1) Akebia trifoliate koiaz Peels are cleaned, is put into drying box, 10-12h is dried at 55-65 DEG C, crushed, cross 35-45 Mesh obtains powder, spare.

2) cellulase of the powder weight 0.6-0.8%, pectase, the 0.1- of 1.5-2.5% are added into powder 0.3% neutral proteinase and the water of 60-70% are mixed evenly, and with ultrasonication 6-9min, stop ultrasound, in Water-bath keeps the temperature 3-5h at 42-54 DEG C, obtains enzymolysis liquid.

3) aluminium chloride of the enzymolysis liquid weight 2-5% is added in Xiang Shangshu enzymolysis liquid, and pH is adjusted to 5.5-6.5, After standing 12-24h, it is centrifuged 10-15min under 2000-2400r/min, filters to isolate filtrate, obtains primary sedimentation；To filtrate The middle calcium chloride that the filtrate weight 3-8% is added, and pH is adjusted to 8-10, after standing 6-8h, in 2000-2400r/min Lower centrifugation 10-15min, filtering, obtains secondary precipitation；The primary sedimentation and secondary precipitation are mixed, mixed precipitation is obtained.

4) by above-mentioned mixed precipitation acid dissolution at liquid, and the ethyl acetate of addition equivalent extracts into the liquid, The volumetric concentration of ethyl acetate is 40-60%, obtains extract liquor.

5) ultrafiltration, crossflow velocity 2-2.5m/ are carried out to above-mentioned extract liquor with the PES film that molecular cut off is 1-3 ten thousand S, operation temperature are 35-45 DEG C, and transmembrane pressure 0.1-0.2MPa is concentrated filtrate after ultrafiltration, dry, obtain three Ye Mu Logical skin polyphenol powder.

Further, the step (2) digest when, be added into powder the powder weight 0.6% cellulase, 0.3% pectase, 0.3% neutral proteinase and 70% water are mixed evenly.

Further, the power of ultrasonic wave is 150-250W, frequency 20-35KHz in the step (2).

Further, acid is food-grade nitric acid or food grade hydrochloric acid in the step (4).

Further, the temperature being concentrated in the step (5) is no more than 75 DEG C, and dry is vacuum drying.

(3) beneficial effect

The present invention compared with the existing technology, has the advantages that

1. carrying out broken wall to Akebia trifoliate koiaz Peels by ultrasound-assisted enzymolysis, precipitating extracts polyphenol, and membrane filter technique is removed A series of technological means such as miscellaneous keep product yield high, purity is high, and operating method is simple, and equipment requirement is low, and extraction cost is low, It is suitble to large-scale production.

2. in enzymolysis step, enzyme activity is can be improved in ultrasonic wave, in addition cellulase, pectase and neutral proteinase collaboration increase Effect accelerates enzyme reaction rate, improve the broken and dissolution degree of mango chrotoplast, enable polyphenol substance most, degree greatly releases Put, ooze out, and pectase and neutral proteinase also by cell protein and pectolysis at small molecule object, matter, reduce The content of protein and pectin, and then reduce the combination of polyphenols and protein, polysaccharide, it further improves free more The content of phenol substance.

3. in settling step, using aluminium chloride and calcium chloride co-precipitation, being first settled out most of polyphenol object with aluminium chloride Matter, then precipitated again with calcium chloride, polyphenol precipitating can be made completely, improve yield and purity；In ultrafiltration, concentration and drying steps In, ultrafiltration is carried out to extract liquor with PES film, can further remove impurity, improves purity.

Specific embodiment

In order to make the objectives, technical solutions, and advantages of the present invention clearer, following embodiment carries out the present invention It is further described.It should be appreciated that the specific embodiments described herein are merely illustrative of the present invention, it is not used to limit The present invention.

The extraction separation method of Akebia trifoliate koiaz Peels polyphenol in the present invention, comprises the following steps that

1) Akebia trifoliate koiaz Peels are cleaned, is put into drying box, 10-12h is dried at 55-65 DEG C, crushed, cross 35-45 Mesh obtains powder, spare.

2) cellulase of the powder weight 0.6-0.8%, pectase, the 0.1- of 1.5-2.5% are added into powder 0.3% neutral proteinase and the water of 60-70% are mixed evenly, and with ultrasonication 6-9min, stop ultrasound, in Water-bath keeps the temperature 3-5h at 42-54 DEG C, obtains enzymolysis liquid.

3) aluminium chloride of the enzymolysis liquid weight 2-5% is added in Xiang Shangshu enzymolysis liquid, and pH is adjusted to 5.5-6.5, After standing 12-24h, it is centrifuged 10-15min under 2000-2400r/min, filters to isolate filtrate, obtains primary sedimentation；To filtrate The middle calcium chloride that the filtrate weight 3-8% is added, and pH is adjusted to 8-10, after standing 6-8h, in 2000-2400r/min Lower centrifugation 10-15min, filtering, obtains secondary precipitation；The primary sedimentation and secondary precipitation are mixed, mixed precipitation is obtained.

4) by above-mentioned mixed precipitation acid dissolution at liquid, and the ethyl acetate of addition equivalent extracts into the liquid, The volumetric concentration of ethyl acetate is 40-60%, obtains extract liquor.

5) ultrafiltration, crossflow velocity 2-2.5m/ are carried out to above-mentioned extract liquor with the PES film that molecular cut off is 1-3 ten thousand S, operation temperature are 35-45 DEG C, and transmembrane pressure 0.1-0.2MPa is concentrated filtrate after ultrafiltration, dry, obtain three Ye Mu Logical skin polyphenol powder.

Wherein, the step (2) digest when, be added into powder the powder weight 0.6% cellulase, 0.3% Pectase, 0.3% neutral proteinase and 70% water be mixed evenly；The power of ultrasonic wave is in the step (2) 150-250W, frequency 20-35KHz；Acid is food-grade nitric acid or food grade hydrochloric acid in the step (4)；In the step (5) The temperature of concentration is no more than 75 DEG C, and dry is vacuum drying.