CN103012544B - A kind of method extracting saponin and polysaccharide from tea seed grouts - Google Patents

A kind of method extracting saponin and polysaccharide from tea seed grouts Download PDF

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CN103012544B
CN103012544B CN201210590251.0A CN201210590251A CN103012544B CN 103012544 B CN103012544 B CN 103012544B CN 201210590251 A CN201210590251 A CN 201210590251A CN 103012544 B CN103012544 B CN 103012544B
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tea seed
saponin
seed grouts
macroporous resin
polysaccharide
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覃江克
黎国庆
叶高杰
杨峰
黄�俊
陆海南
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Guangxi Normal University
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Abstract

The invention provides a kind of method taking water as solvent and extract saponin, polysaccharide from degreasing tea seed grouts.Comprise following processing step: (1) by tea seed grouts grind into powder, and carries out degreasing with sherwood oil; (2) with the saponin in water extraction degreasing tea seed grouts powder and polysaccharide; (3) extracting solution concentrates and regulates pH, adopts macroporous resin adsorption saponin, ethanol elution purifying saponin; (4) effluent liquid obtains polysaccharide with macroporous resin adsorption impurity again.Water of the present invention extracts, green non-pollution; Make eluent with ethanolic soln, boiling point is low, easily recycles and reuses, and produces originally low; Production technique is simple, easy to operate, is applicable to industrialization and produces, can obtain highly purified saponin and polysaccharide.

Description

A kind of method extracting saponin and polysaccharide from tea seed grouts
Technical field
The present invention relates to a kind of method extracting saponin and polysaccharide from tea seed grouts, belong to the extractive technique field of tea seed grouts activeconstituents.
Background technology
Oil tea ( camelliaoleiferaAbel.) be a kind of perennial woody oil crops, be one of four large woody edible oil source seeds equally celebrated for their achievements with oil palm, olive, coconut, have advantageous resources advantage in China, be mainly distributed in the ground such as Guangxi, Jiangxi, Hunan.Camellia oleifera fruit is mainly used in squeezing tea oil, and the residue after squeezing tea oil is tea seed grouts, tea bran withered also known as oil tea etc.Tea seed grouts contain a large amount of food fibres, and the various active composition such as saponin, polysaccharide, polyphenol.
Saponin is the general name extracting the saponins compound obtained from Camellia plant of theaceae, therefore also known as saponin(e, its complex structure, is made up of aglucon, sugared body, organic acid three part.In tea seed grouts, the content of saponin reaches about 15%, is the main products of current camellia oleifera fruit deep processing.Saponin, except having the kinds of surface activity such as emulsification, decontamination, foaming, also has the effects such as significant anti-inflammatory, analgesia, sterilization, desinsection and Promoting plant growth, is widely used in the fields such as daily use chemicals, medicine, food.At present, the extraction purification research about the saponin in tea seed grouts is more, as Ultra filtration membrane technology, organic solvent extractionprocess, flocculation agent method, recrystallization method, silica gel column chromatography etc., all there is deficiency in various degree.
Vegetable polysaccharides is dewatered by monose and is formed in plant, and a class of non-starch, non-cellulose has the general name of the high molecular polymer of physiologically active.Vegetable polysaccharides has antitumor, hypoglycemic, anticoagulation, antithrombotic, pharmacological action and the plurality of health care functions such as anti-oxidant, anti-ageing, antiviral, is one of the hot fields of current food, field of medicaments research.Not mature enough about the technology extracting polysaccharide in cake of camellia oleifera seeds at present, be difficult to amplify.
At present in the method for deproteination, the toxic chemical such as conventional Sevag reagent (mixed solution of chloroform and propyl carbinol), trifluoroacetic acid cause protein denaturation and reach the object of deproteination, and deproteination often needs more than 5 ~ 6 times, agents useful for same is expensive, consumption is large, poisonous, reclaim difficulty, is difficult to industrialization and amplifies and easily cause the problems such as dissolvent residual.
Summary of the invention
The object of this invention is to provide a kind of working condition gentle, safety non-toxic, simple to operate, take water as the method that solvent extracts saponin and polysaccharide from tea seed grouts.
The object of the invention is achieved through the following technical solutions.
From tea seed grouts, extract a method for saponin and polysaccharide, can be realized by following two kinds of approach.
1, from tea seed grouts, extract a method for saponin and polysaccharide, comprise the following steps:
(1) in tea seed grouts, add sherwood oil, make the mass volume ratio of tea seed grouts and sherwood oil be 1:(6 ~ 12) kg/L, extracts, is separated sherwood oil and obtains degreasing tea seed grouts at 65 ~ 90 DEG C;
(2) in degreasing tea seed grouts, add water, make the mass volume ratio of degreasing tea seed grouts and water be 1:(8 ~ 16) kg/L, lixiviate at 60 ~ 90 DEG C, filter to obtain filter residue and filtrate, filter residue and drying obtains food fibre;
(3) step (2) gained filtrate is concentrated into 1/3 ~ 1/2 of original volume, adjust ph is 2 ~ 6 make albumen precipitation, and centrifugation obtains aqueous phase and precipitation, and precipitation drying obtains albumen;
(4) by step (3) gained aqueous phase AB-8 macroporous resin adsorption 2 ~ 5h, collect effluent liquid I, by AB-8 macroporous resin successively Yong Shui ﹑ 0.1 ~ 0.2%NaOH Shui Rong Ye ﹑ 10 ~ 30% ethanolic soln wash-out, finally resolve with the ethanolic soln of 70 ~ 95%, collect desorbed solution, Nong Suo ﹑ drying obtains saponin;
(5) effluent liquid I in step (4) is adopted D3520 macroporous resin adsorption 2 ~ 4h, collect effluent liquid II, effluent liquid II is carried out alcohol precipitation or drying, obtains oil tea polysaccharide.
2, from tea seed grouts, extract a method for saponin and polysaccharide, comprise the following steps:
(1) in tea seed grouts, add sherwood oil, make the mass volume ratio of tea seed grouts and sherwood oil be 1:(6 ~ 12) kg/L, extracts, is separated sherwood oil and obtains degreasing tea seed grouts at 65 ~ 90 DEG C;
(2) in degreasing tea seed grouts, add water, make the mass volume ratio of degreasing tea seed grouts and water be 1:(8 ~ 16) kg/L, lixiviate at 60 ~ 90 DEG C, filter to obtain filter residue and filtrate, filter residue and drying obtains food fibre;
(3) step (2) gained filtrate is concentrated into 1/3 ~ 1/2 of original volume, adjust ph is 2 ~ 6 make albumen precipitation, and centrifugation obtains aqueous phase and precipitation, and precipitation drying obtains albumen;
(4) by step (3) gained aqueous phase D3520 macroporous resin adsorption 2 ~ 4h, collect effluent liquid A, by D3520 macroporous resin successively Yong Shui ﹑ 10 ~ 20% ethanolic soln wash-out, finally resolve with the ethanolic soln of 60 ~ 80%, collect desorbed solution, Nong Suo ﹑ drying obtains oil tea polyphenol;
(5) the effluent liquid A in step (4) is adopted AB-8 macroporous resin adsorption 2 ~ 5h, collect effluent liquid B, effluent liquid B is carried out alcohol precipitation or drying, obtains oil tea polysaccharide;
(6) AB-8 macroporous resin Yong Shui ﹑ 0.1 ~ 0.2%NaOH, 10 ~ 30% ethanolic soln wash-outs successively, finally resolve with the ethanolic soln of 70 ~ 95%, and collect desorbed solution, Nong Suo ﹑ drying obtains saponin.
In above-mentioned steps (1), extract 1 ~ 3 time at 65 ~ 90 DEG C, each 1 ~ 4h.
In above-mentioned steps (2), at 60 ~ 90 DEG C, extract 2 ~ 4h.
In above-mentioned steps (3), adjust ph uses hydrochloric acid or sulfuric acid.
Advantage of the present invention and positively effect: (1) adopts water as solvent extraction saponin and polysaccharide, avoid organic solvent method extract bring costly, the deficiency such as poor stability.(2) macroporous resin has the advantages such as treatment capacity is large, reusable, adopts macroporous resin adsorption to carry out effective isolation and purification to saponin and polysaccharide, improves production efficiency and reduce energy consumption.(3) the present invention first utilizes iso-electric point sedimentation to remove Partial Protein, recycling macroporous resin removes all the other albumen and decolours, macroporous resin can reuse, not only solve traditional method amplify difficulty, noxious solvent residual, affect the bottleneck problems such as product biological activity, also simplify operation, reduction production cost.(4) working condition is gentle, and safety non-toxic is simple to operate, is applicable to suitability for industrialized production.
Accompanying drawing explanation
Fig. 1 is present invention process schema.
Embodiment
Below in conjunction with specific embodiment, the present invention is further elaborated.
The present invention take water as Extraction solvent, adopts macroporous adsorbent resin extraction separation and purification from tea seed grouts to obtain saponin and polysaccharide.In experimentation, have chosen LSA-700B, AB-8, D101, D941 and D3520 macroporous resin and investigate its adsorption to albumen and polyphenol.Result shows: D3520 macroporous resin has well absorption and analytic ability to albumen and polyphenol, but it is weak to the adsorptive power of saponin and polysaccharide, can be used for the albumen in planar water extract and enrichment polyphenol, therefore adopt the albumen (albumen is the principal element affecting purity of polysaccharide) in D3520 macroporous resin Polysaccharide removing and reach the object of enrichment polyphenol.AB-8 macroporous resin has good selective adsorption to saponin, and adsorptive capacity is large, analytic ability strong, therefore selects AB-8 to carry out purifying saponin.
Utilize these two kinds of resins to come separation and purification saponin and polysaccharide, can be realized by following two kinds of methods:
Method one: first with water the compositions such as saponin, polysaccharide, albumen, polyphenol are extracted in the lump and obtain Aqueous extracts, Aqueous extracts is concentrated into 1/3 ~ 1/2 of original volume, regulate pH=2 ~ 6 to make most of albumen precipitation, centrifugation obtains aqueous phase and precipitation.Utilize AB-8 macroporous resin to carry out enrichment absorption to the saponin in aqueous phase, after removing the impurity such as pigment during wash-out, just obtain high purity saponin; Recycling D3520 macroporous resin comes adhesion protein, polyphenol etc., and in the effluent liquid of these two kinds of macroporous resin adsorption, main oil-containing tea polysaccharide, just obtains oil tea polysaccharide through alcohol precipitation or spraying dry.
Method two: first with water, the constituents extraction such as saponin, polysaccharide, albumen, polyphenol are out obtained Aqueous extracts, Aqueous extracts is concentrated into 1/3 ~ 1/2 of original volume, regulate pH=2 ~ 6 to make most of albumen precipitation, centrifugation obtains aqueous phase and precipitation.Utilize D3520 macroporous resin to adsorb and enrichment the albumen in aqueous phase and polyphenol, during wash-out, just obtain oil tea polyphenol; Recycling AB-8 macroporous resin carries out enrichment absorption to saponin, just obtains high purity saponin after removing the impurity such as pigment during wash-out, and in the effluent liquid of these two kinds of macroporous resin adsorption, main oil-containing tea polysaccharide, just obtains oil tea polysaccharide through alcohol precipitation or spraying dry.
Embodiment 1
Oil tea young grouts powder 50g adds sherwood oil, and the mass volume ratio (g/mL) making tea seed grouts powder and sherwood oil is 1:7, at 75 DEG C, extract twice, each 1h, and filter, united extraction liquid, concentration and recovery sherwood oil obtains degreasing tea seed grouts.Water is added to degreasing tea seed grouts, the mass volume ratio (g/mL) making degreasing tea seed grouts and water is 1:16,2h is extracted at 70 DEG C, filter, residue is dried can obtain food fibre, Aqueous extracts is concentrated into original volume 1/3, and add 5% hydrochloric acid and regulate pH=3.5, the albumen precipitated is removed in centrifugation.Aqueous portion, with AB-8 macroporous resin adsorption 3h, collects effluent liquid I.Used by AB-8 macroporous resin water ﹑ 0.15%NaOH ﹑ 20% ethanolic soln wash-out to remove impurity successively, then resolve with 80% ethanolic soln, collect desorbed solution, concentrated, drying can obtain refining saponin, yield 8.6%, and purity is 97%.Effluent liquid I after AB-8 macroporous resin adsorption D3520 macroporous resin adsorption 3h, collects effluent liquid II, adds the dehydrated alcohol of 4 times of volumes in effluent liquid II, stir, leave standstill, drying can obtain oil tea polysaccharide, yield 1.04%, its glucose equivalent is 0.3763.Residue after water extraction is dried can obtain food fibre.
Embodiment 2:
Oil tea young grouts powder 500g adds sherwood oil, and the mass volume ratio (g/mL) making tea seed grouts powder and sherwood oil is 1:9, at 85 DEG C, extract twice, each 2h, and filter, united extraction liquid, concentration and recovery sherwood oil obtains degreasing tea seed grouts.Water is added to degreasing tea seed grouts, the mass volume ratio (g/mL) making degreasing tea seed grouts and water is 1:12,3h is extracted at 85 DEG C, filter, residue is dried can obtain food fibre, Aqueous extracts is concentrated into original volume half, and add 5% hydrochloric acid and regulate pH=4, the albumen precipitated is removed in centrifugation.Aqueous portion, with AB-8 macroporous resin adsorption 3.5h, collects effluent liquid I.Used by AB-8 macroporous resin water ﹑ 0.15%NaOH ﹑ 20% ethanolic soln wash-out to remove impurity successively, then resolve with 80% ethanolic soln, collect desorbed solution, concentrated, drying can obtain refining saponin, yield 8.3%, purity 96.3%.Effluent liquid I after AB-8 macroporous resin adsorption D3520 macroporous resin adsorption 3.5h, collects effluent liquid II, adds the dehydrated alcohol of 4 times of volumes in effluent liquid II, stir, leave standstill, drying can obtain oil tea polysaccharide, yield 0.98%, its glucose equivalent is 0.3321.Residue after water extraction is dried can obtain food fibre.
Embodiment 3:
Take oil tea young grouts powder 10kg, add sherwood oil, the mass ratio (kg/L) of tea seed grouts and sherwood oil is 1:6, at 80 DEG C, extract twice, each 3h, and filter, united extraction liquid, concentration and recovery sherwood oil obtains degreasing tea seed grouts.Water is added in degreasing tea seed grouts, the mass volume ratio (kg/L) making degreasing tea seed grouts and water is 1:8,4h is extracted at 90 DEG C, filter, residue is dried can obtain food fibre, Aqueous extracts is concentrated into original volume half, and add 5% sulfuric acid and regulate pH=4.5, the albumen precipitated is removed in centrifugation.Aqueous portion, with AB-8 macroporous resin adsorption 4h, collects effluent liquid I.Used by AB-8 macroporous resin water ﹑ 0.15%NaOH ﹑ 20% ethanolic soln wash-out to remove impurity successively, then resolve with 80% ethanolic soln, collect desorbed solution, concentrated, drying can obtain refining saponin, yield 8.1%, purity 95.1%.Effluent liquid I after AB-8 macroporous resin adsorption D3520 macroporous resin adsorption 3.5h, collects effluent liquid II, adds the dehydrated alcohol of 4 times of volumes in effluent liquid II, stir, leave standstill, drying can obtain oil tea polysaccharide, yield 0.7%, its glucose equivalent is 0.3221.Residue after water extraction is dried can obtain food fibre.
Embodiment 4:
Similar to Example 1, unlike first using AB-8 macroporous resin adsorption again by D3520 macroporous resin adsorption.
Take oil tea young grouts powder 50g, add sherwood oil, the mass volume ratio (g/mL) making tea seed grouts powder and sherwood oil is 1:7, at 75 DEG C, extract twice, each 1h, and filter, united extraction liquid, concentration and recovery sherwood oil obtains degreasing tea seed grouts.Water is added to degreasing tea seed grouts, the mass volume ratio (g/mL) making degreasing tea seed grouts and water is 1:16,2h is extracted at 70 DEG C, filter, solid residue can obtain food fibre, Aqueous extracts is concentrated into original volume 1/3, and add 5% hydrochloric acid and regulate pH=3.5, the albumen precipitated is removed in centrifugation.Aqueous portion, with D3520 macroporous resin adsorption 4h, collects effluent liquid (A).Used by D3520 macroporous resin water ﹑ 15% ethanolic soln wash-out to remove impurity successively, then resolve with 70% ethanolic soln, collect desorbed solution, concentrated, drying can obtain oil tea polyphenol, yield 0.64%, and its gallic acid equivalant is 0.08572.Effluent liquid (A) after D3520 macroporous resin adsorption AB-8 macroporous resin adsorption 3h, collect effluent liquid (B), effluent liquid (B) concentrate drying can be obtained oil tea polysaccharide, yield 0.66%, its glucose equivalent is 0.6651; AB-8 macroporous resin uses water ﹑ 0.15%NaOH ﹑ 20% ethanolic soln wash-out to remove impurity successively, then resolves with 80% ethanolic soln, and collect desorbed solution, concentrated, drying can obtain refining saponin, yield 7.6%, and purity is 98.5%.Residue after water extraction is dried can obtain food fibre.
Compared with embodiment 1, the saponin color and luster that embodiment 4 obtains is whiter, and purity is higher.
Polyphenol is the major cause causing saponin color and luster to deepen, and in purifying saponin process, how to remove polyphenol is the principal element that will consider.Adopt method described in embodiment 4 can obtain oil tea polyphenol while wash-out D3520.
The inventive method 1 first uses AB-8 macroporous resin, re-use D-3520 macroporous resin purification, because AB-8 macroporous resin has strong selective adsorption effect to saponin, but it is also adsorbing polyphenols simultaneously, in the process of purifying saponin, available 0.15%NaOH by AB-8 macroporous resin polyphenol remove, due to use be alkali cleaning take off, so time polyphenol cannot reclaim.After 0.15%NaOH wash-out, AB-8 macroporous resin still has a small amount of polyphenol remain, therefore during with 80% ethanolysis graining element, a small amount of polyphenol that resin remains also elute and cause that the color and luster of saponin is relatively poor, purity is relatively little together.
And method 2 first uses D3520 macroporous resin, then use the purifying of AB-8 macroporous resin.Because D3520 macroporous resin is to polyphenol and protein adsorption effect very strong (it also has certain adsorption to saponin), after D3520 macroporous resin is put into Aqueous extracts, most polyphenol in extracting solution are by D3520 macroporous resin adsorption, when wash-out D3520 macroporous resin, owing to not using alkali but using alcohol wash-out, therefore polyphenol can be collected.In addition owing to first using D3520 macroporous resin, it is substantially clean for the polyphenol absorption in Aqueous extracts, when using AB-8 macroporous resin adsorption saponin again, farthest decrease the impact of polyphenol on saponin purifying, such that the color and luster of saponin and purity are all corresponding to be improved.But now, saponin have passed through two step absorption, and loss is wherein become cannot be avoided, and yield is just corresponding reduction also.

Claims (3)

1. from tea seed grouts, extract a method for saponin and polysaccharide, it is characterized in that comprising the following steps:
(1) in tea seed grouts, add sherwood oil, make the mass volume ratio of tea seed grouts and sherwood oil be 1:(6 ~ 12) kg/L, extracts 1 ~ 3 time, each 1 ~ 4h, is separated sherwood oil and obtains degreasing tea seed grouts at 65 ~ 90 DEG C;
(2) in degreasing tea seed grouts, add water, make the mass volume ratio of degreasing tea seed grouts and water be 1:(8 ~ 16) kg/L, lixiviate at 60 ~ 90 DEG C, filter to obtain filter residue and filtrate, filter residue and drying obtains food fibre;
(3) step (2) gained filtrate is concentrated into 1/3 ~ 1/2 of original volume, use hydrochloric acid or sulfuric acid adjust ph to be 2 ~ 6 make albumen precipitation, centrifugation obtains aqueous phase and precipitation, and precipitation drying obtains albumen;
(4) by step (3) gained aqueous phase AB-8 macroporous resin adsorption 2 ~ 5h, collect effluent liquid I, by AB-8 macroporous resin successively Yong Shui ﹑ 0.1 ~ 0.2%NaOH Shui Rong Ye ﹑ 10 ~ 30% ethanolic soln wash-out, finally resolve with the ethanolic soln of 70 ~ 95%, collect desorbed solution, Nong Suo ﹑ drying obtains saponin;
(5) effluent liquid I in step (4) is adopted D3520 macroporous resin adsorption 2 ~ 4h, collect effluent liquid II, effluent liquid II is carried out alcohol precipitation or drying, obtains oil tea polysaccharide.
2. from tea seed grouts, extract a method for saponin and polysaccharide, it is characterized in that comprising the following steps:
(1) in tea seed grouts, add sherwood oil, make the mass volume ratio of tea seed grouts and sherwood oil be 1:(6 ~ 12) kg/L, extracts 1 ~ 3 time, each 1 ~ 4h, is separated sherwood oil and obtains degreasing tea seed grouts at 65 ~ 90 DEG C;
(2) in degreasing tea seed grouts, add water, make the mass volume ratio of degreasing tea seed grouts and water be 1:(8 ~ 16) kg/L, lixiviate at 60 ~ 90 DEG C, filter to obtain filter residue and filtrate, filter residue and drying obtains food fibre;
(3) step (2) gained filtrate is concentrated into 1/3 ~ 1/2 of original volume, use hydrochloric acid or sulfuric acid adjust ph to be 2 ~ 6 make albumen precipitation, centrifugation obtains aqueous phase and precipitation, and precipitation drying obtains albumen;
(4) by step (3) gained aqueous phase D3520 macroporous resin adsorption 2 ~ 4h, collect effluent liquid A, by D3520 macroporous resin successively Yong Shui ﹑ 10 ~ 20% ethanolic soln wash-out, finally resolve with the ethanolic soln of 60 ~ 80%, collect desorbed solution, Nong Suo ﹑ drying obtains oil tea polyphenol;
(5) the effluent liquid A in step (4) is adopted AB-8 macroporous resin adsorption 2 ~ 5h, collect effluent liquid B, effluent liquid B is carried out alcohol precipitation or drying, obtains oil tea polysaccharide;
(6) AB-8 macroporous resin Yong Shui ﹑ 0.1 ~ 0.2%NaOH, 10 ~ 30% ethanolic soln wash-outs successively, finally resolve with the ethanolic soln of 70 ~ 95%, and collect desorbed solution, Nong Suo ﹑ drying obtains saponin.
3. according to claim 1 or 2, from tea seed grouts, extract the method for saponin and polysaccharide, it is characterized in that: in step (2), at 60 ~ 90 DEG C, extract 2 ~ 4h.
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