阅读说明：本技术 一种从山茶花中提取花青素的方法 (A method of extracting anthocyanidin from camellia ) 是由 何安乐 李伟 黄华学 于 2019-08-23 设计创作，主要内容包括：一种从山茶花中提取花青素的方法,包括以下步骤：(1)浸提：采用低共熔溶剂的水溶液对山茶花进行浸提,过滤离心,后得浸提液；(2)超滤：将浸提液过超滤膜,收集透过液；(3)纯化浓缩：将透过液依次上吸附树脂柱Ⅰ和吸附树脂柱Ⅱ,水赶,吸附树脂柱Ⅰ采用线性解吸,吸附树脂柱Ⅱ采用梯度解吸,按解吸液中花青素的含量高低,分别收集解吸液,分别浓缩,得浓缩液；(4)结晶干燥：向浓缩液中分别加入低级醇,进行析晶,固液分离,洗晶,干燥,得不同含量范围的产品。该方法从山茶花中提取的花青素产品,花青素含量高；且整个提取工艺条件温和,具有收率高,环境友好,适宜于产业化生产等特点。(A method of from camellia extract anthocyanidin, comprising the following steps: (1) extract: camellia is extracted using the aqueous solution of eutectic solvent, filter centrifugation, after leaching liquor；(2) ultrafiltration: leaching liquor is crossed into ultrafiltration membrane, collects permeate；(3) purifying concentration: liquid successively upper adsorption resin column I and adsorption resin column II be will transmit through, water is caught up with, and adsorption resin column I is using linear desorption, adsorption resin column II uses gradient desorption, by the content height of anthocyanidin in stripping liquid, collects stripping liquid respectively, it is concentrated respectively, obtains concentrate；(4) crystallizing and drying: being separately added into lower alcohol into concentrate, carries out crystallization, is separated by solid-liquid separation, washes crystalline substance, dry, obtains the product of different content range.The anthocyanidin product that this method is extracted from camellia, anthocyanidin content are high；And entire extracting factor is mild, has high income, environmental-friendly, the features such as being suitable for industrialization production.)

1. a kind of method for extracting anthocyanidin from camellia, which comprises the following steps:

(1) it extracts: camellia being extracted using the aqueous solution of eutectic solvent, filter centrifugation obtains leaching liquor；

(2) ultrafiltration: leaching liquor is crossed into ultrafiltration membrane, collects permeate；

(3) purifying concentration: the permeate that step (2) are collected successively upper adsorption resin column I and adsorption resin column II adsorb resin Column I uses gradient desorption using linear desorption, adsorption resin column II, collects and solves respectively by the content height of anthocyanidin in stripping liquid Imbibition is concentrated respectively, obtains concentrate；

(4) crystallization is dry: it is separately added into lower alcohol into concentrate, carries out crystallization, is separated by solid-liquid separation, washes crystalline substance, it is dry, and it obtains and does not suit The product of green cellulose content.

2. the method for anthocyanidin is extracted from camellia according to claim 1, which is characterized in that described low in step (1) Congruent melting solvent includes hydrogen bond receptor and hydrogen bond donor, and the hydrogen bond receptor is organic salt；Preferably, the organic salt is chlorine Change choline or glycine betaine；Preferably, the hydrogen bond donor is in malic acid, citric acid, proline, tartaric acid and ethanedioic acid It is one or more, it is preferred that the mass ratio of the organic salt and hydrogen bond donor is 1:0.25 ~ 4, more preferably, described to have The mass ratio of machine salt and hydrogen bond donor is 1:0.5 ~ 2；Preferably, in the aqueous solution of the eutectic solvent water mass fraction It is 20% ~ 70%, more preferably, the mass fraction of water is 20% ~ 40% in the aqueous solution of the eutectic solvent.

3. the method according to claim 1 or claim 2 for extracting anthocyanidin from camellia, which is characterized in that in step (1), institute The mass ratio for stating the aqueous solution of camellia and the eutectic solvent is 1:20 ~ 50；Preferably, the camellia with it is described low total The mass ratio of the aqueous solution of molten solvent is 1:30 ~ 40；Preferably, the extraction is extracted many times, and number is 2 ~ 5 times, more Preferably, the number is 3 ~ 4 times；Preferably, the time of the extraction be 3 ~ 8h/ times, more preferably, the extraction when Between be 4 ~ 6h/ times.

4. extracting the method for anthocyanidin described in -3 any one from camellia according to claim 1, which is characterized in that step (3) In, the filler of the adsorption resin column I and adsorption resin column II is selected from D101, XAD-1, XAD-7HP, DM-130, AB-8 and DM- One of 301 type macroreticular resins；Preferably, the ratio of height to diameter of the adsorption resin column I and adsorption resin column II is 10 ~ 3:1；More To be preferred, the ratio of height to diameter of the adsorption resin column I and adsorption resin column II is 7 ~ 4:1.

5. extracting the method for anthocyanidin described in -4 any one from camellia according to claim 1, which is characterized in that step (3) In, the concrete operations of the upper adsorption resin column I and adsorption resin column II are as follows: will transmit through adsorption resin column I on liquid first, work as column I when closing on saturation, connect adsorption resin column I and adsorption resin column II, continues will transmit through on liquid adsorption resin column I to upper prop liquid After 0.5 ~ 1.4 times of weight, upper prop terminates, and water is caught up with；More preferably, continue will transmit through on liquid adsorption resin column I to upper prop 0.6 ~ 0.8 times of liquid weight；Preferably, the water, which is caught up with, uses the amount of water for 3 ~ 5BV, and more preferably, the water is caught up with using water Amount be 3.5 ~ 4BV.

6. extracting the method for anthocyanidin described in -5 any one from camellia according to claim 1, which is characterized in that step (3) In, the adsorption resin column I is referred to using linear desorption, from the methanol or ethyl alcohol mass fraction in methanol or ethanol water It is eluted by 0%, until 60 ~ 80% are increased linearly to, it is furthermore preferred that until increasing linearly to 70%；Preferably, the absorption The methanol of resin column I or the volume of ethanol water are 3 ~ 10BV, it is furthermore preferred that the methanol or ethyl alcohol of the adsorption resin column I The volume of aqueous solution is 4 ~ 6BV；Preferably, it is opened when the anthocyanidin content in the stripping liquid of the adsorption resin column I reaches 0.1% Begin to collect, until starting point second segment when anthocyanidin content reaches 40% ~ 60% in stripping liquid and collecting and merge all desorptions below Liquid；It is furthermore preferred that the anthocyanidin content in the stripping liquid of the adsorption resin column I starts to collect when reaching 0.1%, stripping liquid is straight When anthocyanidin content reaches 45% ~ 50% into stripping liquid, starts point second segment and collect and merge the desorption containing anthocyanidin below Liquid.

7. extracting the method for anthocyanidin described in -6 any one from camellia according to claim 1, which is characterized in that step (3) In, the adsorption resin column II is referred to using gradient desorption, and mass fraction is used to carry out for 55 ~ 80% methanol or ethanol water Stripping liquid is desorbed and collects, it is furthermore preferred that being that mass fraction is used to be desorbed and received for 65 ~ 70% methanol or ethanol water Collect stripping liquid；Preferably, the volume of the methanol or ethanol water is 2.5 ~ 5BV, it is furthermore preferred that the methanol or ethanol water The volume of solution is 3 ~ 4BV；Preferably, the concentration is vacuum and low temperature concentration or reverse osmosis membrane concentration；Preferably, the concentration Brix value to concentrate is 40~70%, and more preferably, the Brix value for being concentrated into concentrate is 50~60%.

8. extracting the method for anthocyanidin described in -7 any one from camellia according to claim 1, which is characterized in that step (4) Described in the volume ratio of lower alcohol and the concentrate be 1 ~ 20:1, more preferably, the lower alcohol and the concentrate Volume ratio is 5 ~ 15:1；Preferably, the temperature of the crystallization is -10 ~ 40 DEG C, more preferably, the temperature of the crystallization is 0 ~ 10℃；Preferably, the time of the crystallization be 4 ~ for 24 hours；More preferably, the time of the crystallization is 6 ~ 12h.

9. extracting the method for anthocyanidin described in -8 any one from camellia according to claim 1, which is characterized in that step (4) In, described wash brilliant is directly to be eluted using low-alcohol solution；Preferably, the lower alcohol is methanol or ethyl alcohol；Preferably, described In low-alcohol solution, the content of lower alcohol is 80% or more, and more preferably, the content of the lower alcohol is 90% or more；It is preferred that , the temperature of the elution is -10 ~ 40 DEG C；More preferably, the temperature of the elution is 0 ~ 10 DEG C.

10. extracting the method for anthocyanidin described in -9 any one from camellia according to claim 1, which is characterized in that described dry Dry is vacuum freeze drying, and temperature is -10~-70 DEG C, and vacuum degree is 5~15Pa.

Technical field

The present invention relates to a kind of methods for extracting anthocyanidin, and in particular to a kind of side that anthocyanidin is extracted from camellia Method.

Background technique

Camellia, also known as camellia have long cultivation history in China, are one of ten great tradition famous flowers of China.Camellia Flower is mainly used for nectar source, ornamental and flowers foods etc., and the report and research to camellia anthocyanidin are very few, and anthocyanidin has antioxygen Change, remove a variety of physiological activity such as free radical and anti-mutation, anthocyanidin is extracted from camellia, improves the comprehensive utilization of camellia Efficiency has boundless development prospect.

CN104761596 discloses a kind of method for preparing anthocyanin standard items, and this method is new with the red of camellia Fresh flower valve (or dry petal) is raw material, extracts 12-24h with acidic ethanol aqueous solution at room temperature, is concentrated under reduced pressure after leaching liquor filtering, It is again mobile phase with acid edible alcohol and water, successively carries out column chromatography through macroreticular resin, gel, filler, also by repeatedly anti- Phase column chromatographic purifying is to obtain high-purity anthocyanin standard substance.This method is related to repeatedly preparing chromatogram purification technique, So treating capacity is smaller, technique requires also higher, it is difficult to accomplish scale production.

Summary of the invention

The technical problems to be solved by the present invention are: overcoming the above deficiency, one kind is provided and extracts anthocyanidin from camellia Method, the method extracts the high income of anthocyanidin product from camellia, and the purity of obtained anthocyanidin product is also high, is It comprehensively utilizes camellia and one new way is provided.

The technical solution adopted by the present invention to solve the technical problems is: a kind of side extracting anthocyanidin from camellia Method, comprising the following steps:

(1) it extracts: camellia being extracted using the aqueous solution of eutectic solvent, leaching liquor is obtained after filter centrifugation；

(2) ultrafiltration: leaching liquor is crossed into ultrafiltration membrane, collects permeate；

(3) purifying concentration: liquid successively upper adsorption resin column I and adsorption resin column II be will transmit through, water is caught up with, and adsorption resin column I uses Linear desorption, adsorption resin column II use gradient desorption, collect stripping liquid respectively by the content of anthocyanidin in stripping liquid, dense respectively Contracting, obtains concentrate；

(4) crystallization is dry: it is separately added into lower alcohol into concentrate, carries out crystallization, is separated by solid-liquid separation, washes crystalline substance, it is dry, and obtain different contain Measure the product of range.

The anthocyanidin product that the present invention is extracted from camellia, product purity is high, and entire extracting factor is mild, has High income, environmental-friendly, the features such as being suitable for industrialization production.

Preferably, in step (1), the eutectic solvent includes hydrogen bond receptor and hydrogen bond donor, and the hydrogen bond receptor is Organic salt.Preferably, the organic salt is choline chloride or glycine betaine, and more preferably, the organic salt is chlorination Choline.

Preferably, the hydrogen bond donor be selected from one of malic acid, citric acid, proline, tartaric acid and ethanedioic acid or A variety of, more preferably, the hydrogen donor is citric acid or proline.

Preferably, the mass ratio of the organic salt and hydrogen bond donor is 1:0.25 ~ 4, more preferably, the organic salt Class and the mass fraction ratio of hydrogen bond donor are 1:0.5 ~ 2.

Preferably, the mass fraction of water is 20% ~ 70% in the aqueous solution of the eutectic solvent, more preferably, described The mass fraction of water is 20% ~ 40% in the aqueous solution of the eutectic solvent.

Eutectic solvent is made of hydrogen bond receptor and hydrogen bond donor, have it is at low cost, be easily-synthesized, biodegradable, height The characteristics such as solubilising and hypotoxicity even nontoxicity are a kind of novel green solvents.

Preferably, the camellia flower variety is any one in Camellia, Yunnan camellia or tea plum, more preferably, The camellia flower variety is Camellia.

Preferably, the mass ratio of the aqueous solution of the camellia and the eutectic solvent is 1:20 ~ 50, is more highly preferred to , the mass ratio of the aqueous solution of the camellia and the eutectic solvent is 1:30 ~ 40.

Preferably, in step (1), the extraction is extracted many times, and number is 2 ~ 5 times, more preferably, the number It is 3 ~ 4 times.

Preferably, the time of the extraction is 3 ~ 8h/ times, and more preferably, the time of the extraction is 4 ~ 6h/ times.

Preferably, in step (3), the filler of the adsorption resin column I and adsorption resin column II be selected from D101, XAD-1, One of XAD-7HP, DM-130, AB-8 and DM-301 type macroreticular resin, more preferably, the filler are D101, XAD-1 Or XAD-7HP type macroreticular resin.

Preferably, the ratio of height to diameter of the adsorption resin column I and adsorption resin column II is 10 ~ 3:1, more preferably, described The ratio of height to diameter of adsorption resin column I and adsorption resin column II is 7 ~ 4:1.

Preferably, the concrete operations of the upper adsorption resin column I and adsorption resin column II are as follows: will transmit through on liquid and adsorb first Resin column I, when column I closes on saturation, connect adsorption resin column I and adsorption resin column II, continuing, which will transmit through, adsorbs resin on liquid After column I to 0.5 ~ 1.4 times of upper prop liquid weight, upper prop terminates；More preferably, continue to will transmit through adsorption resin column I on liquid To 0.6 ~ 0.8 times of upper prop liquid weight, upper prop terminates.

Two macroreticular resin string column upper props, can occur competitive adsorption after I adsorption saturation of column, so that impurity quilt Competition enters column II after desorbing, to achieve the purpose that be further purified.

Preferably, attached resin column I and adsorption resin column II are disconnected after the water is caught up with, the water catch up with use the amount of water for 3 ~ 5BV, more preferably, the water, which is caught up with, uses the amount of water for 3.5 ~ 4BV.

After upper prop, water, which is caught up with, to be effectively sufficiently adsorbed unadsorbed feed liquid, while it is miscellaneous to wash off part Matter.

Preferably, the adsorption resin column I is with using linear desorption, i.e., the mass fraction of methanol or ethanol water is by 0% Start to elute, until 60 ~ 80% are increased linearly to, it is furthermore preferred that until increasing linearly to 70%.

Preferably, the volume of the methanol of the adsorption resin column I or ethanol water is 3 ~ 10BV, it is furthermore preferred that described The methanol of adsorption resin column I or the volume of ethanol water are 4 ~ 6BV.

Preferably, start to collect when the anthocyanidin in the stripping liquid of the adsorption resin column I reaches 0.1%, until stripping liquid When middle anthocyanidin content reaches 40% ~ 60%, starts point second segment and collect and merge all stripping liquids below；It is furthermore preferred that described Anthocyanidin in the stripping liquid of adsorption resin column I starts to collect when reaching 0.1%, until anthocyanidin content reaches 45% in stripping liquid When ~ 50%, starts point second segment and collect and merge the stripping liquid containing anthocyanidin below.

Preferably, the adsorption resin column II uses gradient desorption, uses mass fraction for 55 ~ 80% methanol or ethanol water Solution is desorbed and is collected stripping liquid, it is furthermore preferred that mass fraction is used to be solved for 65 ~ 70% methanol or ethanol water It inhales and collects stripping liquid.

Preferably, the volume of the methanol or ethanol water is 2.5 ~ 5BV, it is furthermore preferred that the methanol or ethanol water The volume of solution is 3 ~ 4BV；

Preferably, in step (4), the concentration is vacuum and low temperature concentration or reverse osmosis membrane concentration, more preferably, the concentration For reverse osmosis membrane concentration.Low temperature concentration is more advantageous to stable anthocyanidin.

Preferably, the Brix value for being concentrated into concentrate is 40~70%；More preferably, described to be concentrated into concentrate Brix value be 50~60%.

Preferably, the volume ratio of the lower alcohol and the concentrate is 1 ~ 20:1, more preferably, the lower alcohol with The volume ratio of the concentrate is 5 ~ 15:1.

Preferably, in step (4), the temperature of the crystallization is -10 ~ 40 DEG C, and more preferably, the temperature of the crystallization is 0~10℃。

Preferably, the time of the crystallization be 4 ~ for 24 hours., more preferably, the time of the crystallization is 6 ~ 12h.

Long-time cryogenic conditions are more advantageous to sufficient crystallising, and the saturation anthocyanidin alcoholic solution of certain high concentration, is crystallization Prerequisite.

Preferably, in step (4), described wash brilliant is directly to be eluted using low-alcohol solution.

Preferably, the lower alcohol is methanol or ethyl alcohol, and more preferably, the lower alcohol is ethyl alcohol.

Preferably, the lower alcohol content of the low-alcohol solution is 80% or more, more preferably, the low-alcohol solution Lower alcohol content be 90% or more.

Preferably, the temperature of the elution is -10 ~ 40 DEG C, and more preferably, the temperature of the elution is 0 ~ 10 DEG C.

Described pair of crystallization elutes the impurity that can rinse out plane of crystal.

Preferably, in step (4), the drying be vacuum freeze drying, temperature be -10~-70 DEG C, vacuum degree be 5~ 15Pa。

The temperature of vacuum freeze drying is low, and the influence to anthocyanidin is small.

The beneficial effects of the present invention are:

(1) upper prop is carried out using string column mode due to this method, and individually desorbed, while using the mode linearly desorbed to macropore Resin column is eluted, and more accurately separates target substance, is finally purified again using the method for crystallization, to make It obtains product purity to further increase, anthocyanidin product can be extracted in the slave camellia of high yield using extracting method of the present invention, obtained The purity of the anthocyanidin product arrived is higher, and yield is higher；

(2) entire extracting factor of the invention is mild, and simple process is environmental-friendly, is suitable for industrialization production.

Specific embodiment

The present invention is further detailed with reference to embodiments.

The raw material camellia that each embodiment uses includes passing through the modes such as vacuum drying, forced air drying, freeze-drying, quick-frozen Fresh camellia, freezing camellia and the dehydration camellia of processing.

The detection method of anthocyanidin content is that plant extracts Bilberry fruit P.E SW/T-2013 is attached in the embodiment of the present invention The measuring method of record A.2.The embodiment of the present invention detects the high performance liquid chromatograph used are as follows: high performance liquid chromatography detection instrument is peace Prompt Lun Science & technology Co., Ltd Agilent technologies production, model: HPLC-1260.

Raw material or chemical reagent used in the embodiment of the present invention are obtained by routine business approach unless otherwise specified .

Macroreticular resin used in the embodiment of the present invention is first activated using preceding, specific method are as follows: by fresh tree The alcohol solution dipping of rouge volume fraction 95% for 24 hours after, with distilled water flushing to no alcohol taste, with 3BV, mass fraction 2.5% Hydrochloric acid solution impregnates 5.5h, with distilled water flushing to neutrality, then, then with the NaOH solution immersion of 3BV, mass fraction 3.5% 5.5h, with distilled water flushing to neutrality.