CN104761426B - The method extracting free amino acid from tobacco bud - Google Patents
The method extracting free amino acid from tobacco bud Download PDFInfo
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- CN104761426B CN104761426B CN201510149914.9A CN201510149914A CN104761426B CN 104761426 B CN104761426 B CN 104761426B CN 201510149914 A CN201510149914 A CN 201510149914A CN 104761426 B CN104761426 B CN 104761426B
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Abstract
The invention discloses a kind of method extracting free amino acid from tobacco bud, PEG polymer quality mark 20%～30%, the aqueous two-phase extraction liquid of inorganic salt mass fraction 15%～25% is become with the solution of inorganic salt with PEG polymer, the free amino acid in tobacco bud is extracted then in conjunction with flash extracter, extracting solution siphon separation final vacuum concentrates, it is concentrated in vacuo to dry after purification at the storng-acid cation exchange resin crossed with HCl treatment, obtains aminoacid sterling.The present invention extracts free amino acid from tobacco bud, and efficiency is high, and method is simple, and reagent cost is low；The present invention uses homogenate extraction to combine aqueous two phase extraction technique and extracts the free amino acid in separate tobacco alabastrum, relatively traditional method, flash extraction need not repeatedly extract process, use solvent is less, time-consuming, the solvent extraction effect that aqueous two phase extraction technique is more single is more preferable, and gained aminoacid purity is usually about 96%.
Description
Technical field
Embodiments of the present invention relate to Field of Tobacco, more specifically, embodiments of the present invention relate to one
The method extracting free amino acid from tobacco bud.
Background technology
Land for growing field crops Nicotiana tabacum L. is in growth course, when cigarette strain is bloomed, needs the fireworks to cigarette strain top to extract, with
Ensure the nutritional need of Nicotiana tabacum L. to greatest extent.China has the fireworks of up to ten thousand tons to be taken as garbage to abandon field every year
Between, this not only causes the waste of resource, also results in the propagation of tobacco diseases.
Aminoacid is the important compound in Nicotiana tabacum L., is the ultimate unit of constitutive protein matter, and aminoacid participates in structure
Become enzyme, hormone, some vitamin, be changed into sugar or fat, play nitrogen balance effect.Tobacco bud dissociates
The aminoacid of state is because participating in the synthesis of tobacco protein, and content is compared with the height in Mature Leaf.From food and nutrition
Angle, the aminoacid composition of protein tobacco and content is quite abundant, equilibrium.Aminoacid and derivant thereof
Extensively the field such as application and medicine, food, agricultural, has higher using value.
At present, from tobacco bud extract free amino acid there is no document report, and document report from Nicotiana tabacum L.
The middle separation free amino acid that extracts is only limitted to traditional method, such as: alcohol extracting method, water extraction, ultrasound wave water carry
Method, ethanol microwave extraction method etc., the research report of its associated extraction is also few.Tian Hao etc. are from producing nicotine and eggplant
Nicotiana tabacum L. waste residue after Buddhist nun's alcohol is raw material, uses acid hydrolysis and activated carbon decolorizing to process and extracts aminoacid.Liu Rongsen
With flue-cured tobacco water-removing leaves as raw material, with distilled water extraction in the water-bath of 50 DEG C, and 732 type cationes are used to hand over
Change post to be purified.Method employed in these reports is roughly the same, i.e. uses diluted acid or ethanol in heating
Or the mode by some physics auxiliary such as ultrasonic repeatedly extracts, involved step is more, required quantity of solvent
Greatly, cost is relatively big, and extraction efficiency is relatively low, and can dissolve in during extracting substantial amounts of polysaccharide, nicotine,
The impurity such as protein, pigment, is unfavorable for the isolated and purified of later stage.
Homogenate extraction is occur in recent years a kind of for plant soft, the novel extraction skill of hard material rapid extraction
Art.Rely on high speed machine shearing force and super dynamic molecule diafiltration techniques, in the presence of room temperature and solvent in the several seconds
The materials such as the root of plant, stem, leaf, flower, fruit are crushed to fine particle, and make effective ingredient reach rapidly
To tissue internal and external equilibrium, reached the purpose of extraction by filtration.This technology can protect plant effective to greatest extent
Composition, is notheated destruction, and solvent load is little, and extraction time is short, and efficiency is high, and this technology is extensively used at present
The extraction of effective ingredient in medical material.Aqueous two phase extraction technique (Aqueous tw o-phase ex t ract ion, AT
PE) refer to two kinds of polymer or a kind of polymer together with the aqueous solution of a kind of salt, polymer with
Immiscible property between polymer or between polymer and salt is formed biphase, is the most noticeable, before having
The new separation technology on way, has operating condition gentleness, place compared with traditional extraction and other isolation technics
Reason amount is big, be prone to continuous operation, targeting advantages of higher, be widely used to biological engineering (including
Extract and separate antibiotic, enzyme, separating and purifying protein matter and extraction other biological active substance) bioanalysis and
The aspects such as metal separation.
Summary of the invention
Instant invention overcomes the deficiencies in the prior art, it is provided that a kind of from tobacco bud, extract free amino acid
Method, extracts the aminoacid in tobacco bud expeditiously, and obtains the aminoacid that purity is higher with expectation..
For solving above-mentioned technical problem, one embodiment of the present invention by the following technical solutions:
A kind of method extracting free amino acid from tobacco bud, it is characterised in that it comprises the following steps:
(1) PEG polymer is joined in the aqueous solution of inorganic salt, be configured to PEG polymer quality and divide
The aqueous two-phase extraction liquid that number is 20%～30%, inorganic salt mass fraction is 15%～25%；
(2) tobacco bud naturally dried is put in flash extracter, according to homogenate extraction feed liquid mass ratio
Add the aqueous two-phase extraction liquid configured for 1:10～15, the interior blade rotating speed at flash extracter is
Extract 30～60s under 25000r/min and room temperature condition, remove residue, obtain filtrate, filtrate is stood split-phase,
In tobacco bud, free amino acid is extracted to upper strata；
(3) draw out by the upper strata extracting solution of aqueous two-phase, described upper strata extracting solution be concentrated in vacuo,
Obtain 5～10ml concentrated solutions；
(4) concentrated solution is poured slowly in the storng-acid cation exchange resin crossed with HCl treatment, makes amino
Acid and resin-bonded, after stopping 10min, first remove impurity with distilled water wash exchange column, then use ammonia
Eluting exchange column, obtains eluent, is concentrated in vacuo to by eluent dry, obtains aminoacid sterling.
Homogenate extraction feed liquid mass ratio refers to fireworks alabastrum and the mass ratio of aqueous two-phase extraction liquid.
In order to obtain more preferable extraction effect, thus extract more aminoacid, described pair of water of step (1)
In phase extract, PEG polymer quality mark is 30%, and inorganic salt mass fraction is 20%.
In order to extract more aminoacid and improve amino acid whose purity, step (1) described PEG polymer is
The mixture of PEG800 and PEG6000, the mass ratio of PEG800 and PEG6000 is 1:1～5.
In order to extract more aminoacid and improve amino acid whose purity, step (1) described inorganic salt solution
Aqueous solution for sodium sulfate.
Further technical scheme is that the time of step (2) described standing is 3h.Stand 3h and can make filter
Liquid split-phase is complete, it is simple to the later stage carries out siphon to upper strata extracting solution.
Further technical scheme is, step (3) is described to be drawn out by the upper strata extracting solution of aqueous two-phase
Mode is siphon.It is simple that the mode using siphon separates upper strata extracting solution not only method, and good separating effect.
Further technical scheme is, the temperature being concentrated in vacuo described in step (3) is 70 DEG C.In this temperature
Under the conditions of, the speed being concentrated in vacuo can either be accelerated, can ensure that again the materialitys such as aminoacid in concentrated solution
Matter is stable.
Further technical scheme is, volume is concentrated solution 20～the 30 of step (4) described distilled water
Times, washing speed is 3ml/min.
Further technical scheme is, the concentration of step (4) described ammonia is 4mol/L, and volume is for concentrating
10～20 times of liquid, elution speed is 3ml/min.
The kind of cleaning mixture or eluent can affect washing or the effect of eluting, and consumption is in addition to impact effect,
Also can directly influence next step is concentrated in vacuo the required time, use above-mentioned cleaning mixture or eluent with
And flow velocity, consumption, both can reach preferably to wash or eluting effect, when saving for vacuum concentration step again
Between, the purity of final gained aminoacid sterling is higher, up to about 96%.
Compared with prior art, one of beneficial effects of the present invention is: the present invention extracts trip from tobacco bud
Isolated amino acid, efficiency is high, and method is simple, and reagent cost is low；The present invention uses homogenate extraction to combine aqueous two-phase
Abstraction technique extracts the free amino acid in separate tobacco alabastrum, and relatively traditional method, homogenate extraction need not many
Secondary extraction process, uses solvent less, time-consuming, the solvent extraction effect that aqueous two phase extraction technique is more single
More preferably, gained aminoacid purity is up to about 96% for fruit.
Detailed description of the invention
In order to make the purpose of the present invention, technical scheme and advantage clearer, below in conjunction with embodiment,
The present invention is further elaborated.Should be appreciated that specific embodiment described herein only in order to
Explain the present invention, be not intended to limit the present invention.
Embodiment 1
1) 10g PEG800 and 50gPEG6000 is joined in the aqueous sodium persulfate solution of 266g 15%, stirring
Make it dissolve, be configured to the aqueous two-phase extraction liquid of 200g.
2) tobacco bud that 20g dries naturally is put in flash extracter, adds step 1) in preparation double
Aqueous phase extract, starts flash extracter, and the interior blade rotating speed at flash extracter is 25000r/min and room
60s is extracted under the conditions of temperature.
3) being filtered to remove residue after extraction, obtain filtrate, stand 3h split-phase, in Nicotiana tabacum L., free amino acid is extracted
Get upper strata.
4) use the mode of siphon, draw out by the upper strata extracting solution of aqueous two-phase, under 70 DEG C of vacuum conditions
Extracting solution is concentrated to 10ml.
5) concentrated solution is poured slowly in the strong acid cation exchange resin column crossed with HCl treatment, makes amino
Acid and resin-bonded, after stopping 15min, first wash friendship with the distilled water of 250ml with the speed of 3ml/min
Change post impurity and remove impurity, then use the ammonia speed eluting exchange column with 3ml/min of 200ml 4mol/L,
Being concentrated in vacuo to dry by the eluent of collection, obtain aminoacid sample 0.372g, amino acid content is detected as
96.0%.
6), in the present embodiment, ion exchange chromatography-1,2,3-indantrione monohydrate post-column derivation method is used with automatic amino acid analyzer
Measure amino acid whose content in sample.
Embodiment 2
1) 25g PEG800 and 50g PEG6000 is joined in the aqueous sodium persulfate solution of 225g 20%, stir
Mix and make it dissolve, be configured to the aqueous two-phase extraction liquid of 300g.
2) tobacco bud that 20g dries naturally is put in flash extracter, adds step 1) in preparation double
Aqueous phase extract, starts flash extracter, and the interior blade rotating speed at flash extracter is 25000r/min and room
45s is extracted under the conditions of temperature.
3) being filtered to remove residue after extraction, obtain filtrate, stand 3h split-phase, in Nicotiana tabacum L., free amino acid is extracted
Get upper strata.
4) use the mode of siphon, draw out by the upper strata extracting solution of aqueous two-phase, under 70 DEG C of vacuum conditions
Extracting solution is concentrated to 8ml.
5) concentrated solution is poured slowly in the strong acid cation exchange resin column crossed with HCl treatment, makes amino
Acid and resin-bonded, after stopping 15min, first wash friendship with the distilled water of 200ml with the speed of 3ml/min
Change post impurity and remove impurity, then use the ammonia speed eluting exchange column with 3ml/min of 150ml 4mol/L,
Being concentrated in vacuo to dry by the eluent of collection, obtain aminoacid sample 0.380g, amino acid content is detected as
94.6%.
6), in the present embodiment, ion exchange chromatography-1,2,3-indantrione monohydrate post-column derivation method is used with automatic amino acid analyzer
Measure amino acid whose content in sample.
Embodiment 3
1) 45g PEG800 and 45g PEG6000 is joined in the aqueous sodium persulfate solution of 210g 25%, stir
Mix and make it dissolve, be configured to the aqueous two-phase extraction liquid of 300g.
2) tobacco bud that 20g dries naturally is put in flash extracter, adds step 1) in preparation double
Aqueous phase extract, starts flash extracter, and the interior blade rotating speed at flash extracter is 25000r/min and room
30s is extracted under the conditions of temperature.
3) being filtered to remove residue after extraction, obtain filtrate, stand 3h split-phase, in Nicotiana tabacum L., free amino acid is extracted
Get upper strata.
4) use the mode of siphon, draw out by the upper strata extracting solution of aqueous two-phase, under 70 DEG C of vacuum conditions
Extracting solution is concentrated to 5ml.
5) concentrated solution is poured slowly in the strong acid cation exchange resin column crossed with HCl treatment, makes amino
Acid and resin-bonded, after stopping 15min, first wash friendship with the distilled water of 100ml with the speed of 3ml/min
Change post impurity and remove impurity, then use the ammonia speed eluting exchange column with 3ml/min of 100ml4mol/L,
Being concentrated in vacuo to dry by the eluent of collection, obtain aminoacid sample 0.366g, amino acid content is detected as
95.9%.
6), in the present embodiment, ion exchange chromatography-1,2,3-indantrione monohydrate post-column derivation method is used with automatic amino acid analyzer
Measure amino acid whose content in sample.
Although reference be made herein to invention has been described for the multiple explanatory embodiment of the present invention, but, should
This understanding, those skilled in the art can be designed that a lot of other amendments and embodiment, these amendment and
Embodiment will fall within spirit disclosed in the present application and spirit.More specifically, public in the application
In the range of opening, the building block of theme composite configuration and/or layout can be carried out multiple modification and improvement.
Except modification that building block and/or layout are carried out with in addition to improving, to those skilled in the art, its
His purposes also will be apparent from.
Claims (7)
1. the method extracting free amino acid from tobacco bud, it is characterised in that it comprises the following steps:
(1) PEG polymer is joined in the aqueous solution of inorganic salt, be configured to PEG polymer quality and divide
The aqueous two-phase extraction liquid that number is 20%～30%, inorganic salt mass fraction is 15%～25%；Described PEG
Polymer is the mixture of PEG800 and PEG6000, and the mass ratio of PEG800 and PEG6000 is
1:1～5；The aqueous solution of described inorganic salt is the aqueous solution of sodium sulfate；
(2) tobacco bud naturally dried is put in flash extracter, according to homogenate extraction feed liquid mass ratio
Add the aqueous two-phase extraction liquid configured for 1:10～15, the interior blade rotating speed at flash extracter is
Extract 30～60s under 25000r/min and room temperature condition, remove residue, obtain filtrate, filtrate is stood
Split-phase, in tobacco bud, free amino acid is extracted to upper strata；
(3) draw out by the upper strata extracting solution of aqueous two-phase, described upper strata extracting solution be concentrated in vacuo,
Obtain 5～10ml concentrated solutions；
(4) concentrated solution is poured slowly in the storng-acid cation exchange resin crossed with HCl treatment, makes amino
Acid and resin-bonded, after stopping 10min, first remove impurity with distilled water wash exchange column, then use
Ammonia eluting exchange column, obtains eluent, is concentrated in vacuo to by eluent dry, obtains aminoacid sterling.
The method extracting free amino acid from tobacco bud the most according to claim 1, it is characterised in that
In step (1) described aqueous two-phase extraction liquid, PEG polymer quality mark is 30%, inorganic salt quality
Mark is 20%.
The method extracting free amino acid from tobacco bud the most according to claim 1, it is characterised in that
The time of step (2) described standing is 3h.
The method extracting free amino acid from tobacco bud the most according to claim 1, it is characterised in that
The described mode drawing out by the upper strata extracting solution of aqueous two-phase of step (3) is siphon.
The method extracting free amino acid from tobacco bud the most according to claim 1, it is characterised in that
The temperature being concentrated in vacuo described in step (3) is 70 DEG C.
The method extracting free amino acid from tobacco bud the most according to claim 1, it is characterised in that
Volume is concentrated solution 20～30 times of step (4) described distilled water, washing speed is 3ml/min.
The method extracting free amino acid from tobacco bud the most according to claim 1, it is characterised in that
The concentration of step (4) described ammonia is 4mol/L, and volume is 10～20 times of concentrated solution, elution speed
For 3ml/min.
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| CN108159283B (en) * | 2018-01-29 | 2020-09-11 | 新昌县勤勉生物医药科技有限公司 | Extraction method and application of compound free amino acids of dendrobe leaves |
| CN111876451B (en) * | 2020-08-10 | 2022-05-20 | 武汉轻工大学 | Method for extracting amino acid from chicken manure |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| EP0472143A1 (en) * | 1990-08-23 | 1992-02-26 | Hoechst Schering AgrEvo GmbH | Process for the extractive separation of 1-phosphinothricin and l-glutamic acid in aqueous solution |
| CN1532180A (en) * | 2002-08-14 | 2004-09-29 | 技源科技(中国)有限公司 | Extracting method for natural amino acid mixture with rich 4-hydroxy-isolecine |
| CN102167669A (en) * | 2011-03-14 | 2011-08-31 | 金泳霖 | Technology for extracting amino acids from residual medicine dregs generated in production of erythromycin |
-
2015
- 2015-03-31 CN CN201510149914.9A patent/CN104761426B/en active Active
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| EP0472143A1 (en) * | 1990-08-23 | 1992-02-26 | Hoechst Schering AgrEvo GmbH | Process for the extractive separation of 1-phosphinothricin and l-glutamic acid in aqueous solution |
| CN1532180A (en) * | 2002-08-14 | 2004-09-29 | 技源科技(中国)有限公司 | Extracting method for natural amino acid mixture with rich 4-hydroxy-isolecine |
| CN102167669A (en) * | 2011-03-14 | 2011-08-31 | 金泳霖 | Technology for extracting amino acids from residual medicine dregs generated in production of erythromycin |
Non-Patent Citations (4)
| Title |
|---|
| 双水相体系萃取天冬氨酸转氨酶;陈琪 等;《南京工业大学学报》;20040130;第26卷(第1期);19-23 * |
| 双水相萃取技术及其在药物提取分离中的应用近况;戈延茹 等;《中国现代应用药学杂志》;20090830;第26卷(第8期);623-627 * |
| 植物中游离氨基酸的提取、纯化及分析方法;刘荣森 等;《河南科技大学学报自然科学版》;20070630;第28卷(第3期);76-79 * |
| 烟草中游离氨基酸的测定;王娟 等;《安徽农业科学》;20081230;第36卷(第26期);11413-11416 * |
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Effective date of registration: 20161223 Address after: 610000, Sichuan, Chengdu, Longquanyi economic and Technological Development Zone, Jackie Chan Road, Longquan, No. 2, paragraph Patentee after: CHINA TOBACCO SICHUAN INDUSTRIAL CO., LTD. Patentee after: CHONGQING CHINA TOBACCO INDUSTRIAL CO., LTD. Address before: 610000 Jackie Chan Road, Chengdu economic and Technological Development Zone, Longquanyi District, Sichuan, China, No. 2, No. Patentee before: China Tobacco Chuanyu Industry Co., Ltd. |
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