CN103242224A - Method for extracting and purifying Leucaena leucocephala leaf mimosine - Google Patents
Method for extracting and purifying Leucaena leucocephala leaf mimosine Download PDFInfo
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- CN103242224A CN103242224A CN201310134483XA CN201310134483A CN103242224A CN 103242224 A CN103242224 A CN 103242224A CN 201310134483X A CN201310134483X A CN 201310134483XA CN 201310134483 A CN201310134483 A CN 201310134483A CN 103242224 A CN103242224 A CN 103242224A
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- mimosine
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- leucaena leucocephala
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Abstract
The invention discloses a method for extracting and purifying Leucaena leucocephala leaf mimosine, which comprises the following steps: acquiring wholly expanded leaves; drying the leaves to constant weight, pulverizing with a pulverizer, and screening; weighing a sample, putting into a plug-bearing bottle, and adding distilled water; keeping the temperature, shaking, standing, and leaching while keeping the temperature; centrifuging, and collecting the supernatant; regulating the pH value of the supernatant to 5 with hydrochloric acid, adding ethanol, evenly mixing, and standing; centrifuging, collecting the supernatant, concentrating with a vacuum pump under reduced pressure, and standing for 3 days; centrifuging, collecting crystals, washing with distilled water, and drying; putting the crystals in a beaker, adding distilled water, and heating until the crystals are completely dissolved; filtering the dissolution solution, and collecting the filtrate; concentrating the filtrate under reduced pressure, and standing at 4 DEG C for 3 days; and centrifuging, collecting, washing with distilled water, and carrying out vacuum drying to constant weight at room temperature to obtain white crystals. The analysis on 550nm wavelength by a spectrometer indicates that the purity of the extracted mimosine reaches 99%, 2.1-2.4g of mimosine can be extracted from every 100g of Leucaena leucocephala leaves, and the extraction rate is 40-70%.
Description
Technical field
The invention belongs to mimosine extractive technique field, relate in particular to the method for a kind of extraction and purifying Leucaena leucocephala (L.) blade mimosine.
Background technology
1936, reported first have mimosine in the Touch-me-notAction Plant plant.The Leucaena leucocephala (L.) plant is rich in polyphenol oxides, chlorophyll, polysaccharide, protein etc., in its seed gross protein value be 30%～32%, mimosine 3%～5%, in the blade gross protein value be 22%～29%, mimosine 3%～6%.Leucaena leucocephala (L.) is world-renowned high-protein feeding tannin plant, because the rich mimosine of blade, cause often losing hair or feathers behind the edible Leucaena leucocephala (L.) of fowl poultry (especially non-ruminant animal), symptoms such as cataract, poor growth and dysgenesia, limited the widespread use of Leucaena leucocephala (L.) in livestock industry.But then, mimosine has many-sided physiologically active, and its some physiologically active is exploited, as: the industrial trichogen etc. that is used as.At present, the price of SILVER REAGENT mimosine is about per 100 milligrams 565 yuan on the market.
Mimosine is that J.Renz in 1936 extracts from Touch-me-notAction Plant and obtains at first, extracts from the Leucaena leucocephala (L.) seed and then starts from R.Yoshida (1944).Nineteen fifty-one, people such as Matsumoto have set up the quick colorimetry of measuring mimosine.Nineteen sixty-five, people such as James measure the mimosine content in different Leucaena leucocephala (L.) germplasm blades and the seed first.1978, Megarrity set up the automatic colorimetry that Leucaena leucocephala (L.) blade mimosine is measured.People such as China Chen Xiu orchid utilize the chlorination iron processes, have measured the mimosine content in new Leucaena leucocephala (L.) and Ken Ning Durham Leucaena leucocephala (L.) branch and the blade.People such as Wu Yanzhen have set up extraction and the pure method of mimosine in the Leucaena leucocephala (L.) seed by the research to Leucaena leucocephala (L.) seed extraction high purity mimosine.
The moiety of Leucaena leucocephala (L.) blade and seed has very big-difference, and its mimosine extraction and purification process are also inequality.The method of extraction and purifying mimosine is not also reported from the Leucaena leucocephala (L.) blade.
Summary of the invention
The purpose of the embodiment of the invention is to provide the method for a kind of extraction and purifying Leucaena leucocephala (L.) blade mimosine, and has high extraction, high purity characteristics.
At the problems referred to above, the embodiment of the invention is achieved in that the method for a kind of extraction and purifying Leucaena leucocephala (L.) blade mimosine, and this method may further comprise the steps:
The first step: gather the 1st to 3 fluffy full exhibition blade on the healthy branch of Leucaena leucocephala (L.), sampling 1.0kg;
Second step: blade is dried to constant weight in 39 ℃～40 ℃, extract with being used for mimosine behind the pulverizer crushing screening;
The 3rd step: take by weighing the sample 100g through crushing screening, put into 1000mL band plug bottle, adding distil water 500mL;
The 4th step: in having swingable water bath with thermostatic control, be incubated, wave earlier, leave standstill insulation lixiviate 16～17h again;
The 5th step: centrifugal, collect supernatant liquor;
The 6th step: with hydrochloric acid supernatant liquor is transferred to pH5, add ethanol, be mixed and under 4 ℃ of conditions, place 2d;
The 7th step: centrifugal collection supernatant liquor, and be evaporated to 20mL with vacuum pump, concentrated solution leaves standstill 3d under 4 ℃ of conditions;
The 8th step: centrifugal collection crystallisate, use distilled water wash, the centrifugal collection crystallisate of difference, vacuum-drying is to constant weight under the room temperature condition;
The 9th step: get crystallisate 2g and put into the 500mL beaker, add 200mL distilled water, heating is dissolving fully directly.In the time of while hot, change lysate over to Filter column that powdered carbon is housed and filter, and collect filtrate;
The tenth step: filtrate decompression is concentrated into 100mL, and under 4 ℃ of conditions, leaves standstill 3d;
The 11 step: centrifugal collection crystallisate, use distilled water wash, room temperature vacuum-drying obtains white crystal to constant weight.
Further, blade is dried to constant weight in 39 ℃～40 ℃, pulverized with pulverizer and be used for mimosine behind 20 mesh sieves and extract.
Further, take by weighing the sample 100g through crushing screening, put into 1000mL band plug triangular flask, adding distil water 500mL.
Further, in having swingable 59 ℃～60 ℃ waters bath with thermostatic control, be incubated, wave 30min earlier, leave standstill insulation lixiviate 16～17h again.
Further, under the 6000r/min condition, centrifugal 10min collects supernatant liquor.
Further, with the hydrochloric acid of 2mol/L supernatant liquor is transferred to pH5, add isopyknic 90% ethanol, be mixed and under 4 ℃ of conditions, place 2d.
Further, centrifugal collection crystallisate, with 4 ℃ of distilled water washs 3 times, centrifugal collection crystallisate respectively, vacuum-drying is to constant weight under the room temperature condition.
Further, centrifugal collection crystallisate, with 4 ℃ of distilled water washs 3 times, room temperature vacuum-drying obtains white crystal to constant weight.
The present invention chooses 550 nano wave length analyses by spectrophotometer, and the mimosine purity of extracting reaches 99%.Can extract mimosine 2.1～2.4g in every 100g Leucaena leucocephala (L.) blade, calculate by Leucaena leucocephala (L.) blade mimosine content 3%～6%, the extraction yield of this method is 40%～70%.
Embodiment
In order to make 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 explaining the present invention, and be not used in restriction the present invention.
The embodiment of the invention provides the method for a kind of extraction and purifying Leucaena leucocephala (L.) blade mimosine, and this method may further comprise the steps, and specifically details are as follows:
One, test materials
Vegetable material
Heat is ground No. 1 Leucaena leucocephala (L.) blade (2～3 monthly age).
Chemical reagent
Distilled water, 2mol/L hydrochloric acid, 90% ethanol, powdered carbon post.
Two, plant and instrument
Pulverizer, band plug triangular flask, swingable water bath with thermostatic control, whizzer, vacuum pump, spectrophotometer.
Three, Leucaena leucocephala (L.) blade mimosine extracts and the purification technique scheme
The first step: gather the 1st to 3 fluffy full exhibition blade on the healthy branch of Leucaena leucocephala (L.), about 1.0kg takes a sample.
Second step: blade is dried to constant weight in 39 ℃～40 ℃, be used for mimosine with pulverizer crushing screening (20 order) back and extract.
The 3rd step: take by weighing the sample 100g through crushing screening, put into 1000mL band plug triangular flask, adding distil water 500mL.
The 4th step: insulation (59 ℃～60 ℃) in having swingable water bath with thermostatic control, wave 30min earlier, leave standstill insulation lixiviate 16～17h again.
The 5th step: under the 6000r/min condition, centrifugal 10min collects supernatant liquor.
The 6th step: the hydrochloric acid with 2mol/L transfers to pH5 with supernatant liquor, adds isopyknic 90% ethanol, is mixed and places 2d under 4 ℃ of conditions.
The 7th step: centrifugal collection supernatant liquor, and be evaporated to 20mL with vacuum pump, concentrated solution leaves standstill 3d under 4 ℃ of conditions.
The 8th step: centrifugal collection crystallisate, with 4 ℃ of distilled water washs 3 times, distinguish centrifugal collection crystallisate, vacuum-drying is to constant weight under the room temperature condition.
The 9th step: get crystallisate 2g and put into the 500mL beaker, add 200mL distilled water, heating is dissolving fully directly.In the time of while hot, change lysate over to Filter column that powdered carbon is housed and filter, and collect filtrate.
The tenth step: filtrate decompression is concentrated into 100mL, and under 4 ℃ of conditions, leaves standstill 3d.
The 11 step: centrifugal collection crystallisate, with 4 ℃ of distilled water washs 3 times, room temperature vacuum-drying is to constant weight, the acquisition white crystal.
The above only is preferred embodiment of the present invention, not in order to limiting the present invention, all any modifications of doing within the spirit and principles in the present invention, is equal to and replaces and improvement etc., all should be included within protection scope of the present invention.
Claims (8)
1. the method for an extraction and purifying Leucaena leucocephala (L.) blade mimosine is characterized in that this method may further comprise the steps:
The first step: gather the 1st to 3 fluffy full exhibition blade on the healthy branch of Leucaena leucocephala (L.), sampling 1.0kg;
Second step: blade is dried to constant weight in 39 ℃～40 ℃, extract with being used for mimosine behind the pulverizer crushing screening;
The 3rd step: take by weighing the sample 100g through crushing screening, put into 1000mL band plug bottle, adding distil water 500mL;
The 4th step: in having swingable water bath with thermostatic control, be incubated, wave earlier, leave standstill insulation lixiviate 16～17h again;
The 5th step: centrifugal, collect supernatant liquor;
The 6th step: with hydrochloric acid supernatant liquor is transferred to pH5, add ethanol, be mixed and under 4 ℃ of conditions, place 2d;
The 7th step: centrifugal collection supernatant liquor, and be evaporated to 20mL with vacuum pump, concentrated solution leaves standstill 3d under 4 ℃ of conditions;
The 8th step: centrifugal collection crystallisate, use distilled water wash, the centrifugal collection crystallisate of difference, vacuum-drying is to constant weight under the room temperature condition;
The 9th step: get crystallisate 2g and put into the 500mL beaker, add 200mL distilled water, heating is dissolving fully directly; In the time of while hot, change lysate over to Filter column that powdered carbon is housed and filter, and collect filtrate;
The tenth step: filtrate decompression is concentrated into 100mL, and under 4 ℃ of conditions, leaves standstill 3d;
The 11 step: centrifugal collection crystallisate, use distilled water wash, room temperature vacuum-drying obtains white crystal to constant weight.
2. the method for extraction as claimed in claim 1 and purifying Leucaena leucocephala (L.) blade mimosine is characterized in that, blade is dried to constant weight in 39 ℃～40 ℃, pulverizes with pulverizer to be used for mimosine behind 20 mesh sieves and to extract.
3. the method for extraction as claimed in claim 1 and purifying Leucaena leucocephala (L.) blade mimosine is characterized in that, takes by weighing the sample 100g through crushing screening, puts into 1000mL band plug triangular flask, adding distil water 500mL.
4. the method for extraction as claimed in claim 1 and purifying Leucaena leucocephala (L.) blade mimosine is characterized in that, is incubated in having swingable 59 ℃～60 ℃ waters bath with thermostatic control, waves 30min earlier, leaves standstill insulation lixiviate 16～17h again.
5. the method for extraction as claimed in claim 1 and purifying Leucaena leucocephala (L.) blade mimosine is characterized in that, under the 6000r/min condition, centrifugal 10min collects supernatant liquor.
6. the method for extraction as claimed in claim 1 and purifying Leucaena leucocephala (L.) blade mimosine is characterized in that, with the hydrochloric acid of 2mol/L supernatant liquor is transferred to pH5, adds isopyknic 90% ethanol, is mixed and places 2d under 4 ℃ of conditions.
7. the method for extraction as claimed in claim 1 and purifying Leucaena leucocephala (L.) blade mimosine is characterized in that, centrifugal collection crystallisate, and with 4 ℃ of distilled water washs 3 times, centrifugal collection crystallisate respectively, vacuum-drying is to constant weight under the room temperature condition.
8. the method for extraction as claimed in claim 1 and purifying Leucaena leucocephala (L.) blade mimosine is characterized in that, centrifugal collection crystallisate, and with 4 ℃ of distilled water washs 3 times, room temperature vacuum-drying obtains white crystal to constant weight.
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN112889828A (en) * | 2021-01-12 | 2021-06-04 | 碧沃丰生物科技(广东)股份有限公司 | Plant alkaloid algaecide and application thereof |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101653470A (en) * | 2009-06-02 | 2010-02-24 | 华宝食用香精香料（上海）有限公司 | Sensitive plant total alkaloid extract and preparing method and application thereof |
| JP2010047487A (en) * | 2008-08-19 | 2010-03-04 | Hipep Laboratories | Method for producing mimosine or acid addition salt thereof and mimosine derivative and method for quality control of mimosine |
| JP2012077058A (en) * | 2010-10-06 | 2012-04-19 | Univ Of Ryukyus | Influenza virus neuraminidase inhibitor |
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2013
- 2013-04-18 CN CN201310134483XA patent/CN103242224A/en active Pending
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP2010047487A (en) * | 2008-08-19 | 2010-03-04 | Hipep Laboratories | Method for producing mimosine or acid addition salt thereof and mimosine derivative and method for quality control of mimosine |
| CN101653470A (en) * | 2009-06-02 | 2010-02-24 | 华宝食用香精香料（上海）有限公司 | Sensitive plant total alkaloid extract and preparing method and application thereof |
| JP2012077058A (en) * | 2010-10-06 | 2012-04-19 | Univ Of Ryukyus | Influenza virus neuraminidase inhibitor |
Non-Patent Citations (4)
| Title |
|---|
| KIYOSHI NOKIHARA等: "Preparative scale isolation, purification and derivatization of mimosine, a non-proteinogenic amino acid", 《AMINO ACIDS》 * |
| 于炎湖: "《饲料毒物学附毒物分析》", 31 October 1996 * |
| 伍颜贞，黎仕钊: "从银合欢种子提取高纯度含羞草素的研究", 《热带农业科学》 * |
| 陈秀兰等: "银合欢含羞草素测定技术的研究", 《热带农业科学》 * |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN112889828A (en) * | 2021-01-12 | 2021-06-04 | 碧沃丰生物科技(广东)股份有限公司 | Plant alkaloid algaecide and application thereof |
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Application publication date: 20130814 |