CN104961844A - Method for preparing chitosan from low-age insect larvae at normal temperature - Google Patents
Method for preparing chitosan from low-age insect larvae at normal temperature Download PDFInfo
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- CN104961844A CN104961844A CN201510435931.9A CN201510435931A CN104961844A CN 104961844 A CN104961844 A CN 104961844A CN 201510435931 A CN201510435931 A CN 201510435931A CN 104961844 A CN104961844 A CN 104961844A
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Abstract
The invention relates to a method for preparing chitosan from low-age insect larvae at normal temperature, which is characterized by directly treating larvae at normal temperature to prepare the chitosan. The method comprises the following steps: protein and fat removal: pressing 3-5-day live insect larvae to obtain maggot skin, adding 3 times by volume of 5% NaOH into the maggot skin, stirring at normal temperature, reacting for 24 hours, removing the protein, preparing a filter cake, washing with water until the pH value is 6.9, and washing with deionized water until the pH value is 7; decalcification: putting the filter cake into hydrochloric acid, immersing at normal temperature for 6 hours, and filtering; deacetylation: adding a 40-50% sodium hydroxide solution, stirring at normal temperature, and reacting for 48 hours; and washing with tap water until the pH value is 6.9, washing with deionized water until the pH value is 7, drying the filter cake, and pulverizing to obtain the chitosan powder. The high-vitality larvae are directly treated at normal temperature, thereby saving the energy and maintaining more bioactivities in the product.
Description
Technical field
The invention belongs to biological technical field, particularly relate to a kind of method utilizing low age insect larvae to prepare chitosan at normal temperatures.
Background technology
Chitosan (chitosan) is also known as chitosan, the premium properties such as the good biocompatibility of this natural polymer, security are good, by all trades and professions extensive concern, achieves major progress in the applied research of the numerous areas such as medicine, food, chemical industry, makeup, water treatment, METAL EXTRACTION and recovery, biochemistry and biomedical engineering.Chitosan is the product that chitin takes off N-ethanoyl; generally speaking, N-ethanoyl slough more than 55% just can be referred to as chitosan, in other words; can dissolve the chitosan of 1% in 1% acetic acid or 1% hydrochloric acid, this chitosan is referred to as chitosan.As the chitosan of industrial goods, N-deacetylation is more than 70%.N-deacetylation is low deacetylation chitosan at 55%-70%, and 70%-85%'s is middle deacetylation chitosan, and 85%-95%'s is chitosan with high deacetylation degree, and 95%-100%'s is chitosan with ultrahigh deacetylation degree.The extremely difficult preparation of chitosan of N-deacetylation 100%.The raw material of the current suitability for industrialized production chitin shrimp mainly in fishery products, crab shell, raw material sources are relatively single, and the insect biological group that to be most species in biology on the earth, quantity maximum, containing abundant chitin in its body wall.To research and develop in insect chitin and derivative thereof both at home and abroad at present.With common Chinese medicine cicada slough, Chinese rice grasshopper, Blaps japanensis yunnanesis and wax insect for raw material, carry out the preparation of the extraction of insect chitin and insect shell glycan, cm-chitosan.By single factor experiment and orthogonal test, inquire into differential responses condition in insect chitin leaching process to the impact removing protein and inorganic salt contained by insect raw material; By single factor experiment, inquired into the impact that differential responses condition is prepared chitosan and cm-chitosan, main result is as follows: 1. the extraction of chitin: after removing protein in insect raw material and inorganic salt respectively, thus isolate chitin.(1) the removing of protein: affect deproteination qualitative factor mainly NaOH concentration, temperature of reaction and the reaction times, by single factor experiment and orthogonal test, determine the optimum test condition removing protein in Chinese medicine cicada slough, Chinese rice grasshopper, Blaps japanensis yunnanesis and wax insect and be respectively 6%NaOH, reaction times 1h, temperature of reaction 90 DEG C; 7%NaOH, reaction times 30min, temperature of reaction 90 DEG C; 7%NaOH, reaction times 5h, temperature of reaction 90 DEG C, 7%NaOH, reaction times 1h, temperature of reaction 90 DEG C.(2) the removing of inorganic salt: impact removes factor mainly HCl concentration and the reaction times of inorganic salt.2. the preparation of insect shell glycan: chitosan is the product of de-acetyl chitin reaction.The principal element affecting de-acetyl chitin reaction is NaOH concentration, and temperature of reaction and reaction times, by single factor experiment, determining the optimum test condition preparing insect shell glycan is: NaOH concentration 50%, temperature of reaction 120 DEG C, reaction times 3h.3. the preparation of insect cm-chitosan: chitosan obtains cm-chitosan after carboxymethylation reaction.The principal element affecting chitosan carboxymethylation reaction is NaOH consumption, NaOH concentration, Mono Chloro Acetic Acid consumption, temperature of reaction, reaction times, pass through single factor experiment, the optimum test condition determining preparation insect cm-chitosan is: NaOH: chitosan (mass ratio) is 3: 1, NaOH concentration 35%, Mono Chloro Acetic Acid: chitosan (mass ratio) is 3: 1, temperature of reaction 70 DEG C, reaction times 2h.As: the method for preparing chitosan that " comparative studies of method for preparing chitosan " Chen Donghui, Chen Liang, Hu Jiazhen (Environmental Science and Engineering institute of Donghua University, Shanghai 200051) is representative.That is, de-acetyl chitin reaction need be carried out in concentrated base medium, heats and effectively can accelerate acetylization reaction, improves concentration of lye and also can improve deacetylation with the prolongation reaction times.But along with the strengthening of deacetylation condition, the degraded of chitin main chain is also more and more serious, and this directly affects again the quality of product.Therefore concentration of lye, temperature and reaction times are all major influence factors.Control deacetylation condition, just can obtain the chitosan of different deacetylation.Above-mentioned research shows, the preparation of current chitosan is with shrimp, crab shell or insect cot for raw material, and adopt high temperature to carry out deacetylation process, this processes expend energy is many, and the chitosan physiologically active produced is poor.
Summary of the invention
The object of the invention is to the deficiency overcoming above-mentioned technology, and provide a kind of method utilizing low age insect larvae to prepare chitosan at normal temperatures, the larva utilizing vitality extremely strong also directly processes at normal temperatures, the biological activity of retained product that can be more.
The present invention for achieving the above object, by the following technical solutions: a kind of method utilizing low age insect larvae to prepare chitosan at normal temperatures, it is characterized in that: select the larva that vitality is extremely strong, directly carry out process at normal temperatures and prepare chitosan, concrete technology step is as follows:
Four, deproteinization and fat: after selecting 3-5 days fresh and alive insect larvaes to clean to weigh, be placed in squeezing machine and squeeze, material after squeezing in polypide is extruded, repeatedly rinse with tap water, obtain maggot skin, in maggot skin, add the NaoH of 5% concentration of 3 times of volumes, intermittently stir at normal temperatures, sustained reaction 24 hours, in reaction process, fat slowly floats caking, with scraper plate, fat is removed, filter subsequently, protein is removed, make filter cake, be washed to pH6.9, then be washed till pH7 with deionized water, and filter;
Five, decalcification: by the salt acid soak of filter cake through 3 times of volumes, concentration 5%, simultaneously stir, at normal temperatures soak time 6 hours, after decalcification, filters;
Six, deacetylation: add 40-50% sodium hydroxide solution in the filter cake after filtration, slowly stir at normal temperatures, fully reaction 48 hours; Use tap water repetitive scrubbing, be washed till pH value 6.9, be then washed till pH7 with deionized water, filter, filter cake is dried and is pulverized, and obtains the Chitosan powder of purity more than 80%.
Described insect larvae comprises lepidopteran, Homoptera and hemipteran larva.
The temperature of described normal temperature is 10 DEG C-40 DEG C.
Beneficial effect: compared with prior art, the larva that the present invention utilizes vitality extremely strong directly processing at normal temperatures, one is save energy, and two is biological activitys of retained product that can be more.
Embodiment
The specific embodiment of the present invention is described in detail below in conjunction with preferred embodiment.
Embodiment,
Present embodiments provide a kind of method utilizing low age insect larvae to prepare chitosan at normal temperatures, select the larva that vitality is extremely strong, directly carry out process at normal temperatures and prepare chitosan, concrete technology step is as follows:
One, deproteinization and fat: after selecting 3-5 days fresh and alive insect larvaes to clean to weigh, be placed in squeezing machine and squeeze, material after squeezing in polypide is extruded, repeatedly rinse with tap water, obtain maggot skin, in maggot skin, add the NaoH of 5% concentration of 3 times of volumes, intermittently stir at normal temperatures, sustained reaction 24 hours, in reaction process, fat slowly floats caking, with scraper plate, fat is removed, filter subsequently, protein is removed, make filter cake, be washed to pH6.9, then be washed till pH7 with deionized water, and filter;
Two, decalcification: by the salt acid soak of filter cake through 3 times of volumes, concentration 5%, simultaneously stir, at normal temperatures soak time 6 hours, after decalcification, filters;
Three, deacetylation: add 40-50% sodium hydroxide solution in the filter cake after filtration, slowly stir at normal temperatures, fully reaction 48 hours; Use tap water repetitive scrubbing, be washed till pH value 6.9, be then washed till pH7 with deionized water, filter, filter cake is dried and is pulverized, and obtains the Chitosan powder of purity more than 80%.
Described insect larvae comprises lepidopteran, Homoptera and hemipteran larva.The temperature of described normal temperature is 10 DEG C-40 DEG C.
Feature of the present invention is: the larva utilizing vitality extremely strong also directly processes at normal temperatures, changes the technique that tradition prepares chitosan completely.Its superiority is:
One, the chitosan product that obtains of this method, has larger advantage, proves through clinical trial, can immediate pain alleviating, hemostasis, anti-infection, promote wound healing.
Two, save the energy, be beneficial to environmental protection.
Three, method is easy, production cost is low, is convenient to suitability for industrialized production.This method significantly improves the overall economic efficiency of related industries.The development of chitosan industry can be promoted, realize all-win.
Above-mentioned detailed description of the method that this kind of insect larvae prepares chitosan at normal temperatures being carried out with reference to embodiment; illustrative instead of determinate; several embodiments can be listed according to institute's limited range; therefore in the change do not departed under general plotting of the present invention and amendment, should belong within protection scope of the present invention.
Claims (3)
1. utilize low age insect larvae to prepare a method for chitosan at normal temperatures, it is characterized in that: select the larva that vitality is extremely strong, directly carry out process at normal temperatures and prepare chitosan, concrete technology step is as follows:
One, deproteinization and fat: after the insect larvae selecting 3-5 days ages fresh and alive is cleaned, be placed in squeezing machine and squeeze, material after squeezing in polypide is extruded, repeatedly rinse with tap water, obtain maggot skin, in maggot skin, add the NaoH of 5% concentration of 3 times of volumes, at normal temperatures, interrupted stirring, sustained reaction 24 hours, in reaction process, fat slowly floats caking, is removed by fat with scraper plate, filter subsequently, protein is removed, makes filter cake, be washed to pH6.9, be washed till pH7 with deionized water again, and filter;
Two, decalcification: by the salt acid soak of filter cake through 3 times of volumes, concentration 5%, simultaneously stir, after soak time decalcification in 12 hours at normal temperatures, filters;
Three, deacetylation: add 40-50% sodium hydroxide solution in the filter cake after filtration, slowly stir at normal temperatures, fully reaction 48 hours; Use tap water repetitive scrubbing, be washed till pH value 6.9, be then washed till pH7 with deionized water, filter, filter cake is dried and is pulverized, and obtains the Chitosan powder of purity more than 80%.
2. the method utilizing low age insect larvae to prepare chitosan at normal temperatures according to claim 1, is characterized in that: described insect larvae comprises lepidopteran, Homoptera and hemipteran larva.
3. the method utilizing low age insect larvae to prepare chitosan at normal temperatures according to claim 1, is characterized in that: the temperature of described normal temperature is 10 DEG C-40 DEG C.
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Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1186811A (en) * | 1997-01-03 | 1998-07-08 | 张�荣 | Method for extracting and preparing chitin and chitosan |
| CN1259525A (en) * | 2000-01-10 | 2000-07-12 | 卢永军 | Method of extracting and preparing chitosan |
| CN101565470A (en) * | 2009-05-31 | 2009-10-28 | 中南林业科技大学 | Method for preparing chitin and chitosan from dendrolimus punctatus larvae |
| CN102050883A (en) * | 2009-11-04 | 2011-05-11 | 长沙赛邦生物科技有限公司 | Method for extracting chitosan from yellow mealworm shell |
| CN102585035A (en) * | 2012-01-18 | 2012-07-18 | 广东省昆虫研究所 | Preparation method for extracting chitosan from hermetia illucens |
-
2015
- 2015-07-22 CN CN201510435931.9A patent/CN104961844A/en active Pending
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1186811A (en) * | 1997-01-03 | 1998-07-08 | 张�荣 | Method for extracting and preparing chitin and chitosan |
| CN1259525A (en) * | 2000-01-10 | 2000-07-12 | 卢永军 | Method of extracting and preparing chitosan |
| CN101565470A (en) * | 2009-05-31 | 2009-10-28 | 中南林业科技大学 | Method for preparing chitin and chitosan from dendrolimus punctatus larvae |
| CN102050883A (en) * | 2009-11-04 | 2011-05-11 | 长沙赛邦生物科技有限公司 | Method for extracting chitosan from yellow mealworm shell |
| CN102585035A (en) * | 2012-01-18 | 2012-07-18 | 广东省昆虫研究所 | Preparation method for extracting chitosan from hermetia illucens |
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Application publication date: 20151007 |
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