CN107929326A - A kind of American-cockroach-extract, preparation method and its application in anti-oxidant adjusting - Google Patents

A kind of American-cockroach-extract, preparation method and its application in anti-oxidant adjusting Download PDF

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CN107929326A
CN107929326A CN201711426932.2A CN201711426932A CN107929326A CN 107929326 A CN107929326 A CN 107929326A CN 201711426932 A CN201711426932 A CN 201711426932A CN 107929326 A CN107929326 A CN 107929326A
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cockroach
american
extract
concentrate
preparation
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李辉
魏永凯
王朋川
张汉超
杨志斌
赵昱
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Dali University
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    • AHUMAN NECESSITIES
    • A23FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23LFOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
    • A23L33/00Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
    • A23L33/10Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61KPREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K35/00Medicinal preparations containing materials or reaction products thereof with undetermined constitution
    • A61K35/56Materials from animals other than mammals
    • A61K35/63Arthropods
    • AHUMAN NECESSITIES
    • A23FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23VINDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
    • A23V2002/00Food compositions, function of food ingredients or processes for food or foodstuffs
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61KPREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K2236/00Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
    • A61K2236/30Extraction of the material
    • A61K2236/33Extraction of the material involving extraction with hydrophilic solvents, e.g. lower alcohols, esters or ketones
    • A61K2236/333Extraction of the material involving extraction with hydrophilic solvents, e.g. lower alcohols, esters or ketones using mixed solvents, e.g. 70% EtOH
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61KPREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K2236/00Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
    • A61K2236/30Extraction of the material
    • A61K2236/35Extraction with lipophilic solvents, e.g. Hexane or petrol ether

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  • Health & Medical Sciences (AREA)
  • Chemical & Material Sciences (AREA)
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  • Medicines Containing Plant Substances (AREA)
  • Coloring Foods And Improving Nutritive Qualities (AREA)

Abstract

The present invention relates to a kind of American-cockroach-extract, preparation method and its application in anti-oxidant adjusting.The American-cockroach-extract of the present invention is to be prepared by the full worm of American cockroach after petroleum ether soaks, then after alcohol extracting-water precipitating.The American-cockroach-extract that the present invention is prepared can be used in the anti-oxidant adjusting of body, have the function that to strengthen antioxidant ability of organism.

Description

A kind of American-cockroach-extract, preparation method and its application in anti-oxidant adjusting
Technical field
The present invention relates to a kind of American-cockroach-extract, and in particular to a kind of American cockroach polyphenoils and preparation method thereof And application.
Background technology
American cockroach (Periplaneta americana Linn.) is commonly called as cockroach, is Insecta Pterigota Blattaria Blattidae Periplaneta insect.American cockroach be used as medicine first recorded in《Sheng Nong's herbal classic》, wherein it is classified as middle product, call " taste:It is salty, cold; Control:Blood stasis heavily fortified point fever and chills, broken accumulation, laryngopharynx close, endogenous cold s.m.p " (Sun Xingyan etc.,《Sheng Nong's herbal classic》, the Commercial Press, 1955:90).Bai Nationality In Yunnan Popular Utilization American cockroach treats disease, and particularly malignant sore, wound reparation, anti-inflammatory etc. has long History." cockroach " sees for its popular name head《A Supplement to the Compendium of Materia Medica》, it is also referred to as stone ginger, sliding worm;There is its different title in each place Method, such as:Oily mother-in-law, tea woman, stove ant etc. steathily.Blattidae (Blattidae) is a huge insect families, on earth Survive 3.2 hundred million years, the biology of same epoch appearance is belonged to dinosaur;Undoubtedly it is that vitality is most strong, most ancient, so far in the world Multiply one of most successful insect assembly.Cockroach is urban health epidemic prevention department and customs quarantine control department keypoint control as pest One of object.However, the today maked rapid progress in science and technology, its various agent for killing, this insect populations are directed in face of thousands of kinds Still very active, this shows that it has some special materials and its distinctive mechanism of action in vivo, this is caused both at home and abroad The concern of some scholars.
The American cockroach preparation of country's approved listing mainly has " rehabilitation is new ", " Xinmailong injection ", " liver dragon note at present Penetrate liquid " etc., it is mainly used for treating gastric ulcer, cervical erosion, ulcerative colitis, chronic congestive heart failure, chronic type b Hepatitis etc..Found in the follow-up study to American cockroach:Have with alcohol water mixed solvent extraction American cockroach polypide external anti- The effect of herpes simplex virus (Chinese invention patent ZL200810060885.9);It is less than 5000 dalton (i.e. with filter opening< Ultrafiltration membrane 5KDa) is separated, and can obtain new anti-herpes simplex virus and broad spectrum antibiotic activity position (Chinese invention patent ZL201210137422.4, Chinese invention patent ZL 201210137410.1).
The content of the invention
It is prepared into it is an object of the present invention to provide a kind of preparation method of American-cockroach-extract, and by this method The American-cockroach-extract arrived.
Another object of the present invention is that providing above-mentioned American-cockroach-extract is preparing with antioxidation Application in food or medicine.
It is also another object of the present invention to provide a kind of anti-oxidant conditioning agent containing above-mentioned American-cockroach-extract.
In order to achieve the above object, technical scheme is as follows:
A kind of preparation method of American-cockroach-extract, comprises the following steps:
(1) the full worm of American cockroach is taken out after being soaked with petroleum ether, and drying to constant weight at 40~70 DEG C, and U.S. is obtained after smashing The big Lian coarse powder in continent;
(2) the American cockroach coarse powder is extracted with alcoholic solvent at 70 DEG C~80 DEG C, collects extracting solution;By described in Extracting solution is concentrated into the first concentrate that density is 1.10~1.20, and the alcoholic solvent is methanol or ethanol;
(3) first concentrate is mixed with the water of 6~10 times of volumes, the insulated and stirred at 70~80 DEG C, stood After filter, obtained filtrate is concentrated into the second concentrate that relative density is 1.05~1.10, second concentrate is America Big Lian extracts.
Preferably, the weight of the step 2) alcoholic solvent is 4~8 times of the American cockroach coarse powder weight.
Preferably, the number of extraction is 1~3 time in the step 2), and the time extracted every time is 4~8h, is obtained every time Extracting solution merge.
Preferably, the temperature of step (2) described concentration is 70~80 DEG C.
Preferably, second concentrate further includes the processing step of decoloration, drying.
It is further preferred that second concentrate is decolourized with activated carbon.
It is furthermore preferred that the pH value of the activated carbon decolorizing is 4.0~6.0.
Present invention also offers the American-cockroach-extract that the above method is prepared.
Present invention additionally comprises above-mentioned American-cockroach-extract answering in food or medicine with antioxidation is prepared With.
Present invention also offers a kind of anti-oxidant conditioning agent, including a effective amount of above-mentioned American-cockroach-extract and edible Auxiliary material.
Compared with prior art, the present invention has the following advantages:
The preparation method of American-cockroach-extract of the present invention, first soaks the full worm of American cockroach with petroleum ether, by pole The constituents extractions such as the small fat-soluble grease type of property, pigment come out, the full worm drying of the American cockroach after petroleum ether immersion treatment Afterwards, again with methanol or ethanol are extracted, and carrying out water after extracting solution concentration sinks, and obtains American cockroach polyphenoils.The present invention's The American-cockroach-extract product yield and antioxidation biology activity that method is prepared are obtained for larger raising, and are easy to Operate, be easy to large-scale production operation.
The American-cockroach-extract that the present invention is prepared carries out research discovery, the American-cockroach-extract tool being prepared There are higher DPPH radical scavenging activities and hydroxyl radical free radical scavenging capacity, and be verified by experiments it and can be applied to prepare tool The food or medicine of Wheat Protein.
Brief description of the drawings
Fig. 1 is the full-automatic amino acid analysis chromatogram of American-cockroach-extract Ento-C of the present invention.
Embodiment
The present invention provides a kind of American-cockroach-extract with anti-oxidant adjustment effect, the American-cockroach-extract For product of the full worm of American cockroach after petroleum ether cleans after alcohol extracting-water precipitating.
Specifically, the preparation method of American-cockroach-extract comprises the following steps:First by the full worm petroleum ether of American cockroach Soaked, then alcohol extracting-water precipitating will be carried out after the American cockroach drying and crushing after the immersion, obtain American-cockroach-extract.
In the present invention, American cockroach used is preferably dry American cockroach medicinal material.The present invention carrys out American cockroach Source does not limit, and can use commercial product.
The present invention soaks the full worm of American cockroach with petroleum ether, by the small fat-soluble grease type of polarity in American cockroach, color The constituents extractions such as plain class come out.The present invention is not particularly limited the dosage of petroleum ether, as long as can be whole by the full worm of American cockroach It is immersed in petroleum ether, within protection scope of the present invention.It is preferred that the weight of petroleum ether is the full worm weight of immersion American cockroach More than 4 times of amount, can not have the full worm medicinal material of American cockroach.The present invention is not particularly limited the time of immersion, in order to ensure The effect of petroleum ether immersion, in of the invention, the time of preferably petroleum ether immersion is 12~48h, more preferably 24h.
The full worm of the American cockroach of immersion is filtered, the full worm of the American cockroach after being handled.The present invention is to filtering side Method is not particularly limited, using the customary filtration methods in this area.
The full worm of American cockroach after the immersion is persistently dried at 40~70 DEG C, reaches constant weight to American cockroach.This Preferred drying time is invented as 50~65 DEG C.It can be waved under above-mentioned drying temperature in the case where ensureing American cockroach bioactivity Petroleum ether is sent out, in case petroleum ether impacts extract during extraction.The present invention is not particularly limited drying time, to America Big Lian reaches constant weight.In the present invention, it is preferred to suitable drying time is set according to the difference of drying temperature.It is preferred that dry Time is 1~6h, more preferably 2~4h.
The full worm of American cockroach after drying is crushed, in favor of the extraction of American cockroach active material.The present invention is right Breaking method is not particularly limited, using the conventional smashing method in this area, it is preferred to use high-speed multifunctional pulverizer The full worm of American cockroach after drying is crushed.The present invention is not particularly limited the granularity of crushing, and granularity is smaller more favourable In the extraction of American cockroach active material, American cockroach can be prepared according to the degree routinely crushed in those skilled in the art Powder.It is preferred that the full worm of American cockroach is crushed to below 20 mesh, more preferably 5~15 mesh.American cockroach powder after crushing is American cockroach coarse powder.
In the present invention, American cockroach coarse powder is extracted with methanol or ethanol.In the present invention, the methanol or ethanol Concentration is preferably 70~90%, and more preferably 75~85%.The weight of methanol or ethanol is preferably the 4 of American cockroach coarse powder weight ~8 times, more preferably 5~7 times.In particular American cockroach coarse powder is mixed with methanol or ethanol and is extracted.It is preferred that The time of stirring is 0.5~1h.
The present invention is not particularly limited the device of extraction, using the traditional extraction device in this area.In this hair In bright specific embodiment, preferably extracted in multi-function extractor, be specially Y-TQ-5 type multi-function extractors.The present invention In, the temperature that preferably methanol or ethanol extract American cockroach coarse powder is 70 DEG C~80 DEG C, more preferably 72~77 DEG C.Different Extraction time, the sample yield of acquisition have difference, and those skilled in the art can set suitable extraction time according to actual conditions Number.In the present invention, methanol or ethanol include but not limited to the extraction time of American cockroach 1~3 time, more preferably 2 times, every time The time of extraction is preferably 4~8h, more preferably 5~7h.The liquid phase component extracted every time merges, and obtains extracting solution.
Extracting solution is collected, it is 1.10~1.20 that density is concentrated under reduced pressure into obtained extracting solution, obtains the first concentrate.This In invention, the pressure being concentrated under reduced pressure is preferably -70~-80KPa, and the temperature being preferably concentrated under reduced pressure is 70~80 DEG C, more preferably 73~78 DEG C.Be concentrated under reduced pressure can quick recycling design, reduce concentration time, while do not influence in extracting solution active ingredient again and live Property.
The first obtained concentrate is mixed progress water with water to sink.Water volume multiple is added to receive cream rate and anti-oxidant work to precipitation Property can have an impact.In the present invention, preferably plus water volume is 6~10 times of first volume of the concentrated liquid, is more preferably 7~9 times.By The mixture of one concentrate and water is preferably insulated and stirred at 73~78 DEG C, stands at 70~80 DEG C, carries out water and sinks.This hair In bright, the time preferably stood is 0~24h, more preferably 12h, and liquid and precipitation separating degree are good after standing, and precipitation is soft to be easy to Filtering.
Solution after standing is filtered, preferably pore size filter is 0.45 μm, and obtained filtrate decompression is concentrated into relative density For 1.05~1.10 (70 DEG C), the second concentrate is obtained.It is effective containing American-cockroach-extract of the present invention in second concentrate Active ingredient.
In the present invention, preferably the second concentrate is decolourized, obtains decoloration concentrate.In the present invention, it is preferred to use living Property charcoal decolourizes the second concentrate.It is preferred that the quality of activated carbon is the 0.2~2.0% of above-mentioned second concentrate quality, more Preferably 1.0%.In the present invention, preferably activated carbon is powder-type activated carbon.Specifically, activated carbon is mixed with the second concentrate Mixture is obtained, the pH value of mixture is adjusted to 4.0~6.0, is preferably 5.0, stirring mixture decolourizes.The present invention exchanges The mode of whole pH value is not particularly limited, and in the specific embodiment of the invention, the pH value of mixture is preferably adjusted with acetic acid.This hair It is bright preferably using acetic acid be 1.0M acetum.The present invention is not particularly limited the frequency of stirring, can make activated carbon With the well-mixed stir speed (S.S.) of the second concentrate within protection scope of the present invention.It is preferred that stirring frequency for 200~ 500rpm, more preferably 300rpm.In the present invention, the time preferably decolourized is 10~60min, more preferably 30min.After decoloration Mixture is filtered, obtained filtrate is decoloration concentrate.
In the present invention, by decoloration concentrate drying, the powder of American-cockroach-extract is obtained.In the present invention, the drying Preferably it is freeze-dried, to fully ensure that the activity of American-cockroach-extract.The freeze-drying is specially:It will decolourize dense - 20~-86 DEG C of pre-freezes of contracting liquid warp handle 6~12h, then 36~60h is freeze-dried at -70~-86 DEG C.More preferably decolourize dense Contracting liquid handles 12h in -80 DEG C of pre-freezes, then is freeze-dried 48h at -80 DEG C, obtains American-cockroach-extract.The present invention is prepared into To American-cockroach-extract in active ingredient be mainly amino acid, amino acid classes and ratio are shown in the institute of table 5 in embodiment 6 Row.
Present invention discover that the American-cockroach-extract being prepared has antioxidation activity, can be used for preparing has antioxygen In the food or medicine of changing activity, it can also be developed further into as safely and effectively anti-oxidant conditioning agent.
The American-cockroach-extract being prepared in the anti-oxidant conditioning agent of the present invention containing the above method.Preferably, institute State American-cockroach-extract account for the anti-oxidant conditioning agent mass percentage be 0.01~99.9%, more preferably 1~ 99%, more preferably 5~90%.The anti-oxidant conditioning agent of the present invention further includes in addition to containing American-cockroach-extract Edible or pharmaceutically acceptable auxiliary material or auxiliary element.The present invention does not have special limit to the species and ratio of auxiliary element It is fixed, prepared using the conventional method of this area.
For the object, technical solutions and advantages of the present invention are more clearly understood, with reference to embodiment to the present invention into Row detailed description, but they cannot be interpreted as limiting the scope of the present invention.
In the specific embodiment of the invention, the American-cockroach-extract being prepared is denoted as Ento-C.
Embodiment 1
1) the full worm of suitable American cockroach, is weighed, adds 4000g petroleum ethers (30 DEG C~60 DEG C), (15 DEG C of room temperature per 1000g ~25 DEG C) under soak 24h after, filtering;
2), the full worm of American cockroach after filtering is placed in electric heating constant-temperature blowing drying box, electric heating constant temperature air blast is opened and does Dry case and it is arranged at 40 DEG C and carries out continuing drying, is taken out beautiful (about after 5h) until American cockroach reaches constant weight through after a while The big Lian in continent;
3) American cockroach after drying, is placed into high-speed multifunctional pulverizer and carries out crushing operation, American cockroach crushes After took out 20 mesh sieve nets and obtain American cockroach coarse powder;
4) the American cockroach coarse powder that 1000g, is accurately weighed with electronic balance is put into multi-function extractor, adds 70% second Alcoholic solution 6000g, further makes American cockroach corase meal be mixed with ethanol solution, mixing time about 0.5h by stirring;
5) after, American cockroach solution is mixed, it is 70 DEG C to adjust multi-function extractor temperature adjustment knob design temperature, is opened more Function extractor carries out extraction 8h to American cockroach solution;
6) American cockroach solution is taken out after, extracting, filter residue pours into multi-function extractor, and adds new 70% Ethanol solution 4000g, continues to extract 4h;
7) American cockroach solution is filtered out after, extracting, merges filtrate twice, 70 DEG C are concentrated under reduced pressure into density as 1.10 ~1.20;
8) water of 7 times of volumes, is added into concentrate, 70 DEG C of insulated and stirred 30min, stand 12h, filtering liquid medicine, filtrate It is 1.05~1.10 (70 DEG C) to be concentrated under reduced pressure into relative density;
9) 1.0% active carbon particle, is added into concentrate, 1.0M vinegar acid for adjusting pH value to 5.0, is placed in magnetic agitation 300rpm stirs 30min on device;
10) after, stirring, filter to obtain the decoloration concentrate containing active material by filter paper;
11), decolourize -80 DEG C of pre-freeze processing 12h of concentrate warp, is freeze-dried 48h, obtains Ento-C.
Embodiment 2
1) the full worm of suitable American cockroach, is weighed, adds 5000g petroleum ethers (30 DEG C~60 DEG C), (15 DEG C of room temperature per 1000g ~25 DEG C) under soak 24h after, filtering;
2), the full worm of American cockroach after filtering is placed in electric heating constant-temperature blowing drying box, electric heating constant temperature air blast is opened and does Dry case and be arranged at 50 DEG C carry out continue drying, through after a while (about after 3.5h) until American cockroach reach constant weight, take out American cockroach;
3) American cockroach after drying, is placed into high-speed multifunctional pulverizer and carries out crushing operation, American cockroach crushes After took out 20 mesh sieve nets and obtain American cockroach coarse powder;
4) the American cockroach coarse powder that 1000g, is accurately weighed with electronic balance is put into multi-function extractor, adds 80% second Alcoholic solution 6000g, further makes American cockroach corase meal be mixed with ethanol solution, mixing time about 0.5h by stirring;
5) after, American cockroach solution is mixed, it is 72 DEG C to adjust multi-function extractor temperature adjustment knob design temperature, is opened more Function extractor extracts American cockroach solution, thus by the extraction of 6h;
6) American cockroach solution is filtered out after, extracting, merges filtrate twice, 72 DEG C are concentrated under reduced pressure into density as 1.10 ~1.20;
7) water of 6 times of volumes, is added into concentrate, 72 DEG C of insulated and stirred 30min, stand 12h, filtering liquid medicine, filtrate It is 1.05~1.10 (70 DEG C) to be concentrated under reduced pressure into relative density;
8) 1.5% active carbon particle, is added into concentrate, 1.0M vinegar acid for adjusting pH value to 5.0, is placed in magnetic agitation 300rpm stirs 30min on device;
9) after, stirring, filter to obtain the decoloration concentrate containing active material by filter paper;
10), decolourize -80 DEG C of pre-freeze processing 12h of concentrate warp, is freeze-dried 48h, obtains Ento-C.
Embodiment 3
1) the full worm of suitable American cockroach, is weighed, adds 4000g petroleum ethers (30 DEG C~60 DEG C), (15 DEG C of room temperature per 1000g ~25 DEG C) under soak 24h after, filtering;
2), the full worm of American cockroach after filtering is placed in electric heating constant-temperature blowing drying box, electric heating constant temperature air blast is opened and does Dry case and it is arranged at 60 DEG C and carries out continuing drying, is taken out beautiful (about after 2h) until American cockroach reaches constant weight through after a while The big Lian in continent;
3) American cockroach after drying, is placed into high-speed multifunctional pulverizer and carries out crushing operation, American cockroach crushes After took out 20 mesh sieve nets and obtain American cockroach coarse powder;
4) the American cockroach coarse powder that 1000g, is accurately weighed with electronic balance is put into multi-function extractor, adds 70% first Alcoholic solution 6000g, further makes American cockroach corase meal be mixed with methanol solution, mixing time about 0.5h by stirring;
5) after, American cockroach solution is mixed, it is 75 DEG C to adjust multi-function extractor temperature adjustment knob design temperature, is opened more Function extractor extracts American cockroach solution, thus by the extraction of 5h;
6) American cockroach solution is taken out after, extracting, filter residue pours into multi-function extractor, and adds new 70% Methanol solution 4000g, continues to extract 4h;
7) American cockroach solution is filtered out after, extracting, merges filtrate twice, 75 DEG C are concentrated under reduced pressure into density as 1.10 ~1.20;
8) water of 8 times of volumes, is added into concentrate, 75 DEG C of insulated and stirred 30min, stand 12h, filtering liquid medicine, filtrate It is 1.05~1.10 (70 DEG C) to be concentrated under reduced pressure into relative density;
9) 1.0% active carbon particle, is added into concentrate, 1.0M vinegar acid for adjusting pH value to 6.0, is placed in magnetic agitation 300rpm stirs 30min on device;
10) after, stirring, filter to obtain the decoloration concentrate containing active material by filter paper;
11), decolourize -80 DEG C of pre-freeze processing 12h of concentrate warp, is freeze-dried 48h, obtains Ento-C.
Embodiment 4
1) the full worm of suitable American cockroach, is weighed, adds 4000g petroleum ethers (30 DEG C~60 DEG C), (15 DEG C of room temperature per 1000g ~25 DEG C) under soak 24h after, filtering;
2), the full worm of American cockroach after filtering is placed in electric heating constant-temperature blowing drying box, electric heating constant temperature air blast is opened and does Dry case and be arranged at 70 DEG C carry out continue drying, through after a while (about after 1.5h) until American cockroach reach constant weight, take out American cockroach;
3) American cockroach after drying, is placed into high-speed multifunctional pulverizer and carries out crushing operation, American cockroach crushes After took out 20 mesh sieve nets and obtain American cockroach coarse powder;
4) the American cockroach coarse powder that 1000g, is accurately weighed with electronic balance is put into multi-function extractor, adds 85% first Alcoholic solution 6000g, further makes American cockroach corase meal be mixed with methanol solution, mixing time about 0.5h by stirring;
5) after, American cockroach solution is mixed, it is 80 DEG C to adjust multi-function extractor temperature adjustment knob design temperature, is opened more Function extractor extracts American cockroach solution, thus by the extraction of 4h;
6) American cockroach solution is taken out after, extracting, filter residue pours into multi-function extractor, and adds new 85% Methanol solution 4000g, continues to extract 4h;Repeat this step operation once.
7) American cockroach solution is filtered out after, extracting, merges filtrate three times, 80 DEG C are concentrated under reduced pressure into density as 1.10 ~1.20;
8) water of 10 times of volumes, is added into concentrate, 80 DEG C of insulated and stirred 30min, stand 12h, filtering liquid medicine, filtrate It is 1.05~1.10 (70 DEG C) to be concentrated under reduced pressure into relative density;
9) 1.5% active carbon particle, is added into concentrate, 1.0M vinegar acid for adjusting pH value to 4.0, is placed in magnetic agitation 300rpm stirs 30min on device;
10) after, stirring, filter to obtain the decoloration concentrate containing active material by filter paper;
11), decolourize -80 DEG C of pre-freeze processing 12h of concentrate warp, is freeze-dried 48h, obtains Ento-C.
Embodiment 5
Influence of the parameter setting to extraction efficiency in extraction process
1st, drying temperature
The drying temperature for setting the full worm of American cockroach soaked respectively is 40 DEG C, 50 DEG C, 60 DEG C and 70 DEG C, remaining step With embodiment 1, influence of the drying temperature of the full worm of American cockroach to extract is investigated.It the results are shown in Table 1.
Influence of the different drying temperatures of table 1 to American-cockroach-extract antioxidation activity
The purpose of drying is the petroleum ether that volatilizees in the case where ensureing bioactivity, in case petroleum ether is to extract during extraction Impact, therefore need to investigate drying temperature.It can be seen from Table 1 that at different dry temperature, required drying sample when Between it is also inconsistent.Influence of the different drying temperatures to American-cockroach-extract antioxidation activity is different, preferably selects drying time It is more short better.
2nd, Extracting temperature:
When extracting American cockroach with alcoholic solvent, Extracting temperature is higher, yield is higher, but when temperature is higher than more than 80 DEG C, Extract obtained bioactivity will reduce.
The Extracting temperature for setting the full worm of American cockroach soaked respectively is 70 DEG C, 72 DEG C, 75 DEG C, 77 DEG C and 80 DEG C, its Remaining step investigates influence of the Extracting temperature of the full worm of American cockroach to extract with embodiment 1.It the results are shown in Table 2.
Influence of the different Extracting temperatures of table 2 to American-cockroach-extract antioxidation activity
It can be seen from Table 2 that under different Extracting temperatures, the shadow of the American-cockroach-extract antioxidation activity obtained Ring difference, preferably 72~77 DEG C of Extracting temperature.
3rd, extraction time is preferred
The ethanol of solvent selection 70% during extraction, carries out 1 extraction respectively, and 2 extractions and 3 extractions, remaining step are same Embodiment 1.Investigate influence of the different extraction times to sample yield and activity.It the results are shown in Table 3.
Influence of the 3 ethanol extraction time of table to sample yield and activity
As can be seen from Table 3, the extraction efficiency of ethanol is higher, with the increase of extraction time, the yield of the second concentrate Improve, the antioxidation activity of American-cockroach-extract is not much different.
4th, water sinks the preferred of volume
The second concentrate after concentration, aqueous precipitation, when precipitation, add water volume multiple to receive cream rate and anti-oxidant work to precipitation Property can have an impact, therefore the volume ratio that water sinks is investigated.As can be seen from Table 4, different precipitation multiple, the sample of acquisition Yield has difference.
Influence of the volume ratio that 4 water of table sinks to sample yield and activity
5th, decoloration pH value is preferred
Suction-operated of the pH value to activated carbon has an impact, and decolourizes when pH value is low serious, sample yield is low after decoloration, pH value liter Decolorizing effect can reduce during up to neutral or alkalescence.Therefore investigated, find different pH value, the sample yield of acquisition has difference Not.
Influence of the table 5pH values to activated carbon decolorizing
Embodiment 6
The American-cockroach-extract Ento-C that embodiment 1~4 is prepared carries out amino acid content detection
1 instrument, material and reagent:
S433-D amino-acid analyzers;
Chromatographic condition:LCAK06/Na (hydrolysis amino acid analytical column)
Column temperature:55 DEG C~75 DEG C gradient temperature controls
Elution pump:0.45mL/min;Derivative pump:0.25mL/min
Detector:440nm/570nm dual wavelength detectors;Temperature of reactor:130℃;Sampling volume:50μL
Test sample:The American-cockroach-extract Ento-C that embodiment 1~4 is prepared.
It is prepared by 2 test solutions:Precision weighs Ento-C 1.024g, is respectively placed in the volumetric flask of 50ml, adds a certain amount of Ultra-pure water, mixes, and 53kHz ultrasounds are let cool after being allowed to fully dissolving, are settled to 50ml, are taken supernatant 1ml, 10 on centrifuge, 000rpm/min, centrifuges 5min, takes supernatant up to test solution.
4 assay methods:Take respectively for 50 μ L of sample solution, inject full-automatic amino-acid analyzer, by above-mentioned condition sample introduction, Record chromatogram.
The full-automatic amino acid analysis chromatogram of American-cockroach-extract Ento-C is shown in Fig. 1, and concrete outcome is shown in Table 6.
Find after testing, the total free amino acid content of American-cockroach-extract Ento-C is 29.35%.
Amino acid classes and content in 6 American-cockroach-extract Ento-C of table
Embodiment 7
The anti-DPPH free-radical oxidations verifications of American-cockroach-extract Ento-C
DPPH free radical methods are widely used in the evaluation of the radical scavenging activity of various samples.In ethanol solution, DPPH is a kind of stabilized radical in aubergine, there is strong absorption at 517nm.When there are during antioxidant, resisting in solution Oxidant can provide hydrogen atom to DPPH free radicals and bring it about push, its light absorption value is diminished, color becomes more superficial Bright oxidation resistance is stronger.Therefore, its anti-oxidant work can reflect the Scavenging activity of DPPH free radicals by determination sample The power of property, specific detecting step are as follows:
(1) it is accurate to weigh Ento-C 40mg being prepared according to embodiment 1, the insect biological medicine research of Dali university (sample message refers to the American cockroach activity of fighting against senium position total extract that institute provides:Jiao Chunxiang etc., American-cockroach-extract moderate resistance The preliminary analysis of aging active site antioxidation activity, when treasure's traditional Chinese medical science traditional Chinese medicines, the 6th phase of volume 22 in 2011,1389-1391;Sample Product are numbered:KSL-T) (sample message refers to for 40mg, American cockroach anticancer active constituent:It is turned into osmanthus etc., American cockroach active anticancer Component antioxidation activity in vitro analyze, when treasure's traditional Chinese medical science traditional Chinese medicines, the 9th phase of volume 21 in 2011,2249-2250;Sample number into spectrum:CII- 3) (sample message refers to for 40mg, American-cockroach-extract:Tang Xiaguang etc., American cockroach difference extract are removed free radical and are resisted Lipid peroxidation activity research, food industry science and technology, the 13rd phase of volume 37 in 2016,83-85;Sample number into spectrum:ML-A2) 40mg, It is placed in volumetric flask and is settled to 50mL with absolute ethyl alcohol, shakes up, then sample to be tested is diluted to final concentration of 800 in proportion, 400th, the testing sample solution of 200,100,50,25 μ g/mL.
(2) precision weighs Vit C 50mg and is placed in volumetric flask is settled to 50mL with absolute ethyl alcohol, shakes up, then in proportion Vit C are diluted to the testing sample solution of final concentration of 62.50,31.25,15.63,7.81 μ g/mL.
(3) precision weighs 4.4mg DPPH powder, and with absolute ethyl alcohol dissolving and constant volume is in 50mL volumetric flasks, shakes up, Up to 0.2mmol/LDPPH ethanol solutions, used after room temperature avoid light place 30min.
(4) accurate 3.0mL absolute ethyl alcohols of drawing are mixed with 3.0mL DPPH solution, are shaken up, quiet in 37 DEG C of constant temperature lucifuges After putting 30min, reference is made with absolute ethyl alcohol, absorbance A is measured at 517nm0, each sample replication 3 times, is averaged Value.
(5) precision draws the Ento-C of 3.0mL various concentrations and other testing sample solutions respectively, adds 3.0mL's respectively DPPH solution, measures respective absorbance Ai, and each sample replication 3 times, is averaged.
(6) precision draws the Ento-C of 3.0mL various concentrations and other testing sample solutions again, and respectively plus 3.0mL is anhydrous Ethanol measures respective absorbance Aj, and each sample replication 3 times, is averaged.
According to formula P (%)=1- (Ai-Aj)/A0× 100% calculates the clearance rate P of DPPH.
The results are shown in Table 7.Removings of the Ento-C to DPPH is in preferable dose-effect relationship, and its medium effective concentration is bright Aobvious to be less than other American-cockroach-extracts, prompting the American-cockroach-extract Ento-C of the present invention has significant DPPH free radicals Scavenging capacity, can be developed further into as antioxidant.
The clearance rate of American-cockroach-extract and Vit C prepared by 7 different process of table to DPPH
Sample Medium effective concentration EC50(μg/mL)
Ento-C 106.78
KSL-T 331.27
CII-3 >800.00
ML-A2 573.26
VitC 18.05
Embodiment 8
American-cockroach-extract Ento-C oh resistant free-radical oxidation is verified
OH is a kind of strong oxidizer that can induce and trigger lipid that peroxidization occurs, its content can pass through Phen Method measures indirectly, and the basic principle of the method is:H2O2And Fe2+Generation redox reaction, generates hydroxyl radical free radical, hydroxyl free Base has very high reactivity, but the existing time is shorter, if adding salicylic acid in the reaction system, salicylic acid energy Enough effective seizure hydroxyl radical free radicals, and colored complex is generated, which has certain strong suction under wavelength 510nm Receive.After the sample containing antioxidation activity is added into solution, have the material of antioxidation activity will be with bigcatkin willow in sample Acid competition and hydroxyl radical reaction, so that the content of the colored complex formed in solution changes, its absorbance It accordingly can therewith change, the size of sample oxidation resistance is evaluated by measuring the intensity of variation of its absorbance.Specifically Detecting step is as follows:
(1) it is accurate to weigh Ento-C 40mg, insect biological medicine research institute of Dali university that embodiment 1 is prepared and carry (sample message refers to the American cockroach activity of fighting against senium position total extract of confession:Jiao Chunxiang etc., anti-aging in American-cockroach-extract The preliminary analysis of active site antioxidation activity, when treasure's traditional Chinese medical science traditional Chinese medicines, the 6th phase of volume 22 in 2011,1389-1391;Sample is compiled Number:KSL-T) (sample message refers to for 40mg, American cockroach anticancer active constituent:It is turned into osmanthus etc., American cockroach anticancer active constituent Antioxidation activity in vitro analyze, when treasure's traditional Chinese medical science traditional Chinese medicines, the 9th phase of volume 21 in 2011,2249-2250;Sample number into spectrum:CII-3) (sample message refers to for 40mg, American-cockroach-extract:Tang Xiaguang etc., American cockroach difference extract remove free radical and anti-grease Matter peroxidative activity is studied, food industry science and technology, the 13rd phase of volume 37 in 2016,83-85;Sample number into spectrum:ML-A2) 40mg, puts Be settled to 50mL with absolute ethyl alcohol in volumetric flask, shake up, then by each sample to be tested by be diluted to final concentration of 800 than row, 400th, the testing sample solution of 200,100,50,25 μ g/mL.
(2) precision weighs Vit C 50mg and is placed in volumetric flask is settled to 50mL with absolute ethyl alcohol, shakes up, and then presses than row Vit C are diluted to the testing sample solution of final concentration of 62.50,31.25,15.63,7.81 μ g/mL.
(3) FeSO of 2.0mmol/L is sequentially added in 25mL volumetric flasks4Solution 3.0mL and H l.0mmol/L2O2 Solution 3.0mL, adds the salicylic acid solution 3.0mL of 6.0mmol/L, absolute ethyl alcohol constant volume, shakes up, and takes above-mentioned mixing molten after mixing Liquid 5.0mL and absolute ethyl alcohol 1.0mL, in 37 DEG C of water bath with thermostatic control 15min, makees reference with absolute ethyl alcohol, extinction is measured at 510nm Angle value A0, each sample replication 3 times, is averaged.
(4) Ento-C and other testing sample solution 1.0mL of various concentrations are taken, is separately added into above-mentioned mixed solution 5.0mL, mixes, and in 37 DEG C of water bath with thermostatic control 15min, the survey absorbance A 510nm at, each sample replication 3 times, makes even Average.
According to formula I (%)=(A0-A)/A0× 100% calculates the clearance rate I of OH.
The results are shown in Table 8.Ento-C, which has stronger removing OH certainly, to be shown to the clearance rate measure of OH free radicals By the ability of base, and it is in dose-effect trend.The medium effective concentration that Ento-C removes OH free radicals is big significantly lower than other America Lian extracts, prompting the American-cockroach-extract Ento-C of the present invention has significant hydroxyl radical free radical scavenging capacity, Ke Yijin One step is developed into antioxidant.
American-cockroach-extract and Vit C prepared by 8 different process of table is to OH free radical scavenging activity tables
Sample Half effect concentration EC50(μg/mL)
Ento-C 157.43
KSL-T 281.30
CII-3 >800.00
ML-A2 >800.00
VitC 16.46
Above example can illustrate that American-cockroach-extract disclosed by the invention has an antioxidation activity, extracting method into This is low, easy to operate, efficient, suitable for large-scale production, has a good application prospect.
Embodiment 9
A kind of anti-oxidant conditioning agent, includes the component of following weight percentage:
The American-cockroach-extract Ento-C that embodiment 1 is prepared:40g;Starch:40g;Sodium carboxymethyl starch:3g.
By American-cockroach-extract Ento-C and starch 40g, sodium carboxymethyl starch 3g, mixing, excessively 20 mesh sieve nets are prepared wet Particle, 50 DEG C of dry 3h, whole grain, sieving, fills capsule, anti-oxidant conditioning agent is prepared.The anti-oxidant adjusting being prepared Agent can be used in adjusting antioxidant ability of organism.
The above is only the preferred embodiment of the present invention, it is noted that for the ordinary skill people of the art For member, various improvements and modifications may be made without departing from the principle of the present invention, these improvements and modifications also should It is considered as protection scope of the present invention.

Claims (10)

1. a kind of preparation method of American-cockroach-extract, it is characterised in that comprise the following steps:
(1) the full worm of American cockroach is taken out after being soaked with petroleum ether, and drying to constant weight at 40~70 DEG C, and it is big that America is obtained after smashing Lian coarse powder;
(2) the American cockroach coarse powder is extracted with alcoholic solvent at 70 DEG C~80 DEG C, collects extracting solution;By the extraction Liquid is concentrated into the first concentrate that density is 1.10~1.20, and the alcoholic solvent is methanol or ethanol;
(3) first concentrate is mixed with the water of 6~10 times of volumes, the insulated and stirred at 70~80 DEG C, after standing filtering, Obtained filtrate is concentrated into the second concentrate that relative density is 1.05~1.10, second concentrate carries for American cockroach Take thing.
2. preparation method according to claim 1, it is characterised in that the weight of the step 2) alcoholic solvent is the America 4~8 times of big Lian coarse powder weight.
3. preparation method according to claim 1 or 2, it is characterised in that the number of extraction is 1~3 in the step 2) Secondary, the time extracted every time is 4~8h, and the extracting solution obtained every time merges.
4. preparation method according to claim 1, it is characterised in that the temperature of step (2) described concentration is 70~80 DEG C.
5. preparation method according to claim 1, it is characterised in that second concentrate further includes decoloration, drying Processing step.
6. preparation method according to claim 5, it is characterised in that second concentrate is decolourized with activated carbon.
7. preparation method according to claim 6, it is characterised in that the pH value of the activated carbon decolorizing is 4.0~6.0.
8. the American-cockroach-extract that claim 1~8 any one the method is prepared.
9. application of the anti-extract of American cockroach described in claim 8 in food or medicine with antioxidation is prepared.
A kind of 10. anti-oxidant conditioning agent, it is characterised in that including American-cockroach-extract described in a effective amount of claim 8 and Edible auxiliary materials.
CN201711426932.2A 2017-12-26 2017-12-26 A kind of American-cockroach-extract, preparation method and its application in anti-oxidant adjusting Pending CN107929326A (en)

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Application publication date: 20180420