CN103004979A - Preparation method for natural insect repellent - Google Patents
Preparation method for natural insect repellent Download PDFInfo
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- CN103004979A CN103004979A CN2012105312000A CN201210531200A CN103004979A CN 103004979 A CN103004979 A CN 103004979A CN 2012105312000 A CN2012105312000 A CN 2012105312000A CN 201210531200 A CN201210531200 A CN 201210531200A CN 103004979 A CN103004979 A CN 103004979A
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Abstract
The invention relates to a preparation method for a natural insect repellent. The method comprises the following steps: crushing cuminum grains and sieving by 60-80 mesh sieves to obtain cuminum powder; placing the cuminum powder into a Soxhlet extractor, and taking petroleum ether as a solvent to perform Soxhlet extraction under the water bath condition with the temperature of 40-45 DEG C to obtain the active extracting solution after 6-8 hours; and performing pressure reduction and concentration on the active extracting solution to remove the solvent, so as to obtain the effective extract. The preparation method provided by the invention is simple, the operation is simple, convenient and feasible, and the obtained extract has strong prevention and control effect on major stored grain insects.
Description
Technical field
The present invention relates to food scientific technology and technical field of chemistry and chemical engineering, relate in particular to a kind of preparation method of natural insect-repelling agent.
Background technology
Along with the increase and the minimizing that can utilize the arable land of population, food problem remains one of subject matter of 21 century facing mankind; Grain is as important strategic materials, is of great significance for economic development and social stability also tool.Development along with science and technology; the output of grain is greatly improved; its reserves are also grown with each passing day; yet in the process of grain storage, grain usually can be subject to worm-eaten, goes mouldy, the harm of the plague of rats etc., and the loss that causes is comparatively serious; wherein; the harm of grain storage pest is one of problem comparatively serious in the foodstuff preservation, and the loss that China causes because of insect pest every year is about 3% ~ 5% of total output of grain, and the grain storage of some underdeveloped countries and regions can cause 10% ~ 30% loss every year because of insect.Grain storage pest not only causes mass loss, and the edible quality, processing quality, kind that has also reduced grain storage be with quality and technological quality, has shortened warehouse service life; Propagate simultaneously pathogenic microorganisms, the harm people's is healthy.Therefore, the control grain storage pest of adopting an effective measure, though to the scale silo or to family's grain storage all tool be of great significance.
At present, the main still applied chemistry protective agent of the method for control grain storage pest and fumigant carry out, although these chemicals are very effective, but the grain after stifling not only quality descends to some extent, and the medicament of grain and goods is residual that health is also had certain negative effect, also causes simultaneously insect to develop immunity to drugs and the serious problems such as environmental pollution.Celfume is used very extensive as current main fumigant in China, but Celfume is the important chemical substance that damages the ozone layer, and very large to operating personnel's harm, according to " Montreal Protocol Copenhagen amendment ", All Countries will be eliminated it before 1 day January in 2015.The long-term hydrogen phosphide that uses causes pest resistance to insecticide to strengthen fast, and dosage sharply increases when causing pest resurgence, and is more serious to non-destination object generation toxic action and residual contamination.Enhancing along with development, common people's environmental consciousness and the self health consciousness of integrated pest control (IPM) theoretical enforcement, sustainable agriculture and sustainable plant protection, also day by day urgent to the demand of efficient, low toxicity, low-residual, pollution-free, safe pesticide species in the grain storage process, seek and develop new Stored-grain Protectants and seem more important.
Day by day obtaining today of paying attention in environmental protection, is to realize the Grain Sustainable development, and " green pesticide " of seeking a kind of environment-friendly type is extremely urgent.Plant insecticide is owing to having the following advantages the many plant insecticides of ⑴ to people and animals' low toxicity or nontoxic; ⑵ get nature, easily degraded, and its utilization itself is a kind of circulation of natural plant resource, can not cause the additonal pressure that is difficult to tackle to environment; ⑶ insecticidal action mode is various, some mechanism of action is unique, and selectivity is strong, is difficult for causing that insect develops immunity to drugs; ⑷ Insecticidal Plant Resource is abundant, and kind is numerous; ⑸ generally have certain selective, safer to natural enemy, can not destroy natural, ecological system of defense etc., thereby be considered to the environment amenable broad spectrum activity plant protection product of a class.
Cumin (
Cuminum cyminumL.), also be cumin, the fennel of resting in peace, be Carrot family cumin apium annual herb plant.The cumin seed is listed in GRAS (Generally Recognized As Safe) list by U.S. FDA, and U.S. food spices and extract producer association (FEMA) also allow its extract to use at food flavor, flavouring essence for tobacco, wine and use (No.2343) in the essence.Some active material of exploitation cumin seed will have wide market prospects with its pest repellant that is developed to a kind of green.
Summary of the invention
Technical problem to be solved by this invention provides the preparation method of a kind of method natural insect-repelling agent simple, with low cost.
For addressing the above problem, the preparation method of a kind of natural insect-repelling agent of the present invention is characterized in that: after the cumin seed was pulverized 60 ~ 80 mesh sieves, obtain cumin powder; Described cumin powder is placed apparatus,Soxhlet's, take benzinum as solvent, under 40 ~ 45 ℃ of water bath condition, carry out the Soxhlet extracting, obtain active extract after 6 ~ 8 hours; Described active extract namely gets effective extract after the reduced pressure concentration desolventizing; The mass volume ratio of described cumin powder and described benzinum is 1:20 ~ 1:60.
The condition of described reduced pressure concentration refers to that temperature is 25 ~ 30 ℃, pressure is-0.08 ~-0.07MPa.
The present invention compared with prior art has the following advantages:
1, the inventive method is simple, and is easy to operation; Simultaneously used raw material is cheap and easy to get, and production cost is lower.
2, the present invention's isolated a kind of natural effective extract with anthelmintic activity from the cumin seed, this extract has very strong control effect to topmost several grain storage evils.In walking quickly and keeping away test, stifling test and the test of tagging, this cumin extract has good prevention and control BA to four kinds of Main Stored Grain Insects.Test data shows, this extract has four kinds of Main Stored Grain Insects under the low concentration level well walks quickly and keeps away rate and corrected mortality, and effect lasting stability relatively.
⑴ test material, method and data statistics:
For having a try worm:
Sitophilus zea-mais (
Stiophilus zeamais), confused flour beetle (
Tribolium confusum), lesser grain borer (
Rhizopertha dominica), the flat ostomatid in long angle (
Cryptolestes pusillus) provide by city PCO research institute of Hua Zhong Agriculture University.Place temperature (27 ± 1) ℃ in Cans, relative humidity 75% ± 5% is raised in the dark culturing case.Wherein, sitophilus zea-mais is raised with whole grain wheat; Lesser grain borer is raised with half granulous wheat; Confused flour beetle adopts wheat bran and flour to raise with the ratio mixing of 7:3.All put into baking oven (80 ℃) sterilization 2h before feed uses, water content transfers to (13 ± 1) %, is sub-packed in the Cans.Behind the access adult, place the insectary to raise, 7 ~ 1 days afterwards sieve go adult, until adult of future generation is a large amount of occur after 1 ~ 2 week, sift out adult for examination.In the mensuration process, touch adult afterbody, feeler and the sufficient motionless death that is considered as with pin.
1. walk quickly and keep away test
With acetone the cumin extract is made into respectively 0.001 ul/cm
2, 0.002 ul/cm
2, 0.004 ul/cm
2, 0.008 ul/cm
2, 0.016 ul/cm
2, 0.032ul/cm
26 concentration.The circular filter paper of diameter 9cm is cut into two halves, the general acetone diluted liquid 0.5ml that drips equably the cumin extract, the acetone of second half filter paper dropping equivalent in contrast, in fume hood after acetone volatilizees fully, two filter paper are bonded together with transparent adhesive tape at reverse side (note not staying the slit, get into below the filter paper to prevent from trying worm) after insert in the culture dish with diameter, then on the culture dish inwall, evenly coat polytetrafluoroethylene (PTFE) to prevent that trying worm swashes.Access examination worm is 20 in every ware, and 3 repetitions are established in each processing.Culture dish is moved in the incubator identical with condition of culture, 2h, 4h, 8h, the distribution situation of observing examination worm in the culture dish behind the 12h is calculated the rate of walking quickly and keeping away.
2. fumigating effect test
First filter paper is cut into the filter paper bar of 10cm * 1cm, gets the volumetric flask of 250ml, 20 examinations of access worms in every bottle, according to prerun experimental result set handling dose gradient: the fumigating concentration of sitophilus zea-mais and confused flour beetle is 2.5ul/L, 5ul/L, 10ul/L, 20ul/L and 30ul/L, the fumigating concentration of lesser grain borer are 2.5ul/L, 5ul/L, 10ul/L, 15ul/L and 20ul/L, the fumigating concentration of the flat ostomatid in long angle is 0.31ul/L, 0.63ul/L, 1.25ul/L, 1.25ul/L and 2.5ul/L.The cumin extract of respective amount is added drop-wise to the middle part (control group does not drip extract) of filter paper bar with microsyringe, be bonded at the reagent bottle bottleneck, bottle cap is tightened sealing, 3 repetitions are established in each processing, reagent bottle are moved in the incubator identical with condition of culture, respectively at processing 2h, 4h, 8h observes dead examination borer population behind 12h and the 24h, observe after first bottle cap being opened loose gas 10min during observation again.The calculation correction death rate and LC
50, LC
90
3. the effect test of tagging
According to prerun experimental result set handling dose gradient: the sitophilus zea-mais concentration for the treatment of is 8ml/L, 16 ml/L, 32 ml/L and 64 ml/L, confused flour beetle and lesser grain borer concentration for the treatment of are: 4 ml/L, 8 ml/L, 16 ml/L and 32 ml/L, the concentration for the treatment of of the flat ostomatid in long angle is 8 ml/L, 16 ml/L, 32 ml/L and 48 ml/L.The extract acetone diluted drop of above-mentioned concentration is added on the pronotary of examination worm with microsyringe, sitophilus zea-mais wherein, confused flour beetle and lesser grain borer drip 1ul, and the flat ostomatid in long angle drips 0.5ul, and control group drips the acetone of equivalent.3 repetitions are established in each processing, will try worm after handling and put into culture dish, and move in the incubator identical with condition of culture 2h, 4h, 8h, the borer population of observed and recorded death behind 12h and the 24h, and the calculation correction death rate and LC with the preservative film sealing
50, LC
90
4. data are processed
Utilize SPSS software that data are carried out statistical analysis, and obtain regression equation, coefficient correlation and LC
50, LC
90, multiple ratio adopts the Duncan's duncan's new multiple range method.
Computing formula is as follows:
⑵ result of the test:
1. cumin extract of the present invention is to the avoidance effect of sitophilus zea-mais:
The cumin extract exists sitophilus zea-mais walks quickly and keeps away effect (seeing Table 1), as known from Table 1, the cumin extract has the good effect of walking quickly and keeping away to sitophilus zea-mais under finite concentration, avoidance effect increases along with the rising of extract concentrations, prolongation along with the processing time, walk quickly and keep away effect and also increase, the rate of walking quickly and keeping away is elevated to 0.032 uL/cm from initial 6.67%
2, behind the 12h 60%, compare and reach significant difference with the effect of walking quickly and keeping away of low concentration, illustrate also that simultaneously the cumin extract has preferably stability and continuation to the repellent activity of sitophilus zea-mais.
Table 1 cumin extract is to the avoidance effect of sitophilus zea-mais
2. cumin extract of the present invention is to the avoidance effect of confused flour beetle:
The cumin extract sees Table 2 to the avoidance effect of confused flour beetle, and as can be seen from Table 2, the cumin extract of high concentration is rapider to the effect of confused flour beetle, at 2h, and 0.001uL/cm
2Effect is lower, and the rate of walking quickly and keeping away of confused flour beetle is 0, that is to say not demonstrate to walk quickly and keep away effect, but at 0.032uL/cm
2Under the concentration, avoidance effect reaches 63.33%, 12h and rises to 90%, and is consistent with the effect of sitophilus zea-mais, and the rate of walking quickly and keeping away also raises gradually along with the increase of concentration and the prolongation of time.
Table 2 cumin extract is to the avoidance effect of confused flour beetle
3. cumin extract of the present invention is to the avoidance effect of lesser grain borer:
The cumin extract sees Table 3 to the effect of walking quickly and keeping away of lesser grain borer, and lesser grain borer is lower than confused flour beetle to the sensitivity of cumin extract, but is better than sitophilus zea-mais to the sensitivity of cumin extract, last 0.032 uL/cm that is processing
2, during 12h, the rate of walking quickly and keeping away reaches 73.33%, and effect is significantly.Along with the increase of concentration and the lengthening in processing time, the cumin extract increases the repellent activity of lesser grain borer adult.
Table 3 cumin extract is to the avoidance effect of lesser grain borer
4. cumin extract of the present invention is to growing the avoidance effect of the flat ostomatid in angle:
Table 4 is the cumin extract to the repellent activity result of the flat ostomatid in long angle, and as can be seen from Table 4, in 4 kinds of grain storage pests, the flat ostomatid in long angle is the most responsive to the cumin extract, at beginning 0.001 uL/cm of test
2, during 2h, the rate of walking quickly and keeping away just reaches 23.33%, at 0.016 uL/cm
2, during 4h, it is maximum that the rate of walking quickly and keeping away reaches, and is 100%.At 0.016 uL/cm
2, under the 8h, 0.008 uL/cm
2, under the 12h, the rate of walking quickly and keeping away of the flat ostomatid in long angle has all reached 100%, and to walk quickly and keep away effect particularly remarkable.
Table 4 cumin extract is to the avoidance effect of the flat ostomatid in long angle
Under the condition of walking quickly and keeping away of same concentrations, cumin extract of the present invention has the strongest repellent activity to the flat ostomatid in long angle, and the rate of walking quickly and keeping away will significantly be better than other three kinds for the grain storage pest of examination, and at processing time 4h, concentration is 0.016 uL/cm
2The time, the rate of walking quickly and keeping away is 100%, and under same time and concentration, the rate of walking quickly and keeping away of other 3 kinds examination worms is about 50%, and this may be relevant with the little mechanism of action with the cumin extract of volume of the flat ostomatid in long angle.The size of repellent activity is: the flat ostomatid＞confused flour beetle in long angle＞lesser grain borer＞sitophilus zea-mais.
5. cumin extract of the present invention is to the fumigation action of sitophilus zea-mais:
The cumin extract is as shown in table 5 to the fumigation activity result of sitophilus zea-mais, the cumin extract has obvious fumigating effect to sitophilus zea-mais, the death rate of sitophilus zea-mais rises to 83.33% behind the 12h by test initial 6.67%, the death rate is along with the increase of concentration and the lengthening in processing time, and all there is significant difference in the cumin extract to the effect that fumigation activity increases gradually and various dose is processed of corundum Sitophilusspp. imago.
Table 5 cumin extract is to the fumigation action of sitophilus zea-mais
6. cumin extract of the present invention is to the fumigation action of confused flour beetle:
As can be seen from Table 6, the cumin extract also has an obvious fumigation activity to confused flour beetle, under high concentration, in the 2h, the stifling death rate just arrives 50%, after processing 4h, the death rate had also reached 50% after the dosage of 20 uL/L was processed, later prolongation along with the time, the death rate is also in gradually increase, behind 24h, the death rate of confused flour beetle reaches maximum (95%), all has significant difference between the corrected mortality of the corundum Sitophilusspp. imago that various dose is processed.
Table 6 cumin extract is to the fumigation action of confused flour beetle
7. cumin extract of the present invention is to the fumigation action of lesser grain borer
Table 7 is that the cumin extract is to the fumigation activity result of lesser grain borer, by as seen from Table 7, the stifling sensitivity of the cumin extract of lesser grain borer is than the height of confused flour beetle, at 20.00 uL/L, behind the 24h, the stifling death rate of lesser grain borer is just near 100%, reach 98.33%, when 2.50 uL/L of low concentration, the stifling death rate after 24 is 56.67%, when high concentration 20.00uL/L, the death rate also surpasses half behind the 8h, reach 58.33%, overall trend also is that stifling activity is higher with concentration increase and the prolongation of time.
Table 7 cumin extract is to the fumigation action of lesser grain borer
8. cumin extract of the present invention is to growing the fumigation action of the flat ostomatid in angle:
The flat ostomatid in long angle still is (seeing Table 8) the most responsive in 4 kinds of grain storage pests to the stifling sensitiveness of cumin extract, when stifling dosage was 2.5 uL/L of maximum, the processing time was 2h, and the stifling death rate of the flat ostomatid in long angle is 96.67%, behind stifling 4h, the death rate reaches 100%.After processing 12h, only have that the death rate does not reach 100% under the dosage of 0.31uL/L, under remaining concentration, the death rate is 100%.
Table 8 cumin extract is to the fumigation action of the flat ostomatid in long angle
9. cumin extract of the present invention is to the stifling LC of 4 kinds of grain storage pests
50And LC
90:
Table 9 for the cumin extract by statistical analysis to sitophilus zea-mais, confused flour beetle, the LC that lesser grain borer and 4 kinds of grain storage pests of the long flat ostomatid in angle are fumigated
50And LC
90No matter have in the table 9 and can find out, be LC
50And LC
90All are minimum (LC of the flat ostomatid in long angle
50Be 0.391uL/L, LC
90Be 1.614 uL/L), namely responsive to the fumigation activity of cumin extract.The sensitiveness of sitophilus zea-mais is minimum, LC
50And LC
90All maximum is respectively 3.309 uL/L and 69.117 uL/L, the LC of confused flour beetle
50Be less than the LC of lesser grain borer
50, but its LC
90But large than lesser grain borer.
Table 9 cumin extract is to the stifling LC of 4 kinds of grain storage pests
50And LC
90
All in all, result of the test shows that 4 kinds of grain storage pests of cumin extract all have stronger fumigation activity, but there are differences, wherein to obviously be better than other 3 kinds examination worms to the fumigation activity of the flat ostomatid in long angle, the maximum fumigating concentration of the flat ostomatid in long angle only is 1/120 of the maximum fumigating concentration of sitophilus zea-mais and confused flour beetle, 1/80 of the maximum fumigating concentration of lesser grain borer, but fumigating effect but is better than other 3 kinds of grain storage pests.The stifling LC of the flat ostomatid in long angle
50And LC
90All to be far smaller than sitophilus zea-mais, the LC of confused flour beetle and lesser grain borer
50And LC
90, this shows, if under the same conditions silo is fumigated processing, dead should be the long angle flat ostomatid the most responsive to the cumin extract at first.
10. cumin extract of the present invention is to the action of contace poison of four kinds of grain storage pests
Table 10 ~ 13rd, the variable concentrations of cumin extract be to the action of contace poison result of 4 kinds of grain storage pests, processes 3 average correction death rates in the 24h, and the result shows, the cumin extract all has the effect of tagging preferably to 4 kinds of common grain storage pests, but there are differences.The action of contace poison size is followed successively by the flat ostomatid in long angle, confused flour beetle, lesser grain borer, sitophilus zea-mais.In the test, because the volume of the flat ostomatid in long angle will be much smaller than other 3 kinds for the examination grain storage pest, so dripping in the long flat ostomatid in angle extract amount with it is 0.5ul, be to drip 1/2 of on other 3 kinds examination worms extract amount, but the effect of tagging does not but significantly weaken.4 kinds of grain storage pests are after processing 24h, and corrected mortality has all surpassed 50%, and wherein the death rate of the long flat ostomatid in angle and sitophilus zea-mais is the highest, all above 80%, along with the concentration of tagging increase gradually and the processing time prolongation, the effect of action of contace poison is gradually increasing also.
Table 10 cumin extract is to the action of contace poison of sitophilus zea-mais
Table 11 cumin extract is to the action of contace poison of confused flour beetle
Table 12 cumin extract is to the action of contace poison of lesser grain borer
Table 13 cumin extract is to the action of contace poison of the flat ostomatid in long angle
Integrate and see, the cumin extract is to sitophilus zea-mais, confused flour beetle, and four kinds of grain storage pests of lesser grain borer and confused flour beetle all have good control action, but there are differences between worm and worm, also there are differences between the effect of three kinds of control actions.Three kinds of control effects of the flat ostomatid in long angle all are better than other three kinds examination worms.In three kinds of control actions, as the fumigation activity grain storage pest sitophilus zea-mais of tool with using value, confused flour beetle, the LC behind the lesser grain borer 24h
50Be respectively 3.309 uL/L, 2.065 uL/L, 2.465 uL/L.The LC of the flat ostomatid in long angle
50Be 0.391 uL/L, the LC of the flat ostomatid in long angle that fumigation time is shorter
50Be far smaller than other the 3 kinds examination worms of stifling 24h.This shows, the cumin extract has good quick-acting to the flat ostomatid in long angle, do not need the stifling processing of time, can kill the examination worm, fumigate with the cumin extract under the same conditions, begin to occur death most and dead maximum examination worm is the flat ostomatid in long angle, as strengthening the fumigating concentration of cumin extract, stifling effect will be more obvious, and the effect of effect and other 3 kinds of grain storage pests also can be rapider.
3, products obtained therefrom of the present invention is nontoxic to human-body safety, meets the requirement of environmental protection.
The specific embodiment
Embodiment 1A kind of preparation method of natural insect-repelling agent, the cumin seed pulverized 60 mesh sieves after, obtain cumin powder; The 10g cumin powder is placed apparatus,Soxhlet's, take the 400mL benzinum as solvent, under 40 ℃ of water bath condition, carry out the Soxhlet extracting, obtain active extract after 6 hours; Active extract is 25 ℃ in temperature, and pressure namely gets the 0.5g effective extract under-0.08MPa the condition after the reduced pressure concentration desolventizing.
Embodiment 2A kind of preparation method of natural insect-repelling agent, the cumin seed pulverized 80 mesh sieves after, obtain cumin powder; The 20g cumin powder is placed apparatus,Soxhlet's, take the 600mL benzinum as solvent, under 45 ℃ of water bath condition, carry out the Soxhlet extracting, obtain active extract after 8 hours; Active extract is 30 ℃ in temperature, and pressure namely gets the 1.5g effective extract under-0.07MPa the condition after the reduced pressure concentration desolventizing.
Embodiment 3A kind of preparation method of natural insect-repelling agent, the cumin seed pulverized 70 mesh sieves after, obtain cumin powder; The 15g cumin powder is placed apparatus,Soxhlet's, take the 500mL benzinum as solvent, under 43 ℃ of water bath condition, carry out the Soxhlet extracting, obtain active extract after 7 hours; Active extract is 27 ℃ in temperature, and pressure namely gets the 1.1g effective extract under-0.076MPa the condition after the reduced pressure concentration desolventizing.
Embodiment 4A kind of preparation method of natural insect-repelling agent, the cumin seed pulverized 70 mesh sieves after, obtain cumin powder; The 18g cumin powder is placed apparatus,Soxhlet's, take the 550mL benzinum as solvent, under 44 ℃ of water bath condition, carry out the Soxhlet extracting, obtain active extract after 6.5 hours; Active extract is 29 ℃ in temperature, and pressure namely gets the 1.3g effective extract under-0.073MPa the condition after the reduced pressure concentration desolventizing.
Embodiment 5A kind of preparation method of natural insect-repelling agent, the cumin seed pulverized 70 mesh sieves after, obtain cumin powder; The 10g cumin powder is placed apparatus,Soxhlet's, take the 600mL benzinum as solvent, under 44 ℃ of water bath condition, carry out the Soxhlet extracting, obtain active extract after 6.5 hours; Active extract is 29 ℃ in temperature, and pressure namely gets the 0.52g effective extract under-0.073MPa the condition after the reduced pressure concentration desolventizing.
Embodiment 6A kind of preparation method of natural insect-repelling agent, the cumin seed pulverized 70 mesh sieves after, obtain cumin powder; The 10g cumin powder is placed apparatus,Soxhlet's, take the 200mL benzinum as solvent, under 44 ℃ of water bath condition, carry out the Soxhlet extracting, obtain active extract after 6.5 hours; Active extract is 29 ℃ in temperature, and pressure namely gets the 0.49g effective extract under-0.073MPa the condition after the reduced pressure concentration desolventizing.
Above-mentioned
Embodiment 1 ~ 6In apparatus,Soxhlet's be that the happy glass apparatus of Shanghai sorrow factory produces.
Claims (2)
1. the preparation method of a natural insect-repelling agent is characterized in that: after the cumin seed was pulverized 60 ~ 80 mesh sieves, obtain cumin powder; Described cumin powder is placed apparatus,Soxhlet's, take benzinum as solvent, under 40 ~ 45 ℃ of water bath condition, carry out the Soxhlet extracting, obtain active extract after 6 ~ 8 hours; Described active extract namely gets effective extract after the reduced pressure concentration desolventizing; The mass volume ratio of described cumin powder and described benzinum is 1:20 ~ 1:60.
2. the preparation method of a kind of natural insect-repelling agent as claimed in claim 1, it is characterized in that: the condition of described reduced pressure concentration refers to that temperature is 25 ~ 30 ℃, pressure is-0.08 ~-0.07MPa.
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103952463A (en) * | 2013-07-16 | 2014-07-30 | 中国农业大学 | Specific primer based kit for identifying five common cryptoleste stored grain insects |
| CN103952463B (en) * | 2013-07-16 | 2016-01-13 | 中国农业大学 | A kind of test kit identifying 5 kinds of flat ostomatids of common grain storage based on special primer |
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