CN108935037A - A kind of control method of artificial growth dendrobium candidum persticide residue - Google Patents
A kind of control method of artificial growth dendrobium candidum persticide residue Download PDFInfo
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01G—HORTICULTURE; CULTIVATION OF VEGETABLES, FLOWERS, RICE, FRUIT, VINES, HOPS OR SEAWEED; FORESTRY; WATERING
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- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
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- A01G—HORTICULTURE; CULTIVATION OF VEGETABLES, FLOWERS, RICE, FRUIT, VINES, HOPS OR SEAWEED; FORESTRY; WATERING
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Abstract
The present invention relates to a kind of control methods of artificial growth dendrobium candidum persticide residue, including controlling the application method of dendrobium candidum and the detection method of carbendazim pesticide, in 3~June, by on more clever bacterium pesticide sprayings to the blade face of dendrobium candidum of configured certain concentration, which is 500mg/l~1000mg/l.Dendrobium candidum cannot be picked at once early period in sprinkling, several days after spraying, the degradation rate of more clever bacterium reaches 85%~90% in the cauline leaf of dendrobium candidum, can be picked.The pesticide residue in dendrobium candidum is effectively reduced in the present invention, guarantees the effect of dendrobium candidum final product quality, and the survival rate of dendrobium candidum can be improved, to improve yield.
Description
Technical field
The present invention relates to technical field of pesticide, in particular to a kind of controlling party of artificial growth dendrobium candidum persticide residue
Method.
Background technique
Dendrobium candidum is also known as iron sheet orchid, bracketplant, Yunnan iron sheet, iron sheet bracketplant, is that orchid family Dendrobium perennial herb is planted
Object, dendrobium candidum are the top grades of Shihu " medicinal materials, are known as the laudatory title of " gold in Chinese mesona-medicine ", are used as medicine with stem, contain dendrobium nobile
The effective components such as alkali, alkaloid have nourishing Yin and clearing heat, promoting fluid production and nourishing stomach, throat soothing improving eyesight and other effects, exempt from antitumor, raising human body
Epidemic disease power, treatment enterogastric diseases.Anti-aging, anti-oxidant, platelet aggregation-against, reduction blood glucose, treatment cataract etc. have
There is good efficacy.
Pesticide multi-residues refer to using do not divided/degraded in the regular period after pesticide and remained in organism, soil
The general name of earth, water body, pesticide substance, degradation product and toxic metabolite in atmosphere and impurity.People have eaten in contaminated
After herbal medicine, remaining in pesticide therein will be gathered in vivo, slowly because of its metabolism, finally will lead to acute or chronic poisoning,
Generate serious consequence.
It due to dendrobium candidum medical value with higher, is widely used among the health care of human body, therefore, is planting
The residual quantity that pesticide is controlled when dendrobium candidum is particularly important.But at present in artificial growth dendrobium candidum field, for agriculture
The research of the control method of medicine residual quantity is relatively fewer.
In the prior art, a part be by during plantation do not use pesticide, thus to finished pesticide residual quantity into
Row control, as disclosed the dendrobium candidum planting patterns of non agricultural chemical residuum in one in Chinese patent CN106588410B.Pass through choosing
Sterilization treatment is carried out with specific matrix, and to matrix, the use of pesticide is reduced and avoid, to reduce pesticide in dendrobium candidum
Residual quantity.
Another part is then by controlling persticide residue in finished product process, such as Chinese patent
CN107396989A.The cauline leaf just picked is cleaned by clear water first in process, ozone sterilization is being carried out, into one
Step control processing temperature, to remove pesticide residue while guaranteeing the medical value of dendrobium candidum.
Summary of the invention
The object of the present invention is to provide a kind of control methods of artificial growth dendrobium candidum persticide residue, and this method can
Dendrobium candidum illness during plantation effectively is prevented, and can be free of when picking, in cauline leaf or containing a small amount of pesticide
Residual.
Above-mentioned technical purpose of the invention has the technical scheme that
A kind of control method of artificial growth dendrobium candidum persticide residue, application method and more bacterium including control dendrobium candidum
The detection method of clever pesticide,
1) application method of the control dendrobium candidum, specifically includes the following steps:
1.: compound concentration is the carbendazim pesticide of 500mg/l~1000mg/l, the carbendazim that wherein mass percent is 99.8%
Purchased from the Institute for the Control of Agrochemicals of the Ministry of Agriculture,PRC；
2.: dendrobium candidum fast growing period to the above-mentioned carbendazim pesticide of its foliage spray；
3.: after spraying several days after carbendazim pesticide, it just can be carried out picking；
2) detection method of the carbendazim pesticide, comprising the following steps:
1. weighing 1~5g dendrobium candidum is ground up, sieved or obtains dendrobium candidum powder, which is placed in homogeneous cup, and be added 5
~8mL water, expands dendrobium candidum powder sufficiently, then addition 8~12mL acetone and 8~12mL n-hexane, 12000~
16000r/min homogeneous 1~3 minute, centrifugation, then again with 3~6mL acetone and 8~12mL n-hexane homogeneous twice, in merging
Clear liquid acutely shakes 1~5 minute in the tool plug graduated cylinder added with 8~12mL saturated sodium chloride solution, stands 15~30 minutes,
Layering；
2. drawing upper organic phase supernatant crosses anhydrous sodium sulfate, rotary evaporation to 4~6mL is transferred to glass tube nitrogen and is blown to closely
Dry, add 5~7mL ethyl acetate: hexamethylene is 1:1 dissolution.Sample filtering is purified to gel permeation chromatography GPC, collects 9.5
~17.5 minutes effluxes, 38 DEG C~42 DEG C rotary evaporations to 4~6mL are transferred to glass tube nitrogen and are blown to close dry, 1~3mL of addition
Acetone；
3. activating Carb-NH2 column with 3~7mL acetone+methylene chloride (1+1) in advance, liquid to be clean, which is added, leaves it dropwise,
With 3~7mL acetone+methylene chloride (1+1) elution, collection elution liquid nitrogen, which is blown to, closely to be done, and 1~3mL n-hexane is added；By Fu Luoli
Silica column is successively eluted with 3~7mL acetone+n-hexane, 3~7mL n-hexane in advance, is then washed with 3~7mL × 2 acetone+n-hexane
De-, collection eluent nitrogen, which is blown to, closely to be done, and is settled to 0.5~1.5mL with n-hexane；
4. being detected using gas chromatograph-mass spectrometer (GC-MS) GC-MS to sample.
By the foliage-spray carbendazim pesticide in dendrobium candidum, illness machine can be controlled during Growth of Dendrobium candidum
Rate to improve the survival rate of dendrobium candidum, and can effectively reduce or remove the agriculture in mature dendrobium candidum cauline leaf
Medicine residual quantity.
Using specific carbendazim Pesticides Testing method of the invention, pesticide in dendrobium candidum cauline leaf both can be effectively extracted
Residual, and impurity interference can be completely removed, it is easy, time saving, result is accurate.And this method is to the rate of recovery of carbendazim pesticide
75.6%~125.4%, repeatability meets Detecting Pesticide requirement 3.98%~9.54%.
Further preferably, the spraying time is 3 annual~June, which is the fast growing period of dendrobium candidum；Institute
Stating in 3. step just can be carried out picking after application 20-40 days.
During this period of time, dendrobium candidum is in fast growing period, and own metabolism is vigorous, can quickly absorb and agriculture of degrading
Medicine.And it is picked within 20-40 days after administration, is to guarantee that dendrobium candidum adequately can be absorbed and be degraded by own metabolism
More spirit bacterium pesticides further ensure that in the cauline leaf of dendrobium candidum without containing more clever bacterium pesticides.
Further preferably, phase before the application, more clever bacterial contents in dendrobium candidum cauline leaf were in rising trend, at the 2nd~3 day
Reach obtained the maximum absorption.
Several hours after referring to the early period after carrying out application operation, that is, application application early period of the present invention
Or second day.
It does so, can effectively play the drug effect of more clever bacterium, play the role of preventing dendrobium candidum illness.
Further preferably, it after the dendrobium candidum picking to be detected, is cleaned with clear water, then by fresh dendrobium candidum
Being dried in drying box to moisture content is 5%~12.0%, wherein drying temperature is 60~70 DEG C.
It does so, convenient for the operation that dendrobium candidum is pulverized, and is dried within the scope of the drying temperature,
While fresh dendrobium candidum can be dried, destruction is not generated to the medicinal ingredient of dendrobium candidum.
Further preferably, in the GC-MS detection method, the testing conditions of GC chromatography are as follows:
Chromatographic column selects Agilent DB-17ms(30m × 0.25mm × 0.25 μm)；
Column temperature program heating: 58~60 DEG C of 1~minutes of holding rise to 155~165 DEG C with 28~31 DEG C/min, then with 4~6 DEG C/
Min is warming up to 190~300 DEG C, is kept for 8~12 minutes；Column flow rate: 1~1.5mL/min；
Injector temperature: 220~260 DEG C；0.5~1.5 μ L of sample volume；Splitless injecting samples；Carrier gas: nitrogen, purity >=
99.999%。
It does so, simultaneously, analysis time can shortened with improving the accuracy tested and analyzed.
Further preferably, in the GC-MS detection method, the testing conditions of MS chromatography are as follows:
Ion source: the source EI, 65~75eV；Ion source temperature: 200~250 DEG C；Interface temperature: 250~300 DEG C；
Mensuration mode: Salbutamol Selected Ion Monitoring (SIM).Every kind of target compound selects 1~2 quota ion respectively, and 2-3 qualitative
Ion.Every group detection in need ion according to the sequencing of retention time, detected respectively at times.
It does so, can be further improved the accuracy of detection.
Further preferably, the cultivation medium formula of the dendrobium candidum are as follows: the bottom is the broken brick and Hua Gang of 8~12cm
The mixture of rubble, middle layer be 6~8cm lignite mixed fermentation after bark, broken wood shavings, upper layer be 6~8cm broken wood shavings,
The chopping section of fir tree branch and the mixture of fern root.
It does so, the infiltration gas permeability of matrix bottom can be improved, the moisturizing for increasing the fertility and deep layer of middle layer is protected
Outlet capacity reduces dendrobium candidum in the breeding process so that the gas permeability of matrix, fertile degree, water retention property be made to be improved
Rotten rate.
Further preferably, the fermentation process of the bark in the cultivation matrix of the dendrobium candidum and/or broken wood shavings are as follows:
1. bark and/or the pretreatment of broken shavings raw material: admixing weight in the bark and/or broken wood shavings that parts by weight are 80~94.5%
Dried poultrymanure or birds and livestocks faces that part is 3~9% and urea or sulphur ammonium that parts by weight are 1~5% are measured, then adds parts by weight again
The fermentation assistant that lime and parts by weight for 1~5% are 0.5~1%；After above-mentioned substance is stirred, pour into certain
The water of amount seals heap 2~3 days, makes the 45%~55% of moisture content raw material total content, covers covering, and it is small to place 6~12 naturally
When；.
2. fermentation: pretreated fermentation material being piled to the raft of 1~1.5m high again, upper covering is covered above, carries out
It ferments for the first time, the 5th~7 day after for the first fermentation, so that fermentation temperature is reached 45~50 degree, the surface of fermentation material goes out
First time overturning is carried out when existing white hypha, continues to ferment after moving, when accumulative temperature of fermenting reaches 80~90 degree, into
Second of overturning of row, enters second after second of overturning and ferments the phase；Before second ferments, make the control of fermentation material water content 55
~65% or so, when water deficient, can teeming 3~6% comprehensively urea liquid；Deposit thickness is suitably increased again, again heap
At raft, continue to ferment, can stop substantially when accumulative temperature of fermenting reaches 80~100 DEG C through temperature 2~3 weeks of 55~70 DEG C
Only ferment；
3. product handling system: after fermentation, tunning being tiled by 10 ㎝ left and right thicknesses, is dried, is allowed to water content and reaches 35
~45%, finished product is stored at aeration-drying.
The script in matrix can be made the bark of fertility and/or broken wood shavings do not have certain fertility by doing so, and
The fertility of the bark and/or broken wood shavings that are fermented using this method is more permanent, and action time is longer, can effectively reduce due to matrix
Do not have the even withered risk of dendrobium candidum depauperation caused by fertility, and the frequency of replacement matrix can be reduced,
Reduce expense of human.
Further preferably, the fermentation assistant is Gymboree bark fermentation assistant and carrys out chaff according to the dilution proportion of 1:2.5
It mixes thoroughly, is made into increment microbial inoculum.
It does so, can make the functional microorganism in Gymboree bark fermentation assistant that bark is sufficiently fermented and divided
Solution can be removed effectively to ingredients such as the harmful carbolic acid of plant growth, tannic acids, largely Shangdi enhancing tunning
Physical property, commodity performance and economic value.
Further preferably, the bark in the cultivation matrix of the dendrobium candidum be 10~13% oak bark of parts by weight, 20~
24.5% Qi ebon skin, 30~34% pine barks, the mixture in 20~23% Chinese fir barks.
It does so, the type of the microorganism and bacterium colony in fermentation process can be increased, to improve the fertility of fermentation bark.
Further preferably, more clever bacterium are sprayed on the blade face of dendrobium candidum, and a part is by Foliage Absorption, another part quilt
Naturally degradation.
It does so, absorption of the dendrobium candidum to carbendazim can be effectively reduced.
Further preferably, phase before the application, more clever bacterial contents in dendrobium candidum cauline leaf were in rising trend, at the 2nd~3 day
Reach obtained the maximum absorption.
It does so, can effectively play the drug effect of more clever bacterium, play the role of preventing dendrobium candidum illness.
Further preferably, the dendrobium officinale culture medium, which should have, is retained again not ponding, resistance to rot, and guarantees ventilation etc.
Characteristic.
It does so, the survival rate of dendrobium candidum can be improved, avoid rotten.
Further preferably, the cultivation matrix of the dendrobium candidum be broken brick, charcoal, lignite, oak bark, Qi ebon skin,
One or more of pine bark, the pinecone, broken wood shavings, the chopping section of fir tree branch and the fibrous root of trees, fern root.
It does so, can further guarantee the survival rate of dendrobium candidum.
Further preferably, the degradation rate of the dendrobium candidum after administration the 30th day, more spirit bacterium reaches 98%~99%.
It does so, it can be ensured that more clever bacterial contents in the fresh dendrobium candidum cauline leaf after picking are few or do not have.
In conclusion the invention has the following advantages:
1, carbendazim pesticide is sprayed on the blade face of dendrobium candidum, since the edible part of dendrobium candidum is cauline leaf, so more bacterium
Spirit with the edible position of dendrobium candidum being contacted, and blade has breathing and absorption, can be in order to inhaling
Receive carbendazim and carbendazim of effectively degrading；
2, it is picked after application 30 days, dendrobium candidum is sprayed at the carbendazim on blade face by itself metabolism, essence
Pesticide effectively degrades, and makes to be free of in finished product or remain containing a small amount of carbendazim.
3, using specific detection method, as much as possible can make in pesticide dissolution and the solvent in dendrobium candidum, agriculture
The rate of recovery of medicine 75.6%~125.4%, repeatability meets Detecting Pesticide requirement 3.98%~9.54%.And it is mentioning
It takes pesticide in dendrobium candidum remaining simultaneously, and is completely removed impurity interference, it is easy, time saving, result is accurate.
4, it is configured with the matrix of special formulation in the present invention, bark and/broken wood shavings are especially subjected to specific fermentation hand
Section enriches the microbe colony of bark after fermentation, improves the fertility of bark, and obtained matrix have highly-breathable,
Humectant and fertility are more permanent.
5, in the matrix of dendrobium candidum, different types of bark is selected, carries out mixed fermentation according to certain part by weight, respectively
A bark can mutually promote getting fat in the microorganism that sweathouse generates, and the nutritional ingredient in matrix be improved, to promote iron sheet
The healthy growth of dendrobium nobile.
Specific embodiment
Embodiment 1:
It in 3~June, is that the phase is not before the application on more clever bacterium pesticide sprayings to the blade face of dendrobium candidum of 500mg/l by concentration
Dendrobium candidum can be picked, after administration the 30th day, using pesticide detection device to the pesticide in dendrobium candidum cauline leaf
Remnants are detected, it is found that the degradation rate of more clever bacterium is 98.5%~99%.
A kind of control method of artificial growth dendrobium candidum persticide residue, including control dendrobium candidum application method and
The detection method of carbendazim pesticide,
1) application method of the control dendrobium candidum, specifically includes the following steps:
1.: compound concentration is the carbendazim pesticide of 500mg/l, and the carbendazim that wherein mass percent is 99.8% is purchased from the Ministry of Agriculture
Agriculture chemical examination institution；
2.: 3~June dendrobium candidum fast growing period to the above-mentioned carbendazim pesticide of its foliage spray；
3.: after spraying 30 after carbendazim pesticide days, it just can be carried out picking.
2) detection method of the carbendazim pesticide, comprising the following steps:
1.: it weighs 2g dendrobium candidum and is ground up, sieved or obtains dendrobium candidum powder, which is placed in homogeneous cup, and 6mL is added
Water expands dendrobium candidum powder sufficiently, then addition 10mL acetone and 10mL n-hexane, and 15000r/min homogeneous 1 minute, from
Then the heart uses 5mL acetone and 10mL n-hexane homogeneous twice again, merge supernatant in the tool added with 10mL saturated sodium chloride solution
It fills in graduated cylinder, acutely concussion 2 minutes, stands 20 minutes, layering.
2.: it draws upper organic phase supernatant and crosses anhydrous sodium sulfate, rotary evaporation to 5mL is transferred to glass tube nitrogen and is blown to
Close dry, add 6mL ethyl acetate: hexamethylene is 1:1 dissolution.Sample filtering is purified to GPC, is collected 9.5~17.5 minutes and is flowed
Liquid out, 40 DEG C of rotary evaporations to 5mL are transferred to glass tube nitrogen and are blown to close dry, addition 2mL acetone.
3.: Carb-NH2 column is activated with 5mL acetone+methylene chloride (1+1) in advance, liquid to be clean, which is added, stays it dropwise
Under, it is eluted with 5mL × 2 acetone+methylene chloride (1+1), collection elution liquid nitrogen, which is blown to, closely to be done, and 2mL n-hexane is added.By Fu Luoli
Silica column is successively eluted with 5mL acetone+n-hexane (1+9), 5mL n-hexane in advance, be added it is above-mentioned to be clean, with the acetone of 5mL × 2+
N-hexane (1+9) elution, collection eluent nitrogen, which is blown to, closely to be done, and is settled to 1mL with n-hexane.
4.: sample is detected using GC-MS.
The testing conditions of GC chromatography:
Chromatographic column selects Agilent DB-17ms(30m × 0.25mm × 0.25 μm)；
Column temperature program heating: 60 DEG C are kept for 1 minute, are risen to 160 DEG C with 30 DEG C/min, then be warming up to 295 DEG C with 5 DEG C/min, are protected
It holds 10 minutes；Column flow rate: 1.2mL/min；
Injector temperature: 250 DEG C；1 μ L of sample volume；Splitless injecting samples；Carrier gas: nitrogen, purity >=99.999%.
The testing conditions of MS chromatography are as follows:
Ion source: the source EI, 70eV；Ion source temperature: 230 DEG C；Interface temperature: 280 DEG C；
Mensuration mode: Salbutamol Selected Ion Monitoring (SIM).Every kind of target compound selects 1 quota ion respectively, 2-3 it is qualitative from
Son.Every group detection in need ion according to the sequencing of retention time, detected respectively at times.
The cultivation medium formula of dendrobium candidum are as follows: the bottom is the broken brick of 8cm and the mixture of Hua Gang rubble, and middle layer is
Bark, broken wood shavings after the lignite mixed fermentation of 8cm, upper layer are the broken wood shavings of 8cm, the chopping section of fir tree branch and fern root
Mixture.
The fermentation process of bark and/or broken wood shavings in the cultivation matrix of dendrobium candidum are as follows:
1. bark and/or the pretreatment of broken shavings raw material: admixing parts by weight in the bark and/or broken wood shavings that parts by weight are 80% is
9% dried poultrymanure or birds and livestocks faces and parts by weight be 5% urea or sulphur ammonium, then add again parts by weight be 5% lime with
And the fermentation assistant that parts by weight are 1%；After above-mentioned substance is stirred, a certain amount of water is poured into, heap is sealed 3 days, makes moisture
Content is the 50% of raw material total content.
2. fermentation: pretreated fermentation material being piled to the raft of 1~1.5m high again, upper covering is covered above, carries out
It ferments for the first time, the 5th day after for the first fermentation, so that fermentation temperature is reached 50 degree, white occurs in the surface of fermentation material
First time overturning is carried out when mycelia, continues to ferment after moving, and when accumulative temperature of fermenting reaches 90 degree, is turned over for the second time
, enter second of fermentation phase after second of overturning；Before second ferments, make the control of fermentation material water content 65% or so, moisture
When insufficient, can comprehensively teeming 6% urea liquid.Deposit thickness is suitably increased again, piles raft again, continues to ferment, is passed through
70 DEG C of temperature 2~3 weeks can stop fermenting substantially when accumulative temperature of fermenting reaches 100 DEG C.
3. product handling system: after fermentation, tunning being tiled by 10 ㎝ left and right thicknesses, is dried, is allowed to water content and reaches
To 45%, finished product is stored at aeration-drying.
Embodiment 2:
It in 3~June, is that the phase is not before the application on more clever bacterium pesticide sprayings to the blade face of dendrobium candidum of 750mg/l by concentration
Dendrobium candidum can be picked, after administration the 30th day, using pesticide detection device to the pesticide in dendrobium candidum cauline leaf
Remnants are detected, it is found that the degradation rate of more clever bacterium is 97%~98.6%.
A kind of control method of artificial growth dendrobium candidum persticide residue, including control dendrobium candidum application method and
The detection method of carbendazim pesticide,
1) application method of the control dendrobium candidum, specifically includes the following steps:
1.: compound concentration is the carbendazim pesticide of 750mg/l, and the carbendazim that wherein mass percent is 99.8% is purchased from the Ministry of Agriculture
Agriculture chemical examination institution；
2.: 3~June dendrobium candidum fast growing period to the above-mentioned carbendazim pesticide of its foliage spray；
3.: after spraying 30 after carbendazim pesticide days, it just can be carried out picking.
2) detection method of the carbendazim pesticide, comprising the following steps:
1.: it weighs 2g dendrobium candidum and is ground up, sieved or obtains dendrobium candidum powder, which is placed in homogeneous cup, and 6mL is added
Water expands dendrobium candidum powder sufficiently, then addition 10mL acetone and 10mL n-hexane, and 15000r/min homogeneous 1 minute, from
Then the heart uses 5mL acetone and 10mL n-hexane homogeneous twice again, merge supernatant in the tool added with 10mL saturated sodium chloride solution
It fills in graduated cylinder, acutely concussion 2 minutes, stands 20 minutes, layering.
2.: it draws upper organic phase supernatant and crosses anhydrous sodium sulfate, rotary evaporation to 5mL is transferred to glass tube nitrogen and is blown to
Close dry, add 6mL ethyl acetate: hexamethylene is 1:1 dissolution.Sample filtering is purified to GPC, is collected 9.5~17.5 minutes and is flowed
Liquid out, 40 DEG C of rotary evaporations to 5mL are transferred to glass tube nitrogen and are blown to close dry, addition 2mL acetone.
3.: Carb-NH2 column is activated with 5mL acetone+methylene chloride (1+1) in advance, liquid to be clean, which is added, stays it dropwise
Under, it is eluted with 5mL × 2 acetone+methylene chloride (1+1), collection elution liquid nitrogen, which is blown to, closely to be done, and 2mL n-hexane is added.By Fu Luoli
Silica column is successively eluted with 5mL acetone+n-hexane (1+9), 5mL n-hexane in advance, be added it is above-mentioned to be clean, with the acetone of 5mL × 2+
N-hexane (1+9) elution, collection eluent nitrogen, which is blown to, closely to be done, and is settled to 1mL with n-hexane.
4.: sample is detected using GC-MS.
The testing conditions of GC chromatography:
Chromatographic column selects Agilent DB-17ms(30m × 0.25mm × 0.25 μm)；
Column temperature program heating: 60 DEG C are kept for 1 minute, are risen to 160 DEG C with 30 DEG C/min, then be warming up to 295 DEG C with 5 DEG C/min, are protected
It holds 10 minutes；Column flow rate: 1.2mL/min；
Injector temperature: 250 DEG C；1 μ L of sample volume；Splitless injecting samples；Carrier gas: nitrogen, purity >=99.999%.
The testing conditions of MS chromatography are as follows:
Ion source: the source EI, 70eV；Ion source temperature: 230 DEG C；Interface temperature: 280 DEG C；
Mensuration mode: Salbutamol Selected Ion Monitoring (SIM).Every kind of target compound selects 1 quota ion respectively, 2-3 it is qualitative from
Son.Every group detection in need ion according to the sequencing of retention time, detected respectively at times.
The cultivation medium formula of dendrobium candidum are as follows: the bottom is the broken brick of 10cm and the mixture of Hua Gang rubble, middle layer
For bark, the broken wood shavings after the lignite mixed fermentation of 7cm, upper layer is the chopping section and fern root of the broken wood shavings of 7cm, fir tree branch
Mixture.
The fermentation process of bark and/or broken wood shavings in the cultivation matrix of dendrobium candidum are as follows:
1. bark and/or the pretreatment of broken shavings raw material: admixing parts by weight in the bark and/or broken wood shavings that parts by weight are 88% is
Then the urea or sulphur ammonium that 6% dried poultrymanure or birds and livestocks faces and parts by weight is 3% add the lime that parts by weight are 2.5% again
And the fermentation assistant that parts by weight are 0.5%；After above-mentioned substance is stirred, a certain amount of water is poured into, heap is sealed 3 days, makes
Moisture content is the 50% of raw material total content.
2. fermentation: pretreated fermentation material being piled to the raft of 1~1.5m high again, upper covering is covered above, carries out
It ferments for the first time, the 5th day after for the first fermentation, so that fermentation temperature is reached 50 degree, white occurs in the surface of fermentation material
First time overturning is carried out when mycelia, continues to ferment after moving, and when accumulative temperature of fermenting reaches 90 degree, is turned over for the second time
, enter second of fermentation phase after second of overturning；Before second ferments, make the control of fermentation material water content 65% or so, moisture
When insufficient, can comprehensively teeming 6% urea liquid.Deposit thickness is suitably increased again, piles raft again, continues to ferment, is passed through
70 DEG C of temperature 2~3 weeks can stop fermenting substantially when accumulative temperature of fermenting reaches 100 DEG C.
3. product handling system: after fermentation, tunning being tiled by 10 ㎝ left and right thicknesses, is dried, is allowed to water content and reaches
To 45%, finished product is stored at aeration-drying.
Embodiment 3:
In 3~June, it is on more clever bacterium pesticide sprayings to the blade face of dendrobium candidum of 1000mg/l by concentration, before the application the phase
Dendrobium candidum cannot be picked, after administration the 30th day, using pesticide detection device to the agriculture in dendrobium candidum cauline leaf
Medicine remnants are detected, it is found that the degradation rate of more clever bacterium is 96%~98%.
A kind of control method of artificial growth dendrobium candidum persticide residue, including control dendrobium candidum application method and
The detection method of carbendazim pesticide, it is characterised in that:
1) application method of the control dendrobium candidum, specifically includes the following steps:
1.: compound concentration is the carbendazim pesticide of 1000mg/l, and the carbendazim that wherein mass percent is 99.8% is purchased from the Ministry of Agriculture
Agriculture chemical examination institution；
2.: 3~June dendrobium candidum fast growing period to the above-mentioned carbendazim pesticide of its foliage spray；
3.: after spraying 30 after carbendazim pesticide days, it just can be carried out picking.
2) detection method of the carbendazim pesticide, comprising the following steps:
1.: it weighs 2g dendrobium candidum and is ground up, sieved or obtains dendrobium candidum powder, which is placed in homogeneous cup, and 6mL is added
Water expands dendrobium candidum powder sufficiently, then addition 10mL acetone and 10mL n-hexane, and 15000r/min homogeneous 1 minute, from
Then the heart uses 5mL acetone and 10mL n-hexane homogeneous twice again, merge supernatant in the tool added with 10mL saturated sodium chloride solution
It fills in graduated cylinder, acutely concussion 2 minutes, stands 20 minutes, layering.
2.: it draws upper organic phase supernatant and crosses anhydrous sodium sulfate, rotary evaporation to 5mL is transferred to glass tube nitrogen and is blown to
Close dry, add 6mL ethyl acetate: hexamethylene is 1:1 dissolution.Sample filtering is purified to GPC, is collected 9.5~17.5 minutes and is flowed
Liquid out, 40 DEG C of rotary evaporations to 5mL are transferred to glass tube nitrogen and are blown to close dry, addition 2mL acetone.
3.: Carb-NH2 column is activated with 5mL acetone+methylene chloride (1+1) in advance, liquid to be clean, which is added, stays it dropwise
Under, it is eluted with 5mL × 2 acetone+methylene chloride (1+1), collection elution liquid nitrogen, which is blown to, closely to be done, and 2mL n-hexane is added.By Fu Luoli
Silica column is successively eluted with 5mL acetone+n-hexane (1+9), 5mL n-hexane in advance, be added it is above-mentioned to be clean, with the acetone of 5mL × 2+
N-hexane (1+9) elution, collection eluent nitrogen, which is blown to, closely to be done, and is settled to 1mL with n-hexane.
4.: sample is detected using GC-MS.
The testing conditions of GC chromatography:
Chromatographic column selects Agilent DB-17ms(30m × 0.25mm × 0.25 μm)；
Column temperature program heating: 60 DEG C are kept for 1 minute, are risen to 160 DEG C with 30 DEG C/min, then be warming up to 295 DEG C with 5 DEG C/min, are protected
It holds 10 minutes；Column flow rate: 1.2mL/min；
Injector temperature: 250 DEG C；1 μ L of sample volume；Splitless injecting samples；Carrier gas: nitrogen, purity >=99.999%.
The testing conditions of MS chromatography are as follows:
Ion source: the source EI, 70eV；Ion source temperature: 230 DEG C；Interface temperature: 280 DEG C；
Mensuration mode: Salbutamol Selected Ion Monitoring (SIM).Every kind of target compound selects 1 quota ion respectively, 2-3 it is qualitative from
Son.Every group detection in need ion according to the sequencing of retention time, detected respectively at times.
The cultivation medium formula of dendrobium candidum are as follows: the bottom is the broken brick of 12cm and the mixture of Hua Gang rubble, middle layer
For bark, the broken wood shavings after the lignite mixed fermentation of 6cm, upper layer is the chopping section and fern root of the broken wood shavings of 6cm, fir tree branch
Mixture.
The fermentation process of bark and/or broken wood shavings in the cultivation matrix of dendrobium candidum are as follows:
1. bark and/or the pretreatment of broken shavings raw material: admixing parts by weight in the bark and/or broken wood shavings that parts by weight are 94.5 is
3% dried poultrymanure or birds and livestocks faces and parts by weight be 1% urea or sulphur ammonium, then add again parts by weight be 1% lime with
And the fermentation assistant that parts by weight are 0.5%；After above-mentioned substance is stirred, a certain amount of water is poured into, heap is sealed 3 days, makes water
Dividing content is the 50% of raw material total content.
2. fermentation: pretreated fermentation material being piled to the raft of 1~1.5m high again, upper covering is covered above, carries out
It ferments for the first time, the 5th day after for the first fermentation, so that fermentation temperature is reached 50 degree, white occurs in the surface of fermentation material
First time overturning is carried out when mycelia, continues to ferment after moving, and when accumulative temperature of fermenting reaches 90 degree, is turned over for the second time
, enter second of fermentation phase after second of overturning；Before second ferments, make the control of fermentation material water content 65% or so, moisture
When insufficient, can comprehensively teeming 6% urea liquid.Deposit thickness is suitably increased again, piles raft again, continues to ferment, is passed through
70 DEG C of temperature 2~3 weeks can stop fermenting substantially when accumulative temperature of fermenting reaches 100 DEG C.
3. product handling system: after fermentation, tunning being tiled by 10 ㎝ left and right thicknesses, is dried, is allowed to water content and reaches
To 45%, finished product is stored at aeration-drying.
Comparative example 1:
A kind of control method of artificial growth dendrobium candidum persticide residue, application method and more bacterium including control dendrobium candidum
The detection method of clever pesticide,
1) application method of the control dendrobium candidum, specifically includes the following steps:
1.: compound concentration is the carbendazim pesticide of 500mg/l, and the carbendazim that wherein mass percent is 99.8% is purchased from the Ministry of Agriculture
Agriculture chemical examination institution；
2. the fast growing period of dendrobium candidum sprays above-mentioned carbendazim pesticide to its matrix in 3~June；
3.: after spraying 30 after carbendazim pesticide days, it just can be carried out picking；
2) sample of comparative example is detected using the Pesticides Testing technology of patent CN106198510A, 5g is taken to wait for test sample
Product shred into 1cm2Fragment be placed on 10ml and pH=7.5 phosphoric acid it is slow in liquid impregnate 10 minutes obtained prepare liquids, 1ml is waited for
It surveys drop and is added to well, stand 10 minutes, react prepare liquid sufficiently with the BuCh enzyme being loaded in piece.
Bending part is bent to sample-adding portion direction, keeps well and substrate hole corresponding, protrusion is opposite with first flap
It answers, while pressing sample-adding portion and substrate portion, the pressing force that protrusion applies first flap makes first flap fracture, described
The groove sidewall of protrusion and the scale slot abuts against.
Sample-adding portion and substrate portion closed state 3~8 minutes are kept, so that prepare liquid is in contact with substrate piece, prepare liquid and bottom
Acetylthiocholine iodide in object piece sufficiently reacts, and developing solution is made.
Substrate portion is lifted, makes sample-adding portion and substrate part from developing solution obtained passes through the first gear to fracture in step S3
Piece flow into permeable tank simultaneously be in contact with osmosizing portion, main part is infiltrated through according to osmosis, according to main part discoloration range and
Corresponding graduated scale reads scale, and reading scale is pesticide residue content
Comparative example 2:
A kind of control method of artificial growth dendrobium candidum persticide residue, application method and more bacterium including control dendrobium candidum
The detection method of clever pesticide, it is characterised in that:
1) application method of the control dendrobium candidum, specifically includes the following steps:
1.: compound concentration is the carbendazim pesticide of 500mg/l, and the carbendazim that wherein mass percent is 99.8% is purchased from the Ministry of Agriculture
Agriculture chemical examination institution；
2.: in 3~June, the fast growing period of dendrobium candidum carries out root irrigation to it using above-mentioned carbendazim pesticide；
3.: after spraying 30 after carbendazim pesticide days, it just can be carried out picking；
2) sample of comparative example is detected using the Pesticides Testing technology of patent CN106198510A, 5g is taken to wait for test sample
The slow middle liquid of phosphoric acid that the fragment that product shred into 1cm2 is placed on 10ml and pH=7.5 impregnates 10 minutes obtained prepare liquids, and 1ml is waited for
It surveys drop and is added to well, stand 10 minutes, react prepare liquid sufficiently with the BuCh enzyme being loaded in piece.
Bending part is bent to sample-adding portion direction, keeps well and substrate hole corresponding, protrusion is opposite with first flap
It answers, while pressing sample-adding portion and substrate portion, the pressing force that protrusion applies first flap makes first flap fracture, described
The groove sidewall of protrusion and the scale slot abuts against.
Sample-adding portion and substrate portion closed state 3~8 minutes are kept, so that prepare liquid is in contact with substrate piece, prepare liquid and bottom
Acetylthiocholine iodide in object piece sufficiently reacts, and developing solution is made.
Substrate portion is lifted, makes sample-adding portion and substrate part from developing solution obtained passes through the first gear to fracture in step S3
Piece flow into permeable tank simultaneously be in contact with osmosizing portion, main part is infiltrated through according to osmosis, according to main part discoloration range and
Corresponding graduated scale reads scale, and reading scale is pesticide residue content.
It by comparison, is administered in Examples 1 to 3 using blade face of the carbendazim pesticide to dendrobium candidum, and using special
Fixed GC-MS detection method carries out pesticide residue determination to dendrobium candidum, finds the recycling through GC-MS detection method to carbendazim pesticide
For rate 75.6%~125.4%, repeatability meets Detecting Pesticide requirement 3.98%~9.54%.And after application 30 days
Dendrobium candidum carry out the measurement of persticide residue, it is found that degradation of pesticide rate of the carbendazim in dendrobium candidum cauline leaf is 98.2%
~99.9%.
And application and pouring root are carried out using matrix of the carbendazim pesticide to dendrobium candidum respectively in comparative example 1~2
Processing, and pesticide residue determination is carried out to dendrobium candidum using the Pesticides Testing technology of patent CN106198510A, discovery uses
This method to the rate of recovery of carbendazim pesticide 50%~60% or so, far below GC-MS detection method of the invention.And to application
Dendrobium candidum after 30 days carries out the measurement of persticide residue, it is found that degradation of pesticide rate of the carbendazim in dendrobium candidum cauline leaf is
85%~90%, persticide residue is larger, seriously affects the product quality of dendrobium candidum.
This specific embodiment is only explanation of the invention, is not limitation of the present invention, those skilled in the art
Member can according to need the modification that not creative contribution is made to the present embodiment after reading this specification, but as long as at this
All by the protection of Patent Law in the scope of the claims of invention.
Claims (10)
1. a kind of control method of artificial growth dendrobium candidum persticide residue, it is characterised in that: including control dendrobium candidum
The detection method of application method and carbendazim pesticide,
1) application method of the control dendrobium candidum, specifically includes the following steps:
1.: compound concentration is the carbendazim pesticide of 500mg/l~1000mg/l；
2.: dendrobium candidum fast growing period to the above-mentioned carbendazim pesticide of its foliage spray；
3.: after spraying several days after carbendazim pesticide, it just can be carried out picking；
2) detection method of the carbendazim pesticide, comprising the following steps:
1. weighing 1~5g dendrobium candidum is ground up, sieved or obtains dendrobium candidum powder, which is placed in homogeneous cup, and be added 5
~8mL water, expands dendrobium candidum powder sufficiently, then addition 8~12mL acetone and 8~12mL n-hexane, 12000~
16000r/min homogeneous 1~3 minute, centrifugation, then again with 3~6mL acetone and 8~12mL n-hexane homogeneous twice, in merging
Clear liquid acutely shakes 1~5 minute in added with 8~12mL saturated sodium chloride solution, stands 15~30 minutes, layering；
2. drawing upper organic phase supernatant crosses anhydrous sodium sulfate, rotary evaporation to 4~6mL is transferred to glass tube nitrogen and is blown to closely
It is dry, add 5~7mL ethyl acetate: hexamethylene dissolution；Sample filtering is purified to gel permeation chromatography GPC, collect 9.5~
17.5 minutes effluxes, 38 DEG C~42 DEG C rotary evaporations to 4~6mL are transferred to glass tube nitrogen and are blown to close dry, addition 1~3mL third
Ketone；
3. in advance with 3~7mL acetone+methylene chloride activation column, liquid to be clean, which is added, leaves it dropwise, with 3~7mL acetone+
Dichloromethane eluent collects elution liquid nitrogen and is blown to close dry, addition 1~3mL n-hexane；Florisil silica column is successively used into 3~7mL
Acetone+n-hexane, 3~7mL n-hexane elute in advance, are then eluted with 3~7mL × 2 acetone+n-hexane, collect eluent nitrogen
It is blown to and closely does, be settled to 0.5~1.5mL with n-hexane；
4. being detected to sample.
2. a kind of control method of artificial growth dendrobium candidum persticide residue according to claim 1, it is characterised in that:
The spraying time is 3 annual~June, which is the fast growing period of dendrobium candidum；3. in application 20- in the step
It just can be carried out picking after 40 days.
3. a kind of control method of artificial growth dendrobium candidum persticide residue according to claim 2, it is characterised in that:
Phase before the application, the carbendazim content in dendrobium candidum cauline leaf is in rising trend, reaches obtained the maximum absorption at the 2nd~3 day.
4. a kind of control method of artificial growth dendrobium candidum persticide residue according to claim 1, it is characterised in that:
After the dendrobium candidum picking, cleaned with clear water, then by fresh dendrobium candidum, drying to moisture content is in drying box
5%~12.0%, wherein drying temperature is 60~70 DEG C.
5. a kind of control method of artificial growth dendrobium candidum persticide residue according to claim 1, it is characterised in that:
In the GC-MS detection method, the testing conditions of GC chromatography are as follows:
Chromatographic column selects Agilent DB-17ms(30m × 0.25mm × 0.25 μm)；
Column temperature program heating: 58~60 DEG C of 1~minutes of holding rise to 155~165 DEG C with 28~31 DEG C/min, then with 4~6 DEG C/
Min is warming up to 190~300 DEG C, is kept for 8~12 minutes；Column flow rate: 1~1.5mL/min；
Injector temperature: 220~260 DEG C；0.5~1.5 μ L of sample volume；Splitless injecting samples；Carrier gas: nitrogen, purity >=
99.999%。
6. a kind of control method of artificial growth dendrobium candidum persticide residue according to claim 1, it is characterised in that:
In the GC-MS detection method, the testing conditions of MS chromatography are as follows:
Ion source: the source EI, 65~75eV；Ion source temperature: 200~250 DEG C；Interface temperature: 250~300 DEG C；
Mensuration mode: Salbutamol Selected Ion Monitoring (SIM),
Every kind of target compound selects 1~2 quota ion respectively, 2-3 qualitative ions,
Every group detection in need ion according to the sequencing of retention time, detected respectively at times.
7. a kind of control method of artificial growth dendrobium candidum persticide residue according to claim 4, it is characterised in that:
The cultivation medium formula of the dendrobium candidum are as follows: the bottom is the broken brick of 8~12cm and the mixture of Hua Gang rubble, and middle layer is
Bark, broken wood shavings after the lignite mixed fermentation of 6~8cm, upper layer be the broken wood shavings of 6~8cm, fir tree branch chopping section and
The mixture of fern root.
8. a kind of control method of artificial growth dendrobium candidum persticide residue according to claim 8, which is characterized in that
The fermentation process of bark and/or broken wood shavings in the cultivation matrix of the dendrobium candidum are as follows:
1. bark and/or the pretreatment of broken shavings raw material: admixing weight in the bark and/or broken wood shavings that parts by weight are 80~94.5%
Dried poultrymanure or birds and livestocks faces that part is 3~9% and urea or sulphur ammonium that parts by weight are 1~5% are measured, then adds parts by weight again
The fermentation assistant that lime and parts by weight for 1~5% are 0.5~1%；After above-mentioned substance is stirred, pour into certain
The water of amount seals heap 2~3 days, makes the 45%~55% of moisture content raw material total content；
2. fermentation: pretreated fermentation material being piled to the raft of 1~1.5m high again, upper covering is covered above, carries out first
Secondary fermentation the 5th~7 day after for the first fermentation, makes fermentation temperature reach 45~50 degree, the surface of fermentation material occurs white
First time overturning is carried out when color mycelia, continues to ferment after moving, and when accumulative temperature of fermenting reaches 80~90 degree, carries out the
Secondary overturning enters second of fermentation phase after second of overturning；Before second ferments, make the control of fermentation material water content 55~
65% or so, when water deficient, can teeming 3~6% comprehensively urea liquid；Deposit thickness is suitably increased again, is piled again
Raft continues to ferment, and can stop substantially when accumulative temperature of fermenting reaches 80~100 DEG C through temperature 2~3 weeks of 55~70 DEG C
Fermentation；
3. product handling system: after fermentation, tunning being tiled by 10 ㎝ left and right thicknesses, is dried, is allowed to water content and reaches 35
~45%, finished product is stored at aeration-drying.
9. a kind of control method of artificial growth dendrobium candidum persticide residue according to claim 8, it is characterised in that:
The fermentation assistant is Gymboree bark fermentation assistant and carry out chaff and mix thoroughly according to the dilution proportion of 1:2-3, is made into increment microbial inoculum.
10. a kind of control method of artificial growth dendrobium candidum persticide residue according to claim 9, feature exist
In: bark in the cultivation matrix of the dendrobium candidum be 10~13% oak bark of parts by weight, 20~24.5% Qi ebon skins, 30~
Mixture in 34% pine bark, 20~23% Chinese fir barks.
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| CN110243984A (en) * | 2019-07-24 | 2019-09-17 | 泰山医学院 | A kind of strawberry pesticide residues detection method |
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