CN100520373C - Quick detection paper card for pesticide residue in vegetable and preparation method therof - Google Patents
Quick detection paper card for pesticide residue in vegetable and preparation method therof Download PDFInfo
- Publication number
- CN100520373C CN100520373C CNB2004100536119A CN200410053611A CN100520373C CN 100520373 C CN100520373 C CN 100520373C CN B2004100536119 A CNB2004100536119 A CN B2004100536119A CN 200410053611 A CN200410053611 A CN 200410053611A CN 100520373 C CN100520373 C CN 100520373C
- Authority
- CN
- China
- Prior art keywords
- enzyme
- sheet
- preparation
- card
- pesticide residue
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Expired - Fee Related
Links
Landscapes
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
Abstract
The present invention provides a quick reflection test card and making method of pesticide residue, which can adapt the acetylcholinesterase with extracting the serum of human as enzyme piece and measure the sensibility of organophosphorus and carbamate pesticide ranged from 0.001 to 1.2mg/kg, wherein the quick reflection test card is a rather sensitive enzyme source. This invention discloses a good heat-stability enzyme source, which lasts 7-8 hours by adding gelatin stabler or trehalose in more or less 90 deg.c circumstances.
Description
Technical field
The invention belongs to pesticide residue analysis, specifically, the present invention relates to a kind of vegetable pesticide residue speed and survey paper card and preparation method thereof
Background technology
China is a large agricultural country, and proportion of crop planting is very abundant, except cereal crops, oil plant, fibre crops, sugared material, melon and fruit, medicinal material cigarette, also has large-area fruit, vegetables, grassland, forest etc.Therefore China also is that an agricultural chemicals is used big country.The invention of agricultural chemicals and use have improved agricultural productive force undoubtedly greatly, but annual generation sprinkling chemical pesticide control disease and pest still is can not no requisite measure.Raising along with people's quality of life, strong day by day to safety, nutrition, free of contamination vegetables, fruit cry, and the pesticide dosage on the current fruit, vegetables greatly, often, the abuse phenomenon is serious, therefore residues of pesticides exceed standard and cause that the acute poisoning incident occurs repeatedly, residues of pesticides can also accumulate in human body and cause slow poisoning, even cause: three cause (teratogenesis, carcinogenic, mutagenesis), in addition because the residual report that causes agricultural products in China to be refused abroad of exceeding standard of farming is also of common occurrence.Therefore in modern agricultural production, how the fast detecting residual toxicity has become the active demand of food security and people's dietetic hygiene.
The testing of residues of pesticides on the fruits and vegetables, in the institutionalization already of many countries, the external at present residues of pesticides of using in the technology for detection fruits and vegetables such as immunoassay, supercritical fluid extraction and supercritical fluid chromatography, capillary zone electrophoresis, the degree of accuracy of these discrete testing technology and precision are all very high.But need to be equipped with expensive instrument and equipment, the length that expends time in, expense height; And the concentration of production of many western countries commodity vegetables, local residual toxicity check unit has ample time and detects.And the fruits and vegetables production of China disperses relatively, promptly adopt promptly and sell, can not analyze with analysis instruments such as gas chromatographies a large amount of samples of sampling observation, therefore external habitual pesticide residue analysis method does not have operability in China, can not be suitable for China's small peasant economy and promptly adopt the field quick detection of promptly selling.The scientific worker of China has also made number of research projects from different angles in recent years, tries hard to seek to organophosphorus and the high cholinesterase of carbamate chemicals for agriculture detection sensitivity.What use was more at present is the cholinesterase that extracts from sea life, animal blood serum, housefly, Beijing Agricultural College and Beijing analytical instrument research institute have early developed pesticide residue rapid detector and used key reagents-high active enzyme through several years research, testing process is roughly 1, sampling 2, extract 3, cultivate 4, colour developing 5, Instrument measuring.Vegetables the 3rd phase of calendar year 2001 " comparison test of garden stuff pesticide residue method for quick " discloses organophosphorus and the higher CHE of carbamic acid lipid Pesticides Testing sensitivity, disclosed quick measuring card method of testing is simultaneously: take by weighing sample---adds the ultrasonic 2-3min of phosphate buffer and---sample extracting solution is added to 40 ℃ of reflections of quick measuring card white tablet position---the static about 3min of room temperature---pesticide residue rapid detector 10min, naked eyes Direct observation change color (measuring blank sample simultaneously in contrast).Detection sensitivity, detection reappearance see the following form
Detection sensitivity
As can be seen from the above table: acephatemet 1.2mg/kg, quick measuring card can detect; DDVP 0.1mg/kg, quick measuring card can detect; Rogor 2.0mg/kg, quick measuring card can detect; Cypermethrin 30mg/kg, quick measuring card can detect.But this quick measuring card also must be equipped with a pesticide residue rapid detector, just can detect, and this quick measuring card can only detect organophosphorus pesticide.
Summary of the invention
The high sensitivity, and the enzyme source of Heat stability is good that the purpose of this invention is to provide a kind of fast detecting organophosphorus and carbamate chemicals for agriculture residual hazard, and enzyme preservation technology at normal temperatures.
Another object of the present invention provides a kind of speed easy, the fast detecting pesticide residue amount and surveys paper card and preparation method thereof.
A first aspect of the present invention, provide a kind of high sensitivity, and Heat stability is good the enzyme source.The acetylcholinesterase that extracts from different animal bodies (hereinafter to be referred as enzyme) is to the malicious banned pesticides of various height, detection sensitivity exists sizable difference, some cholinesterase just responds to the agricultural chemicals of denier, and some cholinesterase is also reactionless to the agricultural chemicals of toxic dose, (cholinesterase is generally represented with inhibiting rate % the Pesticides Testing susceptibility) is material with several animal bodies such as animal organ such as the head of horse serum, human serum, beef liver, honeybee head, housefly or serum, extracts the highly sensitive enzyme of screening source---acetylcholinesterase according to a conventional method.Through experiment screening, the inhibiting rate of the enzyme that the enzyme that the enzyme that human serum of the present invention extracts extracts than prior art horse serum, housefly brain extract is obviously high, is high sensitive enzyme source, and high sensitive enzyme provided by the invention is the acetylcholinesterase of human serum extraction.Use enzyme preparation of the present invention, detect various agricultural chemicals lowest detectable limit results such as table 1.
Table 1 human serum enzyme preparation of the present invention is to various agricultural chemicals lowest detectable limit
| The agricultural chemicals Chinese name | English common name | Toxicity | Lowest detectable limit mg/kg |
| Acephatemet | methamidophos | High poison | 0.8 |
| Flolimat | omethoote | High poison | 1.2 |
| Metrifonate | trichlorphon | High poison | 0.25 |
| DDVP | dichlorovos | Poison | 0.10 |
| Carbofuran | carbofuran | High poison | 0.001 |
| Methomyl | methomyl | High poison | 0.08 |
| Carbaryl | carboryl | High poison | 0.08 |
Result of the present invention is as follows with the comparative result as a result of " comparison test of garden stuff pesticide residue method for quick ":
Acephatemet: prior art lowest detectable limit 1.2mg/kg; Lowest detectable limit 0.8mg/kg of the present invention
DDVP: prior art lowest detectable limit 0.1mg/kg; Lowest detectable limit 0.1mg/kg of the present invention
Rogor: prior art lowest detectable limit 2.0mg/kg; Lowest detectable limit 1.2mg/kg. of the present invention
Metrifonate: lowest detectable limit 0.25mg/kg. of the present invention
Carbofuran: lowest detectable limit 0.001mg/kg. of the present invention
Methomyl: lowest detectable limit 0.08mg/kg. of the present invention
Carbaryl: lowest detectable limit 0.08mg/kg. of the present invention
As can be seen from the table, specific sensitivity is significantly high mutually for the enzyme preparation of enzyme preparation of the present invention and prior art.And the quick measuring card of prior art can only detect organophosphorus pesticide, and the present invention not only can detect organophosphorus pesticide but also can detect carbamate chemicals for agriculture.Susceptibility reaches 0.001～1.2mg/kg.Further specify good effect of the present invention.
The highly sensitive human serum enzyme powder PH8.0 that the present invention is screened, 0.1M phosphate buffer is diluted to 0.5～1.5 unit enzyme liquid respectively, adding 0.5～1.% gelatin or 0.5～1.% trehalose then in enzyme liquid mixes, dripping enzyme liquid (about 30 μ l) is on the qualitative filter paper of 1.0cm at diameter, put drying at room temperature, make the enzyme sheet.The enzyme sheet is placed on climatic chamber (adjustable temperature) handles, then the thermal stability result such as the table 2 of tested enzyme.
The thermally-stabilised test of table 2 enzyme sheet
| Time (min) | Temperature ℃ | The enzymatic activity mark+ |
| 0 | 84 | +++ |
| 30 | 70 | +++ |
| 60 | 95 | +++ |
| 90 | 90 | +++ |
| 120 | 88 | +++ |
| 150 | 90 | +++ |
| 180 | 88 | +++ |
| 240 | 90 | +++ |
| 300 | 92 | +++ |
| 360 | 90 | +++ |
| 420 | 90 | +++ |
| 480 | 90 | ++ |
Above test result shows: the enzyme of the present invention's screening, our experiments show that, under 90 ℃ of left and right sides conditions, enzymatic activity can keep 5-6 hours, in enzyme liquid of the present invention, add stabilizing agent 0.5～1.% gelatin or the mixing of 0.5～1.% trehalose, under 90 ℃ of left and right sides conditions, human serum enzymatic activity of the present invention can keep 7-8 hours, had solved the preservation technology under the enzyme normal temperature.The present invention is a kind of enzyme source of Heat stability is good, has solved present domestic exoenzyme and has tied up to the problem that can only preserve a few hours under the normal temperature, further illustrates good effect of the present invention.
A second aspect of the present invention, best enzyme concentration is provided, with PH8.0,0.1M it is 0.5,0.75,1,1.25,1.5 commensurates not that phosphate buffer dilutes human serum enzyme of the present invention, methamidophos pesticide concentration is 5mg/kg, through experiment relatively, preferred concentration is 0.5 unit, and better concentration is 0.75 unit.
A third aspect of the present invention, best concentration of substrate is provided, to make mixed liquor (PH5.5) in the 100ml distilled water adding 0.0448g potassium ferricyanide (potassium ferricyanide) and 0.0715g potassium ferrocyanide (potassium ferrocyanide) salt-mixture, earlier drip (30 a μ l) mixed liquor respectively on 3 filter paper, drip (about 30 a μ l) ethychlozate ester 180mg/10ml acetone more respectively, ethychlozate ester 270mg/10ml acetone, ethychlozate ester 360mg/10ml acetone is on 3 filter paper, dry back and enzyme reaction, preferable concentration of substrate is 270mg/10ml, and best concentration of substrate is 180mg/10ml.
The fourth aspect of we's invention, inventor's sero-enzyme and agricultural chemicals the best use of time are provided, and the enzyme sheet that will contain the human serum enzyme solutions immerses acephatemet solution, takes out the back and acts on 2,5,10,15 minutes respectively with substrate, preferably be 5 minutes action time, and be 8 minutes best action time.
A fifth aspect of the present invention provides the preparation method and the using method of quick measuring card.Extract the acetylcholinesterase in the human serum according to a conventional method, freeze drying is the enzyme powder, use PH8,0.1M inventor's sero-enzyme powder of phosphate buffer dilution refrigeration drying, in enzyme solutions, add assistant agent 0.5～1.0% gelatin or 0.5～1.0% trehalose, the enzyme solutions for preparing is added drop-wise to makes the enzyme sheet on the qualitative filter paper, it is stand-by to place drying at room temperature; Drip the potassium ferricyanide and the potassium ferrocyanide mixed liquor for preparing on qualitative filter paper, drip the acetone soln of the ethychlozate ester 180mg-360/10ml for preparing again, it is standby to make substrate sheet placement drying at room temperature.The agricultural chemicals or the vegetable juice of one (30 μ l) are added drop-wise on the enzyme sheet, moistening 30 seconds, static 8 minutes of room temperature, or the enzyme sheet put into contain certain density pesticidal solutions or vegetable juice, flooded equally 30 seconds, the enzyme sheet is fished for, put room temperature static 8 minutes, substrate is folded on the enzyme sheet, clutched 3 minutes with hand, open the enzyme sheet, observe the colour developing situation.
In order to further specify good effect of the present invention, the speed of the present invention's preparation is surveyed sensitivity result such as the table 3 that paper card detects 8 kinds of agricultural chemicals.
Table 3 quick measuring card of the present invention is to the sensitivity of 8 kinds of Pesticides Testing
| Pesticide name | Detection limit (mg/kg) | ADI(mg/kg) | Male rat per os LD 50(mg/kg) |
| Acephatemet | 5.0 * | 4 * | 20～30 |
| Metrifonate | 5.0 | 10 | 560 |
| DDVP | 1.0 | 4 | 56～80 |
| Flolimat | 5.0 | 10 | 320～380 |
| Parathion-methyl | 1.0 | 20 | 9～25 |
| The furans pellet | 0.2 | 10 | 8～14 |
| Methomyl | 0.5 | 30 | 17～24 |
| Carbaryl | 0.5 | 10 | 246～283 |
As can be seen from Table 3, speed of the present invention is surveyed paper card needn't be by instrument, to the detectability of common pesticides on the vegetables all in 0.2～5mg/kg scope, all propose in the World Health Organization (WHO) (WHO), the World Food Programme (FAO) every day everyone per kilogram of body weight to the permission intake (ADI) of agricultural chemicals below the specified standard, therefore be determined as negative sample according to quick measuring card and can not cause acute hazard human body, for the prevention pesticide poisoning, forbid that pesticide-tainted vegetable comes into the market to play supervisory role.
Comprehensive experimental result of the present invention, specific sensitivity is significantly high mutually for the enzyme preparation of human serum acetylcholinesterase and prior art.Susceptibility reaches 0.001～1.2mg/kg.And the enzyme source that human serum acetylcholinesterase of the present invention is a kind of Heat stability is good, under 90 ℃ of left and right sides conditions, enzymatic activity can keep 5-6 hours, in enzyme liquid of the present invention, add stabilizing agent 0.5～1.% gelatin or the mixing of 0.5～1.% trehalose, under 90 ℃ of left and right sides conditions, human serum enzymatic activity of the present invention can keep 7-8 hours, solve present domestic exoenzyme and tied up to the problem that to preserve a few hours under the normal temperature, surveying paper card with the speed of human serum enzyme preparation of the present invention needn't be by instrument, to the detectability of common pesticides on the vegetables all in 0.2～5mg/kg scope, all at the World Health Organization (WHO) (WHO), the World Food Programme (FAO) proposes every day, everyone per kilogram of body weight was to the permission intake (ADI) of agricultural chemicals below the specified standard, for the prevention pesticide poisoning, forbid that pesticide-tainted vegetable comes into the market to play supervisory role.The present invention is practical, and simple to operate quick, once only measures and needs 12 minutes.With low cost, be a kind of product that wide market outlook are arranged
Further set forth the present invention below in conjunction with specific embodiment, should be understood that these embodiment only are used to the present invention to be described rather than to be used to limit the scope of the invention.
The screening in embodiment 1, enzyme source
With animal organs such as the head of horse serum, human serum, beef liver, honeybee head, housefly or serum is material, extracts, screens highly sensitive enzyme source---acetylcholinesterase according to a conventional method.Respectively the brain of freezing horse serum, human serum, beef liver, honeybee brain, housefly is smashed to pieces, in 1:9 (W/V) ratio adding distil water, homogenate 3 minutes, with 2000-3000r/min centrifugal 10 minutes, supernatant behind filter paper filtering, freeze drying, packing is standby, put-20 ℃ frozen. be the enzyme source.
1) reagent preparation:
Enzyme liquid preparation: use PH8., the 0.1M phosphate buffer respectively the dried horse serum enzyme of dilution refrigeration powder, go into sero-enzyme powder, beef liver enzyme powder, honeybee brain enzyme powder, housefly brain enzyme powder,, be mixed with 0.75 unit.
Pesticide concentration: acephatemet is the red 0.002ppm of 0.8ppm, furans.
Substrate: acetylthiocholine iodide.
Developer: dinitrobenzoic acid, test concentrations are 0.08mg.
2.) instrument: CL-I remains of pesticide analyzer, special-purpose spectrophotometer 410nm.
By special-purpose spectrophotometer, utilize enzyme to suppress method above-mentioned 5 kinds of acetylcholinesterases are carried out The selection result such as table 4.
5 kinds of acetylcholinesterases of table 4 to the agricultural chemicals susceptibility relatively
As can be seen from Table 4, the cholinesterase that extracts from various animal organs or serum is to the sensitivity difference of agricultural chemicals, and the inhibiting rate of the enzyme that the enzyme that the enzyme that human serum of the present invention extracts extracts than prior art horse serum, housefly brain extract is obviously high, is the responsive enzyme of height source,
Embodiment 2: the selection of enzyme concentration
1) enzyme preparation preparation: with the cryodesiccated human serum enzyme of the present invention powder PH8.0, the 0.1M phosphate buffer is diluted to 0.5 respectively, 0.75.1.0,1.25,1.5 commensurate not.Dripping one (about 30 μ l) is on the qualitative filter paper of 1.0cm at diameter, makes the enzyme sheet.
2) substrate: the 0.0448g potassium ferricyanide (potassium ferricyanide), 0.0715g potassium ferrocyanide (potassium ferrocyanide) are mixed, and after adding 100ml distilled water (PH5.5) mixed, adding one (about 30 μ l) became 1.0cm on the limit
2Qualitative filter paper on, drip 30 μ l ethychlozate ester 180mg/10ml acetone solns again, make the substrate sheet.
3) acephatemet concentration is 0.1mg/ μ l.
4) distilled water is selected in contrast for use
With five kinds of enzyme concentration enzyme sheets and agricultural chemicals acephatemet reaction result such as table 3
Five kinds of enzyme concentrations of table 5 to the acephatemet reaction relatively
*+++full contrast color (the normal hydrolysis of substrate for enzymatic activity does not contain agricultural chemicals) is complete blue
++ part colour developing (the enzyme-to-substrate reaction is not exclusively) pale blue
+ colour developing poor (enzyme-to-substrate Low Response) is extremely light blue
-complete (enzyme is suppressed by agricultural chemicals entirely) white that suppresses
The enzyme concentration contrast of 0.75 unit develops the color as can be seen from Table 5, can be suppressed by 0.1mg/ μ l methamidophos pesticide again, and contrast is obvious with the inhibition aberration, so 0.75 unit is best enzyme concentration, 0.5 unit is a preferred concentration.
Embodiment 3: the selection of best concentration of substrate
1) preparation of substrate sheet: the 0.0448g potassium ferricyanide (potassium ferricyanide), 0.0715g potassium ferrocyanide (potassium ferrocyanide) mixed add 100ml distilled water, getting one of mixed liquor (about 30 μ l PH5.5), to drip 3 length of sides respectively earlier be 1.0cm
2On the qualitative filter paper, and then respectively with (about 30 a μ l) ethychlozate ester 180mg/10ml acetone solution, ethychlozate ester 270mg/10ml acetone solution, ethychlozate ester 360mg/10ml acetone solution on 3 qualitative filter papers, the stand-by substrate sheet that becomes of drying at room temperature..
2) preparation of enzyme sheet: with the present invention freezing go into the enzyme powder PH8.0 that serum extracts, the 0.1M phosphate buffer is diluted to 0.5 respectively, 0.75. commensurate not.Dropping is on the qualitative filter paper of 1.0cm, to make the enzyme sheet at diameter.
With the enzyme sheet of 0.5 unit, 0.75 unit respectively with substrate sheet (ethychlozate ester 180mg/10ml acetone, ethychlozate ester 270mg/10ml acetone, ethychlozate ester 360mg/10ml acetone) reaction result such as table 6.
The colour developing of table 6.3 kind of concentration of substrate and enzyme reaction relatively
++ ++ complete blue (enzyme-to-substrate reacts), develop the color (in 50 seconds);
+++complete blue (enzyme-to-substrate reacts), colour developing is slow (above 50 seconds; )
++ inhomogeneous colour developing (being mottled), colour developing fast (in 50 seconds);
+ inhomogeneous colour developing (being mottled), colour developing slow (above 50 seconds).
0.75 unit enzyme concentration as can be seen from Table 4, ethychlozate ester 180mg/10ml acetone concentration of substrate, complete blue (enzyme-to-substrate reacts) develops the color, colour developing is fast,, 0.5 unit enzyme concentration, ethychlozate ester 180mg/10ml acetone concentration of substrate, complete blue (enzyme-to-substrate reacts) colour developing is slow slightly, so ethychlozate ester 180/10ml acetone is optium concentration.
Embodiment 4: enzyme and the selection of agricultural chemicals the best use of time
The enzyme sheet (0.75 unit) that inventor's sero-enzyme preparation is made immerses in the 5mg/kg acephatemet aqueous solution, about 30 seconds, takes out and places 8 minutes, distinguish superimposed 2,5,10,15 minutes with embodiment 3 described substrate sheets then, observe enzyme sheet colour developing situation, distinguish and whether suppress result such as table 5.
The comparison of table 7 enzyme and action of agricultural chemicals different time
+ (agricultural chemicals does not reach inhibition to enzyme) is complete blue;-(agricultural chemicals suppresses enzyme) suppressed look
As can be seen from Table 7.The Best Times of enzyme and action of agricultural chemicals is 8 minutes.
Embodiment 5: the preparation of quick measuring card and use
1, the preparation of quick measuring card
A) extraction of enzyme: with the human serum is material, in 1:9 (W/V) ratio adding distil water, homogenate 3 minutes, with 2000-3000r/min centrifugal 10 minutes, supernatant behind filter paper filtering, freeze drying, packing is standby, puts-20C is frozen. be the enzyme source.
B) preparation of enzyme sheet: use PH8.0,0.1M phosphate buffer dilution the present invention becomes 0.75 unit enzyme liquid from the acetylcholinesterase freeze drying enzyme powder that human serum extracts, adding 1% gelatin in enzyme liquid mixes, the enzyme drop for preparing is added on the qualitative filter paper that diameter is 1.0cm, and the placement drying at room temperature is stand-by.
C) preparation of substrate sheet: the 0.0448g potassium ferricyanide (potassium ferricyanide), 0.0715g potassium ferrocyanide (potassium ferrocyanide) are mixed, add 100ml distilled water, getting one of the potassium ferricyanide and potassium ferrocyanide mixed liquor (about 30 μ l PH5.5) dropping earlier is 1.0cm in the length of side
2On the qualitative filter paper, and then the 180mg ethychlozate ester mixed with the 10ml acetone soln, drip 30 μ l ethychlozate ester acetone solns again on qualitative filter paper, it is stand-by to put drying at room temperature then.
D) preparation of quick measuring card: the thick card lining paper of rectangle of getting wide 2cm, long 5cm, the round enzyme sheet that a) prepares is bonded at the top of paper card with double faced adhesive tape, with b) preparation square substrate sheet be bonded at the following of paper card with double faced adhesive tape, the enzyme sheet and the substrate of quick measuring card is folded inverted, do not contact stand-by mutually.The quick measuring card for preparing is put into little plastic bag sealing, and it is stand-by to put into refrigerator (-20 ℃ frozen).
2, the use of quick measuring card
A) agricultural chemicals of 30 μ l amount or dish juice are added drop-wise on the enzyme sheet moistening 30 seconds;
B) the enzyme sheet was put room temperature static 8 minutes;
C) the substrate sheet is superimposed on the enzyme sheet, clutched 3 minutes with hand;
D) open the enzyme sheet, observe the colour developing situation.
Complete blue, no agricultural chemicals exists, and light blue or white have agricultural chemicals or exceed standard the agricultural chemicals existence.
In order to further specify good effect of the present invention, the speed of the embodiment of the invention 5 preparations is surveyed paper card, the vegetable field in national some areas detects investigation, result such as table 8.
Table 8 quick measuring card detects investigation result at the vegetable field of national some areas
| Tested vegetable species | Use enzyme sheet number | The sampling position | Testing result | Organophosphorus pesticide concentration (mg/kg) |
| Pakchoi | 2 * | Jiangsu | Positive | 0.4 |
| Green vegetables | 2 * | Jiang Qiao | Positive | 0.05 |
| Hangzhoupro Chinese cabbage | 2 | Zhejiang | Positive | 0.12 |
| Chinese little greens | 2 | Shanghai | Positive | 0.38 |
| Water spinach | 2 | Cang Qiao | Positive | 0.15 |
| Cucumber | 2 | Nanhui | Positive | 0.40 |
| Cabbage | 2 | Shandong | Positive | 0.18 |
| Green vegetables | 2 | Shanghai | Positive | 0.6 |
| Pakchoi | 2 | Kingsoft | Positive | 0.79 |
| Cabbage | 2 | Shandong | Positive | 2.10 |
| Cabbage | 2 | Shanghai | Positive | 1.90 |
As can be seen from Table 8, use speed of the present invention to survey paper card detection vegetables susceptibility and reach 0.05～2.10mg/kg, and simple to operate quick, once only measure and need 12 minutes.(0.5 yuan) with low cost, and need be by the remains of pesticide detector, just can judge clearly whether the remains of pesticide on the edible vegetable exceeds standard, it is not only applicable to family edible vegetable is detected, and also will carry out units such as rapid screening monitoring, health and epidemic prevention station, agricultural system Plant Quarantine Center to market surveillance, field vegetables the detection of vegetable pesticide residue is brought great convenience.Also applicable to the mensuration of organophosphorus pesticide, carbamate chemicals for agriculture in water, soil and other agricultural product, the present invention is practical, is a kind of product that wide market outlook are arranged.
Claims (9)
1, a kind of vegetable pesticide residue speed is surveyed paper card, described pesticide in vegetable is selected from organophosphorus or carbamate chemicals for agriculture, wherein said speed is surveyed paper card and is comprised lining paper, substrate sheet and enzyme sheet, it is characterized in that the enzyme in the described enzyme sheet is the acetylcholinesterase that extracts in the human serum
Wherein, diameter is the enzyme liquid that contains on the described enzyme sheet of 1.0cm through the described acetylcholinesterase of 0.5-1.0 unit of drying at room temperature.
2, vegetable pesticide residue speed as claimed in claim 1 is surveyed paper card, it is characterized in that described enzyme sheet is the acetylcholinesterase freeze drying powder pH8.0 that will extract in the human serum, 0.1M phosphate buffer is diluted to 0.5-1.0 unit's enzyme liquid, and the described enzyme drop of 30 microlitres is added in makes after qualitative filter paper that diameter is 1.0cm and the drying at room temperature.
3, vegetable pesticide residue as claimed in claim 1 speed is surveyed paper card, it is characterized in that being added with in the solution of the acetylcholinesterase that the human serum that is used for preparing described enzyme sheet extracts 0.5～1.5% stabilizing agent.
4, vegetable pesticide residue as claimed in claim 3 speed is surveyed paper card, it is characterized in that being added with in the solution of the acetylcholinesterase that the human serum that is used for preparing described enzyme sheet extracts 1% stabilizing agent.
5, described vegetable pesticide residue speed as claimed in claim 3 is surveyed paper card, it is characterized in that described stabilizing agent is selected from gelatin or trehalose.
6, vegetable pesticide residue speed as claimed in claim 1 is surveyed the preparation method of paper card, and wherein said pesticide in vegetable is selected from organophosphorus or carbamate chemicals for agriculture, said method comprising the steps of:
1) extraction of enzyme: with the human serum is material, and in human serum: distilled water is the ratio adding distil water of 1:9 (W/V), homogenate 3 minutes, with 2000-3000r/min centrifugal 10 minutes, supernatant behind filter paper filtering, freeze drying, packing is standby, put-20 ℃ frozen, be the enzyme source;
2) preparation of enzyme sheet: use pH8.0,0.1M the acetylcholinesterase freeze drying enzyme powder that phosphate buffer dilution human serum extracts becomes 0.5-1.0 unit enzyme liquid, the enzyme drop for preparing is added on the round qualitative filter paper that diameter is 1.0cm, and the placement drying at room temperature is stand-by;
3) preparation of substrate sheet: the 0.0448g potassium ferricyanide, 0.0715g potassium ferrocyanide are mixed, add 100ml distilled water, get one of the potassium ferricyanide of pH5.5 and potassium ferrocyanide mixed liquor, 30 μ 1, dripping in the length of side earlier is 1.0cm
2On the qualitative filter paper, and then 180-270mg ethychlozate ester mixed with the 10ml acetone soln, the ethychlozate ester acetone soln that drips one 30 μ l again is on qualitative filter paper, and it is stand-by to put drying at room temperature then;
4) preparation of quick measuring card: the thick card lining paper of rectangle of getting wide 2cm, long 5cm, with 2) the round enzyme sheet for preparing is bonded at the top of paper card with double faced adhesive tape, with 3) the square substrate sheet of preparation is bonded at the below of paper card with double faced adhesive tape, the enzyme sheet and the substrate of quick measuring card is folded inverted, do not contact mutually, the quick measuring card for preparing is put into little plastic bag sealing, put into refrigerator,-20 ℃ frozen, stand-by.
7, vegetable pesticide residue speed as claimed in claim 6 is surveyed the preparation method of paper card, it is characterized in that the enzyme liquid that described enzyme liquid is 0.5-0.75 unit.
8, vegetable pesticide residue speed as claimed in claim 7 is surveyed the preparation method of paper card, it is characterized in that the enzyme liquid that described enzyme liquid is 0.75 unit.
9, vegetable pesticide residue speed as claimed in claim 6 is surveyed the preparation method of paper card, it is characterized in that when enzyme liquid is the enzyme liquid of 0.5 unit the optium concentration of ethychlozate ester acetone soln is 180mg ethychlozate ester/10ml acetone on the substrate sheet.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CNB2004100536119A CN100520373C (en) | 2004-08-10 | 2004-08-10 | Quick detection paper card for pesticide residue in vegetable and preparation method therof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CNB2004100536119A CN100520373C (en) | 2004-08-10 | 2004-08-10 | Quick detection paper card for pesticide residue in vegetable and preparation method therof |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN1734257A CN1734257A (en) | 2006-02-15 |
| CN100520373C true CN100520373C (en) | 2009-07-29 |
Family
ID=36076768
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CNB2004100536119A Expired - Fee Related CN100520373C (en) | 2004-08-10 | 2004-08-10 | Quick detection paper card for pesticide residue in vegetable and preparation method therof |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN100520373C (en) |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108398426A (en) * | 2018-03-15 | 2018-08-14 | 鲁东大学 | A kind of simple rapid detection for pesticide residue device and its application |
Families Citing this family (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101832986A (en) * | 2010-04-26 | 2010-09-15 | 浙江师范大学 | Kit for detecting pesticide by using thin layer chromatography-enzymatic inhibition method and preparation method of cholinesterase bar |
| CN104535563B (en) * | 2014-12-25 | 2017-09-26 | 浙江大学 | It is a kind of to dry spraying droplet density and detection architecture, preparation method and the detection method of size be advantageously used under wet environment |
| CN105044101A (en) * | 2015-07-31 | 2015-11-11 | 复旦大学 | Quick detection card for pesticide residues based on naked eye visual colorimetric determination |
| CN105928929A (en) * | 2016-04-20 | 2016-09-07 | 郑州大学 | Rapid detection point card for pesticide residues and method for detecting content of pesticide residues by using detection point card |
| CN106191212B (en) * | 2016-08-08 | 2019-11-19 | 陈英就 | The preparation method of fast detecting reagent kit for toxicity of pesticide residue, the preparation method of stabilizer and cholinesterase enzyme powder |
| CN108169220B (en) * | 2017-11-24 | 2021-02-12 | 武汉市农业科学院 | Horse serum cholinesterase-based pesticide residue rapid detection card and production process and use method thereof |
| CN109884041B (en) * | 2019-02-14 | 2021-04-06 | 广州奕昕生物科技有限公司 | Pesticide residue short-term test card |
| CN111272726B (en) * | 2020-05-08 | 2021-03-09 | 北京中检葆泰生物技术有限公司 | Method for detecting pesticide residue in food |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US5624831A (en) * | 1994-03-28 | 1997-04-29 | Vu Khue; Nguyen | Immobilized acetylcholinesterase stabilized by a film of gelatin or albumin containing trehalose |
| CN1224160A (en) * | 1998-11-19 | 1999-07-28 | 山东农业大学 | Method for fast testing farm chemicals contamination in vegetables and fruits |
-
2004
- 2004-08-10 CN CNB2004100536119A patent/CN100520373C/en not_active Expired - Fee Related
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US5624831A (en) * | 1994-03-28 | 1997-04-29 | Vu Khue; Nguyen | Immobilized acetylcholinesterase stabilized by a film of gelatin or albumin containing trehalose |
| CN1224160A (en) * | 1998-11-19 | 1999-07-28 | 山东农业大学 | Method for fast testing farm chemicals contamination in vegetables and fruits |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108398426A (en) * | 2018-03-15 | 2018-08-14 | 鲁东大学 | A kind of simple rapid detection for pesticide residue device and its application |
| CN108398426B (en) * | 2018-03-15 | 2021-03-23 | 鲁东大学 | Simple pesticide residue rapid detection device and application thereof |
Also Published As
| Publication number | Publication date |
|---|---|
| CN1734257A (en) | 2006-02-15 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Veneziano et al. | Determination of carbendazim, thiabendazole and thiophanate-methyl in banana (Musa acuminata) samples imported to Italy | |
| Knežević et al. | Screening of fresh fruit and vegetables for pesticide residues on Croatian market | |
| Chen et al. | Evaluation of pesticide residues in fruits and vegetables from Xiamen, China | |
| Kaaya et al. | The effect of storage time and agroecological zone on mould incidence and aflatoxin contamination of maize from traders in Uganda | |
| Salazar et al. | Determination of the antioxidant activity of plants from Northeast Mexico | |
| Berny et al. | Evaluation of the toxicity of imidacloprid in wild birds. A new high performance thin layer chromatography (HPTLC) method for the analysis of liver and crop samples in suspected poisoning cases | |
| CN100520373C (en) | Quick detection paper card for pesticide residue in vegetable and preparation method therof | |
| Suárez-Jacobo et al. | Pesticide residues in orange fruit from citrus orchards in Nuevo Leon State, Mexico | |
| Di Stefano et al. | Determination of aflatoxins and ochratoxins in Sicilian sweet wines by high-performance liquid chromatography with fluorometric detection and immunoaffinity cleanup | |
| CN108535249B (en) | Pesticide residue fast detection point card | |
| Ritenour et al. | Ethanol effects on the ripening of climacteric fruit | |
| CN101503731B (en) | Fast detecting reagent kit for toxicity of pesticide residue | |
| CN105928929A (en) | Rapid detection point card for pesticide residues and method for detecting content of pesticide residues by using detection point card | |
| CN201749086U (en) | Rapid test card for detecting pesticide residue | |
| CN100545638C (en) | Residual toxicity detects with solid enzyme reagent and preparation method thereof and using method | |
| Hongsibsong et al. | Blood cholinesterase activity levels of farmers in winter and hot season of Mae Taeng District, Chiang Mai Province, Thailand | |
| CN202221412U (en) | Test paper strip for quickly detecting residual quantity of farm chemical in food | |
| CN102399850A (en) | Rapid detection card for detecting pesticide residues, preparation method and application thereof | |
| CN106053459A (en) | Domestic pesticide residue detecting instrument and method | |
| Xu et al. | Comparison of four commercial enzymatic assay kits for the analysis of organophosphate and carbamate insecticides in vegetables | |
| Lebailly et al. | Urine mutagenicity and lymphocyte DNA damage in fruit growers occupationally exposed to the fungicide captan | |
| Aikpokpodion et al. | Appraisal of pesticide residues in kola nuts obtained from selected markets in Southwestern, Nigeria | |
| US20040115755A1 (en) | Method and apparatus for detecting response to damage and diagnostic method therefor | |
| Chiu et al. | Rapid Bioassay of Pesticide Residues (RBPR) on Fruits and Vegetable | |
| CN106191212B (en) | The preparation method of fast detecting reagent kit for toxicity of pesticide residue, the preparation method of stabilizer and cholinesterase enzyme powder |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C14 | Grant of patent or utility model | ||
| GR01 | Patent grant | ||
| C17 | Cessation of patent right | ||
| CF01 | Termination of patent right due to non-payment of annual fee |
Granted publication date: 20090729 Termination date: 20130810 |