CN106770227A - The detection method and test strips of AMOZ in a kind of dairy products - Google Patents
The detection method and test strips of AMOZ in a kind of dairy products Download PDFInfo
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- CN106770227A CN106770227A CN201611079660.9A CN201611079660A CN106770227A CN 106770227 A CN106770227 A CN 106770227A CN 201611079660 A CN201611079660 A CN 201611079660A CN 106770227 A CN106770227 A CN 106770227A
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/75—Systems in which material is subjected to a chemical reaction, the progress or the result of the reaction being investigated
- G01N21/77—Systems in which material is subjected to a chemical reaction, the progress or the result of the reaction being investigated by observing the effect on a chemical indicator
- G01N21/78—Systems in which material is subjected to a chemical reaction, the progress or the result of the reaction being investigated by observing the effect on a chemical indicator producing a change of colour
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N1/00—Sampling; Preparing specimens for investigation
- G01N1/28—Preparing specimens for investigation including physical details of (bio-)chemical methods covered elsewhere, e.g. G01N33/50, C12Q
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/17—Systems in which incident light is modified in accordance with the properties of the material investigated
- G01N21/25—Colour; Spectral properties, i.e. comparison of effect of material on the light at two or more different wavelengths or wavelength bands
- G01N21/31—Investigating relative effect of material at wavelengths characteristic of specific elements or molecules, e.g. atomic absorption spectrometry
- G01N21/314—Investigating relative effect of material at wavelengths characteristic of specific elements or molecules, e.g. atomic absorption spectrometry with comparison of measurements at specific and non-specific wavelengths
- G01N21/3151—Investigating relative effect of material at wavelengths characteristic of specific elements or molecules, e.g. atomic absorption spectrometry with comparison of measurements at specific and non-specific wavelengths using two sources of radiation of different wavelengths
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Abstract
The invention discloses a kind of detection method of AMOZ in dairy products, comprise the following steps：First, experiment reagent is made：Measure the concentrated hydrochloric acid that 9 milliliters of concentration are 2mol/L, plus deionized water is settled to 120 milliliters, weigh 10 grams of dipotassium hydrogen phosphates, plus 400 ml deionized water dissolve and be settled to 400 milliliters, to 80 grams of sodium chloride of addition in 400 milliliters of dipotassium hydrogen phosphate solutions, fully dissolve standby, weigh 3.5 grams of NaOH, with deionized water dissolving and be settled to 100 milliliters.The present invention is determined by using the specificity that AMOZ connection exempts from kit by carrying out cross reaction experiment with corresponding material, the content of the furaltadone in dairy products is determined by the size of absorbance, detection method used in the present invention, it is simple to operate, it is cheap, and detection time is short, the requirement of enterprise and mechanism is disclosure satisfy that.
Description
Technical field
The present invention relates to technical field of food detection, in specially a kind of dairy products the detection method of AMOZ and
Test strips.
Background technology
Furaltadone, also known as furazolidone, is Nitrofuran metabolites, is a kind of artificial synthesized antimicrobial, in yellow powder
Or crystalline powder, molecular weight is 225.16,245-258 degrees Celsius of fusing point, meets caustic digestion, and color is gradually deepened under high light,
The antibacterial activity for finding 5- nitrofurans earliest is the biosynthesis and Microbiology Experiment that nineteen forty-four is carried out in the U.S. and Germany
In, the homologue such as subsequent furaltadone is listed in succession, and find this medicine efficacy stability suppress or kill various Gram-positives and
Negative bacteria, has certain effect to some protozoons, fungi, because its drug effect high price is cheap, is widely used in medicine and herding etc.
Industry.
AOZ, chemical name：3- amino -2- oxazolidones, are furaltadone Metabolic residue things main in vivo, warp
The analyte hydroxyethylhydrazine of hydrochloric acid in gastric juice generation is hypertoxic class material, there is carcinogenic danger to human body, therefore from nineteen ninety-five, European Union, day
This.The developed countries such as the U.S. start to forbid such antiseptic to be used in edible livestock and poultry and aquatic livestock, and strict implement pair
The food fish of input, shrimp and birds carry out residue detection, the Ministry of Agriculture from 2 months 2002 issue associated documents, also to furan
Mutter its ketone purposes and addition consumption done related limitation.
Therefore, in order to improve the security of food, the AMOZ in food is needed to detect, and it is existing
AMOZ detection method, or being exactly expensive, or being exactly cumbersome operating process, detection time is long,
The purpose of Site Detection can not be moved, the requirement of enterprise and mechanism is not reached much.
The content of the invention
（One）The technical problem of solution
In view of the shortcomings of the prior art, the invention provides the detection method and test paper of AMOZ in a kind of dairy products
Bar, solves the problems, such as the detection method price of AMOZ in existing dairy products, process is cumbersome and detection time is long.
（Two）Technical scheme
To achieve the above object, the present invention provides following technical scheme：The detection method of AMOZ in a kind of dairy products,
Comprise the following steps：
First, experiment reagent is made：The concentrated hydrochloric acid that 9 milliliters of concentration are 2mol/L is measured, plus deionized water is settled to 120 milliliters,
Weigh 10 grams of dipotassium hydrogen phosphates, plus 400 ml deionized waters dissolve and are settled to 400 milliliters, it is molten to 400 milliliters of dipotassium hydrogen phosphates
In liquid add 80 grams of sodium chloride, fully dissolve standby, weigh 3.5 grams of NaOH, with deionized water dissolving and be settled to 100 milli
Rise；
2nd, the accurate dairy food sample for weighing 150-200 milliliters is in 250 milliliters of centrifuge tube, and to being sequentially added in centrifuge tube
10 ml deionized waters, the concentrated hydrochloric acid of 2 milliliters of 2mol/L and 60 milliliters of o-nitrobenzaldehydes, abundant whirling motion to dairy products disperse,
The insulating box that temperature is 36 degree is then placed within to be incubated one hour, it is then molten to 8 milliliters of bufferings are sequentially added in dairy soln again
Liquid, 100 milliliters of sodium hydroxide solutions and 8 milliliters of carboxylate solutions, at room temperature, acutely rock 2 minutes, are subsequently adding 2 millis
Rise hexane, abundant whirling motion 15 seconds adds 2 milliliters of sample diluting liquids, low speed whirling motion 5 minutes, filter off upper strata hexane and in
Between impurity, take 150 milliliters of sample solutions and detected, this liquid be A liquid；
3rd, the accurate brown sugar slurry samples for weighing 50-100 milliliters are in 250 milliliters of centrifuge tube, and to being sequentially added in centrifuge tube
10 ml deionized waters, the concentrated hydrochloric acid of 2 milliliters of 2mol/L and 60 milliliters of o-nitrobenzaldehydes, abundant whirling motion to brown sugar slurry point
Dissipate, be then placed within the insulating box that temperature is 36 degree and be incubated one hour, it is then slow to sequentially adding 8 milliliters in red syrup solution again
Solution, 100 milliliters of sodium hydroxide solutions and 8 milliliters of carboxylate solutions are rushed, at room temperature, is acutely rocked 2 minutes, be subsequently adding
2 milliliters of hexanes, abundant whirling motion 15 seconds adds 2 milliliters of sample diluting liquids, low speed whirling motion 5 minutes, filter off upper strata hexane and
Interstitial impurity, takes 150 milliliters of sample solutions and is detected, this liquid is B liquid；
4th, S1, by required ELIAS strip insertion ELISA Plate frame, after untapped ELIAS strip is sealed with valve bag, immediately
It is stored in 3-7 degrees Celsius of environment, and records the position of each standard sample；
S2,100 microlitres of each standard sample solution are separately added into corresponding standard sample sample wells；
S3,100 microlitres of AMOZ enzyme conjugates of addition in each plate hole；
S4, cover plate film is covered, gently vibrate enzyme mark version about ten seconds, fully mixed, 23-28 degrees Celsius of the reaction of lucifuge at room temperature
45 minutes；
S5, cover plate film is opened, outwell solution in plate hole, 3 milliliters of wash operating solutions are added in every hole, soaked 30 minutes, weighed successively
Multiple operation is twice；
S6, outwell solution in plate hole, ELISA Plate be inverted on blotting paper, blot, added in every hole immediately 10 milliliters of A liquid and
10 milliliters of B liquid, are sufficiently mixed, and mixed liquor was used in 3 minutes, it is to avoid contained using metalware；
S7, cover plate film is covered, gently vibrate ELISA Plate ten seconds, fully mixed, 23-28 degrees Celsius of the reaction of lucifuge at room temperature 30
Minute；
S8, cover plate film is opened, 2 milliliters of terminate liquids are added in each plate hole, gently vibrated ten seconds, fully mixed, use ELIASA
Absorbance is detected under 400 nanometers of dual wavelength, 600 nanometers, is as a result read in 5 minutes after termination；
S9, the mean absorbance values by each sample, divided by zero standard（Concentration is the sample of 0ppb）Absorbance, is multiplied by 100, can be with
Obtain the percentage of the corresponding absorbance of each sample；Bring the percentage of sample absorbance into standard curve（Calibration curve formula
It is the absorbance of the zero standards of absorbance * 100/ of sample）, the corresponding concentration of sample can be drawn, multiplied by with the dilution of respective sample times
Number, side obtains actual drug content in sample.
Preferably, each reagent before must fully mix, and before experiment, should first open biochemical cultivation case
The temperature needed for, and check whether biochemical cultivation case temperature is normal.
Preferably, during the extraction medicine, whirling motion or vibration should try one's best fully, after addition sample is weak solution, answer low speed
Whirling motion 2 minutes, above operation will directly affect splendid attire and testing result.
Preferably, during the mix reagent, bubble should be avoided the occurrence of, unspent microplate should be sealed, and be placed in 2-8 and taken the photograph
Family name's degree is preserved.
Preferably, the test strips include paper slip body, and one end of the paper slip body is fixedly connected with bow strip, described
Anticorrosive coat is provided with the outer wall of paper slip body.
（Three）Beneficial effect
The invention provides the detection method and test strips of AMOZ in a kind of dairy products.Possesses following beneficial effect：
（1）, the present invention exempts from the specificity of kit by using AMOZ connection is carried out by with corresponding material
Cross reaction is tested come what is determined, and the content of the furaltadone in dairy products is determined by the size of absorbance, and the present invention is made
Detection method, it is simple to operate, it is cheap, and detection time is short, disclosure satisfy that the requirement of enterprise and mechanism.
Brief description of the drawings
Fig. 1 is test strips structure schematic diagram of the present invention.
In figure：1 paper slip body, 2 bow strips, 3 anticorrosive coats.
Specific embodiment
Below in conjunction with the accompanying drawing in the embodiment of the present invention, the technical scheme in the embodiment of the present invention is carried out clear, complete
Site preparation is described, it is clear that described embodiment is only a part of embodiment of the invention, rather than whole embodiments.It is based on
Embodiment in the present invention, it is every other that those of ordinary skill in the art are obtained under the premise of creative work is not made
Embodiment, belongs to the scope of protection of the invention.
As shown in figure 1, the present invention provides a kind of technical scheme：The detection method of AMOZ in a kind of dairy products,
Comprise the following steps：
First, experiment reagent is made：The concentrated hydrochloric acid that 9 milliliters of concentration are 2mol/L is measured, plus deionized water is settled to 120 milliliters,
Weigh 10 grams of dipotassium hydrogen phosphates, plus 400 ml deionized waters dissolve and are settled to 400 milliliters, it is molten to 400 milliliters of dipotassium hydrogen phosphates
In liquid add 80 grams of sodium chloride, fully dissolve standby, weigh 3.5 grams of NaOH, with deionized water dissolving and be settled to 100 milli
Rise, each reagent before experiment, should first open biochemical cultivation case to required temperature, and examine must fully be mixed before
Whether normal look into biochemical cultivation case temperature；
2nd, the accurate dairy food sample for weighing 150-200 milliliters is in 250 milliliters of centrifuge tube, and to being sequentially added in centrifuge tube
10 ml deionized waters, the concentrated hydrochloric acid of 2 milliliters of 2mol/L and 60 milliliters of o-nitrobenzaldehydes, abundant whirling motion to dairy products disperse,
The insulating box that temperature is 36 degree is then placed within to be incubated one hour, it is then molten to 8 milliliters of bufferings are sequentially added in dairy soln again
Liquid, 100 milliliters of sodium hydroxide solutions and 8 milliliters of carboxylate solutions, at room temperature, acutely rock 2 minutes, are subsequently adding 2 millis
Rise hexane, abundant whirling motion 15 seconds adds 2 milliliters of sample diluting liquids, low speed whirling motion 5 minutes, filter off upper strata hexane and in
Between impurity, take 150 milliliters of sample solutions and detected, this liquid be A liquid；
3rd, the accurate brown sugar slurry samples for weighing 50-100 milliliters are in 250 milliliters of centrifuge tube, and to being sequentially added in centrifuge tube
10 ml deionized waters, the concentrated hydrochloric acid of 2 milliliters of 2mol/L and 60 milliliters of o-nitrobenzaldehydes, abundant whirling motion to brown sugar slurry point
Dissipate, be then placed within the insulating box that temperature is 36 degree and be incubated one hour, it is then slow to sequentially adding 8 milliliters in red syrup solution again
Solution, 100 milliliters of sodium hydroxide solutions and 8 milliliters of carboxylate solutions are rushed, at room temperature, is acutely rocked 2 minutes, be subsequently adding
2 milliliters of hexanes, abundant whirling motion 15 seconds adds 2 milliliters of sample diluting liquids, low speed whirling motion 5 minutes, filter off upper strata hexane and
Interstitial impurity, takes 150 milliliters of sample solutions and is detected, this liquid is B liquid, and when extracting medicine, whirling motion or vibration should try one's best fully,
After adding sample and being weak solution, low speed whirling motion is answered 2 minutes, above operation will directly affect splendid attire and testing result；
4th, S1, by required ELIAS strip insertion ELISA Plate frame, after untapped ELIAS strip is sealed with valve bag, immediately
It is stored in 3-7 degrees Celsius of environment, and records the position of each standard sample；
S2,100 microlitres of each standard sample solution are separately added into corresponding standard sample sample wells；
S3,100 microlitres of AMOZ enzyme conjugates of addition in each plate hole；
S4, cover plate film is covered, gently vibrate enzyme mark version about ten seconds, fully mixed, 23-28 degrees Celsius of the reaction of lucifuge at room temperature
45 minutes；
S5, cover plate film is opened, outwell solution in plate hole, 3 milliliters of wash operating solutions are added in every hole, soaked 30 minutes, weighed successively
Multiple operation is twice；
S6, outwell solution in plate hole, ELISA Plate be inverted on blotting paper, blot, added in every hole immediately 10 milliliters of A liquid and
10 milliliters of B liquid, are sufficiently mixed, and mixed liquor was used in 3 minutes, during mix reagent, should avoid the occurrence of bubble, unspent
Microplate should be sealed, and be placed in 2-8 degrees Celsius of preservation, it is to avoid contain using metalware；
S7, cover plate film is covered, gently vibrate ELISA Plate ten seconds, fully mixed, 23-28 degrees Celsius of the reaction of lucifuge at room temperature 30
Minute；
S8, cover plate film is opened, 2 milliliters of terminate liquids are added in each plate hole, gently vibrated ten seconds, fully mixed, use ELIASA
Absorbance is detected under 400 nanometers of dual wavelength, 600 nanometers, is as a result read in 5 minutes after termination；
S9, the mean absorbance values by each sample, divided by zero standard（Concentration is the sample of 0ppb）Absorbance, is multiplied by 100, can be with
Obtain the percentage of the corresponding absorbance of each sample；Bring the percentage of sample absorbance into standard curve（Calibration curve formula
It is the absorbance of the zero standards of absorbance * 100/ of sample）, the corresponding concentration of sample can be drawn, multiplied by with the dilution of respective sample times
Number, side obtains actual drug content in sample, the test strip of AMOZ in dairy products, including paper slip body 1, paper slip
One end of body 1 is fixedly connected with bow strip 2, and anticorrosive coat 3 is provided with the outer wall of paper slip body 1.
Can to sum up obtain, the present invention by using AMOZ connection exempt from kit specificity be by with it is corresponding
Material carries out cross reaction experiment come what is determined, and the content of the furaltadone in dairy products is determined by the size of absorbance, this
The used detection method of invention, it is simple to operate, it is cheap, and detection time is short, disclosure satisfy that the requirement of enterprise and mechanism.
It should be noted that herein, such as first and second or the like relational terms are used merely to a reality
Body or operation make a distinction with another entity or operation, and not necessarily require or imply these entities or deposited between operating
In any this actual relation or order.And, term " including ", "comprising" or its any other variant be intended to
Nonexcludability is included, so that process, method, article or equipment including a series of key elements not only will including those
Element, but also other key elements including being not expressly set out, or also include being this process, method, article or equipment
Intrinsic key element.In the absence of more restrictions, the key element limited by sentence "including a ...", it is not excluded that
Also there is other identical element in process, method, article or equipment including the key element.
Although an embodiment of the present invention has been shown and described, for the ordinary skill in the art, can be with
Understanding can carry out various changes, modification, replacement to these embodiments without departing from the principles and spirit of the present invention
And modification, the scope of the present invention be defined by the appended.
Claims (5)
1. in a kind of dairy products AMOZ detection method, it is characterised in that：Comprise the following steps：
Make experiment reagent：The concentrated hydrochloric acid that 9 milliliters of concentration are 2mol/L is measured, plus deionized water is settled to 120 milliliters, weighs
10 grams of dipotassium hydrogen phosphates, plus 400 ml deionized waters dissolve and are settled to 400 milliliters, in 400 milliliters of dipotassium hydrogen phosphate solutions
80 grams of sodium chloride are added, fully dissolves standby, weigh 3.5 grams of NaOH, with deionized water dissolving and be settled to 100 milliliters；
The accurate dairy food sample for weighing 150-200 milliliter in 250 milliliters of centrifuge tube, and to sequentially adding 10 in centrifuge tube in the least
Deionized water, the concentrated hydrochloric acid of 2 milliliters of 2mol/L and 60 milliliters of o-nitrobenzaldehydes are risen, abundant whirling motion to dairy products disperses, then
Be placed on insulating box that temperature is 36 degree and be incubated one hour, then again to sequentially added in dairy soln 8 milliliters of cushioning liquid,
100 milliliters of sodium hydroxide solutions and 8 milliliters of carboxylate solutions, at room temperature, acutely rock 2 minutes, are being subsequently adding 2 milliliters just
Ethane, abundant whirling motion 15 seconds adds 2 milliliters of sample diluting liquids, low speed whirling motion 5 minutes, filter off upper strata hexane and in be mixed with
Matter, takes 150 milliliters of sample solutions and is detected, this liquid is A liquid；
The accurate brown sugar slurry samples for weighing 50-100 milliliters in 250 milliliters of centrifuge tube, and to sequentially adding 10 in centrifuge tube
Ml deionized water, the concentrated hydrochloric acid of 2 milliliters of 2mol/L and 60 milliliters of o-nitrobenzaldehydes, abundant whirling motion to brown sugar are starched and disperseed,
It is then placed within insulating box that temperature is 36 degree and is incubated one hour, then again to sequentially adding 8 milliliters of bufferings in red syrup solution
Solution, 100 milliliters of sodium hydroxide solutions and 8 milliliters of carboxylate solutions, at room temperature, acutely rock 2 minutes, are subsequently adding 2
Milliliter hexane, abundant whirling motion 15 seconds adds 2 milliliters of sample diluting liquids, low speed whirling motion 5 minutes, filter off upper strata hexane and
Interstitial impurity, takes 150 milliliters of sample solutions and is detected, this liquid is B liquid；
S1, by required ELIAS strip insertion ELISA Plate frame, after untapped ELIAS strip is sealed with valve bag, preserve immediately
In 3-7 degrees Celsius of environment, and record the position of each standard sample；
S2,100 microlitres of each standard sample solution are separately added into corresponding standard sample sample wells；
S3,100 microlitres of AMOZ enzyme conjugates of addition in each plate hole；
S4, cover plate film is covered, gently vibrate enzyme mark version about ten seconds, fully mixed, 23-28 degrees Celsius of the reaction of lucifuge at room temperature
45 minutes；
S5, cover plate film is opened, outwell solution in plate hole, 3 milliliters of wash operating solutions are added in every hole, soaked 30 minutes, weighed successively
Multiple operation is twice；
S6, outwell solution in plate hole, ELISA Plate be inverted on blotting paper, blot, added in every hole immediately 10 milliliters of A liquid and
10 milliliters of B liquid, are sufficiently mixed, and mixed liquor was used in 3 minutes, it is to avoid contained using metalware；
S7, cover plate film is covered, gently vibrate ELISA Plate ten seconds, fully mixed, 23-28 degrees Celsius of the reaction of lucifuge at room temperature 30
Minute；
S8, cover plate film is opened, 2 milliliters of terminate liquids are added in each plate hole, gently vibrated ten seconds, fully mixed, use ELIASA
Absorbance is detected under 400 nanometers of dual wavelength, 600 nanometers, is as a result read in 5 minutes after termination；
S9, the mean absorbance values by each sample, divided by zero standard（Concentration is the sample of 0ppb）Absorbance, is multiplied by 100, can be with
Obtain the percentage of the corresponding absorbance of each sample；Bring the percentage of sample absorbance into standard curve（Calibration curve formula
It is the absorbance of the zero standards of absorbance * 100/ of sample）, the corresponding concentration of sample can be drawn, multiplied by with the dilution of respective sample times
Number, side obtains actual drug content in sample.
2. in a kind of dairy products according to claim 1 AMOZ detection method, it is characterised in that：Each reagent
Must fully be mixed before, and before experiment, biochemical cultivation case to required temperature should be first opened, and check biochemical culture
Whether case temperature is normal.
3. in a kind of dairy products according to claim 1 AMOZ detection method, it is characterised in that：Extract medicine
During thing, whirling motion or vibration should try one's best fully, after adding sample to be weak solution, answer low speed whirling motion 2 minutes, and above operation is by direct shadow
Ring and contain and testing result.
4. in a kind of dairy products according to claim 1 AMOZ detection method, it is characterised in that：Mixing examination
During agent, bubble should be avoided the occurrence of, unspent microplate should be sealed, be placed in 2-8 degrees Celsius of preservation.
5. in a kind of dairy products according to claim 1 AMOZ detection method, it is characterised in that：The detection
Method includes paper slip body with reagent strip（1）, the paper slip body（1）One end be fixedly connected with bow strip（2）, the paper slip
Body（1）Outer wall on be provided with anticorrosive coat（3）.
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Cited By (1)
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| CN109632427A (en) * | 2019-01-28 | 2019-04-16 | 广州食源生物科技有限公司 | A kind of pre-treating method that Nitrofuran metatolites quickly detect and its detection method |
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|---|---|---|---|---|
| CN109632427A (en) * | 2019-01-28 | 2019-04-16 | 广州食源生物科技有限公司 | A kind of pre-treating method that Nitrofuran metatolites quickly detect and its detection method |
| CN109632427B (en) * | 2019-01-28 | 2021-02-09 | 广州食源生物科技有限公司 | Pretreatment method for rapid detection of nitrofuran metabolites and detection method thereof |
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