CN106770736A - A kind of chicken Ractopamine detection method and kit - Google Patents
A kind of chicken Ractopamine detection method and kit Download PDFInfo
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- CN106770736A CN106770736A CN201611091157.5A CN201611091157A CN106770736A CN 106770736 A CN106770736 A CN 106770736A CN 201611091157 A CN201611091157 A CN 201611091157A CN 106770736 A CN106770736 A CN 106770736A
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- chicken
- ractopamine
- ethyl acetate
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- detection method
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/02—Column chromatography
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- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
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- General Health & Medical Sciences (AREA)
- General Physics & Mathematics (AREA)
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- Pathology (AREA)
- Meat, Egg Or Seafood Products (AREA)
Abstract
The invention discloses a kind of chicken Ractopamine detection method and kit, it is related to technical field of food safety detection.The chicken Ractopamine detection method and kit, comprise the following steps, by chicken, ethyl acetate and solution of potassium carbonate configuration testing solution, the methanol solution number point of the Ractopamine standard items after being made, being injected separately into GC MS carries out chromatogram, finally crosses peak height method or area-method is depicted as canonical plotting.The chicken Ractopamine detection method and kit, by to test solution system match somebody with somebody and GC MS use, the result for detecting is set to draw canonical plotting, so not only facilitate the observation of people, and more easily chicken further can be contrasted and reference, it is convenient for people to use and such curve map can effectively reduce error.
Description
Technical field
The present invention relates to technical field of food safety detection, specially a kind of chicken Ractopamine detection method and reagent
Box.
Background technology
Ractopamine is a kind of β agonists medicine, is one kind of clenbuterol hydrochloride, can be used as animal feed addition
Thing, is used to encourage pig, ox, turkey and bears muscle, reduces body fat, it is a large amount of eat the meats containing Rct opamine residue or
During internal organ, poisoning symptom may be triggered, to this medicine aquaculture the scope of application and security countries in the world regulation not to the utmost
It is identical.
Ractopamine is used when in commercial chicken, and commercial chicken can expeditiously produce the high-quality measured chicken of lean meat percentage
Meat, the pricing system of lean meat trunk result in the further selection to the lean meat speed of growth, carcass lean meat percentage and the price of deed, be good for
The implementation of health, nutrition and device management strategies is provided to improve lean meat growth efficiency as much as possible, but Ractopamine exists
Use in whole world poultry husbandry is all illegal.
But the detection of existing chicken Ractopamine is not accurate enough and directly perceived, people are caused to enter to commercial chicken
The analysis and research of one step, have so had a strong impact on the work of people, reduce the work quality of staff.
The content of the invention
（One）The technical problem of solution
In view of the shortcomings of the prior art, the invention provides a kind of chicken Ractopamine detection method and kit, solve
The detection of chicken Ractopamine is not accurate enough and intuitively problem.
（Two）Technical scheme
To realize object above, the present invention is achieved by the following technical programs：
A kind of chicken Ractopamine detection method, it is characterised in that comprise the following steps：
S1：The test solution of 5-6ml is configured by chicken, ethyl acetate and solution of potassium carbonate；
S2：The methanol solution number point of the 0.025~0.5mg/L of Ractopamine standard items after being made, is injected separately into GC-MS
Carry out chromatogram；
S3：Canonical plotting is depicted as by peak height method or area-method.
Preferably, in the S1 test solution compounding method following steps：
A1：10-20 grams of chicken is first taken out with cutting tool, degreasing and cleaning then are carried out to chicken, then chicken is placed
In pulper, the distilled water of 50-100ml is added, chicken is blended, blend the time for 3-5 minutes, produce mixed liquor；
A2：Then the mixed liquor of 5-10ml is extracted, is put into centrifuge, added 10-20 ethyl acetate and 1-2mL potassium carbonate is molten
Liquid, the concentration of solution of potassium carbonate is 4mol/L, centrifugation 10-20 minutes after homogeneous；
A3：Then ethyl acetate layer is collected, 20-30mL ethyl acetate is added in centrifuge tube in residue, after vibrating 5-10 minutes,
By above-mentioned similarity condition centrifugation, the ethyl acetate layer obtained by merging is concentrated under conditions of 20-30 degrees Celsius, removed
Remove ethyl acetate；
A4：The dissolving of 30-40mL acetonitriles is added in residue, is moved into separatory funnel, add 30-40mL acetonitrile saturation n-hexanes,
Vibration, discards n-hexane layer, repeats 2-3 times；
A5：Acetonitrile layer is concentrated under 20-30 degrees celsius, removes acetonitrile, and the dissolving of 1-2mL methyl alcohol is added in residue, is made
Contain the methanol solution of 0.025~0.5mg/L of Ractopamine standard items in 5-6ml.
Preferably, the degreasing and cleaning process of the A1 chicken are as follows：
B1：First the chicken block of well cutting is placed on screen pack, it is the wine of 70%-90% then to draw concentration by glue head dropper
Essence, alcohol capacity is 20-30ml, then the surface of slow cleaning chicken, until alcohol washes are complete, the fat on removal chicken
Fat；
B2：Then the chicken that cleaning is completed is placed on and is dried on plate, dried 5-10 minutes；
B3：Then the chicken for drying is prevented on filter, the distilled water of 20-30ml is then drawn by colloid dropper, it is right
The chicken for drying is cleaned, and scavenging period is 2-3 minutes；
B4：Chicken after cleaning is placed on blotting paper, carries out drying treatment, flash-off time is 10-20 minutes.
Preferably, in the A2 centrifugal treating, it is necessary to the use of to rotating speed be 3000 rpms of centrifuges of rotating speed.
A kind of chicken Ractopamine kit, including reagent box body, the top of the reagent box body pass through rotating shaft
Lid is connected with, the inside of the reagent box body is installed with rack for test tube, and the lid is near reagent box body
Side is installed with drawing board, and the front surface of the reagent box body is installed with deck, and the front end of the lid is fixed
Buckle is installed.
（Three）Beneficial effect
The invention provides a kind of chicken Ractopamine detection method and kit.Possesses following beneficial effect：
（1）, the chicken Ractopamine detection method and kit, matched somebody with somebody by the system to test solution and GC-MS use, make
The result for detecting can draw canonical plotting, so not only facilitate the observation of people, and can be more convenient
Chicken is further contrasted and reference, and such curve map can effectively reduce error, it is convenient for people to use.
（2）, the chicken Ractopamine detection method and kit, by the setting to degreasing and cleaning process, can
Fat in effective removal chicken, so improves the accuracy to Ractopamine detection in chicken, improves the effect of experiment
Really.
Brief description of the drawings
Fig. 1 is the structural representation of chicken Ractopamine kit of the present invention.
In figure：1 reagent box body, 2 lids, 3 rack for test tubes, 4 drawing boards, 5 decks, 6 buckles.
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.
Embodiment one
A kind of chicken Ractopamine detection method, comprises the following steps:
S1：The test solution of 5ml is configured by chicken, ethyl acetate and solution of potassium carbonate；
The compounding method following steps of test solution in S1：
A1：10 grams of chicken is first taken out with cutting tool, degreasing and cleaning is then carried out to chicken, first the chicken block of well cutting
It is placed on screen pack, the alcohol that concentration is 70% is then drawn by glue head dropper, alcohol capacity is 20ml, then slowly
The surface of chicken is cleaned, until alcohol washes are complete, then the fat on removal chicken be placed on the chicken that cleaning is completed and dry
On plate, dry 5 minutes, then the chicken for drying is prevented on filter, the distillation of 20ml is then drawn by colloid dropper
Water, the chicken to drying is cleaned, and scavenging period is 2 minutes, and the chicken after cleaning is placed on blotting paper, is dried in the air
Dry-cure, flash-off time is 10 minutes, and then chicken is placed in pulper, adds the distilled water of 50ml, and chicken is carried out
Blend, blend the time for 3-5 minutes, produce mixed liquor；
A2：Then the mixed liquor of 5ml is extracted, is put into centrifuge, centrifugal treating is, it is necessary to be 3000 rpms using to rotating speed
The centrifuge of rotating speed, adds 10 ethyl acetate and 1mL solution of potassium carbonate, and the concentration of solution of potassium carbonate is 4mol/L after homogeneous,
Centrifugation 10 minutes；
A3：Then ethyl acetate layer is collected, 20mL ethyl acetate is added in centrifuge tube in residue, after vibrating 5 minutes, by above-mentioned
Similarity condition centrifugation, the ethyl acetate layer obtained by merging is concentrated under conditions of 20 degrees Celsius, removes acetic acid second
Ester；
A4：The dissolving of 30mL acetonitriles is added in residue, is moved into separatory funnel, add 30mL acetonitrile saturation n-hexanes, vibration is abandoned
N-hexane layer is removed, is repeated 2 times；
A5：Acetonitrile layer is concentrated under 20 degrees celsius, removes acetonitrile, and the dissolving of 1mL methyl alcohol is added in residue, is made in 5ml
Methanol solution containing Ractopamine standard items 0.025mg/L；
S2：The methanol solution number point of the Ractopamine standard items 0.025mg/L after being made, being injected separately into GC-MS is carried out
Chromatogram；
S3：Canonical plotting is depicted as by peak height method or area-method.
Embodiment two
A kind of chicken Ractopamine detection method, comprises the following steps:
S1：The test solution of 5.5ml is configured by chicken, ethyl acetate and solution of potassium carbonate；
The compounding method following steps of test solution in S1：
A1：15 grams of chicken is first taken out with cutting tool, degreasing and cleaning is then carried out to chicken, first the chicken block of well cutting
It is placed on screen pack, the alcohol that concentration is 80% is then drawn by glue head dropper, alcohol capacity is 25ml, then slowly
The surface of chicken is cleaned, until alcohol washes are complete, then the fat on removal chicken be placed on the chicken that cleaning is completed and dry
On plate, dry 7.5 minutes, then the chicken for drying is prevented on filter, the steaming of 25ml is then drawn by colloid dropper
Distilled water, the chicken to drying is cleaned, and scavenging period is 2.5 minutes, and the chicken after cleaning is placed on blotting paper, is entered
Row dries treatment, and flash-off time is 15 minutes, and then chicken is placed in pulper, the distilled water of 75ml is added, to chicken
Blended, blended the time for 4 minutes, produced mixed liquor；
A2：Then the mixed liquor of 7.5ml is extracted, is put into centrifuge, centrifugal treating is, it is necessary to be 3000 revolutions per minute using to rotating speed
The centrifuge of clock rotating speed, adds 15 ethyl acetate and 1.5mL solution of potassium carbonate, and the concentration of solution of potassium carbonate is for after homogeneous
4mol/L, centrifugation 15 minutes；
A3：Then ethyl acetate layer is collected, 25mL ethyl acetate is added in centrifuge tube in residue, after vibrating 7.5 minutes, by upper
Similarity condition centrifugation is stated, the ethyl acetate layer obtained by merging is concentrated under conditions of 25 degrees Celsius, remove acetic acid second
Ester；
A4：The dissolving of 35mL acetonitriles is added in residue, is moved into separatory funnel, add 35mL acetonitrile saturation n-hexanes, vibration is abandoned
N-hexane layer is removed, is repeated 2 times；
A5：Acetonitrile layer is concentrated under 25 degrees celsius, removes acetonitrile, and the dissolving of 1.5mL methyl alcohol is added in residue, is made
Contain the methanol solution of Ractopamine standard items 0.263mg/L in 5.5ml；
S2：The methanol solution number point of the Ractopamine standard items 0.263mg/L after being made, being injected separately into GC-MS is carried out
Chromatogram；
S3：Canonical plotting is depicted as by peak height method or area-method.
Embodiment three
A kind of chicken Ractopamine detection method, comprises the following steps:
S1：The test solution of 6ml is configured by chicken, ethyl acetate and solution of potassium carbonate；
The compounding method following steps of test solution in S1：
A1：20 grams of chicken is first taken out with cutting tool, degreasing and cleaning is then carried out to chicken, first the chicken block of well cutting
It is placed on screen pack, the alcohol that concentration is 90% is then drawn by glue head dropper, alcohol capacity is 30ml, then slowly
The surface of chicken is cleaned, until alcohol washes are complete, then the fat on removal chicken be placed on the chicken that cleaning is completed and dry
On plate, dry 10 minutes, then the chicken for drying is prevented on filter, the distillation of 30ml is then drawn by colloid dropper
Water, the chicken to drying is cleaned, and scavenging period is 3 minutes, and the chicken after cleaning is placed on blotting paper, is dried in the air
Dry-cure, flash-off time is 20 minutes, and then chicken is placed in pulper, adds the distilled water of 100ml, and chicken is carried out
Blend, blend the time for 5 minutes, produce mixed liquor；
A2：Then the mixed liquor of 10ml is extracted, is put into centrifuge, centrifugal treating is, it is necessary to be 3000 revolutions per minute using to rotating speed
The centrifuge of clock rotating speed, adds 20 ethyl acetate and 2mL solution of potassium carbonate, and the concentration of solution of potassium carbonate is 4mol/ after homogeneous
L, centrifugation 20 minutes；
A3：Then ethyl acetate layer is collected, 30mL ethyl acetate is added in centrifuge tube in residue, after vibrating 10 minutes, by upper
Similarity condition centrifugation is stated, the ethyl acetate layer obtained by merging is concentrated under conditions of 30 degrees Celsius, remove acetic acid second
Ester；
A4：The dissolving of 40mL acetonitriles is added in residue, is moved into separatory funnel, add 40mL acetonitrile saturation n-hexanes, vibration is abandoned
N-hexane layer is removed, is repeated 3 times；
A5：Acetonitrile layer is concentrated under 30 degrees celsius, removes acetonitrile, and the dissolving of 2mL methyl alcohol is added in residue, is made in 6ml
Methanol solution containing Ractopamine standard items 0.5mg/L；
S2：The methanol solution number point of the Ractopamine standard items 0.5mg/L after being made, being injected separately into GC-MS carries out color
Spectrum；
S3：Canonical plotting is depicted as by peak height method or area-method.
Embodiment one, embodiment two and ethyl acetate layer is removed by point liquid in embodiment three are so not only thorough
But also more facilitate, the separatory funnel in wherein A4 is placed in oscillator, is carried out concussion and is removed n-hexane layer, in the method
The use of GC-MS is High Performance Liquid Chromatography/Mass Spectrometry instrument.
Fig. 1 is referred to, the present invention provides a kind of chicken Ractopamine kit, including reagent box body 1, kit sheet
The top of body 1 is connected with lid 2 by rotating shaft, and the inside of reagent box body 1 is installed with rack for test tube 3, and lid 2 is close to
The side of reagent box body 1 is installed with drawing board 4, and the front surface of reagent box body 1 is installed with deck 5, lid 2
Front end is installed with buckle 6.
Buckle 6 can be effectively fixed on deck 5, facilitated in the use of people, rack for test tube 3 and be provided with ethyl acetate
Reagent, alcohol reagent, distilled water, solution of potassium carbonate, acetonitrile saturation hexane solution and methanol solution, drawing board 4 can make one
Quickly and easily draw figure.
It should be noted that herein, term " including ", "comprising" or its any other variant be intended to non-row
His property is included, so that process, method, article or equipment including a series of key elements not only include those key elements, and
And also include other key elements being not expressly set out, or also include for this process, method, article or equipment institute are intrinsic
Key element.In the absence of more restrictions.By sentence " including one ... the key element for limiting, it is not excluded that including
Also there is other identical element in the process of the key element, method, article or equipment ".
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. a kind of chicken Ractopamine detection method, it is characterised in that comprise the following steps：
S1：The test solution of 5-6ml is configured by chicken, ethyl acetate and solution of potassium carbonate；
S2：The methanol solution number point of the 0.025~0.5mg/L of Ractopamine standard items after being made, is injected separately into GC-MS
Carry out chromatogram；
S3：Canonical plotting is depicted as by peak height method or area-method.
2. a kind of chicken Ractopamine detection method according to claim 1, it is characterised in that：Test molten in the S1
The compounding method following steps of liquid：
A1：10-20 grams of chicken is first taken out with cutting tool, degreasing and cleaning then are carried out to chicken, then chicken is placed
In pulper, the distilled water of 50-100ml is added, chicken is blended, blend the time for 3-5 minutes, produce mixed liquor；
A2：Then the mixed liquor of 5-10ml is extracted, is put into centrifuge, added 10-20 ethyl acetate and 1-2mL potassium carbonate is molten
Liquid, the concentration of solution of potassium carbonate is 4mol/L, centrifugation 10-20 minutes after homogeneous；
A3：Then ethyl acetate layer is collected, 20-30mL ethyl acetate is added in centrifuge tube in residue, after vibrating 5-10 minutes,
By above-mentioned similarity condition centrifugation, the ethyl acetate layer obtained by merging is concentrated under conditions of 20-30 degrees Celsius, removed
Remove ethyl acetate；
A4：The dissolving of 30-40mL acetonitriles is added in residue, is moved into separatory funnel, add 30-40mL acetonitrile saturation n-hexanes,
Vibration, discards n-hexane layer, repeats 2-3 times；
A5：Acetonitrile layer is concentrated under 20-30 degrees celsius, removes acetonitrile, and the dissolving of 1-2mL methyl alcohol is added in residue, is made
Contain the methanol solution of 0.025~0.5mg/L of Ractopamine standard items in 5-6ml.
3. a kind of chicken Ractopamine detection method according to claim 1, it is characterised in that：The A1 chicken goes
Fat and cleaning process are as follows：
B1：First the chicken block of well cutting is placed on screen pack, it is the wine of 70%-90% then to draw concentration by glue head dropper
Essence, alcohol capacity is 20-30ml, then the surface of slow cleaning chicken, until alcohol washes are complete, the fat on removal chicken
Fat；
B2：Then the chicken that cleaning is completed is placed on and is dried on plate, dried 5-10 minutes；
B3：Then the chicken for drying is prevented on filter, the distilled water of 20-30ml is then drawn by colloid dropper, it is right
The chicken for drying is cleaned, and scavenging period is 2-3 minutes；
B4：Chicken after cleaning is placed on blotting paper, carries out drying treatment, flash-off time is 10-20 minutes.
4. a kind of chicken Ractopamine detection method according to claim 2, it is characterised in that：In the A2 at centrifugation
Reason is, it is necessary to be 3000 rpms of centrifuges of rotating speed using to rotating speed.
5. a kind of chicken Ractopamine kit, including reagent box body（1）, it is characterised in that：The reagent box body（1）
Top lid is connected with by rotating shaft（2）, the reagent box body（1）Inside be installed with rack for test tube（3）, institute
State lid（2）Near reagent box body（1）Side be installed with drawing board（4）, the reagent box body（1）Front surface
It is installed with deck（5）, the lid（2）Front end be installed with buckle（6）.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201611091157.5A CN106770736A (en) | 2016-12-01 | 2016-12-01 | A kind of chicken Ractopamine detection method and kit |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201611091157.5A CN106770736A (en) | 2016-12-01 | 2016-12-01 | A kind of chicken Ractopamine detection method and kit |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN106770736A true CN106770736A (en) | 2017-05-31 |
Family
ID=58914238
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201611091157.5A Pending CN106770736A (en) | 2016-12-01 | 2016-12-01 | A kind of chicken Ractopamine detection method and kit |
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Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN202486133U (en) * | 2012-03-21 | 2012-10-10 | 李同立 | Integrated detection tool kit |
| CN204039403U (en) * | 2014-04-17 | 2014-12-24 | 中国人民解放军军事医学科学院军事兽医研究所 | Portable animal pathogenic micro-organism rapid detection box |
| CN104872692A (en) * | 2015-04-24 | 2015-09-02 | 安徽卫食园肉类食品有限公司 | Preparation method of smoked and preserved meat |
| CN105647857A (en) * | 2016-04-08 | 2016-06-08 | 安徽农业大学 | Separation and purification method for skeletal muscle satellite cells in goats |
-
2016
- 2016-12-01 CN CN201611091157.5A patent/CN106770736A/en active Pending
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN202486133U (en) * | 2012-03-21 | 2012-10-10 | 李同立 | Integrated detection tool kit |
| CN204039403U (en) * | 2014-04-17 | 2014-12-24 | 中国人民解放军军事医学科学院军事兽医研究所 | Portable animal pathogenic micro-organism rapid detection box |
| CN104872692A (en) * | 2015-04-24 | 2015-09-02 | 安徽卫食园肉类食品有限公司 | Preparation method of smoked and preserved meat |
| CN105647857A (en) * | 2016-04-08 | 2016-06-08 | 安徽农业大学 | Separation and purification method for skeletal muscle satellite cells in goats |
Non-Patent Citations (3)
| Title |
|---|
| IN KYUNG SUNG ET AL.: "Development and Application of a Method for Rapid and Simultaneous", 《KOREAN J. FOOD SCI. AN.》 * |
| JIANG JINQING ET AL.: "Preparation and Identification of anti-Ractopamine Monoclonal antibodies", 《INTERNATIONAL CONFERENCE ON LIFE SCIENCE AND TECHNOLOGY》 * |
| 应永飞 等: "气相色谱-质谱法测定猪肉中的莱克多巴胺", 《中国兽药杂志》 * |
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