CN107505417A - The detection method of clenbuterol hydrochloride in fresh meat and meat products - Google Patents
The detection method of clenbuterol hydrochloride in fresh meat and meat products Download PDFInfo
- Publication number
- CN107505417A CN107505417A CN201710028489.7A CN201710028489A CN107505417A CN 107505417 A CN107505417 A CN 107505417A CN 201710028489 A CN201710028489 A CN 201710028489A CN 107505417 A CN107505417 A CN 107505417A
- Authority
- CN
- China
- Prior art keywords
- sample
- mobile phase
- detection method
- solution
- volume fraction
- 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.)
- Granted
Links
Classifications
-
- 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
- G01N30/04—Preparation or injection of sample to be analysed
- G01N30/06—Preparation
-
- 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
Landscapes
- Physics & Mathematics (AREA)
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Analytical Chemistry (AREA)
- Biochemistry (AREA)
- General Health & Medical Sciences (AREA)
- General Physics & Mathematics (AREA)
- Immunology (AREA)
- Pathology (AREA)
- Extraction Or Liquid Replacement (AREA)
- Investigating Or Analysing Biological Materials (AREA)
Abstract
The invention discloses the detection method of clenbuterol hydrochloride in fresh meat and meat products, it is related to the detection method of illegal additive in food, the detection method is：Sample pre-treatments are first carried out, obtain sample solution, then gained sample solution is determined using high performance liquid chromatography；Wherein, the sample is fresh meat or meat products；The sample pre-treatments include digesting, extract, purify, concentrate, redissolve and filtering, and the Extraction solvent of the extraction step is at least one of dichloromethane, ethyl acetate, hexamethylene, dimethyl sulfoxide (DMSO).Enzymolysis is combined with the organic solvent of water, extracts reagent in the sample pre-treatments of the present invention, the processing time of sample can be shortened and improve clenbuterol hydrochloride recovery rate.
Description
Technical field
The present invention relates to the detection method of illegal additive in food, more particularly to clenbuterol hydrochloride in fresh meat and meat products
Detection method.
Background technology
Clenbuterol hydrochloride be having of referring to that aquaculture illegally uses promote growth of animal, improve lean meat percentage containing β-adrenal gland by
A kind of material of body activator, clenbuterol hydrochloride include Clenbuterol, salbutamol, Ractopamine, bambuterol, tartaric acid A Fu
Tens kinds of compounds such as special sieve and phenolethanolamine A.Research finds that long-term use of clenbuterol hydrochloride can be such that it accumulates in animal body, people
Class eat clenbuterol hydrochloride exceed safety value animal meat after, it may appear that tremble, palpitaition, stress, have a headache, exert one's utmost effort, vomitting and
The symptoms such as weakness of limbs, or even threat to life.Illegal use of many countries all to clenbuterol hydrochloride in food animal including China
Implement strict supervision, since two thousand two, China with regard to successively issue multiple communique files to prevent clenbuterol hydrochloride
It is illegal to use.
In order to strengthen the supervision to clenbuterol hydrochloride medicine, domestic and international scientific research personnel is to clenbuterol residue amount in meat and meat products
Detection method, which has been carried out, to be extensively studied, and existing detection method has high performance liquid chromatography (HPLC), liquid phase chromatogram-mass spectrometry combination
Usage (LC-MS), chromatography of gases-MS (GC-MS), immunoassay (ELISA), colloid gold immune test strip
And Capillary Electrophoresis (EC) etc. (DIGFA).At present, testing agency predominantly detects Clenbuterol, salbutamol, the Lay in clenbuterol hydrochloride
Gram three kinds of materials of dopamine；And existing detection method is longer to the processing time of fresh meat and meat products sample, clenbuterol hydrochloride carries
Take efficiency low, the detection to clenbuterol content produces certain influence.
The content of the invention
Present invention aims to overcome that insufficient existing for existing clenbuterol hydrochloride detection method, and when providing a kind of processing of sample
Between in short, fresh meat and meat products that clenbuterol hydrochloride recovery rate is high clenbuterol hydrochloride detection method, to achieve the above object, the present invention adopts
The technical scheme taken is：The detection method of clenbuterol hydrochloride, the detection method are in raw fresh meat and meat products：Before first carrying out sample
Processing, sample solution is obtained, then institute's sample solution is determined using high performance liquid chromatography；Wherein, the sample is fresh meat or meat
Product, sample pre-treatments comprise the following steps:
(1) digest：The sample blended is weighed, buffer solution and enzyme are added into sample, centrifuges, takes after digesting 10~30min
Supernatant produces sample enzymolysis liquid；The buffer solution is ammonium acetate solution, and the enzyme includes β-glucuronic acid glycosides peptase and aryl
Sulfuric acid lipase；
(2) extract：Ratio according to every gram of sample addition 6~10mL extracts reagent adds into gained sample enzymolysis liquid to be carried
Reagent is taken, 5~10min of ultrasonic extraction, produces sample extracting solution；Wherein, the extracts reagent is made up of water and organic solvent, water
Volume ratio with organic solvent is 1:3~1:5, and the organic solvent is dichloromethane, ethyl acetate, hexamethylene and dimethyl
At least one of sulfoxide；
(3) purify：Water absorbing agent and salting-out agents are added into gained sample extracting solution, is centrifuged after mixing, takes supernatant to produce
Sample purification liquid；The mass ratio of the water absorbing agent and sample is 1:2~2:3, the mass ratio of the salting-out agents and sample is 1:1~
2:1；
Preferably water absorbing agent is Na2SO4, salting-out agents NaCl, salt ionic concentration in sample extracting solution can be increased, with drop
Low clenbuterol hydrochloride is dissolved in aqueous phase, is dissolved in so as to improve clenbuterol hydrochloride in the organic solvent of extracts reagent；Na2SO4With very strong
Water imbibition, further increase clenbuterol hydrochloride and be dissolved in the organic solvent of extracts reagent；NaCl can promote organic solvent to enter with water
Row layering, improves clenbuterol hydrochloride separative efficiency.
(4) concentrate：Nitrogen blows blows concentration by gained sample purification liquid progress nitrogen, obtains concentrate；
(5) redissolve：Using concentrate obtained by the mixed liquor dissolving step (4) being made up of acetic acid aqueous solution and ethanol, obtain
Sample redissolves liquid；The volume fraction of acetic acid is 0.15%~0.25% in the acetic acid aqueous solution, acetic acid aqueous solution and ethanol
Volume ratio is 1:1~1:3；
(6) filter：Redissolved to the sample in liquid and add adsorbent, centrifuged after mixing, take supernatant, reuse filter membrane mistake
Filter, produces sample solution；Liquid is redissolved to sample and first takes adsorption-edulcoration, then through organic filter membrane filtration, clenbuterol hydrochloride can be purified
Purity, avoid influenceing follow-up high performance liquid chromatography measure；Use 3~5min of supersonic wave cleaning machine processing sample solution；It is super
Sound wave cleaning machine can remove the bubble in sample solution, avoid influenceing follow-up high performance liquid chromatography measure.
The detection method of the present invention includes sample pre-treatments and high performance liquid chromatography determines；The sample pre-treatments include enzyme
Solution, extraction, purification, concentration, redissolution and filtering, the pre-treatment of inventive samples first add extraction afterwards using enzymolysis, enzymolysis
Reagent, wherein, water can increase the degree of scatter of sample, fully contact, be able to sample enzymolysis liquid so as to improve organic solvent
Improve extraction effect；Meanwhile enzymolysis and water, organic solvent extraction be combined with each other, when the extraction of sample is effectively shortened in collaboration
Between.
As the improvement of above-mentioned technical proposal, in the step (1), enzymolysis time 20min.
Preferably due to β-glucuronic acid glycosides peptase/sulfuric acid lipase optimum activity condition is：PH=5.2, temperature 37
DEG C, therefore the pH value of ammonium acetate buffer is 5.2, temperature is 37 DEG C.
As the improvement of above-mentioned technical proposal, in the step (2), the Extraction solvent is dichloromethane；Dichloromethane
With preferably extraction effect.
As the improvement of above-mentioned technical proposal, in the step (2), organic solvent is ethyl acetate and dimethyl sulfoxide (DMSO)
Mixed liquor, and the volume ratio of ethyl acetate and dimethyl sulfoxide (DMSO) is 2:1, the mixed liquor of ethyl acetate and dimethyl sulfoxide (DMSO) have compared with
Excellent extraction effect.
As the improvement of above-mentioned technical proposal, in the step (2), the ratio of 9mL extracts reagents is added according to every gram of sample
Example adds extracts reagent into gained sample enzymolysis liquid, and in the extracts reagent, the volume ratio of water and organic solvent is 1:4；
Excessive moisture and a small amount of organic solvent will influence the extraction effect of clenbuterol hydrochloride.
As the improvement of above-mentioned technical proposal, in the step (3), the mass ratio of water absorbing agent and sample is 3:5, salting-out agents
Mass ratio with sample is 1.5:1.Appropriate water absorbing agent, salting-out agents can improve clenbuterol hydrochloride and be dissolved in the organic molten of extracts reagent
In agent, excessive salt ion can be avoided to influence follow-up high performance liquid chromatography measure again.
As the improvement of above-mentioned technical proposal, in the step (5), the volume fraction of acetic acid is in acetic acid aqueous solution
0.20%, the volume ratio of acetic acid aqueous solution and ethanol is 1:2.It is higher to extract the dissolved efficiency of clenbuterol hydrochloride, and more matching is efficient
Liquid chromatogram gradient elution program.
Further improved as above-mentioned technical proposal, the condition of the high performance liquid chromatography measure is：
Chromatographic column is：C18Chromatographic column, column length 250mm, column internal diameter 4.6mm, particle diameter are 5 μm；
Detection wavelength is：243nm；
Flow velocity is：0.5mL/mim；
Sample size is：10μL；
Mobile phase is made up of mobile phase A and Mobile phase B, and mobile phase A is 0.05mol/ml ammonium acetate solutions, and mobile phase
Containing the acetic acid that volume fraction is 0.1% in A, Mobile phase B is ethanol.Under the condition determination, clenbuterol hydrochloride has obvious
Absworption peak.
Further improved as above-mentioned technical proposal, the gradient elution program of the high performance liquid chromatography is:
When time is 0min, the volume fraction of mobile phase A is 75% in mobile phase, and the volume fraction of Mobile phase B is 25%；
When time is 5min, the volume fraction of mobile phase A is 60% in mobile phase, and the volume fraction of Mobile phase B is 40%；
When time is 8min, the volume fraction of mobile phase A is 60% in mobile phase, and the volume fraction of Mobile phase B is 40%；
When time is 13min, the volume fraction of mobile phase A is 40% in mobile phase, and the volume fraction of Mobile phase B is
60%；
When time is 15min, the volume fraction of mobile phase A is 40% in mobile phase, and the volume fraction of Mobile phase B is
60%.In the gradient elution program, Clenbuterol, salbutamol and Ractopamine easily distinguish.
Further improved as above-mentioned technical proposal, the standard items of the high performance liquid chromatography measure are Ke Lunte
Sieve, salbutamol and Ractopamine, the solvent of standard solution is ethanol.Standard solution is injected into high performance liquid chromatography
Instrument, with peak area (Y) for ordinate, the concentration (X, μ g/mL) of standard items mixed liquor is abscissa, draws standard curve.
The beneficial effects of the present invention are：The present invention provides the detection method of clenbuterol hydrochloride in a kind of fresh meat and meat products,
The detection method of the present invention includes sample pre-treatments and high performance liquid chromatography determines；Sample pre-treatments include enzymolysis, extract, be net
Change, concentrate, redissolving and filtering, extracts reagent are water and organic solvent, the enzymolysis of sample and water, organic solvent extraction are combined
Extraction time can be shortened and improve the extraction efficiency of clenbuterol hydrochloride.Water absorbing agent (such as Na is used in purification process2SO4) and salting-out agents
(such as NaCl), clenbuterol hydrochloride can be made to be adequately dissolve in the organic solvent of extracts reagent.The present invention also improves high-efficient liquid phase color
The condition of measure is composed, makes to be dissolved in acetic acid aqueous solution and molten middle clenbuterol hydrochloride is mixed with ethanol (Clenbuterol, salbutamol and Rec are more
Bar amine) it can be separated by mobile phase, fast and effeciently detected.
Embodiment
For the object, technical solutions and advantages of the present invention are better described, below in conjunction with specific embodiment to the present invention
It is described further.
Embodiment 1
The detection method of clenbuterol hydrochloride in a kind of fresh meat, the detection method of the embodiment are：Sample pre-treatments are first carried out, are obtained
Gained sample solution is determined to sample sample liquid, then using high performance liquid chromatography；Wherein sample pre-treatments comprise the following steps:
(1) digest：Weigh the live fresh pork 1.0g blended, buffer solution and enzyme added into sample, digest after 10min from
The heart, supernatant is taken to produce sample enzymolysis liquid；The buffer solution is ammonium acetate solution, the enzyme include β-glucuronic acid glycosides peptase and
Aryl sulphatase；
(2) extract：Extracts reagent is added into gained sample enzymolysis liquid, ultrasonic extraction 5min, produces sample extracting solution；Its
In, the extracts reagent is made up of water and organic solvent, and the organic solvent is dichloromethane, water 1.5mL, and dichloromethane is
4.5mL；
(3) purify：Na is added into gained sample extracting solution2SO4And NaCl, centrifuged after mixing, take supernatant to produce sample
Scavenging solution；The Na2SO4For 0.5g, NaCl 1.0g；
(4) concentrate：Nitrogen blows blows concentration by gained sample purification liquid progress nitrogen, obtains concentrate；
(5) redissolve：Using concentrate obtained by the mixed liquor dissolving step (4) being made up of acetic acid aqueous solution and ethanol, obtain
Sample redissolves liquid；The volume fraction of acetic acid is 0.15% in the acetic acid aqueous solution, and the volume ratio of acetic acid aqueous solution and ethanol is
1:1；
(6) filter：Redissolved to the sample and adsorbent C is added in liquid18, centrifuged after mixing, take supernatant, reuse filter membrane
Filtering, produces sample solution.
The present embodiment high performance liquid chromatography measure condition be：
Chromatographic column is：C18Chromatographic column, column length 250mm, column internal diameter 4.6mm, particle diameter are 5 μm；
Detection wavelength is：243nm；
Flow velocity is：0.5mL/mim；
Sample size is：10μL；
Mobile phase is made up of mobile phase A and Mobile phase B, and mobile phase A is 0.05mol/ml ammonium acetate solutions, and mobile phase
Containing the acetic acid that volume fraction is 0.1% in A, Mobile phase B is ethanol.
The gradient elution program of the high performance liquid chromatography of the present embodiment is shown in Table 1：
Table 1 is gradient elution program
The preparation of this implementation standard items mixed liquor:
Accurate weighing standard items Clenbuterol, salbutamol and Ractopamine, dissolved with ethanol and prepare standard items mixing
The concentration of liquid is 0,0.05,0.1,0.2 and 0.4 μ g/mL.
Embodiment 2
The detection method of clenbuterol hydrochloride in a kind of fresh meat, the detection method are：Sample pre-treatments are first carried out, it is molten to obtain sample
Liquid, then gained sample solution is determined using high performance liquid chromatography；Wherein sample pre-treatments comprise the following steps:
(1) digest：Weigh the live fresh pork 1.0g blended, buffer solution and enzyme added into sample, digest after 20min from
The heart, supernatant is taken to produce sample enzymolysis liquid；The buffer solution is ammonium acetate solution, the enzyme include β-glucuronic acid glycosides peptase and
Aryl sulphatase；
(2) extract：Extracts reagent is added into gained sample enzymolysis liquid, ultrasonic extraction 8min, produces sample extracting solution；Its
In, the extracts reagent is made up of water and organic solvent, and the organic solvent is ethyl acetate and dimethyl sulfoxide (DMSO), and water is
1.8mL, ethyl acetate 4.8mL, dimethyl sulfoxide (DMSO) 2.4mL；
(3) purify：Na is added into gained sample extracting solution2SO4And NaCl, centrifuged after mixing, take supernatant to produce sample
Scavenging solution；The Na2SO4For 0.6g, NaCl 2.0g；
(4) concentrate：Nitrogen blows blows concentration by gained sample purification liquid progress nitrogen, obtains concentrate；
(5) redissolve：Using concentrate obtained by the mixed liquor dissolving step (4) being made up of acetic acid aqueous solution and ethanol, obtain
Sample redissolves liquid；The volume fraction of acetic acid is 0.20% in the acetic acid aqueous solution, and the volume ratio of acetic acid aqueous solution and ethanol is
1:2；
(6) filter：Redissolved to the sample and adsorbent C is added in liquid18, centrifuged after mixing, take supernatant, reuse filter membrane
Filtering, produces sample solution.
The high performance liquid chromatography measure of the present embodiment and the preparation of standard items mixed liquor are the same as embodiment 1.
Embodiment 3
The detection method of clenbuterol hydrochloride in a kind of fresh meat, the detection method of the embodiment are：Sample pre-treatments are first carried out, are obtained
Gained sample solution is determined to sample solution, then using high performance liquid chromatography；Wherein sample pre-treatments comprise the following steps:
(1) digest：Weigh the live fresh pork 1.0g blended, buffer solution and enzyme added into sample, digest after 30min from
The heart, supernatant is taken to produce sample enzymolysis liquid；The buffer solution is ammonium acetate solution, and the enzyme includes β-glucuronic acid glycosides peptase and virtue
Base sulfuric acid lipase；
(2) extract：Extracts reagent is added into gained sample enzymolysis liquid, ultrasonic extraction 10min, produces sample extracting solution；
Wherein, the extracts reagent is made up of water and organic solvent, and the organic solvent is hexamethylene, water 1.7mL, and hexamethylene is
8.3mL；
(3) purify：Na is added into gained sample extracting solution2SO4And NaCl, centrifuged after mixing, take supernatant to produce sample
Scavenging solution；The Na2SO4For 0.67g, NaCl 2.0g；
(4) concentrate：Nitrogen blows blows concentration by gained sample purification liquid progress nitrogen, obtains concentrate；
(5) redissolve：Using concentrate obtained by the mixed liquor dissolving step (4) being made up of acetic acid aqueous solution and ethanol, obtain
Sample redissolves liquid；The volume fraction of acetic acid is 0.25% in the acetic acid aqueous solution, and the volume ratio of acetic acid aqueous solution and ethanol is
1:3；
(6) filter：Redissolved to the sample and adsorbent C is added in liquid18, centrifuged after mixing, take supernatant, reuse filter membrane
Filtering, produces sample solution.
The high performance liquid chromatography measure of the present embodiment and the preparation of standard items mixed liquor are the same as embodiment 1.
It is with a collection of, Clenbuterol, salbutamol and Rec DOPA in fresh meat to implement 1~3 live fresh pork determined
Amine content is shown in Table 2, is average value containing numerical quantity.
Table 2 is the content of Clenbuterol, salbutamol and Ractopamine
Embodiment 4
The checking of clenbuterol content measure
Stability test
Take with a fresh meat, the pre-treatment of sample carried out using the detection method for implementing 1~3, after processing respectively 0,
2nd, 4,6 and 8h is measured using high performance liquid chromatography.The relative standard deviation (RSD) of clenbuterol hydrochloride measure content is shown in Table in 0~8h
3, relative standard deviation's Pass Test requirement.
Table 3 is the relative standard deviation that Clenbuterol, salbutamol and Ractopamine determine content
| Detection method | Clenbuterol | Salbutamol | Ractopamine |
| Embodiment 1 | 2.68% | 5.37% | 4.35% |
| Embodiment 2 | 2.79% | 5.12% | 4.65% |
| Embodiment 3 | 2.82% | 5.08% | 4.57% |
Average recovery is tested
Clenbuterol, salbutamol and Ractopamine standard mixed liquor are added to the negative live fresh pork without clenbuterol hydrochloride
In (quality 1.0g), it is 5,10 and 20 μ g/g to control mark-on amount.Sample pre-treatments are carried out using the detection method for implementing 1~3,
It is measured after processing using high performance liquid chromatography.The recovery of standard addition of clenbuterol hydrochloride is shown in Table 4, and mark-on reclaims rate score is average
Value, the rate of recovery are of a relatively high.
Table 4 is the mark-on reclaims amount of Clenbuterol, salbutamol and Ractopamine
In summary, detection method stability of the invention is good, and the rate of recovery is high；And spend the time not grow relatively, Ke Yiying
Detection for Clenbuterol, salbutamol and Ractopamine in fresh meat and meat products；Meanwhile find that embodiment 2 uses
Detection method measure clenbuterol content it is of a relatively high, reason is that the extraction efficiency of the detection method is higher.
Finally, it should be noted that above example to illustrate technical scheme rather than to the present invention protect
The limitation of scope, although being explained in detail with reference to preferred embodiment to the present invention, one of ordinary skill in the art should manage
Solution, technical scheme can be modified or replaced on an equal basis, without departing from technical solution of the present invention essence and
Scope.
Claims (10)
1. the detection method of clenbuterol hydrochloride in fresh meat and meat products, it is characterised in that the detection method is：Before first carrying out sample
Processing, sample solution is obtained, then gained sample solution is determined using high performance liquid chromatography；Wherein, the sample be fresh meat or
Meat products, sample pre-treatments comprise the following steps:
(1) digest：The sample blended is weighed, buffer solution and enzyme are added into sample, is centrifuged after digesting 10~30min, takes supernatant
Liquid produces sample enzymolysis liquid；The buffer solution is ammonium acetate solution, and the enzyme includes β-glucuronic acid glycosides peptase and aromatic sulfuric acid
Lipase；
(2) extract：The ratio that 6~10mL extracts reagents are added according to every gram of sample adds extraction examination into gained sample enzymolysis liquid
Agent, 5~10min of ultrasonic extraction, produces sample extracting solution；Wherein, the extracts reagent is made up of water and organic solvent, water and is had
The volume ratio of solvent is 1:3~1:5, and the organic solvent is dichloromethane, ethyl acetate, hexamethylene and dimethyl sulfoxide (DMSO)
At least one of；
(3) purify：Water absorbing agent and salting-out agents are added into gained sample extracting solution, is centrifuged after mixing, takes supernatant to produce sample
Scavenging solution；The mass ratio of the water absorbing agent and sample is 1:2~2:3, the mass ratio of the salting-out agents and sample is 1:1~2:1；
(4) concentrate：Nitrogen blows blows concentration by gained sample purification liquid progress nitrogen, obtains concentrate；
(5) redissolve：Using concentrate obtained by the mixed liquor dissolving step (4) being made up of acetic acid aqueous solution and ethanol, sample is obtained
Redissolve liquid；The volume fraction of acetic acid is 0.15%~0.25% in the acetic acid aqueous solution, the volume of acetic acid aqueous solution and ethanol
Than for 1:1~1:3；
(6) filter：Redissolved to sample obtained by step (5) in liquid and add adsorbent, centrifuged after mixing, take supernatant, reuse filter
Membrane filtration, produce sample solution.
2. detection method as claimed in claim 1, it is characterised in that in the step (1), enzymolysis time 20min.
3. detection method as claimed in claim 1, it is characterised in that in the step (2), organic solvent is dichloromethane.
4. detection method as claimed in claim 1, it is characterised in that in the step (2), organic solvent be ethyl acetate and
The mixed liquor of dimethyl sulfoxide (DMSO), and the volume ratio of ethyl acetate and dimethyl sulfoxide (DMSO) is 2:1.
5. detection method as claimed in claim 1, it is characterised in that in the step (2), 9mL is added according to every gram of sample
The ratio of extracts reagent adds extracts reagent into gained sample enzymolysis liquid, and in the extracts reagent, water and organic solvent
Volume ratio is 1:4.
6. detection method as claimed in claim 1, it is characterised in that in the step (3), the mass ratio of water absorbing agent and sample
For 3:5, the mass ratio of salting-out agents and sample is 1.5:1.
7. detection method as claimed in claim 1, it is characterised in that in the step (5), the body of acetic acid in acetic acid aqueous solution
Fraction is 0.20%, and the volume ratio of acetic acid aqueous solution and ethanol is 1:2.
8. the detection method as described in any one of claim 1~7, it is characterised in that the bar of the high performance liquid chromatography measure
Part is：
Chromatographic column is：C18Chromatographic column, column length 250mm, column internal diameter 4.6mm, particle diameter are 5 μm；
Detection wavelength is：243nm；
Flow velocity is：0.5mL/mim；
Sample size is：10μL；
Mobile phase is made up of mobile phase A and Mobile phase B, and mobile phase A is 0.05mol/ml ammonium acetate solutions, and in mobile phase A
Containing the acetic acid that volume fraction is 0.1%, Mobile phase B is ethanol.
9. detection method as claimed in claim 8, it is characterised in that the gradient elution program of the high performance liquid chromatography is:
When time is 0min, the volume fraction of mobile phase A is 75% in mobile phase, and the volume fraction of Mobile phase B is 25%；
When time is 5min, the volume fraction of mobile phase A is 60% in mobile phase, and the volume fraction of Mobile phase B is 40%；
When time is 8min, the volume fraction of mobile phase A is 60% in mobile phase, and the volume fraction of Mobile phase B is 40%；
When time is 13min, the volume fraction of mobile phase A is 40% in mobile phase, and the volume fraction of Mobile phase B is 60%；
When time is 15min, the volume fraction of mobile phase A is 40% in mobile phase, and the volume fraction of Mobile phase B is 60%.
10. detection method as claimed in claim 7, it is characterised in that the standard items of the high performance liquid chromatography measure are gram
Lun Teluo, salbutamol and Ractopamine, the solvent of standard solution is ethanol.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201710028489.7A CN107505417B (en) | 2017-01-16 | 2017-01-16 | Detection method of clenbuterol in fresh meat and meat product |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201710028489.7A CN107505417B (en) | 2017-01-16 | 2017-01-16 | Detection method of clenbuterol in fresh meat and meat product |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN107505417A true CN107505417A (en) | 2017-12-22 |
| CN107505417B CN107505417B (en) | 2020-08-04 |
Family
ID=60679298
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201710028489.7A Active CN107505417B (en) | 2017-01-16 | 2017-01-16 | Detection method of clenbuterol in fresh meat and meat product |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN107505417B (en) |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109612819A (en) * | 2018-11-28 | 2019-04-12 | 温州医科大学 | Based on pretreatment technology and its application associated with enzymatic hydrolysis auxiliary and solidification floating drop micro-extraction |
| CN111426678A (en) * | 2020-05-22 | 2020-07-17 | 合肥学院 | Method for detecting residual antibiotics in duck meat by using Raman instrument based on raspberry-shaped gold substrate |
-
2017
- 2017-01-16 CN CN201710028489.7A patent/CN107505417B/en active Active
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109612819A (en) * | 2018-11-28 | 2019-04-12 | 温州医科大学 | Based on pretreatment technology and its application associated with enzymatic hydrolysis auxiliary and solidification floating drop micro-extraction |
| CN111426678A (en) * | 2020-05-22 | 2020-07-17 | 合肥学院 | Method for detecting residual antibiotics in duck meat by using Raman instrument based on raspberry-shaped gold substrate |
Also Published As
| Publication number | Publication date |
|---|---|
| CN107505417B (en) | 2020-08-04 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN103760269B (en) | A kind of detection method of residue of veterinary drug | |
| CN103472160B (en) | Liquid chromatography-mass spectrometry method for rapidly detecting content of beta-agonists and sample pretreatment method | |
| CN101007072B (en) | Quality-control method of a traditional Chinese medicine 'Xuebijing' injection | |
| CN107290470B (en) | A kind of method of sulfamido and quinolones medicament relict in quick measurement egg | |
| CN104306745A (en) | Quality control method for rhizoma gastrodiae capsule | |
| CN107782835B (en) | A kind of method of efficient liquid phase detection fleabane flower ingredient | |
| CN106404932A (en) | Method for detecting beta-receptor agonist residual quantity in animal derived food | |
| CN109633012B (en) | Identification method of Zhejiang ophiopogon root | |
| CN110824032B (en) | Fingerprint identification method for bombyx batryticatus and counterfeit bombyx batryticatus | |
| CN102081077A (en) | Determination method of residual quantity of five sulfonamides in animal foods | |
| CN103543218A (en) | Method for measuring tetracycline antibiotic residue in protein-rich sample | |
| CN105974018B (en) | Method based on Multifunctional cleanup column-high performance liquid chromatography detection sitotoxismus flavine | |
| CN110907550B (en) | Method for simultaneously detecting contents of six components in infant lung-ventilating and cough-relieving granules | |
| CN107505417A (en) | The detection method of clenbuterol hydrochloride in fresh meat and meat products | |
| CN103235117A (en) | Beta 2-acceptor stimulant ELISA kit and usage method and application thereof | |
| CN108614052A (en) | The method for detecting gelsemine in biological sample, koumine and hexa-gelsemicine simultaneously based on two-dimensional liquid chromatography | |
| CN103217498B (en) | Method for detecting dicyandiamide in milk powder with LC-MS (liquid chromatography/mass spectrometry) and sample preparation method | |
| CN101961472B (en) | Detection method for lung-soothing syrup | |
| CN111426779A (en) | Method for determining related substances of pharmaceutical preparation containing acetaminophen, dextromethorphan hydrobromide and phenylephrine hydrochloride | |
| CN103267824B (en) | Method for detecting wind-cold cold granules | |
| CN102706984A (en) | Method for determining ephedrine hydrochloride content in lung-clearing inflammation pill by high-performance liquid phase | |
| CN109521123A (en) | A kind of application of PMP-HPLC method in garden ginsent and Ginseng under Forest identify | |
| CN107402277A (en) | The method of quality control of compound balloonflower root ephedrine syrup | |
| CN105181875B (en) | Method and sample pretreatment method for detecting content of chloramphenicol in silkworm faeces | |
| CN106198821A (en) | A kind of method detecting the residual of sulfa antibiotics in milk |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant |