CN106525800A - Quantitative detection method of salbutamol - Google Patents
Quantitative detection method of salbutamol Download PDFInfo
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Abstract
The invention discloses a quantitative detection method of salbutamol. The method comprises the following steps: carrying out a refluxing reaction on 3,4,9,10-perylenetetracarboxylic dianhydride and sodium hydroxide in an aqueous solution at 95-100 DEG C for 5-6 h, and acidifying the obtained reaction solution to prepare 3,4,9,10-tetracarboxylic acid, wherein a molar ratio of the 3,4,9,10-perylenetetracarboxylic dianhydride to the sodium hydroxide is 1:(3-5), and the concentration of the 3,4,9,10-perylenetetracarboxylic dianhydride in the aqueous solution is 0.04-0.06 mol/L. The quantitative detection method of salbutamol has the advantages of low cost, high detection precision, and simplicity in operation.
Description
Technical field
The present invention relates to a kind of detection method, specifically a kind of albuterol quantitative detecting method.
Background technology
Albuterol belongs to beta-receptor agonist, the nutritional labeling in animal body can be made to be shifted from fat to muscle, performance
Go out repartitioning function effect, and then the substance metabolism of regulation and control animal body, strengthen steatolysiss, promote protein synthesis, significantly carry
High carcass lean meat percentage and the price of deed.But, beta-receptor agonist accumulates residual easily in animal tissue, particularly internal organs, its
The symptoms such as muscular tremor, arrhythmia, headache can be caused after food chain enters human body, severe patient may threat to life.Mirror
In above reason, the beta-receptor agonist such as albuterol, Clenbuterol, Ractopamine was classified as in 1997 and is forbidden by China
Medicine used in feedstuff and cultivation domestic animal drinking water.But, still there is culturist by its illegal use in animal husbandry at present,
Cause drug residue, so as to cause serious harm to consumer health.It is saturating that Chinese patent CN104251898 discloses a kind of film
The method that analysis-Liquid Chromatography-Tandem Mass Spectrometry determines various beta-receptor agonist residuals in livestock products, comprises the following steps that：(1) sample
Product pre-treatment：The sample that 5g is crushed accurately is weighed, and adds 20ml0.02mol/L ammonium acetate buffers (pH4- to be tuned into acetic acid
5), mix, ultrasound 20 minutes, 8000 leave the heart 5 minutes, supernatant is moved in bag filter, bag filter immersion is filled into 400-
In the brown beaker of 500ml0.02mol/L ammonium acetate buffers (pH4-5), rotor is put into, beaker is placed on magnetic stirring apparatuss
Dialysed, speed of agitator drives bag filter to rotate to be formed to be vortexed in cup, and bag filter does not sink to being defined, opaque on cup outer housing
Sack, time are 6h.Bag filter is taken out after the completion of dialysis, the dialysis solution in beaker is adjusted to into pH to 9-10, move into the pyriform of 1L
In shunting funnel, 40ml solvent (hexamethylene is added:Ethyl acetate=4:1), extract 2 times, combining extraction liquid is rotated, use
0.1% aqueous formic acid (v/v) constant volume is determined for HPLC-MS/MS and is analyzed into 1.0ml；(2) HPLC-MS/MS is determined：Using outer
Mark method determines the content of beta-receptor agonist, and liquid-phase chromatographic column adopts Thermo Hypersil Gold (2.1mm × 100mm, 3 μ
M), filler is selected from Octadecylsilane bonded phase silica gel, and mobile phase is 0.1% aqueous formic acid A- acetonitrile B, and flow velocity is 0.2ml/min,
Condition of gradient elution is：T=0min, 96%A, 4%B；T=2min, 96%A, 4%B；T=5min, 77%A, 23%B；T=
12min, 5%A, 95%B；T=28min, 44%A, 56%B；T=24min, 5%A, 95%B；T=24.1min, 96%A,
4%B；T=32min, 96%A, 4%B；Mass Spectrometry Conditions are：Electron spray ionisation source cation scanning ESI+, multiple-reaction monitoring pattern
MRM；Electron spray voltage：5.5kV；Ion source temperature：550 DEG C, atomization gas pressure GSI：40psi；Secondary air speed GS2：
60psi, gas curtain atmospheric pressure CUR：40psi；Collision cell entrance potential EP：10V；Collision cell exit potential CXP：13V, monitoring analysis
The ion pair of thing carries out drug residue confirmatory analysis.Said method first carries out sample pre-treatments using film dialysis, adopts again afterwards
Liquid Chromatography-Tandem Mass Spectrometry determines various beta-receptor agonist in livestock products, however, above-mentioned detection method has sample pre-treatments
It is complicated, detection process is loaded down with trivial details, analysis cost is high, detection cycle length, expensive equipment, the problems such as be difficult to operation, it is difficult to carry out scene
Operation, is restricted in actual applications.
The content of the invention
It is an object of the invention to provide a kind of albuterol quantitative detecting method, to solve to propose in above-mentioned background technology
Problem.
For achieving the above object, the present invention provides following technical scheme：
A kind of albuterol quantitative detecting method, by 3,95 DEG C in aqueous of 4,9,10- tetracarboxylic acid dianhydrides and sodium hydroxide
Back flow reaction 5h-6h at～100 DEG C, then reactant liquor is acidified, you can 3,4,9,10- tetracids are obtained；Described 3,4,9,
10- tetracarboxylic acid dianhydrides are 1 with the ratio of the amount of the material of sodium hydroxide：3～5；3,4,9,10- tetracarboxylic acid dianhydrides are in aqueous solution
In concentration be 0.04～0.06mol/L；Solution used by acidifying is hydrochloric acid solution, its with 3,4,9,10- tetracarboxylic acid dianhydrides
The ratio of the amount of material is 1：20～30；By to 3, in the compound system of 4,9,10- tetracids/cetyl trimethylammonium bromide
The albuterol of variable concentrations is added, by the line style relation between albuterol concentration and fluorescent quenching ratio (F0-F)/F0
Detection by quantitative goes out the concentration of albuterol.
As further scheme of the invention：F0, F are respectively 3,4,9,10- tetracids/cetyl trimethylammonium bromide
Compound system add the fluorescence intensity level before and after albuterol at the 488nm.
As further scheme of the invention：Also comprise the steps：A. multigroup 3 are configured, 4,9,10- tetracid/ten
The composite solution of six alkyl trimethyl ammonium bromides：Respectively by 3,4,9,10- tetra- aqueous acids and cetyl trimethyl bromination
Aqueous ammonium mixes, and reacts 2～5 minutes；B. concentration range is sequentially added in 0～60Um in above-mentioned composite solutionSand
Butylamine alcohol-water solution and the Na2HPO4-Na3PO4 buffer solution that pH value is 9.0～10.6, and deionized water is diluted to 10mL,
Continue stirring reaction 20～30 minutes, the fluorescence intensity of measurement system；C. with albuterol concentration as X-axis, corresponding fluorescence is quenched
Ratio of going out (F0-F)/F0 maps for Y-axis, obtains the linear side between albuterol concentration and fluorescent quenching ratio (F0-F)/F0
Journey, the albuterol concentration according to corresponding to linear equation can calculate any fluorescence intensity F；Described 3,4,9,10- tetracids
2.75 × 10-7mol is respectively with the final concentration of cetyl trimethylammonium bromideWith 2.5 × 10-6mol；The husky fourth
The final concentration of amine alcohol is respectively 0,1.0Um、2.0Um、2.75Um、3.5Um、4.7Um、5.5UmWith
6.0Um。
Compared with prior art, the invention has the beneficial effects as follows：Albuterol quantitative detecting method of the present invention has cost
Low, accuracy of detection is high, simple operation and other advantages.
Specific embodiment
Below the technical scheme in the embodiment of the present invention is clearly and completely described.
In the embodiment of the present invention, a kind of albuterol quantitative detecting method, by 3,4,9,10- tetracarboxylic acid dianhydrides and hydrogen-oxygen
Change back flow reaction 5h-6h at 95 DEG C in aqueous～100 DEG C of sodium, then reactant liquor is acidified, you can be obtained 3,4,9,10-
Tetracid；Described 3,4,9,10- tetracarboxylic acid dianhydrides are 1 with the ratio of the amount of the material of sodium hydroxide：3～5；3,4,9,10-
Tetracarboxylic acid dianhydride concentration in aqueous is 0.04～0.06mol/L；, solution used by acidifying is hydrochloric acid solution, its with 3,4,
The ratio of the amount of the material of 9,10- tetracarboxylic acid dianhydrides is 1：20～30；By to 3,4,9,10- tetracids/cetyl front three
The albuterol of variable concentrations is added in the compound system of base ammonium bromide, by albuterol concentration and fluorescent quenching ratio (F0-
F the line style relation detection by quantitative between)/F0 goes out the concentration of albuterol.F0, F are respectively 3,4,9,10- tetracids/hexadecane
The compound system of base trimethylammonium bromide adds the fluorescence intensity level before and after albuterol at the 488nm.Also comprise the steps：
A. multigroup 3 are configured, the composite solution of 4,9,10- tetracids/cetyl trimethylammonium bromide：Respectively by 3,4,9,10- tetra-
Aqueous acid and cetyl trimethylammonium bromide aqueous solution, react 2～5 minutes；B. in above-mentioned composite solution successively
Concentration range is added in 0～60UmAlbuterol aqueous solutions and pH value be 9.0～10.6 Na2HPO4-Na3PO4 bufferings
Solution, and deionized water is diluted to 10mL, continues stirring reaction 20～30 minutes, the fluorescence intensity of measurement system；C. with sand
Butylamine determining alcohol is X-axis, and corresponding fluorescent quenching ratio (F0-F)/F0 is that Y-axis is mapped, and obtains albuterol concentration sudden with fluorescence
Linear equation between ratio of going out (F0-F)/F0, the husky butylamine according to corresponding to linear equation can calculate any fluorescence intensity F
Determining alcohol；Described 3,4,9,10- tetracids are respectively 2.75 × 10-7mol with the final concentration of cetyl trimethylammonium bromideWith 2.5 × 10-6mol；The final concentration of the albuterol is respectively 0,1.0Um、2.0Um、2.75Um、
3.5Um、4.7Um、5.5UmAnd 6.0Um。
Embodiment 1：
Albuterol quantitative detecting method of the present invention, by 3,4,9,10- tetracarboxylic acid dianhydrides and sodium hydroxide in aqueous 95
DEG C～100 DEG C at back flow reaction 5h, then reactant liquor is acidified, you can 3,4,9,10- tetracids are obtained；Described 3,4,9,
10- tetracarboxylic acid dianhydrides are 1 with the ratio of the amount of the material of sodium hydroxide：3；3,4,9,10- tetracarboxylic acid dianhydrides are in aqueous
Concentration be 0.04～0.06mol/L；, solution used by acidifying is hydrochloric acid solution, its with 3, the thing of 4,9,10- tetracarboxylic acid dianhydrides
The ratio of the amount of matter is 1：20；By to 3, adding not in the compound system of 4,9,10- tetracids/cetyl trimethylammonium bromide
With the albuterol of concentration, quantitatively examined by the line style relation between albuterol concentration and fluorescent quenching ratio (F0-F)/F0
Measure the concentration of albuterol.F0, F are respectively 3, and the compound system of 4,9,10- tetracids/cetyl trimethylammonium bromide adds
Enter the fluorescence intensity level at the 488nm before and after albuterol.Also comprise the steps：A. multigroup 3 are configured, 4,9,10- tetracids/
The composite solution of cetyl trimethylammonium bromide：Respectively by 3,4,9,10- tetra- aqueous acids and cetyl trimethyl bromine
Change aqueous ammonium mixing, react 2 minutes；B. concentration range is sequentially added in 0～60Um in above-mentioned composite solutionHusky fourth
Amine alcohol aqueous solution and the Na2HPO4-Na3PO4 buffer solution that pH value is 9.0, and deionized water is diluted to 10mL, continues stirring
Reaction 20 minutes, the fluorescence intensity of measurement system；C. with albuterol concentration as X-axis, corresponding fluorescent quenching ratio (F0-F)/
F0 maps for Y-axis, obtains the linear equation between albuterol concentration and fluorescent quenching ratio (F0-F)/F0, according to linear side
Journey can calculate the albuterol concentration corresponding to any fluorescence intensity F；Described 3,4,9,10- tetracids and cetyl front three
The final concentration of base ammonium bromide is respectively 2.75 × 10-7molWith 2.5 × 10-6mol；The final concentration of the albuterol point
Not Wei 0,1.0Um、2.0Um、2.75Um、3.5Um、4.7Um、5.5UmAnd 6.0Um。
Embodiment 2：
Albuterol quantitative detecting method of the present invention, by 3,4,9,10- tetracarboxylic acid dianhydrides and sodium hydroxide in aqueous 95
DEG C～100 DEG C at back flow reaction 6h, then reactant liquor is acidified, you can 3,4,9,10- tetracids are obtained；Described 3,4,9,
10- tetracarboxylic acid dianhydrides are 1 with the ratio of the amount of the material of sodium hydroxide：5；3,4,9,10- tetracarboxylic acid dianhydrides are in aqueous
Concentration be 0.04～0.06mol/L；, solution used by acidifying is hydrochloric acid solution, its with 3, the thing of 4,9,10- tetracarboxylic acid dianhydrides
The ratio of the amount of matter is 1：30；By to 3, adding not in the compound system of 4,9,10- tetracids/cetyl trimethylammonium bromide
With the albuterol of concentration, quantitatively examined by the line style relation between albuterol concentration and fluorescent quenching ratio (F0-F)/F0
Measure the concentration of albuterol.F0, F are respectively 3, and the compound system of 4,9,10- tetracids/cetyl trimethylammonium bromide adds
Enter the fluorescence intensity level at the 488nm before and after albuterol.Also comprise the steps：A. multigroup 3 are configured, 4,9,10- tetracids/
The composite solution of cetyl trimethylammonium bromide：Respectively by 3,4,9,10- tetra- aqueous acids and cetyl trimethyl bromine
Change aqueous ammonium mixing, react 5 minutes；B. concentration range is sequentially added in 0～60Um in above-mentioned composite solutionHusky fourth
Amine alcohol aqueous solution and the Na2HPO4-Na3PO4 buffer solution that pH value is 10.6, and deionized water is diluted to 10mL, continues to stir
Mix reaction 30 minutes, the fluorescence intensity of measurement system；C. with albuterol concentration as X-axis, corresponding fluorescent quenching ratio (F0-
F)/F0 maps for Y-axis, obtains the linear equation between albuterol concentration and fluorescent quenching ratio (F0-F)/F0, according to linear
Equation can calculate the albuterol concentration corresponding to any fluorescence intensity F；Described 3,4,9,10- tetracids and cetyl three
The final concentration of methyl bromide ammonium is respectively 2.75 × 10-7molWith 2.5 × 10-6mol；The final concentration of the albuterol
Respectively 0,1.0Um、2.0Um、2.75Um、3.5Um、4.7Um、5.5UmAnd 6.0Um。
Albuterol quantitative detecting method, by 3,95 DEG C in aqueous of 4,9,10- tetracarboxylic acid dianhydrides and sodium hydroxide
Back flow reaction 5h-6h at～100 DEG C, then reactant liquor is acidified, you can 3,4,9,10- tetracids are obtained；Described 3,4,9,
10- tetracarboxylic acid dianhydrides are 1 with the ratio of the amount of the material of sodium hydroxide：3～5；3,4,9,10- tetracarboxylic acid dianhydrides are in aqueous solution
In concentration be 0.04～0.06mol/L；, solution used by acidifying is hydrochloric acid solution, its with 3,4,9,10- tetracarboxylic acid dianhydrides
The ratio of the amount of material is 1：20～30；By to 3, in the compound system of 4,9,10- tetracids/cetyl trimethylammonium bromide
The albuterol of variable concentrations is added, by the line style relation between albuterol concentration and fluorescent quenching ratio (F0-F)/F0
Detection by quantitative goes out the concentration of albuterol.F0, F are respectively 3,4,9,10- tetracids/cetyl trimethylammonium bromide it is compound
System adds the fluorescence intensity level before and after albuterol at the 488nm.Also comprise the steps：A. multigroup 3,4,9,10- are configured
The composite solution of tetracid/cetyl trimethylammonium bromide：Respectively by 3,4,9,10- tetra- aqueous acids and cetyl three
Methyl bromide aqueous ammonium mixes, and reacts 2～5 minutes；B. concentration range is sequentially added in 0～60Um in above-mentioned composite solutionAlbuterol aqueous solutions and pH value be 9.0～10.6 Na2HPO4-Na3PO4 buffer solution, and deionized water dilution
To 10mL, continue stirring reaction 20～30 minutes, the fluorescence intensity of measurement system；C. it is with albuterol concentration as X-axis, corresponding
Fluorescent quenching ratio (F0-F)/F0 maps for Y-axis, obtains between albuterol concentration and fluorescent quenching ratio (F0-F)/F0
Linear equation, the albuterol concentration according to corresponding to linear equation can calculate any fluorescence intensity F；Described 3,4,9,10-
Tetracid is respectively 2.75 × 10-7mol with the final concentration of cetyl trimethylammonium bromideWith 2.5 × 10-6mol；Institute
The final concentration for stating albuterol is respectively 0,1.0Um、2.0Um、2.75Um、3.5Um、4.7Um、5.5UmAnd 6.0Um。
Embodiment 3：
Albuterol quantitative detecting method of the present invention, by 3,4,9,10- tetracarboxylic acid dianhydrides and sodium hydroxide in aqueous 95
DEG C～100 DEG C at back flow reaction 6h, then reactant liquor is acidified, you can 3,4,9,10- tetracids are obtained；Described 3,4,9,
10- tetracarboxylic acid dianhydrides are 1 with the ratio of the amount of the material of sodium hydroxide：4；3,4,9,10- tetracarboxylic acid dianhydrides are in aqueous
Concentration be 0.05mol/L；, solution used by acidifying is hydrochloric acid solution, its with 3, the amount of the material of 4,9,10- tetracarboxylic acid dianhydrides
Ratio be 1：25；By to 3, adding variable concentrations in the compound system of 4,9,10- tetracids/cetyl trimethylammonium bromide
Albuterol, sand is gone out by the line style relation detection by quantitative between albuterol concentration and fluorescent quenching ratio (F0-F)/F0
The concentration of butylamine alcohol.F0, F are respectively 3, and the compound system of 4,9,10- tetracids/cetyl trimethylammonium bromide adds husky fourth
Fluorescence intensity level before and after amine alcohol at the 488nm.Also comprise the steps：A. multigroup 3 are configured, 4,9,10- tetracids/hexadecane
The composite solution of base trimethylammonium bromide：Respectively by 3,4,9,10- tetra- aqueous acids and cetyl trimethylammonium bromide water
Solution mixes, and reacts 4 minutes；B. concentration range is sequentially added in 0～60Um in above-mentioned composite solutionAlbuterol water
Solution and the Na2HPO4-Na3PO4 buffer solution that pH value is 10.6, and deionized water is diluted to 10mL, continues stirring reaction
25 minutes, the fluorescence intensity of measurement system；C. with albuterol concentration as X-axis, corresponding fluorescent quenching ratio (F0-F)/F0 is
Y-axis is mapped, and obtains the linear equation between albuterol concentration and fluorescent quenching ratio (F0-F)/F0, can according to linear equation
Calculate the albuterol concentration corresponding to any fluorescence intensity F；Described 3,4,9,10- tetracids and cetyl trimethyl bromine
The final concentration for changing ammonium is respectively 2.75 × 10-7molWith 2.5 × 10-6mol；The final concentration of the albuterol is respectively
0、1.0Um、2.0Um、2.75Um、3.5Um、4.7Um、5.5UmAnd 6.0Um。
It is obvious to a person skilled in the art that the invention is not restricted to the details of above-mentioned one exemplary embodiment, Er Qie
In the case of spirit or essential attributes without departing substantially from the present invention, the present invention can be realized in other specific forms.Therefore, no matter
From the point of view of which point, embodiment all should be regarded as exemplary, and be nonrestrictive, the scope of the present invention is by appended power
Profit is required rather than described above is limited, it is intended that all in the implication and scope of the equivalency of claim by falling
Change is included in the present invention.Moreover, it will be appreciated that although this specification is been described by according to embodiment, not each
Embodiment only includes an independent technical scheme, and this narrating mode of description is only this area for clarity
Technical staff should using description as an entirety, the technical scheme in each embodiment can also Jing it is appropriately combined, form this
Art personnel may be appreciated other embodiment.
Claims (3)
1. a kind of albuterol quantitative detecting method, it is characterised in that by 3,4,9,10- tetracarboxylic acid dianhydrides are existed with sodium hydroxide
Back flow reaction 5h-6h at 95 DEG C～100 DEG C in aqueous solution, then reactant liquor is acidified, you can it is obtained 3,4,9,10- tetra-
Acid；Described 3,4,9,10- tetracarboxylic acid dianhydrides are 1 with the ratio of the amount of the material of sodium hydroxide：3～5；3,4,9,10- tetramethyls
Acid dianhydride concentration in aqueous is 0.04～0.06mol/L；, solution used by acidifying is hydrochloric acid solution, and which is with 3,4,9,10-
The ratio of the amount of the material of tetracarboxylic acid dianhydride is 1：20～30；By to 3,4,9,10- tetracids/cetyl trimethyl bromination
The albuterol of variable concentrations is added in the compound system of ammonium, by albuterol concentration and fluorescent quenching ratio (F0-F)/F0
Between line style relation detection by quantitative go out the concentration of albuterol.
2. albuterol quantitative detecting method according to claim 1, it is characterised in that F0, F are respectively 3,4,9,10-
The compound system of tetracid/cetyl trimethylammonium bromide adds the fluorescence intensity level before and after albuterol at the 488nm.
3. albuterol quantitative detecting method according to claim 1, it is characterised in that also comprise the steps：A. match somebody with somebody
Multigroup 3 are put, the composite solution of 4,9,10- tetracids/cetyl trimethylammonium bromide：Respectively by 3,4,9,10- tetra- sour waters
Solution and cetyl trimethylammonium bromide aqueous solution, react 2～5 minutes；B. sequentially add in above-mentioned composite solution
Concentration range is in 0～60UmAlbuterol aqueous solutions it is molten with Na2HPO4-Na3PO4 that pH value is 9.0～10.6 buffering
Liquid, and deionized water is diluted to 10mL, continues stirring reaction 20～30 minutes, the fluorescence intensity of measurement system；C. with husky fourth
Amine alcohol concentration is X-axis, and corresponding fluorescent quenching ratio (F0-F)/F0 is that Y-axis is mapped, and obtains albuterol concentration and fluorescent quenching
Linear equation between ratio (F0-F)/F0, the albuterol according to corresponding to linear equation can calculate any fluorescence intensity F
Concentration；Described 3,4,9,10- tetracids are respectively 2.75 × 10-7mol with the final concentration of cetyl trimethylammonium bromide
With 2.5 × 10-6mol；The final concentration of the albuterol is respectively 0,1.0Um、2.0Um、2.75Um、
3.5Um、4.7Um、5.5UmAnd 6.0Um。
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|---|---|---|---|
| CN201611025686.5A CN106525800A (en) | 2016-11-22 | 2016-11-22 | Quantitative detection method of salbutamol |
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Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104792762A (en) * | 2015-05-06 | 2015-07-22 | 江西省农业科学院农产品质量安全与标准研究所 | Method for quantitatively detecting salbutamol by using CdTe quantum dots |
| CN105928912A (en) * | 2016-04-14 | 2016-09-07 | 安徽师范大学 | Heparin detection method |
-
2016
- 2016-11-22 CN CN201611025686.5A patent/CN106525800A/en active Pending
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104792762A (en) * | 2015-05-06 | 2015-07-22 | 江西省农业科学院农产品质量安全与标准研究所 | Method for quantitatively detecting salbutamol by using CdTe quantum dots |
| CN105928912A (en) * | 2016-04-14 | 2016-09-07 | 安徽师范大学 | Heparin detection method |
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