Solid phase extraction method for detection of β-agonist agonist in animal-derived foods (PolybaseTM MCX)
First, the purpose of the experiment
In this study, solid phase extraction was used as a pretreatment method for pork samples, and LC-MS/MS method was used as a detection method. The method can simplify the pretreatment process of pork samples, save the use of organic solvents, and is easy to operate.
Second, the experimental target
Five β-receptor agonist substances: clenbuterol (CAS: 37148-27-9), salbutamol (CAS: 18559-94-9), ractopamine (CAS: 97825-25-7), terbuta Lin (CAS: 23031-25-6), Tobbutro (CAS: 41570-61-0).
Third, the scope of application
The method is applicable to the detection and confirmation of β-agonist LC-MS/MS in animal-derived foods.
Fourth, the reference standard
Recommended national standard GB/T 21313-2007 Method for the determination of β-agonist residues in animal-derived foods by liquid chromatography-mass spectrometry/mass spectrometry
V. Experimental materials
PolybaseTM MCX Solid Phase Extraction Column 6mL/200mg.
Sixth, experimental methods
1, sample extraction
Weigh 10.0 g of the homogeneous sample (accurate to 0.01 g), add 15 mL of acetic acid-ammonium acetate buffer solution of pH 5.2, homogenate for 1000 min at 1000 r/min, and then add β-glucuronidase/ 100 μL of the aromatic sulfonate solution was hydrolyzed overnight at 37 ° C ± 1 ° C. After taking out the cooling, it was centrifuged at 9500 r/min for 10 min at 10 °C. Take all the supernatant and purify it.
2, SPE column purification
(1) Activation: Activation was carried out by adding 6 mL of 5% methanolic ammonia, 6 mL of methanol, 6 mL of water, and 6 mL of 0.1 mmol/L perchloric acid (pH 4.0) solution.
(2) Loading and elution: When the solvent liquid level reaches the surface of the column adsorption layer, immediately pour the above-mentioned solution to be purified, pass the column, discard the filtrate, rinse with 2mL methanol, 2mL 2% methanol aqueous solution, and finally use 7mL5 % methanol ammonia eluted.
(3) Re-dissolution: The eluate was blown to near dryness under a slow nitrogen flow condition, dissolved to 1.0 mL with methanol-0.1% formic acid aqueous solution, and vortexed for 1 min for LC-MS/MS measurement.
3. LC-MS/MS conditions
Column: Venusil ASB C18 (2.1 × 150 mm, 5 μm, 100 Å);
Mass spectrometer: API 4000
Mobile phase: A: 10 mM ammonium acetate (0.1% formic acid) solution B: methanol solution;
Table 1 Gradient elution conditions
Time/min | B/% | A/% | 0.00 | 5 | 95 | 4.00 | 50 | 50 | 6.00 | 10 | 90 | 8.00 | 95 | 5 | 8.01 | 5 | 95 | 12.00 | Stop | |
|
Flow rate: 0.2mL/min
Column temperature: 40 ° C
Injection volume: 2μL
Ion source: electrospray (ESI), positive ion mode
Detection method: multiple reaction monitoring (MRM) |
Table 2 mass spectrometer ion source parameters | Source/Gas | | Collision Gas (CAD) | 10 | | Curtain Gas (CUR) | 30 | | Ion Source Gas 1 (GS 1) | 50 | | Ion Source Gas 2 (GS 2) | 50 | | Ion Spray Voltage(IS) | 5500 | | Temperature(TEM) | 550 | | Interface Heater(ihe) | On |
|
Table 3 Table of parent and daughter ion parameters of β-agonists | Substance name | Retention time (min) | Detecting ion pairs | DP | EP | CE | CXP | Salbutamol | 3.86 | 240.1/147.8 | 60 | 10 | 18 | 12 | 240.1/221.9 | 60 | 10 | 12 | 12 | Clenbuter | 5.82 | 277.1/202.9 | 60 | 10 | twenty three | 12 | 277.1/259.0 | 60 | 10 | 15 | 12 | Lake dopamine | 5.45 | 302.1/163.7 | 60 | 10 | twenty three | 12 | 302.1/284.2 | 60 | 10 | 19 | 12 | Terbutaline | 3.77 | 226.0/151.7 | 60 | 10 | 16 | 12 | 226.0/169.8 | 60 | 10 | 12 | 12 | Tobbuto | 6.17 | 227.9/153.8 | 60 | 10 | 16 | 12 | 227.9/171.8 | 60 | 10 | 12 | 12 |
|
Seven, the experimental results
1. 0.5 ppb pork matrix spiked recovery results: The recovery rates of the five analytes were all greater than 80%, and the RSD value was less than 5, meeting the standard requirements. Table 4 Results of 0.5ppb spike recovery experiment in pork | name | 1(%) | 2(%) | 3 (%) | The average recovery rate(%) | RSD (%) | Salbutamol | 88.62 | 82.79 | 81.45 | 84.29 | 4.52 | Clenbuter | 85.32 | 88.29 | 86.59 | 86.73 | 1.72 | Lake dopamine | 86.56 | 88.33 | 81.36 | 85.42 | 4.24 | Terbutaline | 81.95 | 80.95 | 87.26 | 83.39 | 4.06 | Tobbuto | 82.64 | 82.56 | 88.14 | 84.45 | 3.79 |
|
2, blank sample added β-receptor agonist analyte chromatogram ![]()
![]()
![]()
|
Related Products PolybaseTM MCX Solid Phase Extraction Column Related applications | Numbering | Data name | Data category | | MCX-003-1 | Solid phase extraction method for detection of β-agonist agonist in animal-derived foods (PolybaseTM MCX) | Application manual | | MCX-002-1 | Solid phase extraction method for MCX malachite green residue detection in aquatic products (PolybaseTM MCX) | Application manual | | MCX-001-1 | Solid phase extraction method for detecting papaverine residues in hot pot sauce (PolybaseTM MCX) | Application manual | | FLORISIL-001-1 | Solid phase extraction method for detection of multiple pesticide residues in fruits and vegetables (SilibaseTM Florisil) ![comma]() | Application manual | | HLB001-1 | Solid phase extraction method for the detection of quinolones in animal-derived foods (PolybaseTM HLB) ![comma]() | Application manual | | NH2 -002-1 | Solid phase extraction method for detection of multiple pesticide residues in vegetables (SilibaseTM NH2) ![comma]() | Application manual | | NH2- 001-1 | Solid phase extraction method for the detection of aldicarb and its metabolites in ginger (SilibaseTM NH2) | Application manual | | Carb-GCB-001-1 | Solid phase extraction method for detection of multiple pesticide residues in vegetables (SilibaseTM Carb-GCB) | Application manual | | Carb-PSA-001-1 | Solid phase extraction method for detection of organophosphorus pesticide residues in vegetables (SilibaseTM Carb-GCB-PSA) ![comma]() | Application manual | | Carb-GCB-NH2-001-1 | Solid phase extraction method for detection of multiple pesticide residues in vegetables (SilibaseTM Carb-GCB-NH2) ![comma]() | Application manual | | C18-002-1 | Solid phase extraction method for detection of organophosphorus pesticide residues in vegetables (SilibaseTM C18) ![comma]() | Application manual | | C18-001-1 | Solid phase extraction method for detection of chloramphenicol in animal-derived foods (SilibaseTM C18) ![comma]() | Application manual | | C8-SCX-003-1 | Solid phase extraction method for the detection of nitrofurans in animal-derived foods (SilibaseTM C8-SCX) ![comma]() | Application manual | | C8-SCX-001-1 | Solid phase extraction method for detection of clenbuterol hydrochloride in animal-derived foods (SilibaseTM C8-SCX) ![comma]() | Application manual | | AZO-001-1 | Solid phase extraction method for the inhibition of azo dye detection in textiles (SilibaseTM azo dye detection column) ![comma]() | Application manual | | ALN-002-1 | Solid phase extraction method for detection of sulfonamides in animal-derived foods (SilibaseTM ALN) | Application manual | | ALN-001-1 | Solid phase extraction method for detection of Sudan red dye in food (SilibaseTM ALN) ![comma]() | Application manual | | SLE-004-1 | Rapid extraction method for detection of non-steroidal anti-inflammatory drugs in serum ![comma]() | Application manual | | SLE-002-1 | Rapid extraction method for detection of tricyclic antidepressants in urine ![comma]() | Application manual | | SLE-001-1 | Rapid extraction method for detection of ibuprofen in serum ![comma]() | Application manual |
|
![Download this document]()
|
|
Non Contact Termometer
Non Contact Termometer,Smart thermometer,ear thermometer,best baby thermometer,forehead thermometer
Shenzhen Urion Technology Co., Ltd. , https://www.urionhealth.com