Solid phase extraction method for detection of MCX malachite green residue in aquatic products (PolybaseTM MCX)
First, the purpose of the experiment
In this study, solid phase extraction was used as a pretreatment method for the determination of papaverine residues in hot pot sauce, and LC-MS/MS was used as a detection method. The method can simplify the pretreatment process of the hot pot sauce sample, save the use of organic solvent, and is easy to operate.
Second, the experimental target
Malachite Green Standard: Malachite Green (CAS: 569-64-2).
Third, the scope of application
The method is applicable to the detection and confirmation of malachite green LC-MS/MS in fresh aquatic products and their products.
Fourth, the reference standard
Recommended national standard "Determination of malachite green and crystal violet residues in aquatic products GB/T 19857-2005".
V. Experimental materials
PolybaseTM MCX Solid Phase Extraction Column 3mL/60mg.
Sixth, experimental methods
1, sample extraction
Weigh 5.0g sample (accurate to 0.01g) of homogenous sample in 50mL centrifuge tube, add 10mL acetonitrile, 10000r/min homogenate for 30s, add 5g acid alumina, shake for 2min, centrifuge at 4000r/min for 10min, The supernatant was transferred to a 125 mL separatory funnel, and 2 mL of diethylene glycol, 3 mL of potassium borohydride solution was added to the separatory funnel, and shaken for 2 min.
Another 50mL centrifuge tube was added 10mL acetonitrile, the homogenizer head was washed for 10s, the washing liquid was transferred into the previous centrifuge tube, 3mL potassium borohydride solution was added, the precipitate in the centrifuge tube was dispersed with glass and mixed well, vortexed and mixed. The device was shaken for 1 min, allowed to stand for 20 min, centrifuged at 4000 r/min for 10 min, and the supernatant was incorporated into a 125 mL separatory funnel.
Continue to add 1.5mL hydroxylamine hydrochloride solution, 2.5mL p-toluenesulfonic acid solution, 5.0mL ammonium acetate buffer solution in the centrifuge tube, shake for 2min, then add 10mL acetonitrile, continue to shake for 2min, centrifuge at 4000r/min for 10min, the supernatant is incorporated. The above operation was repeated once in a 125 mL separatory funnel.
Add 20 mL of dichloromethane to the separatory funnel, stopper, shake vigorously for 2 min, let stand for stratification, transfer the lower layer solution to 250 mL chicken heart bottle, continue to add 5 mL acetonitrile, 10 mL dichloromethane, shake in the separatory funnel After shaking for 2 min, the whole solution was transferred to a 50 mL centrifuge tube, centrifuged at 4000 r/min for 10 min, and the lower layer solution was combined into a 250 mL chicken heart bottle, evaporated to near dryness at 45 ° C, and the residue was dissolved with 2.5 mL of acetonitrile. .
2, SPE column purification
(1) Activation: Add 5 mL of acetonitrile to the MCX composite column.
(2) Loading and elution: The above-mentioned stock solution was passed through a column and eluted by adding 3 mL of acetonitrile-ammonium acetate (volume ratio: 1:1).
(3) Concentration and concentration: The eluate was collected, and the volume of acetonitrile was adjusted to 3 mL, and passed through a 0.45 um filter for liquid chromatography-mass spectrometry.
3. LC-MS/MS conditions
Column: XR-ODSIIIC18 (2.1 × 150 mm, 2.2 μm, 100 Å);
Mass spectrometer: API 4000
Mobile phase: A: acetonitrile (0.1% formic acid); B: 0.1% formic acid water column: XR-ODSIIIC18 (2.1 × 150 mm, 2.2 μm, 100 Å);
Mass spectrometer: API 4000
Mobile phase: A: acetonitrile (0.1% formic acid); B: 0.1% formic acid water
Table 1 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 2 Malachite green and internal standard parent ion and daughter ion parameter table
| Time/min | Retention time (min) | Detecting ion pairs | DP | EP | CE | CXP |
| Malachite green | 4..95 | 329.0/313.0 | 70 | 10 | 35 | 12 |
| 329.0/208.0 | 70 | 10 | 35 | 12 | ||
| Malachite green -D5 | 5.11 | 334.0/318.0 | 86 | 10 | 35 | 12 |
Seven, the experimental results
1, Wuchang fish substrate 10μg / L malachite green plus standard recovery results:
The average recovery of the three parallel test samples was greater than 80%, and the relative standard deviations of the parallel samples were less than 5%, which met the standard requirements. (See Table 3)
Table 3 The recovery result of 10μg/L malachite green in Wuchang fish matrix
| name | 1(%) | 2(%) | 3 (%) | The average recovery rate(%) | RSD (%) |
| Malachite green | 93.26 | 93.34 | 96.06 | 94.89 | 1.53 |
2. Adding the chromatogram of aldicarb and its metabolites to the blank sample
Related Products
PolybaseTM MCX Solid Phase Extraction Column
Related applications
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