Solid phase extraction method for detection of papaverine residues in hot pot sauce (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

Five papaverine standards: morphine (CAS: 57-27-2), codeine (CAS: 76-57-3), ibaine (CAS: 115-37-7), papaverine (CAS: 58 -74-2), Nastin (CAS: 912-60-7).

Third, the scope of application

The method is suitable for LC-MS/MS detection and confirmation of papaverine, morphine, narcotine, codeine and thebaine in hot pot food such as hot pot sauce, soup, seasoning oil and solid seasoning powder.

Fourth, the reference standard

Shanghai local standard "DB31/2010-2012 hot pot food in the determination of papaverine, morphine, narcotine, codeine and thebaine liquid chromatography-tandem mass spectrometry."

V. Experimental materials

PolybaseTM MCX Solid Phase Extraction Column 3mL/60mg.

Sixth, experimental methods

1, sample extraction

Weigh 2g sample (accurate to 0.01g) in a 50mL centrifuge tube, add 5mL water, shake to evenly disperse, add 15mL acetonitrile, vortex for 1min, add 6g anhydrous magnesium sulfate and 1.5g anhydrous sodium acetate The powder was mixed, shaken for 2 min, centrifuged at 4000 r/min for 10 min, and the supernatant was to be purified.

2, SPE column purification

(1) Activation: 5 mL of methanol and 5 mL of 0.1 M hydrochloric acid were sequentially added to the MCX composite column cartridge to activate.
(2) Loading and elution: The above supernatant was passed through a column, washed successively with 5 mL of water, 5 mL of methanol water (1+1), vacuumed for 10 min under vacuum, and finally eluted with 4 mL of methanol ammonia (4%). .
(3) Concentration and concentration: The eluate was collected, and the nitrogen was blown to near dryness, and the volume was adjusted to 2 mL with acetonitrile, and passed through a 0.22 μm 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

Table 1 Mobile phase gradient elution procedure

Column temperature: 35 ° C
Injection volume: 5 μL
Ion source: electrospray (ESI), positive ion mode detection method: multiple reaction monitoring (MRM)

Table 2 mass spectrometer ion source parameters

Table 3 Papaverine waiting for the precursor ion and daughter ion parameter table

Table 4 Recovery results of hot pot sauce base 25μg/kg

2. Add a mass spectrum of residues such as papaverine to the blank sample.

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