Method for detecting capsaicin compound in grease
A technology of capsaicin and oil, which is applied in the direction of measuring devices, analysis materials, instruments, etc., can solve the problems of long time-consuming, complicated pre-treatment, and sensitivity that cannot meet the detection of refined oils, etc., and achieve the effect of shortening the detection time and reducing the detection cost
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Embodiment 1
[0015] Example 1 Sample pretreatment scheme
[0016] 1. Preparation of immunomagnetic beads:
[0017] (1) Take 1ml of magnetic beads (10mg / ml, product number MS300 / Carboxyl, Japan JSR Company), place the magnetic beads in a magnetic field, collect the magnetic beads, and discard the supernatant;
[0018] (2) Resuspend the magnetic beads with 1ml PBS (phosphate buffered saline, pH 7.4), add 5mg EDC (1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride), Shake at room temperature for 15 minutes;
[0019] (3) Place the magnetic beads in a magnetic field, collect the magnetic beads, and remove the supernatant; resuspend the particles with 1ml PBS (pH7.4); repeat this step once;
[0020] (4) Add 20 mg of anti-capsaicin antibody and mix at room temperature for 2 hours;
[0021] (5) add blocking agent, continue to react for 2h;
[0022] (6) Place the magnetic beads in a magnetic field, collect the magnetic beads, and remove the supernatant; resuspend the particles with 1ml...
Embodiment 2
[0035] The influence of immunomagnetic bead blocking agents on the detection results: Based on Example 1, different blocking agents (see Table 1) were used to block the magnetic beads. Different blocking agents had a significant impact on the detection sensitivity, the detection limit of 5-aminovaleric acid was the best, and that of deep-water fish gelatin was the worst. Particles sealed with Tris are prone to aggregation in oil-water mixtures, especially in a magnetic field. After aggregation, it is difficult to disperse, making subsequent detection operations impossible.
[0036] Table 1 Effect of blocking agent on the results
[0037] sealant Working concentration of blocking agent (final concentration) Lower detection limit (μg / kg) Remark BSA (Bovine Serum Albumin) 1% (W / V) 60 Deep-water fish gelatin (Sigma company) 0.2% (W / V) 240 PEG20000 (polyethylene glycol 20000) 1% (W / V) 30 Tris (trishydroxymethylaminomethane) 20mM ...
Embodiment 3
[0039] The influence of immunomagnetic bead reconstitution solution on the detection results: Based on Example 1, the influence of different reconstitution solutions (Table 2) on the detection results was compared. It can be seen from the table that the reconstitution solution added with hydrolyzed milk protein is The increase in detection sensitivity has been significantly improved.
[0040] Table 2 Effect of complex solution on detection results
[0041] Numbering formula Lower detection limit (μg / kg) 1 20mM Tris-HCl, 1% BSA, pH7.5 6 2 PBS, 1%BSA, pH7.4 24 3 20mM Tris-HCl, pH7.5 3 4 PBS, 1% hydrolyzed milk protein, pH7.4 1.5
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