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Abstract: Capsaicin ((trans) 8-methyl-N-vanillyl-6-nonenamide) is the active ingredient of pepper. Capsaicin and some related compounds are called capsaicin, and they are secondary metabolites produced by capsicum.
1 Scope:
Method for determining capsaicinoids in chili peppers and chili products and expression of spiciness by high performance liquid chromatography.
The detection limit of this method: capsaicin and dihydrocapsaicin are both 1mg / Kg, and the linear range is 1mg / L-150mg / L.
2. Instruments
Liquid chromatograph, chromatographic column, column oven, ultrasonic cleaner, filter, filter membrane, nitrogen blower, liquid injection needle 3. Sample extraction: add 25ml of methanol-tetrahydrofuran mixed solvent to the sample and keep it fresh The membrane was sealed, a few small holes were pierced with a needle, and then extracted under an ultrasonic cleaner for 30 min under a bath condition of 60 ° C. Filter with filter paper to collect the filtrate, then add 25ml of methanol-tetrahydrofuran mixed solvent to the filter residue together with the filter paper, use an ultrasonic cleaner to extract for 10min, repeat twice, combine the filtrate collected by filtration three times, and concentrate at 70 ° C with a nitrogen blower To 10ml-20ml, then make up to 50ml with methanol-tetrahydrofuran mixed solvent, filter through a filter (0.45µm microporous filter membrane) and then perform chromatographic analysis. According to the total amount of capsaicinoids in the sample and the sensitivity of the detector, the dilution factor can be adjusted appropriately if necessary.
4. Chromatographic reference conditions:
Column: Kromasil C-18 (4.6mm * 250mm 5um)
Mobile phase: methanol: water = 65: 35
Injection volume: 10 µl
Flow rate: 1.0 ml / min
UV detection wavelength: 280nm
Column oven temperature: 30 ℃
5. Results:
According to the chromatographic conditions, HPLC analysis was performed, and the retention time of the chromatographic peaks of the standard substances was used for qualitative analysis; according to the standard curve of capsaicin and dihydrocapsaicin and the peak areas in the samples were quantified.
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