A method for determining capsaicinoids in pepper and pepper products by high performance liquid chromatography and a method for expressing the degree of hotness.
The detection limit of the method: capsaicin and dihydrocapsaicin are both 1 mg/Kg , and the linear range is 1 mg/L-150 mg/L .
Liquid chromatograph , column , column oven, ultrasonic cleaner, filter, filter, nitrogen blower , liquid phase syringe
3. Sample extraction:
25 ml of a methanol - tetrahydrofuran mixed solvent was added to the sample, sealed with a plastic wrap, and several small holes were needled, and then extracted at 60 Â° C for 30 min using an ultrasonic cleaner . Filtered with filter paper, collected the filtrate, and then added 25 ml of methanol - tetrahydrofuran mixed solvent together with filter paper, and extracted with ultrasonic cleaner for 10 min , repeated twice, and the filtrates collected by three filtrations were combined and concentrated by a nitrogen blowing instrument at 70 Â° C. to 10ml-20ml, and then with methanol - tetrahydrofuran mixed solvent volume to 50ml, through a filter (0.45 m microporous filtration membrane) filtration chromatography analysis. Depending on the total amount of capsaicinoids in the sample and the sensitivity of the detector, the dilution factor can be adjusted as necessary.
4. Chromatographic reference conditions:
Â Â Â Column: Kromasil C-18 (4.6mm*250mm 5um)
Â Â Â Mobile phase: methanol: water = 65 : 35
Â Â Â Injection volume: 10 Î¼l
Â Â Â Flow rate: 1.0 ml/min
Â Â Â UV detection wavelength: 280nm
Â Â Â Column oven temperature: 30 Â°C
Â Â Â HPLC analysis was carried out according to the chromatographic conditions, and the retention time of the chromatographic peak of the standard substance was qualitatively determined; the peak area in the sample was quantified based on the capsaicin, dihydrocapsaicin standard curve and the sample.
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