Method for detecting caffeic acid content in tobacco
A technology of caffeic acid and content, applied in measuring devices, instruments, scientific instruments, etc., can solve the problem of no patent for the determination method of caffeic acid, and achieve the effect of less organic reagents, strong accuracy and high sensitivity
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Embodiment 1
[0023] Example 1 Determination method example
[0024] First, prepare the standard solution: accurately weigh 10 mg of the caffeic acid standard, dilute to 10 mL with acetonitrile, absorb 1 mL and dilute to 10 mL with acetonitrile to make a standard solution, and set aside;
[0025] Then prepare the test sample solution:
[0026] (1) Prepare fresh tobacco leaf samples: take liquid nitrogen samples, then put the fresh tobacco samples into a freeze dryer to freeze-dry until constant weight (moisture-free), and then grind them into powder for later use;
[0027] (2) Prepare the cured tobacco leaf sample: the cured tobacco leaf is crushed and passed through a 40-mesh sieve, and set aside;
[0028] (3) Extraction of caffeic acid: take 0.6g of fresh tobacco leaf powder or 0.05g of roasted tobacco leaf powder, weigh it accurately, add 30mL of 4mol / L NaOH and shake on a shaker for 4h, add 4mol / L HCl to adjust after suction filtration pH to 2, then extracted 3 times with 20mL eth...
Embodiment 2
[0033] Embodiment 2 Linear relationship investigation
[0034] Take 0.5, 1, 2, 3, 5, 10, 20, 30, and 50 μL of the above-mentioned standard solution respectively, measure according to the chromatographic conditions in Example 1, and record the chromatogram and peak area. Taking the concentration of each standard substance as the abscissa and the peak area as the ordinate to calculate, the regression equation of caffeic acid is y=8.16×10 3 x-1.82×10 3 , R 2 =0.9998, the linear range is 1.113~56.236μg / mL.
[0035] Precision test: Accurately draw the test sample solution according to the above chromatographic conditions and continuously inject 10 times, the injection volume is 2 μL, record the chromatogram, and the RSD of the peak area of caffeic acid is 0.13%, the result shows that the precision of the instrument is good.
[0036] Repeatability test: Take the same batch of samples, parallel 10 samples according to the preparation method of the test product, inject continuou...
Embodiment 3
[0038] Embodiment 3 The selection of detection wavelength
[0039] When exploring the chromatographic conditions of the test sample solution, the full-wavelength scanning was carried out at 190-400nm, among which there were maximum absorptions at 233 and 324nm, and the peak shape at 320nm was better, and the baseline noise was small, so 320nm was selected as the maximum detection wavelength.
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