Method for extracting and identifying flavonoid metabolites in plum blossom petals
The technology of an extraction method and an identification method is applied in the field of extraction and identification of flavonoid metabolites in plum blossom petals, which can solve the problems of sample waste, high impurities and low purity of the extract, and achieve the effect of less impurities and high purity
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[0030] The preparation method of rutin standard curve in the embodiment of the present invention is:
[0031] (1) Accurately weigh 0.010 g of rutin standard substance, add 60% ethanol extractant to dissolve to 5 ml, and prepare a standard solution with a mass concentration of 2 mg / ml.
[0032] (2) Pipette 25 μl, 50 μl, 100 μl, 150 μl, and 200 μl of the standard solution into 5 test tubes, add 5% NaNO 2 Solution 100μl, shake well and let stand for 6min, add mass fraction 10% Al(NO 3 ) 3 100 μl of the solution, shake well and stand for 6 minutes; add 1000 μl of 4% NaOH solution, add water to 2.5ml, shake well and let stand for 15 minutes.
[0033] (3) the rutin standard solution of different mass concentrations is at 510nm with UV-4802 double-beam scanning type ultraviolet-visible spectrophotometer, with distilled water as blank reference, measure its absorbance, by absorbance value and rutin standard solution concentration Draw a standard curve.
[0034] (4) Find the linear...
Embodiment 1
[0035] Example 1 Extraction and identification analysis of plum blossom petal flavonoids
[0036] This embodiment provides a method for extracting and quantitatively analyzing flavonoids in plum blossom petals, and the specific steps are as follows:
[0037] (1) Take the petals of 'Baixu Zhusha' in full bloom stage as the test material, freeze the fresh plum blossom samples in liquid nitrogen, weigh 0.015-0.025g freeze-dried samples of plum blossom petals with an electronic balance, put them in 2ml of liquid nitrogen quick-frozen samples and grind them In the tube, two steel balls are added to each tube, and the grinding equipment is used for automatic grinding.
[0038] (2) Add the extracting agent with a material-to-liquid ratio of 1:8000 (g / μl), shake it evenly with a vortex instrument, and use an ultrasonic cleaner to sonicate at 25°C; the extracting agent in this example is 60% ethanol, The ultrasonic power of ultrasonic-assisted extraction was 200W, the ultrasonic time ...
Embodiment 2
[0052] Example 2 Extraction and Quantitative Analysis of Flavonoids from Plum Blossom Petals
[0053] The method adopted in this embodiment is the same as that of Example 1, the only difference being that in step (2) of this embodiment, the extraction agent is 2% formic acid methanol, the ultrasonic power of ultrasonic-assisted extraction is 200W, and the ultrasonic time is 30min. Ice cubes were added during this period so that the temperature did not exceed 25°C. The obtained flavonoid extraction rate was 12.9553%. Use Q-TOF-LC-MS liquid chromatography-mass spectrometry instrument to obtain mass spectrometry information, positive mode ion chromatogram see image 3 , negative mode ion chromatogram see Figure 4 . image 3 and Figure 4 The middle peak type is clear and the number of peaks is large, which proves that the flavonoid content in the extract extracted in this embodiment is high and rich in types.
[0054] Metabolites were identified by comparing the m / z values ...
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