A kind of fingerprint detection method of white ginseng bacteria
A technology of fingerprint spectrum and detection method, applied in the field of fingerprint spectrum detection of white ginseng bacteria, can solve the problem of high cost, and achieve the effects of simple operation, good peak shape and objective conclusion.
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Embodiment 1
[0030] (1) Screen different chromatographic columns to investigate the number of peaks, resolution and peak shape. (area normalization method, peaks below 0.05% are not counted)
[0031]
[0032] From the results, MCI GEL SCA04 (4.6×150mm) has the most detected peaks and the best resolution, so MCI GELSCA04 was selected
[0033] It is used to develop the detection method of HPLC characteristic fingerprint of white ginseng bacteria.
[0034] (2) Screen different mobile phases to investigate the number of peaks, resolution and peak shape. (area normalization method, peaks below 0.05% are not counted)
[0035]
[0036] From the results, the detection peaks of 5% methanol-water and methanol-0.02N sodium carbonate solution (5:95) systems are basically the same, and the resolution is optimal, so 5% methanol-water is selected as the HPLC characteristic of developing white ginseng bacteria. Mobile phase salts for fingerprint detection.
[0037] (3) Screen different wavelengt...
Embodiment 2
[0041] 1 Methodological investigation 1.1 Chromatographic conditions: MCI GEL SCA04 (4.6×150mm) chromatographic column was used, methanol was used as mobile phase A, 5% methanol-water was used as mobile phase B, gradient elution, flow rate 1.2ml / min, column temperature 30°C, the detection wavelength is 230nm; the number of theoretical plates should not be less than 3000 based on the peak of schizophyllin.
[0042] Gradient elution table:
[0043]
[0044] 1.2 Processing of samples and reference substances
[0045] (1) Preparation of the test solution: Freeze-dry the white ginseng bacteria, crush them, take 5-20g, weigh them accurately, add 80ml of 10% acetic acid to soak for 24h, add 55% methanol for ultrasonic extraction for 1 hour, let cool, and then Weigh the weight, add methanol to make up the weight, shake well, filter, and take the filtrate to get it;
[0046] (2) Preparation of reference substance solution 1: Take 6 mg of schizophyllin reference substance, put it i...
Embodiment 3
[0054] 1. Instrument
[0055] Agilent1260 high performance liquid chromatography (Agilent, USA); PB-10 pH meter (Sartorius), BP-211D analytical electronic balance (Sartorius).
[0056] 2. Test solution
[0057] Methanol (Sigma Company, chromatographically pure), and water was Wahaha purified water.
[0058] 3. Reference substance
[0059] Schizophyllan, valine, isoleucine, leucine, threonine, methionine, lysine, phenylalanine, tryptophan, cysteine, serine, glycine, Tyrosine, Aspartic Acid, Asparagine, Glutamic Acid, Glutamine, Alanine, Arginine, Histidine, Proline.
[0060] 4. Sample
[0061] White ginseng fungus (wild, picked from dead wood on Taoyuan Mountain in Changde).
[0062] 5. Inject each reference substance separately for positioning.
[0063] 6. Establishment of fingerprints and marking of common peaks
[0064] The white ginseng bacterium is prepared need testing solution by the method under "1.2" item in embodiment 2 respectively, is analyzed by the chromato...
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