Method for simultaneously detecting seven flavones substances in hops
A technology for flavonoids and substances, which is applied in the field of chemical analysis and detection, can solve the problems of no efficient and accurate method, etc., and achieves the effects of saving time, simple operation and good separation effect.
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Embodiment 17
[0019] The standard sample detection of embodiment 17 kinds of flavonoids
[0020] Accurately weigh 0.0010g of flavonoid standard substance and fully dissolve it with chromatographic grade methanol, dilute to 10mL, dilute to different concentrations as required, prepare for injection and store in sample bottle for analysis.
[0021] Reverse-phase liquid chromatography analysis conditions:
[0022] Chromatographic column: Zorbax Eclipse XDB-C18 (4.6×250mm, 5m);
[0023] Injection volume 10μL, flow rate 0.8mL / min, column temperature 30℃;
[0024] Detection wavelength: 280nm and 370nm;
[0025] Mobile phase: organic phase B is acetonitrile, inorganic phase A is aqueous solution containing 0.1% (v / v) formic acid; gradient elution method is adopted, and the elution program is: 0-20min, 25%B-80%B; 20- 25min, 80%B-100%B; 25-28min, 100%B-100%B; 28-30min, 100%B-25%B.
[0026] The obtained standard sample chromatogram is as follows figure 1 As shown, peak 1 is kaempferol 3-O rutino...
Embodiment 2
[0030] The detection of flavonoids in the hop sample of embodiment 2
[0031] After the hop granules are crushed and ground into powder, accurately weigh 1g of hop powder, add 7mL of anhydrous ether, shake for 15min, then ultrasonicate for 5min, centrifuge at 1500×g for 10min, remove the supernatant, then add 20mL of 50% methanol solution, shake for 15min, and then ultrasonicate for 60min. Centrifuge at 1500×g for 10 min, filter the supernatant through a 0.45 μm microporous membrane, store in a sample bottle, and inject for analysis.
[0032] Reverse-phase liquid chromatography analysis conditions:
[0033] Chromatographic column: Zorbax Eclipse XDB-C18 (4.6×250mm, 5m);
[0034] Injection volume 10μL, flow rate 0.8mL / min, column temperature 30℃;
[0035] Detection wavelength: 280nm and 370nm;
[0036] Mobile phase: organic phase B is acetonitrile; inorganic phase A is aqueous solution containing 0.1% (v / v) formic acid. Using gradient elution method, the elution program is:...
Embodiment 3
[0038] Stability and reproducibility assessment of the detection method of embodiment 3
[0039] Get the hop sample of known target substance content, add respectively the standard sample of different concentrations processed according to the method of embodiment 1, specifically add standard sample concentration as follows: xanthohumol (80mg / L, 60mg / L, 40mg / L, 20mg / L, 10mg / L, 5mg / L, 2mg / L, 1mg / L, 0.5mg / L, 0.25mg / L), kaempferol 3-O rutinoside (80mg / L, 60mg / L, 40mg / L , 20mg / L, 10mg / L, 5mg / L, 2mg / L, 1mg / L, 0.5mg / L, 0.25mg / L), isoxanthohumol (10mg / L, 8mg / L, 6mg / L, 4mg / L, 2mg / L, 1mg / L, 0.5mg / L, 0.25mg / L), 6-prenyl flavone (5mg / L, 4mg / L, 3mg / L, 2mg / L, 1mg / L , 0.5mg / L, 0.25mg / L, 0.125mg / L), 8-prenyl flavone (5mg / L, 4mg / L, 3mg / L, 2mg / L, 1mg / L, 0.5mg / L , 0.25mg / L, 0.125mg / L), hexamethoxyflavone (5mg / L, 4mg / L, 3mg / L, 2mg / L, 1mg / L, 0.5mg / L, 0.25mg / L, 0.125mg / L L), quercetin (5mg / L, 4mg / L, 3mg / L, 2mg / L, 1mg / L, 0.5mg / L, 0.25mg / L, 0.125mg / L). The prepared mixed solutions with different...
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