| In this article, we have studied the Allium cepa L.var. agrogatum Don. It includes three parts: the extraction process, the purification process and the quality standard of total flavonoids in Allium cepa L.var. agrogatum Don. The study settles a good foundation for exploitation the Allium cepa L.var. agrogatum Don. in order to discover a new drug.1. The extract process of total flavonoids in Allium cepa L.var. agrogatum Don.We have selected the solvent and method first, and then inspected four important factors through experiment of monofactor and orthogonal experiment. Finally, we determined the suitable condition and optimal extraction technology of total flavonoids, it was that 3 times of 70% alcohol,refluxing 3 times and two hours each.2. The purify process of total flavonoids in Allium cepa L.var. agrogatum Don.In this part, we have compared the results of different purify methods: abstraction, precipitation and macroporus resin chromatography et al. the optimal method was macroporus resin chromatography. Then, we have chosen the macroporous resin AB-8 as the optimal resin through static adsorption test and dynamic adsorption test. Because the macroporous resin AB-8 possessed the strongest adsorption ability to the total flavonoids among the 9 resins studied, additionally an easy desorption property.Eventually, we have tested the factors, including the loading concentration, loading velocity, eluant, eluting velocity and adsorbing time et al. and decided the optimal purify process was: the extracted solution (concentration was between 4.0 mg·mL-1～5.5 mg·mL-1) flowed the macroporous resin chromatographic column by 0.5 BV·h-1, absorbed for 6 hours, Then, eluted by distilled water about 10 BV. Finally, eluted by 70% ethanol, collected the ethanol elution between 0.5～4 BV, evaporation to dryness, the extract was obtained. The content of total flavonoids in the extract is above 55%, and the recovery of total flavonoids is above 85%.3. The quality standard of total flavonoids in Allium cepa L.var. agrogatum Don.In the last part, we have studied the quality standard of total flavonoids in Allium cepa L.var. agrogatum Don. We have identified the quercetin, quercetin-4′-O-β-D-glucopyranoside and quercetin-3, 4′-di-O-β-D-glucopyrano -side by TLC, determinated the water content and the residue on ignition. The water content should below 10%, the residue on ignition should below 2.0%.Besides determinated the content of total flavonoids by UV chromatophy, we have also determinated the content of quercetin,quercetin-4′-O-β-D- glucopyranoside and quercetin-3, 4′-di-O-β-D-glucopyranoside by HPLC. The content of total flavonoids should above 55%, and the content of quercetin, quercetin-4′-O-β-D-glucopyranoside and quercetin-3,4′-di-O-β-D-glucopyrano- side should above 15.7%, 18.1% and 19.9%.The article studied the preparation technology of total flavonoids in Allium cepa L.var. agrogatum Don. systematically for the first time. The technology is stable, easy and feasible; the equipment is simple; the content of total flavonoids in the extract is above 55% and the yield of total flavonoids calculated by dry Allium cepa L.var. agrogatum Don. can reach 2.3%. The raw material is just ethanol and macroporous resin, so the cost is low, besides of low-toxicity.It also studied the quality standard of total flavonoids in Allium cepa L.var. agrogatum Don. For the first time, besides established the content determination of total flavonoids, we have determined the content of 3 compounds of flavonoids by HPLC and identified them by TLC for the first time.The study was supported by science and technology office of Jilin Province, it settles a good foundation for exploitation the Allium cepa L.var. agrogatum Don. in order to discovery a new drug. We have applied patent for it. |
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