Study On Preparation Technology And Quality Standard Of Total Flavonoids From Nymphaea Candida

Nymphaea candida Presl is a perennial aquatic herbaceous plant of the genus Nymphaea,and is mainly used in the Uygur medicine for various deseases such as headache,cough and hepatitis.Preparation technique and quality standard of total flavonoids from N.candida(NCTF)were studied on the base of compound analysis.(1)In this study,HSCCC was used to separate and purify the characteristic compounds from NCTF.Isoquercitrin,astragalin,isostrictiniin and nicotiflorin were isolated by two-step separation using solvent fesystem at ethyl acetate:methanol:water(4:1:5,v/v)and ethyl acetate:butanol:water(4:1:5,v/v).Their purities were 73.53%,92.42%,82.36% and 85.65% respectively by HPLC analysis.(2)The single factor tests and orthogonal experimental design method were applied to analyse the influence of each factor in content of nicotiflorin and astragal in the best extraction condition of reflux extraction: 70% ethanol as solvent,ratio of material to liquid(1: 10,g/m L),extract 2 times with 90 min/each time;The content of nicotiflorin and astragalin was taken as the index.(3)Purified technique of NCTF was constrcuted by methods of combination macroporous resin with polyamide resin.The best concentration of macroporous resin was nicotiflorin 0.3220 mg / L,astragalin 0.0865 mg / L,removing impurities at 5 BV30% ethanol,eluenting at 5 BV 50% ethanol.After purification with resin,the polyamide resin was used to enrich and purify again.The optimum technological conditions were as follows: extracts:polyamide in proportion of 1:8,5BV 70% ethanol.The results showed that the content of nicotiflorin and astragalin reached to 116.0mg/g and 58.0 mg/g.The total flavonoid yield was above 4.5% by verification test.(4)The physical and chemical identification and TLC identification method of total flavonoids in N.candida were established.According to Chinese Pharmacopoeia2015 edition fourth,moisture content,residue on ignition,heavy metal and arsenic salts were determined,and preliminary defined that moisture content was not exceed7.0%,residue was not over 4.0%,heavy metals should not exceed twenty ppm,arsenic salt should not exceed 0.5 ppm;The effects of high temperature,high temperature and strong light on the appearance,color and the content of the components of NCTF were studied.The results showed that there was no significant effect on the appearance,color and content of the components of NCTF.