| Hemerocallis spp.,which belongs to Liliaceae,is a perennial ratoon herb.It has been cultivated in China for thousands of years.Meanwhile,it is a kind of functional vegetable with high economic.medicinal and ornamental value.Daylily flower contains colchicine,flavonoids and polyphenols that has anti-oxidant,lipid-lowering,anti-tumor and other effects.At present,researches always concentrated in the composition analysis on the buds of Daylily,however few reports on the differences of the material composition of the daylily flower.In this literature,materials from the Shanxi Academy of Agricultural Germplasm Germplasm Garden were selected to standardize colchicine,total flavonoids,and 9 polyphenols in flower buds of Hemerocallis by high performance liquid chromatography and UV-vis spectrophotometer.In order to find a theoretical basis for the development of Hemerocallis,the test results in this paper are as follows:I.This reaserch optimized the testing system of colchicine by the effects of different chromatographic condition on the test of colchicine content,and compared the colchicine content in 29 dalilies' flower buds.The results showed that HPLC conditions of colchicine content determination in the dalilies' flower buds are Thermo-Syncronis C18(250mm X 4.6mm,5?m)chromatographic column,50%methanol and 50%"Wahaha Purified Water' at a flow rate of 0.8mL/min,column temperature was 25 0C,detection wavelength was at 243nm,and the injection volume was IOuL.The average content of colchicine in flower buds of 29 Hemerocallis accessions was 0.125p?g/g,H.'North yellow' and H.'Datong jin zhen'expressed higher colchicine content among them,which were 0.301 ?g/g and 0.291 ?g/g.H."suqian 2' and H.'huaian 2'expressed inferior quality,which were 0.017pg/g and 0.016?g/g.The recovery rate of average addition standard was between 89.1%to 98.8%.Furthermore,the correlation analysis between colchicine and color was-0.765,showing an extremely remarkable level.2.The orthogonal test was designed by SPSS software to study the extraction process of total flavonoids from the flower buds of Hemerocallis spp.The simple and optimal extraction technology of flavonoids from total flavonoids was optimized,and the content of total flavonoids in 29 flower buds of Valeriana was determined.The results showed that the optimum extraction conditions were as follows:ethanol concentration 70%,ultrasonic time 20 min,ultrasonic frequency 80 kHz,and solid-liquid ratio 1:10.The factors influencing the extraction rate of total flavonoids were:material-liquid ratio>ultrasonic time>ethanol concentration>ultrasonic frequency,relatively.Among the 29 materials,the highest content of total flavonoids was H."Huaian 2',which is 4.84%.The lowest content was "H.'Datong Jin Zhen'".which is 0.80%.Its average content was 3.12%.The spike recovery rate was between 92.25%and 96.51%.3.A method for the simultaneous determination of 9 polyphenols in the flower buds of Echinochloa crusgalli by HPLC was established,which provided technical support for the extraction and application of phenols in Daylily flowers.The preferred chromatographic conditions were:using a Thermo-Syncronis C18(250 mm X 4.6 mm,5 ?m)column,flow rate:0.8 ml/min,temperature:30?,injection volume 10 ?L,mobile phase:0.2%aqueous acetic acid(A)and Methanol(B),gradient elution,elution procedure:0-5 min,10%(B);5?10 min,10?20%(B);10?20 min,20-35%(B);20?25 min,35?45%(B);25?30 min,45%(B);30?35 min,45?60%(B);35?45 min,60?45%(B);45?50 min,45?10%(B).In the range of 190?600nm,the standard product was scanned in 3D.The maximum absorption wavelength range of the nine standard products was 255?370nm.It was able to simultaneously detect and determine the final wavelength of 280nm.Under the established detection system,9 kinds of phenolic compounds were well separated,all the resolution were greater than 1.5,the linear correlation coefficient were greater than 0.99,the linear range was 1 to 100 ?g/mL,and the minimum detection limit was 0.005 to 0.06 ?g/ml.The RSDs of 9 polyphenols were between 0.27%and 1.05%.The RSDs of repeatability of them were less than 2.54%,and the RSD values of retention time and peak areas in the stability test were less than 2.64%and 3.24%.The method can be used as a simultaneous detection of the 9 phenolic compounds.4.The established system was used to detect and analyze the differences in the contents of 9 phenolic compounds such as rutin and chlorogenic acid in 46 species of Hemerocallis,The results showed that gallic acid,rutin and quercetin were detected in the buds of Hemerocallis.The content of gallic acid in H.'Crimson Pirate' was 43.60?g/g which was the highest among 9 phenolic;in the same time,the content of rutin in H.'Bonanza',which was 23.40?g/g,was the highest;Quercetin in the H.'Xiao Hong 74' was reached up to 221.58 Mg/g.The average content of coumaric acid,chlorogenic acid,catechin,vanillic acid,syringic acid and ferulic acid in the buds of 46 species of Valeriana was 8.49 ?g/g,9.11 ?g/g,4.56 ?g/g,1.97?g/g,3.87 ?g/g,and 1.31 ?g/g. |
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