| High Performance Liquid Chromatography(HPLC)as an important modern measurement of analysis and separation is deveopled on the basis of gas Chromatography and classical Liquid Chromatography. Due to its high-speed, accuracy, sensitivity and convenience, it has been widely used in the areas of foodstuff, environment, biology engineering, pharmacy and petroleum so far.The thesis includes two sections-review and research report. In the review section, information about high performance liquid chromatography such as characteristic, the development of chromatography technology and application in the effective elments of herb medicine Eucommia ulmoides Oliv by HPLC analysis and isoflavones research was summarized. In the next research report section, analysis methods of determination of chlorogenic acid and rutin in Eucommia ulmoides Oliv and isoflavones in food were reported. That is:Determination of four isoflavones in soybean milk powder by HPLC; Determination of four isoflavones in different beans by HPLC; Determination of four isoflavones in mung bean sprouts by HPLC.1.Determination of chlorogenic acid and rutin in Eucommia ulmoides Oliv by HPLCTo establish an HPLC method for determination of chlorogenic acid and rutin, and determination chlorogenic acid and rutin in Eucommia ulmoides Oliv from different places. Using a phenomenex C1g column(150 mm×4.6 mm,5μm),the mobile phase of methanol-acetic acid(pH=3.10)(45:55) with elution, the flow rate was 0.8 mL/min, the detection wavelength was at 327 nm, the content of chlorogenic acid and rutin was determined by external standard method. The calibration curves for chlorogenic acid and rutin were linear between 0.1275 and 24.0000μg/mL, between 0.1250 and 25.0000μg/mL(r=0.9996 and r=0.9996 respectively).The average recoveries were 99.23% and 97.13%.RSD were 1.02% and 2.98%. The method is simple, rapid, sensitive and reproducible, it provides scientific proof for quality standard in Eucommia ulmoides Oliv from different places.2.Determination of four isoflavones in soybean milk powder by HPLCTo develop an HPLC method for determination of four isoflavones, daidzin, genistin, daidzein, genistein in soybean milk powder. An phenomenex C18 column(150 mm×4.6 mm,5.0μm)was applied, a gradient mobile phase composing of acetonitrile and 0.2% formic acid was used, and the flow rate was 0.8 mL/min, detective wavelength was set at 260 nm.The best condition of extracted four isoflavones of soybean milk powder was studied.The results showed the best condition was the ethanol concentration 70%,60℃,extraction for 40 min. The results were obtained that quantitative linear range of four isoflavones were 0.74-100.00μg/mL,0.74-200.00μg/mL,0.167～500.00μg/mL,0.198～160.00μg/mL(r>0.9996)respectively, and the average recoveries(n=9)were 100.01%,100.01%,99.48%.99.91% respectively. A specific, sensitive, simple and reproducible HPLC method has been established for determination of four isoflavones in soybean milk powder.3.Determination of four isoflavones in different beans by HPLCTo develop an HPLC method for determination of four isoflavones, daidzin, genistin, daidzein, genistein in different beans. An phenomenex C1g column(150 mm×4.6 mm,5.0 vm)was applied, a gradient mobile phase composing of acetonitrile and 0.2% formic acid was used, and the flow rate was 0.8 mL/min, detective wavelength was set at 260 nm.The best condition of extracted four isoflavones of black beans was studied. The results showed the best condition was the ethanol concentration 70%,60℃,extraction for 40 min. The results were obtained that quantitative linear range of four isoflavones were 0.74～100.00μg/mL,0.74～400.00μg/mL,0.167v500.00μg/mL, 0.198～160.00μg/mL(r>0.9996)respectively, and the average recoveries(n=9)were 99.81%. 99.99%,99.49%,99.89% respectively. A specific, sensitive, simple and reproducible HPLC method has been established for determination of four isoflavones in different beans.4.Determination of four isoflavones in mung bean sprouts by HPLCTo develop an HPLC method for determination of four isoflavones. daidzin, genistin, daidzein, genistein in mung bean sprout. An phenomenex C18 column(150 mm×4.6 mm,5.0μm)was applied, a gradient mobile phase composing of acetonitrile and 0.2% formic acid was used, and the flow rate was 0.8 mL/min, detective wavelength was set at 260 nm.The best condition of extracted four isoflavones of mung bean sprout was studied, the results showed the best condition was the ethanol concentration 60%,55℃,extraction for 60 min with the solid-liquid ratio 1:12(g/mL).The results were obtained that quantitative linear range of four isoflavones were 0.74℃100.00μg/mL,0.74～200.00μg/mL,0.167～500.00μg/mL,0.198～160.00μg/mL(r>0.9996)respectively, and the average recoveries(n=9)were 99.99%,100.01%,100.07%.99.81% respectively. A specific, sensitive, simple and reproducible HPLC method has been established for determination of four isoflavones in mung bean sprouts. It provided scientific bases for the best eating time of mung bean sprout everyday. |
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