| Herba Cistanche, as perennial herbs, distribute in Xinjiang, Inner Mongolia, Shaanxi, Gansu, Ningxia and other places, with kidney-yang, essence and blood, and Runchang purge function.This thesis includes four parts:1. Study on the polysaccharides of Herba CistancheCistanche deserticola polysaccharides are the active ingredients in Herba Cistanche. Polysaccharides were separated by using aqueous alcohol extraction method and the content of crude polysaccharide and soluble polysaccharide was determined with ultraviolet spectrometry. The results showed that the content of crude polysaccharide and soluble polysaccharide in Herba Cistanche is 4.56% and 17.41% separately. Based on the study of the extraction of polysaccharide in Cistanche, polysaccharides content is the highest when 20% ethanol was used, which can go to up to 9.71% of sample and 44.0% of total polysaccharides.2. Study on the Acteoside of Herba CistancheUltrasound-assisted extraction of active ingredients was employed, and acteoside in Herba Cistanche was determined by high performance capillary electrophoresis. After detailed study of the impact of the separation voltage in the range of 8 ~ 25 kV and boric acid - borax buffer solution of pH 8.0 ~ 10.0 on the separation of acteoside, the best separation conditions were as follows: voltage at 8kV, running buffer solution with 30mmol/L of boric acid-borax (pH = 9.0), room temperature, and detection wavelength at 334nm. Acteoside concentration in the range of 0.015 ~ 0.35mg/mL showed good linear relationship. The linear regression equation is y=13946x+560.32 (R2=0.9983),and sample recovery yield was 99.26% with RSD(1.16%) and precision of RSD(0.96%). Through this research, we established the measurement of acteoside by high performance capillary electrophoresis method, and the content of Mullein glycoside in cistanche was determined to be 1.02 ~ 1.04% by the above method.3. Study on the determination of salidroside in Herba CistancheThe use of HPLC and HPCE methods for the determination of salidroside in Herba Cistanche was studied. For HPLC method, column loaded with octadecyl silane bonded silica, mobile phase of acetonitrile: water (1:9), flow rate 1.0mL/min at room temperature, and the detection wavelength at 278nm were employed. The results indicated that salidroside in the range of 0.055 ~ 0.44μg was in good linear relationship with the linear regression equation y=91207x-1097, (R~2=0.9953), and sample recovery yield was 94.8%, RSD = 1.50%, and precision degree of RSD = 0.39%.Through our study, the best conditions for HPCE method were as follows: separation voltage at 8kV, running buffer 30mmol/L of boric acid-borax (pH = 9.0), room temperature, and detection wavelength at 278nm. The results showed salidroside concentration in the range of 0.007 ~ 0.14mg/mL was in good linear relationship with the linear regression equation y=42915x-92.541,(R~2=0.9903). Sample recovery yield was 100.1%, RSD = 1.45%, and precision of RSD = 0.96%.In comparison of separation and determination of salidroside by two methods, the results showed that both high-performance liquid chromatography and capillary electrophoresis can well work on salidroside cistanche separation without significant differences in measurements, and its content ranges from 0.120% to 0.103%.4. Study on the fingerprint of Herba CistancheIn this paper, HPCE method was employed to analyze the ingredient in Cistanche samples. The main active ingredient, the Acteoside of Herba Cistanche, was set as evaluation index to establish the HPCE cistanche fingerprint, and the separation degree and reproducibility are reliable and excellent. The RSD of all relative peak area and the relative retention time are less than 5%. Our research provides a solid scientific basis to the establishment of quality standards of medicinal Cistanche. |
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