Studay On Quality Evaluation Of Rehmanniae Radix&Processing Method Of Rehmanniae Radix Praeparata

Objective:To establish the quality criterion for Rehmanniae Radix,and compare the changes of active ingredients of Rehmanniae Radix Praeparata during the process of steamed for nine times and shined for nine times.To select the best processing method by which the quality of the industrial production of Rehmanniae Radix Praeparata might be close to the production processed by the traditional method,to ensure the clinical efficacy,and create benefits for processing enterprises.Methods:1.To detemine the contents of the indicative compounds of Rehmanniae Radix through literature research and pre-experimental study.An HPLC method was established for simultaneous determination of catalpol,rehmannioside D,leonuride,acteoside,isoacteoside in Rehmanniae Radix.A Synergi^TM Hydro-RP 80?column?250 mm×4.6 mm,4?m?was used,with acetonitrile?A?-0.2%phosphoric acid?B?as the mobile phase in gradient elution mode?0⁸ min,3%A;8⁹ min,3%?4%A;4²5 min,4%A;25³5 min,4%?20%A;35⁵0 min,20%A?.The UV detection wavelength was set at the maximum absorption wavelength,203 nm for the catalpol,rehmannioside D and leonuride,334 nm for the acteoside and isoacteoside.2.The content of total polysaccharides was analyzed by sulfuric acid-phenol method,and detected by UV at 490 nm,the sample solution was prepared by the methods of alcohol extracting-water precipitating.3.Rehmanniae Radix Praeparata was prepared as the traditional process“Nine steamed nine shined”.The contents of 5 glycosides in the samples during each steamed and shined progress was determined by HPLC simultaneously,while total polysaccharides were determined by UV.The contents of 5 glycosides and total polvsaccarides in samples with different steamed and shined were detected and compared.Self-made Rehmanniae Radix Praeparata by nine steamed and nine shined was compared with commercial samples.4.In the lab,Rehmanniae Radix was processed as the traditional method with nine steamed and nine shined.The contents of 5 kinds of glycosides and total polysaccharides were chosen as observation indicators to observe the influence of different heating temperatures and drying temperatures.5.The processing method of Rehmanniae Radix Praeparata was optimized by L9?3⁴?orthogonal test,designed with heating temperature/pressure,heating time and heating times as main factors,and the contents of 5 kinds of glycosides and total polysaccharides as indexes.Result:1.The simultaneous detemination method of catalpol,rehmannioside D,leonuride,acteoside,and isoacteoside in Rehmanniae Radix by HPLC was simple,accurate,and stable,with excellent chormatographic separation and good reproducibility.In the contents determination of Rehmanniae Radix,the linear ranges of catalpol,rehmannioside D,leonuride,acteoside and isoacteoside were0.925¹3.875?g?r=0.999 0?,0.162².430?g?r=0.999 2?,0.168².520?g?r=0.999 5?,0.041⁰.615?g?r=0.999 8?,and 0.038⁰.057?g?r=0.999 0?,respectively.The average recovery rates were 96.43%,97.52%,96.16%,100.48%,and 97.21%,respectively,with RSDs 1.44%,2.56%,1.07%,1.98%,and 3.35%,respectively.In the contents determination of Rehmanniae Radix Praeparata,the linear ranges of catalpol,rehmannioside D,leonuride,acteoside and isoacteoside were0.012⁰.300?g?r=0.999 3?,0.030⁰.75?g?r=0.999 0?,0.010⁰.250?g?r=0.9995?,0.021⁰.525?g?r=0.999 3?,and 0.018⁰.045?g?r=0.999 5?,respectively.The average recovery rates were 98.27%,98.77%,99.01%,99.65%,and 98.54%,respectively,with RSDs 1.42%,2.29%,1.86%,2.96%,and 1.65%,respectively.2.The determination method of total polysaccharides in Rehmanniae Radix by UV was simple,and accurate,stable,with good reproducibility.The linear range was0.005 96⁰.059 6 mg/m L,and the average recovery rate was 100.44%,with RSD2.89%.3.During the traditional processing of Rehmanniae Radix Praeparata,the contents of catalpol,rehmannioside D,leonuride,acteoside,and isoacteoside decreased to much lower levels after the first procedure of steamed and shined,while the content of total polysaccharides show a tendency of increasing first and decreasing afterwards.4.In the industrialized processing courage,the contents of 5 kinds of glycosides above varied,which might be related to the stability of their chemical structures.The contents of catalpol and leonuride decreased significantly,rehmannioside D was basically stable,acteoside decreased,while isoacteoside increased in a certain heating time range.The content of total polysaccharides shows a clear upward trend during the processing.5.According to the results of orthogonal test and compared with the contents in samples by traditional processing method,the optimum industrial processing factors were as flows:Rehmanniae Radix was heated at 125?for 2 times,each time for 2 h.Validation tests confirmed that the process had good repeatability.Conclusion:In this paper,an HPLC method was established for simultaneous determination of 5 kinds of glycosides in Rehmanniae Radix,and a UV method was developed to determine the content of total polysaccharides.The methods are simple,accurate,reproducible and stable,which can be used for evaluation of the quality of Rehmanniae Radix and Rehmanniae Radix Praeparata.The processing method using industrial equipments have been optimized to ensure that quality of Rehmanniae Radix Praeparata can be close to the production processed as traditional method.The validation tests confirmed its good repeatability.