The Study On Quality Standard Improvement Of Rehmanniae Radix Praeparta And Liuwei Dihuang Capsules

Objectes: This topic mainly through the in-depth study of the quality standards of Rehmannia glutinosa and Liuwei Dihuang capsules,to enhance the existing standards and provide a reasonable scientific basis for the quality improvement research of Rehmanniae Radix Praeparta and Liuwei Dihuang capsules.Methods:1.Target component separation: The 50 kg Rehmanniae Radix Praeparta extract was obtained by ethanol reflux.The reference substance was separation and purification by chemical column chromatography,followed by D101 macroporous resin,MCI column and liquid phase.2.The Study on the improvement of quality standard of Rehmanniae Radix and Rehmanniae Radix Praeparta: catalpol and acteoside were used as the TLC identification indicators,and the HP MN254 thin layer plate was used to develop the system with ethyl acetate: methanol: formic acid: water=14:3:1:1.8,and sprayed with 10% sulfuric acid to develop color.Acteoside and isoacteoside were used as TLC identification index components,which were carried out using a mixture of trichloromethane-ethyl acetate-water(13:7:2)as a developing solvent on silica gel GF254,then inspecting with colorating 10% sulfuric acid ethanol and examine in daylight.Samples were separated in MG II(4.6×250 mm,5 ?m)column with the gradient elution of acetonitrile-water(containing 0.1% formic acid)at a flow rate of 1.0 m L/min.The column tempreture was maintained at 30 °C and the detection wavelength was set at 334 nm.The content of four phenylethanoid glycosides of purpureaside C,jionoside A1,acteoside,jionoside B1 was determined by quantitative analysis of multicomponents with single marker(QAMS).Eight phenylethanoid glycosides were identified by LC-MS,and an HPLC fingerprint was established.With acteoside as reference peak and labeled S,the relative retention time and relative peak area of peak in fingerprint were calculated and methodological investigation was carried out.3.The Study on the improvement of quality standard of Liuwei Dihuang Capsules: Based on the existing pharmacopoeia,the thin layer identification and the quantitative control of the Rehmanniae Radix Preparata and Poria of Liuwei Dihuang Capsule was established.Choose acteoside and isoacteoside as the reference substance,the quality of Rehmanniae Radix Preparata was identified by Yantai silica gel GF254 and use mixture of trichloromethane-methanol-water(13:7:2)as a developing solvent.Spray with a 10% sulfuric acid ethanol and observe at daylight.Pachymic acid was selected as a reference material for the identification of Poria in Liuwei Dihuang Capsule,the TLC(thin layer chromatography)identification of pachymic acid was carried out using a mixture of trichloromethane-ethyl acetate-0.1% formic acid(10:2:0.2)as a developing solvent on HP merck GF254,and inspecting with Coloring 10% sulfuric acid ethanol.Choose acteoside as the quantitative index component in Rehmanniae Radix Preparata on an Uni Sil? 5-120 C18-Polar(250 mm×4.6 mm,5 ?m)with acetonitrile-0.1% phosphoric acid at a flow rate of 1.0 m L·min-1.The detecting wavelength was 334 nm and the column temperature was 30?.Pachymaric acid was used as the quantitative index component in Poria cocos.Samples were separated in Chrom Core TM C18(4.6×250 mm,5 ?m)column with the gradient elution of acetonitrile-0.1% phosphoric acid at a flow rate of 1.0 m L/min.The column tempreture was maintained at 30 °C and the detection wavelength was set at 210 nm.Results: 1.Target component separation: 9 compounds were isolated and purified from Rehmanniae Radix Praeparta by chemical column chromatography.And they are identified as eight phenylethanoid glycosides and two iridoid terpenoids by physicochemical properties,spectral and spectral data.Among the phenylethanoid glycosides,they are purpureaside C,echinacoside,jionoside A1,acteoside,jionoside B1,isoacteoside,jionoside D and iridoids are gentiopicroside and swertioside.2.The Study on the improvement of quality standard of Rehmanniae Radix Praeparta: The thin layer chromatograms of the Rehmanniae Radix and the Rehmanniae Radix Praeparta are clear,the band in the chromatogram obtained with test solution corresponds in position and colour to the band in the chromatogram obtained with the reference drug and reference solution on the same Rf value.The method for HPLC determination of Rehmanniae Radix and Rehmanniae Radix Praeparta has been investigated and verified,which has good accuracy and durability,and overcomes the shortcomings of reference standards,which is simple and feasible.The content of the four phenylethanoid glycosides in the 18 batches of Rehmanniae Radix and 17 batches of Rehmanniae Radix Praeparta were determined to be in the range of 0.4504 to 2.5086 mg/g and 0.4644 to 1.9778 mg/g,respectively.The similarity between 18 batches of Rehmanniae Radix and 17 batches of Rehmanniae Radix Praeparta was above 0.9(except one batche of Rehmanniae Radix).3.The Study on the improvement of quality standard of Liuwei Dihuang Capsules: In the thin layer qualitative identification of Liuwei Dihuang Capsules,the information of each sample was identical with that of the reference substance,while the negative sample had no corresponding spots.The two components content determination method has good precision and high accuracy.The content range of acteoside in 25 batch samples of Liuwei Dihuang Capsules was 0.005~0.065 mg/ granule.The content range of pachymaric acid in 25 batch samples of Liuwei Dihuang Capsules was 0.046~0.078 mg/ granule.Conclutions:1.Drafted quality standard for Rehmanniae Radix and Rehmanniae Radix Praeparta,add or improve the current Pharmacopoeia of Rehmanniae Radix and Rehmanniae Radix Praeparta thin layer identification methods,and stipulates that the content of four kinds of phenylethanoid glycosides in Rehmanniae Radix should not be less than 0.08%,and the content of four kinds of phenylethanoid glycosides in Rehmanniae Radix Praeparta should not be less than 0.07%.Compared with before and after treatment,the similarity was high,indicating that the main chemical components were basically no difference between Rehmanniae Radix and Rehmanniae Radix Praeparta.At the same time,a quantitative analysis of multi-components with single marker combined with HPLC fingerprint feature distinguishing method can be successfully used to evaluate the quality of Rehmannia.2.Drafted the internal control draft of Liuwei Dihuang Capsule,which stipulates that the content of acteoside in Liuwei Dihuang Capsule should not be lower than 20?g/granule and the content of pachymaric acid should not be lower than 35?g/granule.