1、Liuwei Dihuang Pills(concentrated pills)六味地黃丸（濃縮丸）Pinyin Liuwei Dihuang WanIgredients Prepared Rehmannia Root 120g; Fructus Macrocarpii 60g; Cortex Moutan Radicis 45g; Rhizoma Dioscoreae Oppositae 60g; Poria 45g; Rhizoma Alismatis 45gProcedure Distillate Cortex Moutan Radicis with water vapo for abs

2、tracting volatility ingredients; Decoct the gruffs, Fructus Macrocarpii 20g, Prepared Rehmannia Root, Poria and Rhizoma Alismatis with water twice, 2 hours for every time, filter, combine the filtrated, concentrated them to extract; pulverize Rhizoma Dioscoreae Oppositae and excess of Fructus Macroc

3、arpii to fine powder, sifting, mix thoroughly, add the above extract and volatility ingredients of Cortex Moutan Radicis, mix thotoughly, make to pills, dry, polishing.Description Brownish to black concentrated pills, taste, sweat, acid, slightly bitter.Identification (1)Microscopical: Epidermal cel

4、ls of pericarp orange-yellow, polygonal in surface view, with somewhat beaded anticlinal walls. Starch granules triangular-ovoid or oblong, 24-40 m in diameter hila short cleft Vshaped. (2)Pulverize 5 g of the pills,heat under reflux with 20 ml of ether for 1 hour, filter and expel ether. Dissolve t

5、he residue in 1 ml of acetone as the test solution. Dissolve paeonol CRS in acetone to prepare a solution containing 1 mg per ml. as the reference solution. Carry out the method for thin layer chromatography (Appendix B), using silica gel G as the coating substance and cyclohexane and ethyl acetate(

6、3:1) as the mobile phase. Apply separately 5-10 l of each of the two solution to the plate. After developing and removal of the plate, dry in air, spray with 5% ferric chloride in ethanol acidified with hydrochloric acid and heat to the spots clear. The spot in the chromatogram obtained with the tes

7、t solution corresponds in position and colour to the spot in the chromatogram obtained with the reference solution.(3)Pulverize 10 g of the pills, dissolve it with 100 ml of water, heat to boil, allow to stand for cool, filter in absorbent cotton, wash the filtrate with two-30 ml quantities of aceti

8、din, combine the filtrate, evaporate the filtrate to dryness, dissolve the residue in 1 ml of methanol as the test solution. Prepare a solution of 4 g of Prepared Rehmannia Root reference drug and 60 ml water, decoct 30 minutes, stand for cool, filter with absorbent cotton, wash the filtrate with tw

9、o-20 ml quantities of acetidin, combine the filtrate, evaporate the filtrate to dryness, dissolve the residue in 1 ml of methanol as the reference solution. Carry out the method for thin layer chromatography(Appendix ), using silica gel G as the coating substance and the lower layer of a mixture of

10、chloroform, xylene and acetidin(1:1), as the mobile phase. Apply separately 3-5l of each of the two solution to the plate. After developing and removal of the plate, dry in air. Spary with 2,4 -dimethylethanolnitrile to spots clear. The spot in the chromatogram obtained with the test solution corres

11、pond in position and colour to the spot in the chromatogram obtained with the reference solution and the reference drug solution.(4)Pulverize 5 g of the pills, dissolve it with 30 ml of water, cool and filter, wash the residue with 30 ml of water, and heat under reflux the residue, with 50 ml of 30%

12、 hydrochloric acid for 1 hour, cool and filter, wash the filtrate with two-25 ml chloroform, evaporate the filtrate to dryness, dissolve the residue in 1 ml of chloroform as the solution. Dissolve the 0.3 g of Rhizoma Dioscoreae Oppositae drug in 50 ml of 30% hydrochloric acid, and make the referenc

13、e solution as the same way above. Carry out the method for thin thin layer chromatography(Appendix B), using silica gel G as the coating substance and mixture of chloroform and acetone （9 : 1.5）as the mobile phase. Apply separately 5 l of each of the test solution and the reference solution to the p

14、late. After the developing and removal the plate, dry in air, examine under the ultraviolet light at 365 nm. The fluorescent spots in the chromatogram obtained with the test solution correspond in position and colour to the spots obtained with the reference drug solution.(5)Pulverize 10 g of the pil

15、ls, dissolve it in 100 ml of water, cool and filter in absorbent cotton, wash the residue with 50 ml of petroleum ether (60-90) third, evaporate the filtrate to dryness, dissolve the residue in 1 ml of petroleum ether (60-90) as the test solution. Decoct the 2 g of Rhizoma Alismatis drug with 50 ml

16、of water, cool and filter in absorbent cotton, and make the reference solution as the same way above. Carry out the method for thin layer chromatography (Appendix B), using silica gel G as the coating substance and mixture of petroleum ether, acetidin and chloroform (2:2:1) as the mobile phase. Appl

17、y separately 10-20 l of test solution and 10 l of reference drug solution to the plate. After developing and removal the plate, dry in air. Spary with 5% phosphomolybdic acid in ethanol and heat at 110 to the spots clear. The spot in the chromatogram obtained with the test solution correspond in pos

18、ition and colour to the spots obtained with the reference drug solution.(6)Pulverize 10 g of the pills, heat under reflux it with 50 ml of ether, cool and filter, evaporate the filtrate to dryness, dissolve the residue in n-hexane as the test solution. Heat under reflux 2 g Poria with 30 ml ether fo

19、r 1 hour, filter, evaporate the filtrate to dryness, dissolve the residue in n-hexane as the reference drug solution. Carry out the method for thin layer chromatography (Appendix B), using silica gel G as the coating substance and mixture of petroleum ether (60-90) and ether (3:2) as the mobile phas

20、e. Apply separately 20 l of test solution and 10 l of reference drug solution to the plate. After developing and removal the plate, dry in air, examine under the ultraviolet light at 365nm. The spot in the chromatogram obtained in the test solution correspond in position and colour to the spot obtai

21、ned with the reference drug solution.Other requirements Comply with the general requirements for pills(Appendix A).Assay Fructus Macrocarpii Carry out the method for high performance liquid chromatography(Appendix VI D).Chromatographic system and system suitability Use octadecylsilane bonded silica

22、gel as the stationary phase and a mixture of tetrahydrofuran, acetonitrile, methanol and 0.05% solution of phosphoric acid (1:8:4:87) as the mobile phase. As detector a spectrophotometer set at 230nm, maintain the column temperature at 40.The number of theoretical plates of the column is not less th

23、an 4000, calculated with reference to the peak of loganin.Reference solution Dissolve a quantity of loganin CRS, accurately weighed, in 50% methanol to prepare a solution containing 20 g per ml.Test solution Pulverize 0.4 g of the pills, accurately weighed, place to a stopper conical flask, add accu

24、rately 50 ml of 50% methanol, stopper tightly and weigh, heat under reflux for 1 hours and cool. Weigh again and replenish the lost weight with 50% methanol, mix well and filter. Messure accurately 10 ml fo the successive filtrate, apply to a column(1 cm in internal diameter) packed with neutral alu

25、minium oxide (100-200 mesh, 4 g). Elute with 50 ml of 40% methanol, collect the eluate, evaporate to dryness and dissolve the residue with 50% methanol, transfer to a 10 ml volumetric flask, dilute with 50% methanol to volume, mix well and filter.Procedure Inject accurately 10 l of the reference sol

26、ution and 10 l of the test solution into the column, and calculate the content.It contains not less than 1.4 mg per 1 g of concentrated pills of loganin(C17H26O10) , referred to Fructus Corni.Cortex Moutan Carry out the method for high performance liquid chromatography (Appendix VI D).Chromatographi

27、c system and system suitability Use octadecylsilane bonded silica gel as the stationary phase and mixture of methanol and water (70:30) as the mobile phase. As detector a spectrophotometer set at 230 nm. The number of theoretical plated of the column is not less than 3500, calculated with the reference to the peak of paeonol. Reference solution Dissolve a quantity of paeonol CRS, accurately weighed, in 50% methanol to prepare a solution containing 20 l per ml.Test solution Pulverize 0.4 g of the pills, weigh accurately, place to a stoppe