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Test method for Centella Asiatica Extract Total Glucosides——( CP2010 )
Jun 22, 2018

Centella Total Glucosidesare the saponin extracts prepared from Centella Asiatica Herba.


【Description】 Faint yellow to faint brownish-yellow powder; odour, odorless; taste, bitter; slightly hygroscopic. Solublein water and ethanol. Insoluble in chloroform and ether.


【Identification】

(1) Place 2 mg, in a test tube, add 1 ml of anhydride acetic, and shake well. Slowly add 1 ml of sulfuric acid along the test tube wall, and a violet-red ring is produced on the two-liquid border.

(2) Dissolve a quantity in ethanol to produce a solution containing 10 mg per ml as the test solution. Weigh separately a quantity of madecassoside CRS and asiaticoside CRS, add ethanol to prepare the solutions containing 10 mg per ml, respectively, use as the reference solutions. Carry out the method for thin layer chromatography (Appendix Ⅳ B). Using silica gel G as the coating substance and the upper layer of a mixture of n-butanol, ethyl acetate and water( 4:1:5)as the mobile phase. Apply separately 5 µl of each of the three solutions to the plate. After developing and removal of the plate, dry in air. Spray with a solution composed of anhydride acetic, sulfuric acid, and anhydrous ethanol (1:1:10), heat at 105℃ to the spots clear. Two bluish-brown spots in the chromatogram obtained with the test solution correspond in position and colour to the spots in the chromatogram obtained with the reference solutions.

 

【Loss on drying】 When dried to constant weight at 105℃, loses not more than 10.0 per cent of its weight (Appendix Ⅸ G).


【Heavy metal and harmful elements】 Carry out the method for determination of lead (Pb),Cadmium(Cd),Arsenic(As),Mercury(Hg),Copper(Cu) (Appendix Ⅸ B). Not more than 5 ppm of lead, not more than 0.3 ppm of codmiun, not more than 2 ppm of Arsenie, not more than 0.2 ppm of Mercury,not more than 20 ppm of Copper.


【Fingerprint】 Carry out the method for high performance liquid chromatography (AppendixVI D) . Chromatographic system and system suitability Use octadecylsilane bonded silica gel as the stationary phase (25cm in length ,4.6mm in inner diameter ,5µm in particle diameter )and a mixture of acetonitrile and 2mmol/L cyclodextrin solution (24:76) as the mobile phase .As detector a sepctrophotometer set at 205 nm. The number of the theoretical

plates of the column is not less than 4000,calulated with the reference to the peak of asiaticoside . Reference solution Weigh accurately Madecassoside CRS and Asiaticoside CRS separately , add methanol to prepare the solutions containing 0.2mg per ml respectively, as the reference solutions . Test solution Carry out the mehod for the test solutionpreparation underAssay . Procedure Inject 10 µl of each of the reference solutions and the test solution into the columnrespectively,and record the chromatogram.


According to the Similarity evaluation system for chromatographic fingerprint of TCM ,calculate the similarity between test solution fingerprint and reference fingerprint ,not less than 0.90.

Integration events Slope sensitivity :1, peak width: 0.1,area reject :20 ,height reject :0.5 .


【Assay】 Carry out the method for high performance liquid chromatography (Appendix VI D) . Chromatographic system and system suitability Use octadecylsilane bonded silica gel as the stationary phase and a mixture of acetonitrile and 2mmol/L cyclodextrin solution

(24:76) as the mobile phase .As detector a sepctrophotometer set at 205 nm. The number of the theoretical plates of the column is not less than 4000,calulated with the reference to the peak ofasiaticoside . Reference solution Weigh accurately Madecassoside CRS and Asiaticoside CRS separately , add methanol to produce the solutions containing 0.2mg per ml respectively as the reference solutions . Test solution Weigh accurately 50 mg to a 50ml of volumetric flask ,dissolve in methanol ,diluteto volume ,shake well ,and filter . Procedure Inject 10 µl of each of the reference solutions and the test solution into the columnrespectively , calculate the content . It contains not less than 55.0 per cent of total content of Madecassoside (C48H78O20 ) and Asiaticoside (C48H78O19 ), calculated with reference to the dried basis .


【Storage】 Preserve in tightly closed containers .

 

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