Asiaticoside content

REAGENTS & INSTRUMENT REQUIRED:

1. High Performance Liquid Chromatography

2. Water HPLC grade

3. Methanol LR grade

4 .Asiaticoside working standard

PROCEDURE:

Standard Preparation :

Weigh accurately about 25mg of in working standard into a 25mL volumetric flask. Add 20mL methanol. Ultrasonicate for 15 minutes. Cool and make up the volume with methanol. Shake well and filter through 0.45μm filter paper.

Sample Preparation :

Weigh accurately about 100mg of sample into a 100 mL volumetric flask.  Add about 20mL methanol. Ultrasonicate for 15 minutes. Cool and make up the volume with methanol. Ensure that the concentration of active ingredient in sample solution is approximately similar to that of std solution. Shake well and filter through 0.45μm filter paper.

For Raw material: Take 10 gm of herb powder(20#) and reflux in methanol for 1hrs. twice

Chromatographic Condition:

Mobile Phase               - Acetonitrile : Water [ 60 : 40 ]                                        

Column                        - Phenomenex luna 5µ, C18 (250mm X 4.6mm)

Mode                           – Isocratic

Injection Volume         – 20 µL

Flow rate                     – 1.0mL /minute

Detector wavelength   – 220nm

Condition                     - RT

Calculation:

                                        Area of sample  X Concentration of std    X   purity of std

  % of  Asiaticoside    =-----------------------------------------------------------------------------------

                                            Area of standard    X  Concentration of sample