Determination of Asiaticoside and Madecasoside 

Determination of Asiaticoside and Madecasoside 

1) Reagents

a. Nanopure water       b. Acetonitrile (HPLC)        

2) Apparatus

HPLC (Shimadzu)            Analytical balance (1/10000)         Ultrasonic cleaner

0.45μm PTFE filter          50ml volumetric flask                Screw cap vials detector(Alltech)

3) HPLC condition

Column:  Kromasil C18 5 mm  4.6′200mm

Mobile Phase: A:Water      B: Acetonitrile 

A:B=67:33

Flow Rate: 0.7ml/min

Injection: 10μl

Drift tube temperature :100.2℃

Gas flow:2.7L/min

Impactor: Off

Gain:2

4) Procedure

Samples Preparation

  Accurately weigh 30mg extraction into 50ml volumetric flask. Add Water to volume and sonicate to mix, filter through a 0.45μm PTFE filter into a screw cap vial and proceed with the analysis.

Standard preparation /calibration

Prepare a standard solution (approximately 0.2mg/ml in Water using a standard of Asiaticoside and Madecasoside ，Sonicate to mix, filter through a 0.45μm PTFE filter into a screw cap vial and proceed with the analysis.

5) Calculation

Determine the peak area’s logarithm of Asiaticoside and Madecasoside the on the sample chromatogram and compare to the corresponding peaks in the chromatogram for the standard  solution.