Page 44 - Drug Discovery and Development: Prospects and Challenges

34 / Drug Discovery and Development: Prospects and Challenges

plate using the TLC to identify the presence of asiaticoside. All fractions
contained asiaticoside except the EtOAc fraction. The TLC profile spots
showed that the Rf value of the asiaticoside in the methanol:chloroform
mobile phase was 0.77, which was similar to the Rf value of the standard
asiaticoside.

HPLC Analysis

The HPLC technique was performed to determine the concentration
of asiaticoside content in each extracted sample. Based on the HPLC
analysis, the concentration of Activated Charcoal Fraction (CA
FAC), Silica Gel Ethyl Acetate Fraction (SGE FAC), Silica Gel Ethyl
Acetate:Methanol Fraction (SGEM FAC), and Silica Gel Methanol
Fraction (SGM FAC) were 10 mg/mL each, while the concentration of
asiaticoside standard was 1 mg/mL. All fractions and standards were
soluble in a methanol-water mixture. The retention time and peak area of
each sample were tabulated in Table 2. The CA FAC displayed the highest
peak area compared to other fractions with the peak value of 412 870 at
21.855 retention time. Therefore, CA FAC was selected for the upscale
production of the hydrogel. Figure 25 shows the HPLC chromatogram
of all fractions including the asiaticoside standard.

Table 2: The retention time and peak area for all samples including
the asiaticoside standard
Retention Peak
Samples
time value
Asiaticoside standard 21.789 832 861
Activated charcoal fraction (CA FAC) 21.855 412 870
Silica gel ethyl acetate fraction (SGE FAC) 22.078 28 888
Silica gel ethyl acetate: methanol fraction 21.144 48 526
(SGEM FAC)
Silica gel methanol fraction (SGM FAC) 21.971 248 906