Centella asiatica (L.) Urb. (Family Apiaceae commonly known as Gotu Kola or Indian Pennywort) has long
been used in the Ayurvedic system of medicine for improving memory and for the treatment
of a variety of ailments [1]. The triterpenoid compounds purportedly represent the
chief pharmacologically active constituents. The triterpenoids, especially asiticoside,
triterpine trisaccharide, are reported as the most active compounds in the plant [2].
A simple and fast method was developed for the quantitative determination of four
triterpenes and their glycosides i.e. asiatic acid (AA), madecassic acid (MA), asiaticoside
(AS) and madecoside (MS) in Centella asiatica and Centella erecta by using high performance thin layer chromatographic method. The separation was achieved
with chloroform: methanol: water: 13.0:6.5:0.5 v/v/v on silica gel 60F254 HPTLC plates. Quantitation was performed with densitometry in absorption-reflection
mode at 600 nm by scanning the HPTLC plates after a color development by anisaldehyde
reagent. The linear regression data for the calibration plots showed a good linear
relationship with r = 0.999, 0.998, 0.997 and 0.998 for asiatic acid, madecassic acid,
asiaticoside and madecoside, respectively. The established method was validated in
terms of LOD and LOQ, linearity. Acknowledgements: This research is funded in part by The United States Department of Agriculture Specific
Cooperative Research Agreement Number 58-6408-6-067 and the FDA/CFSAN grant entitled
Science Based Authentication of Dietary Supplements Number 2 U01 FD 002071-07 References: [1] Shakir JS, et al. (2007), Nat. Prod. Radiance, 6 (2): 158–170. [2] de Paula Reis,
et al. (1996), Revista Brasileira de Farmácia, 77(2): 71–72.
Fig. 1 (A) Centella asiatica, (B) Standard mix, (C) Centella erecta.
Table 1 Validation Parameters.
|
|
Parameter
|
AA
|
MA
|
AS
|
MS
|
|
1
|
Linearity range (ng/spot)
|
200–600
|
200–600
|
100–500
|
100–500
|
|
2
|
Correlation coefficient
|
0.999
|
0.998
|
0.997
|
0.998
|
|
4
|
LOD (ng/spot)
|
30
|
60
|
30
|
30
|
|
5
|
LOQ (ng/spot)
|
180
|
200
|
100
|
100
|
|
6
|
Specificity
|
Specific
|
Specific
|
Specific
|
Specific
|
|
7
|
Regression equation
|
Y=94.580 +8.961 X
|
Y=61.937 +3.124 X
|
Y=22.600 +0.495 X
|
Y=12.773 +0.113 X
|
|
8
|
Rf
|
0.72
|
0.61
|
0.17
|
0.11
|
Table 2 Percentage (w/w) of asiatic acid, madecassic acid, asiticoside, and madecoside in
plant sample.
|
Sample name (Percentage in dry plant material)
|
AA
|
MA
|
AS
|
MS
|
|
C. asiatica
|
0.2
|
0.2
|
3.6
|
2.0
|
|
C.erecta
|
0.1
|
0.1
|
4.5
|
3.1
|
