Biological and chemical characterization of centella asiatica accession (CaFR04)
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Date
2007
Authors
Abas, Rafedah
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Abstract
Centella asiatica, locally known as pegaga contains a diverse array of
triterpenoid compound that have causal effects in various diseases. The major
constituents of pegaga are the four triterpenes, asiatic acid, asiaticoside,
madecassic acid and madecassoside. In Malaysia, there are 14 accessions of
C.asiatica that have been studied previously and four accessions were
identified as potential accessions of pegaga. In the present study, one
accession was selected among the potential accessions and named as a
CaFR04 to investigate the effect of harvesting period and the fertilizer treatment
on the production of triterpenes and the biological activities. The plant samples
were extracted with methanol and ethanol-aqueous and analysed by HPLC. A
high concentration of triterpenes was obtained in both methanol and ethanolaqueous
extract at month two of harvesting and chicken manure as a fertilizer.
Asiaticoside was always found at high concentrations but very low
concentrations of madecassic acid and asiatic acid were detected. CaFR04
shows high antioxidant activity at each month of harvesting and also from every
treatment of fertilizer and control group. Only methanol extract of CaFR04
harvested at month three was active against MCF-7, Vero and WRL-68 cell
lines whereas no activity was detected from each treatments of fertilizer and
control group. The study can be further clarified by comparing on the level of
proteins by using proteomics approach. The protein profile of CaFR04 and the
changes due to the response to the different harvesting period and fertilizer
treatment were obtained using SDS-PAGE and SELDI-MS analysis. The pattern
of the SDS-PAGE gel was similar at each month of harvesting and from each
treatment of fertilizer and control group. The SDS-PAGE is typically focused on
the 15 - 200 kDa range while SELDI-MS is used for analysis of proteins in
particular those of < 40 kDa. The protein peaks obtained from SELDI-MS
analysis varied with different ProteinChip® surfaces. The SDS-PAGE gel profile
of CaFR04 was compared with other accessions. Different pattern of protein
bands was observed. The 31 kDa protein bands found in abundance in CaFR04
were also detected in other accessions, namely CaFR05, CaFR06, CaFR07
and CaFR08 but the 23 kDa protein bands present in CaFR04 were not
detected in other accessions. The 31 kDa and 23 kDa protein bands were
digested with trypsin and the peptides were analysed by MALDI TOF MS. The
31 kDa protein was identified to match the oxygen-evolving enhancer protein 1
from Pisum sativum (garden pea) but no significant match for 23 kDa protein
was found.
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Keywords
Chemical characterization , Centella asiatica