Quality Control Analysis, Standardization And Stability Studies Of Eurycoma Longifolia And Effect Of Its Saponin Content On Solubility And Toxicity Of Eurycomanone

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dc.contributor.authorKhari, Nursyazura
dc.date.accessioned2016-12-05T09:20:03Z
dc.date.available2016-12-05T09:20:03Z
dc.date.issued2014-08
dc.description.abstractQuality control test was done on E. longifolia root, stem and leaf from five different localities in order to determine the range of quality control values. Moisture, ash and acid insoluble ash content of all the root raw material were found to be within acceptable limit set by Standard of Asean Herbal Medicine and Malaysian Herbal Monograph Volume 1. The range of quality control values of the stem and leaf raw material have been set. The percentages of water soluble extracts were higher as compared to alcohol soluble extracts in all parts of the plant. Percentage extractive values by the hot method were higher as compared to the cold method for both water and alcohol soluble extracts. The amounts of Pb, Cd, As and Hg in E. longifolia raw material were found within acceptable limit according to World Health Organization. Extraction method was optimized by different root powder: solvent ratio and extraction time. Results suggest the best extraction conditions at the root powder: water ratio of 1:15 and at 3 reflux cycles each for 4 h. The percentage yield and chemical profile involving qualitative analysis (HPTLC, UV and HPLC) and quantitative analysis (HPLC) were carried out for all the extracts. Analytical spectroscopy and chromatographic methods were used for chemical profiling and standardization of E. longifolia root, stem, leaf water extracts and six registered commercial products. HPLC validated methods give the percentage of eurycomanone for root, stem, leaf and commercial products in a ranges 0.89 ± 0.01% to 3.28 ± 0.01%, 0.23 ± 0.10% to 1.10 ± 0.00%, 0.10 ± 0.00% to 0.34 ± 0.01% and 0.07 ± 0.00% to 0.16 ± 0.00%, respectively. The percentage of total phenolics, glycosaponins, polysaccharides and protein for root ranging from 5.94 ± 0.23% to 10.10 ± 0.08%, 40.12 ± 1.05% to 44.45 ± 0.54%, 25.30 ± 0.15% to 51.30 ± 2.56% and 21.42 ± 0.6% to 44.92 ± 2.5%, respectively. The percentage of total phenolics, glycosaponins, polysaccharides and protein for stem ranging from 4.20 ± 0.13% to 8.76 ± 0.30%, 24.09 ± 1.06% to 47.24 ± 1.07%, 20.53 ± 0.48% to 68.18 ± 0.22% and 12.17 ± 0.6% to 48.20 ± 2.0%, respectively. The percentage of total phenolics, glycosaponins, polysaccharides and protein for leaf ranging from 15.88 ± 0.10% to 22.33 ± 0.28%, 52.26 ± 0.31% to 61.58 ± 0.44%, 7.59 ± 0.04% to 12.59 ± 0.11% and 58.90 ± 1.6% to 81.58 ± 2.3%, respectively. The percentage of total phenolics, glycosaponins, polysaccharides and protein for commercial products ranging from 2.53 ± 0.00% to 3.34 ± 0.01%, 3.31 ± 0.31% to 16.44 ± 0.21%, 5.98 ± 0.54% to 66.17 ± 3.96% and 6.4 ± 0.2% to 7.8 ± 0.3 %, respectively. In accelerated stability study, extracts (BR, BS and BL) were more stable at 30ºC compared to other temperatures studied, and the estimated shelf life (t90) of BR, BS and BL were 0.31, 0.23 and 0.36 months, respectively, at 30ºC. Eurycomanone has higher shelf life in the leaf (BL) as compared to root (BR) and stem (BS). The marker compound followed the first order degradation and its degradation rate was increased by increasing storage temperature and humidity. Therefore, in order to guarantee a longer shelf life of the products, it is recommended to store the products containing E. Longifolia extract at below 30ºC in a tightly closed container. Saponins increase the solubility of eurycomanone through the formation of micelles as indicated by the presence of sub-micron particles. The Critical Micellar Concentration (CMC), was estimated to be 267 μg/mL for the crude root water extract (TA), 57 μg/mL for saponins-rich extract and 847 μg/mL for eurycomanone-rich extract. These results indicate the presence of micelles and show the role of saponins in the formation of these micelles as indicated by the lower CMC value in saponins-rich extract compared to the CMC value of eurycomanone-rich extract. Cytotoxicity effect of eurycomanone-rich extract on HCT 116 colorectal carcinoma cells showed dose dependent growth inhibitory effect with median inhibitory concentration (IC50) of 22.14 ± 1.0 μg/mL, whereas the saponins-rich extract at the same concentration range did not show any cytotoxic effect on the same cells. Cytotoxicity effect of combination saponins and eurycomanone-rich extract on HCT 116 cells was increased in a dose dependent manner. It was found that adding the saponins-rich extract to the eurycomanone-rich extract reduced the cytotoxic effect of the later particularly at low concentration of the saponins-rich extract (25, 50 and 75 μg/mL), and has no effect at higher concentration (100 μg/mL). These findings indicate that the presence of saponins in the E. longifolia extracts protect against the cytotoxic effect of the eurycomanone. Furthermore, these findings indicate that using the crude extracts of E. longifolia could be safer than using pure compounds or concentrated fractions of eurycomanone.en_US
dc.identifier.urihttp://hdl.handle.net/123456789/3245
dc.language.isoenen_US
dc.publisherUniversiti Sains Malaysiaen_US
dc.subjectQuality control values.en_US
dc.subjectE. longifolia root, stem and leaf from five different localities.en_US
dc.titleQuality Control Analysis, Standardization And Stability Studies Of Eurycoma Longifolia And Effect Of Its Saponin Content On Solubility And Toxicity Of Eurycomanoneen_US
dc.typeThesisen_US
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