• Bulletin of the Korean Chemical Society
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• v.29 no.3
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• pp.551-554
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• 2008

An improved HPLC method for the separation of madecassoside isomers (madecassoside and asiaticoside-B) has been developed. The isomers can be separated with high resolution from extracts of Centella asiatica by HPLC using $\beta$-cyclodextrin as a mobile phase added on a $C_{18}$ column. The result shows that the isomers can be separated with a mobile phase consisting of methanol:water (50:50, v/v) with 4 mmol/L $\beta$-CD. To elucidate the mechanism of the separation of madecassoside and asiaticoside-B, this paper studied the separation of their aglycon parts (madecassic acid and terminolic acid), another pair of isomers. The isomers can also be separated with high resolution with a mobile phase consisting of methanol:water (65:35, v/v) with 4 mmol/L $\beta$-CD and the pH of the mobile phase was adjusted to 4. The paper also studied the separation of the two isomers by HPLC using $\alpha$-CD and Glucosyl-$\beta$-CD as a mobile phase additive in order to elucidate the mechanism of the separation process.

• Journal of Pharmaceutical Investigation
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• v.28 no.1
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• pp.15-23
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• 1998

Extract of Centella asiatica(ECA), which is poorly water-soluble extract from the Centella asiatica is known to express excellent wound healing properties. $ECA-{\beta}-cyclodextrin$ $(asiaticoside-{\beta}-cyclodextrin\;and\;genin-{\beta}-cyc1odextrin)$ solid dispersion system, which was prepared by freezedrying method, was formulated as gels containing poloxamer 407 and propylene glycol, and evaluated with respect to their viscosity, stability, skin permeation and drug amount in the skin of hairless mouse. The average molar ratio $asiaticoside-{\beta}-CD$ and $genin-{\beta}-CD$ was 1:1.7 and 1:22, respectively. When the molar ratio of genin and ${\beta}-CD$ was 1:5, madecassic acid made 100% solid dispersion system and asiatic acid about 65%. In dissolution study, >99% of asiaticoside from $asiaticoside-{\beta}-CD$ was dissolved in 5 minutes, and >99% madecassic acid and >64% asiatic acid from $genin-{\beta}-CD$. The apparent viscosity of poloxamer 407 gels with $ECA-{\beta}-CD$ solid dispersion system increased in proportion to poloxamer 407 and propylene glycol concentration. In the accelerated stability test, all $ECA-{\beta}-CD$ poloxamer 407 gels showed that asiaticoside was most stable and madecassic acid stable and asiatic acid similar to stability of gel with free ECA. The permeation amount of asiaticoside in poloxamer gels through hairless mouse skin decreased as the concentration of poloxamer 407 increased. When propylene glycol was added at the level of 10%, the permeation amount of asiaticoside at poloxamer gels through hairless mouse skin increased but from 15% it decreased. The permeation of asiaticoside into the skin of hairless mouse was estimated to be about $0.10\;{\mu}g/cm^2$.

• Archives of Pharmacal Research
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• v.28 no.4
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• pp.502-508
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• 2005

Titrated extract of Centella asiatica (TECA) contains three principal ingredients, asiaticoside (AS), asiatic acid (AA), and madecassic acid (MA). These components are known to be clinically effective on systemic scleroderma, abnormal scar formation, and keloids. However, one problem associated with administration of TECA is its low solubility in aqueous as well as oil medium. In this study, various nonionic surfactants and bile salts as anionic surfactant were tested and screened for solubilizing TECA with a view to developing topical hydrogel type of ointment which is stable physicochemically, and has better pharmacological effects. When TECA was incorporated into various nonionic surfactant systems, labrasol had the most potent capacity for solubilizing TECA. In cases of bile salt systems, Na-deoxycholate (Na-DOC) had foremost solubilizing capacity, even more than labrasol. In differential scanning calorimetric study, the peaks of AA, MA, AS and Na-DOC disappeared at the coprecipitate of $1\%$ TECA and $1\%$ Na-DOC, suggesting the optimum condition of Na-DOC for solubilizing TECA. When the physicochemical stability of hydrogel containing this mixture was assessed, it was stable at room temperature for at least one month. Pharmacologically it significantly decreased the size of wound area at the $9^{th}$ day when applied to the wound area of rat dorsal skin. Taken together, solubility of TECA was dramatically improved by using non ionic and anionic surfactant systems, and Na-DOC was found to be the most effective solubilizer of TECA in formulating a TECA-containing hydrogel typed ointment. Moreover this gel was considered to be applicable to clinical use for wound healing effect.

• Proceedings of the PSK Conference
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• 2002.10a
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• pp.340.1-340.1
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• 2002

Centella asiatica is an herbal plant used on different continents by diverse ancient cultures and tribal groups. Historically. the extract has been used as a wound healing agent, The extract has three different triterpenoid ingredients: asiaticoside. asiatic acid, and madecassic acid. It has been reported that its wound healing activity is associated with the modulation of collagen synthesis in the skin dermis. The wound healing property of the extract has led to its commercial introduction under the trade name, Madecrlssol. As part of our program toward the development of new wound healing agents. structere activity relationship (SAR) studies have been performed by modifying asiatic acid. In this communication. the SAR study of esiatic acid for the development of an efficient woind healing agent is reported.

• Natural Product Sciences
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• v.13 no.1
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• pp.33-39
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• 2007

Eleven polyhydroxyursane triterpenoids (PHUTs) were tested to determine their cytoprotective, immunosuppressive and anti-inflammatory effects. To compare the bioactivities of $19{\alpha}$-hydroxyursane-type triterpenoids {23-hydroxytormentic acid (6), its methyl ester (7), tormentic acid (8), niga-ichigoside $F_1$ (9),euscaphic acid (10) and kaji-ichigoside $F_1$ (11)} of the Rosaceae crude drugs (Rubi Fructus and Rosa rugosae Radix) with PHUTs possessing no $19{\alpha}-hydroxyl$ of Centella asiatica (Umbelliferae), the four PHUTs, asiaticoside (1), madecassoside (2), asiatic acid (3), and madecassic acid (4) were isolated from C. asiatica and 23-hydroxyursolic acid (5) from Cussonia bancoensis. Cytoprotective effects were assessed by measuring cell viabilities against cisplatin-induced cytotoxocity in $LLC-PK_1$, cells (proximal tubule, pig kidney) to determine whether these agents have protective effects against nephrotoxicity caused by cisplatin. The inhibitory effect of 11 PHUTS on nitric oxide (NO) and prostaglandin $E_2\;(PGE_2)$ were evaluated by measuring nitrite accumulation in lipopolysaccharide (LPS)-induced macrophage RAW 264.7 cells, and their anti-inflammatory effects were tested in 12-O-tetradecanoylphorbol-13-acetate (TPA)-induced mouse ear edema model. Six MHUTs (compounds 1, 2, 4, 6, 10, and 11) exhibited higher cell viabilities during cisplatin-induced cytotoxicity testing even at a concentration of $200\;{\mu}g/ml$ than cisplatin only-treated group, suggesting that ese compounds have the potentcytoprotective efffcts. Compounds 1 and 3 of the C. asiatica and niga-ichigoside $F_1$ exhibited no inhibitory effect on NO and/or $PGE_2$ production whereas other PHUTs produced mild to significant NO and/or $PGE_2$ production.The four compounds (2, 5, 9, and 10) potently inhibited mouse ear edema induced by TPA whereas two compounds (1 and 3) had no activity in this test. These results suggest that many PHUTs are potentchemopreventives. Structure-activity relationship (SAR) was also discussed in each assay with regard to the significant role of OHs at the position of 2, 3, 6, 19, and 23 and to the glycoside linkage at the 28-carboxyl.

• Natural Product Sciences
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• v.22 no.1
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• pp.20-24
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• 2016

Pentacyclic triterpenes, mainly, asiatic acid, madecassic acid, asiaticoside, and madecassoside are the active constituents of Centella asiatica. A pentacyclic triterpene enriched C. asiatica extract (PRE) was prepared and standardized to contain a total pentacyclic triterpenes not less than 65% w/w. This work was focused on determination of antiinflammatory, antioxidant, and tyrosinase inhibitory activities of PRE and its stability. The PRE exhibited a satisfactory nitric oxide inhibitory effect, with an $IC_{50}$ value of $64.6{\mu}g/mL$. In addition, the PRE inhibited tyrosinase enzyme activity with an $IC_{50}$ value of $104.8{\mu}g/mL$. In contrast, the PRE possessed only weak antioxidant activity. The PRE was stable over a period of four months when stored as a dried powder but only in a well-closed container protected from light at $4^{\circ}C$. An aqueous alcoholic solution of the PRE was stable at pH values of 5.8 and 7.0, but was not stable at a pH of 8.2. Preparations of the PRE in an aqueous solution should be performed in acidic or neutral conditions.

• The Korean Journal of Food And Nutrition
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• v.36 no.6
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• pp.436-444
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• 2023

To produce an intestinal immunomodulatory beverage containing Centella asiatica extract (CAE), three types of CAE-added beverage prototypes were prepared, and their immunomodulatory activities and marker compounds were analyzed. As a result of the cytotoxicity assessment, all the beverages did not show significant toxicity compared to the control group. Next, the immunomodulatory activities of the beverage prototype were evaluated using the inflammatory model of IL-1β-induced intestinal epithelial cell line. All the samples significantly reduced the production of IL-6, IL-8, and MCP-1 in a CAE concentration-dependent manner. In addition, CAE-added beverages inhibited NO, IL-6, and IL-12 production in LPS-induced RAW 264.7 cells. When the major triterpenoids, as marker compounds for the production of CAE-added beverages, were analyzed by HPLC-DAD, only asiaticoside was detected beyond the limit of quantification, while madecassoside, madecassic acid, and asiatic acid were not detected. The amounts of asiaticoside in CAE-added beverage prototypes were confirmed in No. 1 (19.39 ㎍/mL), 2 (19.25 ㎍/mL), and 3 (19.98 ㎍/mL). In conclusion, the results of this study suggested that CAE-added beverage prototypes induced immunomodulatory effects in the intestinal inflammatory cell line models and asiaticoside could be used as a marker compound for CAE-added beverage production.

• Journal of the Society of Cosmetic Scientists of Korea
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• v.45 no.4
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• pp.363-372
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• 2019

The leaves and stems of Centella asiatica have a long history of their usage as a medicine for the treatment of skin diseases such as ulcers and psoriasis, especially in Asia. Triterpenoids, the active components of Centella asiatica including asiaticoside, madecasosside, asiatic acid and madecassic acid, have shown to inhibit skin inflammation as well as improve skin photoaging. The main objective of this study is to investigate whether the Centella asiatica ripened with lava seawater which is rich in minerals known to be beneficial to human body can provide anti-inflammatory and moisturizing effects to skin. HPLC analysis showed that the concentration of triterpenoids increased further after ripening Centella asiatica with lava seawater. In order to confirm the inflammatory efficacy of the extract of the extract of the ripened Centella asiatica, the production of NO in LPS-activated RAW 264.7 cells and the expression of inflammatory cytokines in PM10 or UVB-induced HaCaT cells were observed. We found that the extract of the ripened Centella asiatica inhibited the expression of NO, IL-6, IL-8, and TNF-a and had higher inhibitory effect compared to the extract of the non-ripened Centella asiatica. In order to confirm the skin moisturizing effect, we investigated the synthesis of HA in HaCaT cells. The result showed HA production was enhanced in a concentration-dependent manner from the ripened group, while there was no efficacy from the non-ripened group. Taken together, it is concluded that the extract of the Centella asiatica ripened with lava seawater was effective in anti-inflammation and moisturization.

• Journal of Life Science
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• v.28 no.8
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• pp.917-922
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• 2018

Centella asiatica is one of the local herbs that is claimed to possess various physiological effects. C. asiatica also accumulates large amounts of pentacyclic triterpenoid saponins known as centelloids. These terpenoids usually include asiatic acid, asiaticoside, madecassoside, and madecassic acids. In the present study, to understand the changes of triterpene glycoside levels in the different shoot tissues of C. asiatica during seasonal cultivation, we investigated the High-Performance Liquid Chromatography (HPLC) analysis via different extraction methods, such as water, 20% ethanol and methanol extracts. Significant increases were observed in the levels of madecassoside and asiaticoside in the leaf extracts by methanol compare with extracts using water or 20% ethanol. Additionally, we also analyzed the various antioxidant activity in the different shoot tissues of C. asiatica using different extracts, such as leaves, petioles and both materials. Among these petiole extracts showed high 2,2-azino-bis (3-ethylbenzothiazoline-6-sulfonic acid) diammonium salt (ABTs) scavenging activity in all extracts, whereas 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging activity exhibited high activity levels in the leaf part using methanol and ethanol extracts. Levels of total phenolics and flavonoid also showed the highest levels in the leaf tissues using all extracts such as methanol, ethanol and water. Our results indicated that the increased levels of triterpene glycoside and antioxidant activity in the leaf parts of the C. asiatica were indicating that useful metabolites were mainly maintained through seasonal cultivation such as madecassoside, asiaticoside.

• The Korean Journal of Food And Nutrition
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• v.33 no.6
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• pp.692-701
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• 2020

Centella asiatica (CA) has been widely used as herbal plants. It is a valuable resources. The aim of the present study was to evaluate physiological activities of solvent extracts from CA cultivated in Chungju, Korea (Good tiger care). After preparing water (cold-water, CA-WE; hot-water, CA-HWE) and EtOH extracts (50% EtOH, CA-50E; 70% EtOH, CA-70E), their total polyphenol, flavonoid, and triterpenoid contents, and anti-oxidant activities, and anti-inflammatory activities were examined and compared. CA-70E showed the most potent anti-oxidant activity based on ABTS radical scavenging activity and reducing ability assays, while CA-50E and CA-70E showed the highest DPPH radical scavenging activity. Among major triterpenoid glycosides present in CA, madecassoside and asiaticoside contents were found to be the highest in CA-70E, and madecassic acid and asiatic acid were the highest in CA-50E. In LPS-stimulated RAW 264.7 cells, CA-70E showed the highest TNF-α inhibitory activity, although CA-50E and CA-70E similarly inhibited nitric oxide production. Ethanol extracts significantly inhibited IL-6 and IL-8 production more than water extracts using TNF-α/IFN-γ-stimulated HaCaT cells, indicating its better inhibitory against skin inflammation. Therefore, Chungju-CA EtOH extract, especially 70% EtOH extract, has high physiologically active ingredients and potent anti-oxidant and anti-inflammatory activities, suggesting its industrial application as a functional material.