• Bulletin of the Korean Chemical Society
• /
• 제29권3호
• /
• pp.551-554
• /
• 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
• /
• 제28권1호
• /
• pp.15-23
• /
• 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
• /
• 제28권4호
• /
• pp.502-508
• /
• 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.

• 대한약학회:학술대회논문집
• /
• 대한약학회 2002년도 Proceedings of the Convention of the Pharmaceutical Society of Korea Vol.2
• /
• pp.340.1-340.1
• /
• 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
• /
• 제13권1호
• /
• pp.33-39
• /
• 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
• /
• 제22권1호
• /
• pp.20-24
• /
• 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.

• 한국식품영양학회지
• /
• 제36권6호
• /
• pp.436-444
• /
• 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.

• 대한화장품학회지
• /
• 제45권4호
• /
• pp.363-372
• /
• 2019

병풀의 잎과 줄기는 오랜 기간 궤양, 건선과 같은 피부질환 치료를 위한 약재로 사용되어왔으며, 병풀의 유효성분인 asiaticoside, madecasosside, asiatic acid, madecassic acid와 같은 triterpenoids는 피부 항염 및 피부 광 노화 개선 등의 효능이 밝혀져 있다. 본 연구의 목적은 인체에 유익한 미네랄을 많이 함유하고 있는 제주용암해수로 병풀을 숙성한 뒤 추출하여 피부 항염 및 보습 효능을 확인하는 것이다. 제주용암해수로 병풀을 숙성한 뒤 추출물을 HPLC에 분석하였을 때 triterpenoids의 농도가 더 증가하는 것을 관찰하였다. 숙성한 병풀 추출물의 항염증 효능을 확인하기 위해 LPS로 활성화된 RAW 264.7 세포에서 NO의 생성과 PM10 혹은 UV에 의해 유도된 HaCaT 세포에서 염증성 사이토카인들의 발현을 확인하였다. 실험 결과, 숙성한 병풀 추출물이 NO와 IL-6, IL-8, TNF-a의 발현을 저해하였고 숙성하지 않은 군에 비해 숙성한 군에서 억제 효능이 더 높은 것을 확인하였다. 또한, 피부 보습 효능을 확인하기 위해 HaCaT 세포에서 HA의 합성을 확인한 결과 숙성하지 않은 군에서는 효능이 없는 것에 반해 숙성한 군에서 농도 의존적으로 HA 생성이 증가하는 것을 확인하였다. 따라서 본 연구 결과를 바탕으로 제주용암해수로 숙성한 병풀 추출물은 항염과 보습에 유효한 소재임을 확인하였고 숙성하지 않은 병풀 추출물보다 효능이 우수할 것으로 판단된다.

• 생명과학회지
• /
• 제28권8호
• /
• pp.917-922
• /
• 2018

본 연구에서는 약용식물인 병풀의 계절적 생육조건의 변화에 따른 다양한 지상부 조직에서 triterpene glycoside 함량을 조사하기 위해, 20% 에탄올, 메탄올, 물을 이용한 3가지 추출방법을 이용하여 HPLC 분석을 수행하였다. 메탄올 추출방법에서 triterpene glycoside의 추출효율이 가장 높았으며, 특히 madecassoside와 asiaticoside의 값이 계절적인 생육조건의 변화동안 높게 확인되었다. 본 연구에서는 추가적으로 항산화활성을 분석하였으며, ABTs 소거활성은 잎자루에서 높게 확인되었으며, DPPH 소거활성은 메탄올과 에탄올로 추출한 잎조직에서만 높은 결과를 보였다. 총 페놀 및 플라보노이드 함량은 잎 조직에서 사용된 물, 에탄올, 메탄올 추출에 의한 결과가 가장 높은 값을 보였다. 본 연구의 결과로서, 병풀 식물체의 잎 조직에서 계절적 생육 조건 동안 triterpene glycoside 및 항산화활성이 증가함은 병풀의 재배시 madecassoside 및 asiaticoside를 포함한 유용한 주요 대사물질들이 식물체내에서 항상성을 위해 유지되며, 이는 향후 보 다 구체적인 생리활성에 대한 연구가 필요할 것으로 생각되는 바이다.

• 한국식품영양학회지
• /
• 제33권6호
• /
• pp.692-701
• /
• 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.