• 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.

• Analytical Science and Technology
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• v.36 no.3
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• pp.128-134
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• 2023

Centella asiatica (C. asiatica) extracts, including asiaticoside and madecassoside, are used in ointments to treat the wound and atopic dermatitis due to their antibacterial and skin-regenerating effects in Asia. Therefore, research on the cultivation and extraction efficiency of C. asiatica is being actively conducted to increase commercialization efficiency. In this study, various deep eutectic solvents (DESs) were prepared and used as the extraction solvents according to the mole ratio between the hydrogen bond acceptor (HBA) and hydrogen bond donor (HBD). And then, the extraction yields in distilled water (DW) and methanol (MeOH), commonly used extraction solvents for C. asiatica, were compared and analyzed by HPLC in the optimized operating condition. As a result, a mixture of DW and DES at a ratio of 3:7 showed about 1.4 times higher extraction efficiency than MeOH only. Conversely, the extraction efficiency in a mixture of MeOH and DES at a ratio of 3:7 was about 6 % lower than that in MeOH only.

• Journal of Veterinary Clinics
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• v.38 no.2
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• pp.56-62
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• 2021

Madecassoside, an active ingredient extracted from Centella asiatica, is used for treatment of various skin disorders in humans. However, the effect of madecassoside on the skin of dogs and cats has not been studied yet. The purpose of this study was to evaluate clinical efficacy of topical madecassoside cream in dogs and cats with skin diseases. A total of twenty-one dogs and ten cats with various skin diseases were included in the study. The 1% topical madecassoside cream was applied to the animal's skin lesion at least once a day for 7 days, and the skin condition was evaluated before the application of madecassoside cream (day 0) and 7 days after the application (day 7). The skin condition was scored by five clinical indices: canine atopic dermatitis extent and severity index-4 (CADESI-4), coat condition, pruritus, scale, and general condition. In dogs, all five clinical indices (CADESI-4, coat condition, pruritus, scale, and general condition) were significantly decreased on day 7 compared to those on day 0 (p < 0.0001, p < 0.05, p < 0.001, p < 0.01, and p < 0.05, respectively). In cats, the CADESI-4 and scale were significantly decreased on day 7 compared to those on day 0 (p < 0.01 and p < 0.05, respectively). No adverse effects were observed during the trial period in the dogs and cats included in this study. The results of this study demonstrate that the topical madecassoside cream is applicable to skin lesions in dogs and cats.

• 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.

• Korean Journal of Medicinal Crop Science
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• v.10 no.5
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• pp.375-378
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• 2002

The seasonal variation of two triterpene glycosides, madecassoside and asisaticoside, in the aerial parts of Centella asiatica from May through October was investigated. In leaves, the highest level of triterpene glycosides content was about 108.1 mg/g in September. In petiols, the content was as low as 19.02 mg/g. According to the experimental results obtained, we found that Centella asiatica leaf can accumulate the triterpene glycosides and September is the optimal season for the collection.

• Journal of the Society of Cosmetic Scientists of Korea
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• v.49 no.2
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• pp.147-157
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• 2023

Centella asiatica extract is widely used as a raw material for cosmetics due to its various effects, but it is difficult to expect penetration into the skin due to its high molecular weight and low solubility. In order to solve these problems, lipid-based liposomes of various types were developed to increase skin absorption. Therefore, in this study, we tried to increase the skin absorption rate by preparing transfersomes using surfactants as edge activators in existing liposomes. Liposome and transfersomes containing Span 80 and Tween 20, 60, 80, and 85, respectively, were prepared using a high-pressure homogenizer, and we evaluated the particle size, polydispersity index, zeta potential, and skin absorption rate. As a result, there was almost no change in the physical properties of particle size, polydispersity index and zeta potential from 25 ℃ to 60 d, and the particle size of transfersomes containing Tween 20, 60, and 80 increased after 60 d at 45 ℃. Madecassoside, main substances of the Centella asiatica extract was used as an standard and madecassoside was measured and calculated when measuring the skin absorption rate using Franz diffusion cells. As a result, formulations containing Tween 20 were the most, whereas formulations containing Span 80 were the least. According to the skin absorption coefficient (Kp) value, all formulations showed 'very fast', and the absorption rate was similar or greater than that of liposomes, except for formulations containing Span 80. Through this, it was confirmed that the larger the HLB value of the nonionic surfactant, the smaller the particle size of the transfersome, and the increased skin absorption rate due to the increased flexibility of the vesicle membrane. Through this study, transfersome using surfactant as an edge activator can be expected to solve local skin problems not only as a cosmetic raw material or product, but also by increasing skin absorption.

• 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.

• Korean Journal of Medicinal Crop Science
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• v.13 no.5
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• pp.233-236
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• 2005

Centella asiatica accumulates large amounts of triterpene saponin, such as centellasaponin, asiaticoside, madecassoside. We examined the effect of two candidates, MJ (Methyl jasmonate) and TDZ (thidiazuron), on asiaticoside production and the accumulation of bAS mRNA associated with asiaticoside biosynthesis in leaves of cultured whole plants. The growth of whole plants treated with 0.1 mM MJ was found to decrease significantly, however, the growth of whole plants treated with 0.1 mM MJ plus 0.025 mg/l TDZ was better than that treated with MJ alone. When MJ alone was added to culture medium, asiaticoside contents in leaves were higher than that of control after 7 days of treatments. The maximum level of bAS $({\beta}-amyrin\;synthsae)$ mRNA in leaves of whole plant treated TDZ and MJ was transiently observed after exposure to 5 days. These results showed the up-regulation of bAS gene by adding TDZ and MJ at the molecular level, however, synergic effects of TDZ and MJ on asiaticoside biosynthesis were not testified.

• 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.

• Plant Biotechnology Reports
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• v.5 no.3
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• pp.283-287
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• 2011

In order to produce centellosides from whole plant cultures of Centella asiatica (L.) Urban, we evaluated the synergistic effects of thidiazuron (TDZ) and methyl jasmonate (MJ) on whole plant growth and centelloside production. After 4 weeks of treatment with 0.025 mg/L of TDZ coupled with 0.1 mM MJ, the production of made-cassoside and asiaticoside from whole plant cultures was estimated to be 2.40- and 2.44-fold, respectively, above that of MJ elicitation alone. When whole plants were treated with a growth regulator and an elicitor, the growth of whole plants, as compared to the controls, did not differ. Additionally, total phytosyterol content in the leaves of whole plants co-treated with MJ and TDZ was 1.08-fold greater than those of MJ alone. These results demonstrate that combined treatments not only stimulate the accumulation of centellosides in the leaves but also inhibit the reduction of phytosterol levels caused by MJ elicitation.