• Bulletin of the Korean Chemical Society
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• 제29권3호
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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.

• 분석과학
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• 제36권3호
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• pp.128-134
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• 2023

병풀(Centella asiatica) 추출물인 asiaticoside와 madecassoside는 항균성 및 피부재생 등의 효과로 인해 아시아 지역에서 치료용 연고에 사용되고 있으며, 병풀의 재배 및 추출효율을 높이기 위한 연구가 상업화를 위해 활발히 진행되고 있다. 본 연구에서는 환경 친화적인 추출 용매인 deep eutectic solvents(DESs)가 병풀에서 asiaticoside와 madecassoside의 추출 효율에 미치는 영향을 관찰하기 위하여 다양한 수소결합 받개(hydrogen bond acceptor, HBA)와 주개(hydrogen bond donor, HBD)의 혼합 몰비(HBA1: HBD2, HBA1:HBD3, HBA1:HBD4, 그리고 HBA1:HBD5)로 제조하여 추출용매로 사용하였며, 용매 추출에서 일반적으로 사용되고 있는 추출용매인 증류수(DW)와 methanol (MeOH)의 추출 결과를 최적화된 HPLC조건으로 분리하여 정량 분석하였다. 그 결과, DW와 DES를 3:7로 혼합하여 사용한 경우가 단일 MeOH를 사용한 경우보다 약 1.4배 높은 추출 효율을 보였다. 반대로 MeOH과 DES를 3:7로 혼합하여 사용하는 경우에서는 단일 MeOH를 사용하는 경우보다 약 6 % 낮은 추출 효율을 보였다.

• 한국임상수의학회지
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• 제38권2호
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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.

• 생명과학회지
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• 제28권8호
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• pp.917-922
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• 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.

• 한국약용작물학회지
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• 제10권5호
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• pp.375-378
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• 2002

5월부터 10월까지 병풀의 지상부에 triterpene glycosides인 madecassoside와 asiaticoside의 시기별 함량변화를 조사하였다. 잎에서는 9월에 약 108.1 mg/g으로 가장 높은 수준의 함량을 보였다. 엽병에서는 19.02 mg/g으로 잎에서 보이는 수준보다는 낮은 수준을 나타냈다. 병풀의 triterpene glycosides는 잎에 주로 축적되며, 가장 적절한 병풀의 수확시기는 9월로 잠적 확인되었다.

• 대한화장품학회지
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• 제49권2호
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• pp.147-157
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• 2023

병풀추출물은 다양한 효능으로 인하여 화장품 원료로 많이 사용되지만, 분자량이 크고 용해도가 낮아 피부에의 흡수 효과를 기대하기 어렵다. 이러한 문제점을 해결하기 위하여 지질 기반 다양한 형태의 리포좀을 개발하여 피부흡수율을 높이고자 하였다. 이에 본 연구에서는 기존 리포좀에 edge activator로서 계면활성제를 사용하여 트렌스퍼좀을 제조함으로써 피부흡수율을 높이고자 하였다. 리포좀과 Span 80 및 Tween 20, 60, 80, 85를 각각 함유한 트렌스퍼좀을 high-pressure homogenizer를 이용하여 제조하였고, 입자크기, 다분산지수, 제타전위, 피부흡수율에 대하여 평가하였다. 그 결과, 25 ℃에서 60 d까지 입자크기, 다분산지수 및 제타전위의 물성변화가 거의 없었으며, 45 ℃에서는 60 d 경과 후에 Tween 20, 60, 80을 함유한 트렌스퍼좀의 입자크기가 증가하였다. Franz diffusion cell을 이용한 피부흡수율은 병풀추출물의 주요 성분 중 하나인 madecassoside를 지표 성분으로 사용하여 계산하였다. 그 결과, Tween 20을 함유한 제형에서 가장 흡수율이 높았으며, 반면에 Span 80을 함유한 제형에서 가장 적었다. 흡수계수(Kp) 값에 의하면 모든 제형에서 'very fast'로 나타났고, Span 80을 함유한 제형을 제외하고 리포좀보다 흡수 속도가 유사하거나 크게 나타났다. 이를 통하여 비이온성 계면활성제의 HLB 값이 클수록 트렌스퍼좀의 입자크기가 작았으며, 베시클 막의 유연성 향상으로 피부흡수율이 증가함을 확인하였다. 본 연구를 통해 edge activator로서 계면활성제를 사용한 트렌스퍼좀은 화장품 원료나 제품으로서뿐만 아니라 피부흡수율 증가로 피부의 국소적 문제 해결 가능성을 기대할 수 있다.

• Natural Product Sciences
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• 제13권1호
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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.

• 한국약용작물학회지
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• 제13권5호
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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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• 제22권1호
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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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• 제5권3호
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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.