[논문]오미자의 모유두세포 증식 활성성분과 반응표면분석을 이용한 추출조건의 최적화

조현대; 정지연; 류화선; 이정노; 박성민

doi:10.15230/scsk.2020.46.4.415

오미자의 모유두세포 증식 활성성분과 반응표면분석을 이용한 추출조건의 최적화
Dermal Papilla Cells Proliferation Constituent of Schisandra chinensis Fruits and Optimization Using Response Surface Methodology 원문보기

大韓化粧品學會誌 = Journal of the society of cosmetic scientists of Korea, v.46 no.4, 2020년, pp.415 - 424

조현대 (충북대학교 약학대학) , 정지연 ((주)코씨드바이오팜 바이오융합연구소) , 류화선 ((주)코씨드바이오팜 바이오융합연구소) , 이정노 ((주)코씨드바이오팜 바이오융합연구소) , 박성민 ((주)코씨드바이오팜 바이오융합연구소)

초록
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본 연구에서는 오미자 열매로부터 모유두세포 증식에 활성을 나타내는 gomisin N을 정제하고 gomisin N의 함량이 높은 추출물을 얻기 위한 최적의 추출조건을 확인하였다. 추출물 및 분획물에 대하여 모유두세포 증식 활성 실험을 진행한 결과, n-hexane 분획물 1 ㎍/mL 처리군에서 약 29%의 증식 활성을 나타내었다. n-Hexane 분획물의 활성성분을 규명하기 위하여 column chromatography를 진행하였으며 2 개의 화합물 gomisin N (1)과 schizandrin (2)를 분리 동정하였다. 분리된 화합물에 대한 모유두세포 증식 활성 실험 결과, gomisin N에서 최대 20%이상의 증식 활성을 나타내었다. 따라서 오미자 열매로부터 최대치의 gomisin N을 얻을 수 있는 최적추출조건을 반응표면분석(RSM)을 사용하여 확인하였다. 최적추출조건을 설정하기 위해 에탄올의 비율, 추출시간, 추출온도를 독립변수로 하여 실험한 결과, 0.95 이상의 결정계수와 0.05 이하의 p-value를 나타내어 모델의 적합성이 확인 되었다. Gomisin N의 최대함량을 위한 최적추출조건은 에탄올 비율 83.8%, 추출온도 80 ℃, 추출시간 8.7 h으로 확인되었고, 최적추출조건에서 gomisin N의 함량은 378,300 ppm으로 예측되었다. 최적추출조건에 대한 실제 검증에서 평균 376,884 ppm으로 예측된 값과 유사한 결과를 확인 할 수 있었다

Abstract ▼ AI-Helper

In the present study, we have refined gomisin N, which represents activity in the proliferation of dermal papilla cells (HFDPCs) from the fruit of Schisandra chinensis (S. chinensis), and have identified optimal extraction conditions for obtaining extracts with high content of gomisin N. The activity of the extracts and fractions was evaluated, and the results indicated approximately 29% proliferation activity in the group treated with 1 ㎍/mL of n-hexane fraction. Column chromatography was used to assess the active ingredient in the n-hexane fraction, and two compounds, namely gomisin N(1) and schisandrin(2), were isolated and identified. When the HFDPCs proliferation activity was tested for the isolated compounds, gomisin N exhibited ≥ 20% proliferation activity. Thus, via response surface methodology (RSM), the optimum extraction conditions to obtain the maximum level of gomisin N from the fruit of S. chinensis were determined, where ethanol proportion, extraction time, and extraction temperature were used as the independent variables. The results revealed coefficient of determination ≥ 0.95 and p-value ≤ 0.05, which confirmed the fit of the model. The optimum extraction conditions to achieve the maximum content of gomisin N were as follows: ethanol proportion 83.8%, extraction temperature 80 ℃, and extraction time 8.7 h. The content of gomisin N using these conditions was predicted as 378,300 ppm, and a mean value close to the predicted value (376,884 ppm) was obtained while validating the aforementioned conditions.

주제어

AI 본문요약
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문제 정의

This study aimed to find a natural material that promotes HFDPCs proliferation to alleviate hair loss. Among the various materials screened, the fruit of S.

제안 방법

The RSM used in this study was the Box-Behnken design (BBD), a model used to determine the secondary regression equation and optimum conditions by setting three ranges for three variables. The extraction solvent, time, and temperature were used as variables to optimize the extraction conditions.
The extraction solvent, time, and temperature were used as variables to optimize the extraction conditions. The range of the three independent variables were encoded into three stages as –1, 0, 1 (min, median, max), and the extraction was carried out based on the conditions determined by BBD. Each response can be expressed as the following quadratic polynomials[15-17].
The analysis of HFDPCs proliferation activity of the two isolated compounds revealed that, while compound 1 exhibited ≥ 20% HFDPCs proliferation activity, compound 2 presented no proliferation activity. To determine the optimum conditions for extracting the highest content of gomisin N from the fruit of S. chinensis using RSM, the independent variables used to design the experiment based on the Box-Behnken design (BBD) were EtOH proportion (50 – 90%), extraction time (3 - 9 h), and extraction temperature (40 - 80 ℃). The optimum extraction conditions based on the RSM were 83.
chinensis fruit extract led to approximately 15% activity of HFDPCs proliferation and the group treated with 1 μg/mL of n-hexane fraction led to approximately 29% activity. To identify the active ingredient in n-hexane fraction, column chromatography was performed to isolate and identify two compounds, namely gomisin N and schisandrin. The analysis of HFDPCs proliferation activity of the two isolated compounds revealed that, while compound 1 exhibited ≥ 20% HFDPCs proliferation activity, compound 2 presented no proliferation activity.

대상 데이터

S. chinensis fruits were purchased from Mungyeong Market, located in Mungyeong, North Gyeongbuk province (Korea). The fruits were dried before use.

이론/모형

For statistical optimization of RSM, MINITAB (Minitab Inc., USA) was used. To confirm the fit of the model, the analysis of variance (ANOVA) was applied, and significance was set to p-value ≤ 0.
The HFDPCs were treated with SCE, SCH, and isolated compounds. The cell proliferation was determined with MTT assay.
The three factors: extraction solvent (X1), extraction temperature (X2), and extraction time (X3), were selected as the variables for determining the optimum extraction conditions. To identify the correlations among these three variables and the content of gomisin N, the 3-level-3-factor Box-Behnken design (BBD) was used. Table 1 summarizes the encoded experimental range for each variable.

성능/효과

chinensis fruit extract and fractions on HFDPCs, the cells were treated with varying concentrations. The results indicated that the group treated with 100 μg/mL of S. chinensis fruit extract led to approximately 15% activity of HFDPCs proliferation and the group treated with 1 μg/mL of n-hexane fraction led to approximately 29% activity. To identify the active ingredient in n-hexane fraction, column chromatography was performed to isolate and identify two compounds, namely gomisin N and schisandrin.

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저자의 다른 논문 :

표제어: PCR

동의어: Packet Collision Rate

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용어 설명: PCR은 세균 특이성이 있는 primer를 이용하여 적은 수의 세균이 있을지라도 쉽게 검출할 수 있는 유용한 방법이며, 이를 이용하여 구강 내 치면세균막이나 타액에서 직접 세균을 검출할 수 있게 되었다[8].

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