[논문]클러스터 분석을 이용한 황련 추출물 항산화 활성 최적화 추출공정

부민민; 현송화; 박수남

doi:10.14478/ace.2018.1065

클러스터 분석을 이용한 황련 추출물 항산화 활성 최적화 추출공정
Optimization of the Extraction Process and Antioxidant Capacity of Coptis chinensis Franch Extract through Cluster Analysis 원문보기

공업화학 = Applied chemistry for engineering, v.29 no.5, 2018년, pp.604 - 612

부민민 (서울과학기술대학교 정밀화학과, 나노바이오화장품연구실, 화장품종합기술연구소) , 현송화 (서울과학기술대학교 정밀화학과, 나노바이오화장품연구실, 화장품종합기술연구소) , 박수남 (서울과학기술대학교 정밀화학과, 나노바이오화장품연구실, 화장품종합기술연구소)

초록
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황련은 여러 가지 질병 치료에 사용되는 효과적인 전통약용식물이다. 하지만 황련 추출물의 생리활성성분 함량에 영향을 미치는 주요 인자 및 생리활성성분과 항산화 효능의 상관관계는 아직 알려지지 않았다. 따라서 본 논문에서는 용매, 온도 및 추출 시간이 황련 추출물의 생리 활성성분의 함량과 항산화 활성에 미치는 영향을 조사하였다. 클라스터를 이용하여 분석한 결과, $35^{\circ}C$에서 추출시간 30 min인 에탄올(50% 에탄올 : 50% 물) 조건으로 추출 시, 생리활성성분 함량 및 항산화 활성이 제일 높으므로 공장에서 사용하기에 가장 적합한 추출조건임을 확인하였다. 다중선형회귀분석 결과, 총 페놀화합물 함량은 DPPH소거 활성에 기여하였으며 총 알칼로이드 함량과 총 플라보노이드 함량은 FRAP활성에 기여하였다. 이상의 결과들로부터 황련의 추출 조건이 생리 활성 화합물의 수율 및 항산화 활성을 조절가능하며, 이는 공업적인 응용에 중요한 정보를 제공할 수 있을 것으로 사료된다.

Abstract ▼ AI-Helper

Coptis chinensis Franch is a valuable traditional oriental medicinal plant used for the treatment of various diseases. The major factors affecting the content of bioactive compounds and the relationship between bioactive compounds and antioxidant capacities of Coptis chinensis Franch were poorly understood. Thus, effects of the solvent, temperature, and extraction time on the extraction yields of bioactive compounds and the antioxidant activity of C. chinensis Franch extracts were investigated in this work. Our cluster analysis indicated that the hydroalcoholic solvent (50% ethanol : 50% water) at $35^{\circ}C$ for 30 min (extract time) was the best extraction condition for a factory use because the highest level of bioactive compounds and antioxidant activities was achieved. Multiple linear regression analysis revealed that total phenolic content (TPC) contributed to the 1,1-diphenyl-2-picrylhydrazyl radical (DPPH) scavenging activity, while both total alkaloid content (TAC) and total flavonoid content (TFC) were responsible for ferric reducing antioxidant power (FRAP) activity. These results indicated that extraction conditions controlled the yield of bioactive compounds and the antioxidant activity of C. chinensis Franch, which can provide important information for the industrial extraction.

주제어

표/그림 (8)

그림 Figure 1. Scheme of C. chinensis Franch extract preparation. Each extract was prepared in triplicate and each replicate was analyzed in triplicate.
그림 Figure 2. Effect of solvent, temperature and extraction time on extraction yield, total alkaloid content, total flavonoid content and total phenolic content per gram extraction powder of C. chinensis Franch.
표 Table 1. Analysis of Variance (ANOVA) Statistics
표 Table 2. Pearson’s Correlation Coefficients of DPPH⋅ Scavenging Ability and FRAP Versus TAC, TPC and TFC
그림 Figure 3. Effect of solvent, temperature and extraction time on antioxidant activity (A. A.) per gram extract powder of the C. chinensis Franch.
표 Table 3. Multiple Linear Regression Models of DPPH⋅ Scavenging Ability and FRAP with TAC, TPC and TFC
그림 Figure 4. Dendrogram from a cluster analysis of the extraction operating conditions considering the total alkaloid, flavonoid, phenolic content, and their antioxidant capacity per gram dry weight of plant as variables.
표 Table 4. Mean Values of the Bioactive Compounds and Antioxidant Activity of Several Clusters

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제안 방법

The extracts’ radical scavenging ability was analyzed according to the method described in a previous study[15], with some modifications.
To evaluate the influence of the solvent, temperature and extraction time on the extraction yields of bioactive components as well as their anti-oxidative activity, a factorial experiment (5 × 4 × 4) was designed.
We could ignore the extraction time since this did not significantly affect the yield of TAC, TPC and DPPH⋅ scavenging activity based on statistical analysis.

대상 데이터

Berberine chloride form, quercetin, gallic acid, α-tocopherol, Folin-Ciocalteu reagent, DPPH⋅ (2,2-Diphenyl-1-picrylhydrazyl), sodium carbonate monohydrate, iron(III) chloride hexahydrate, ferric chloride, TPTZ (2,4,6-tripyridyl-s-triazine), and sodium acetate trihydrate were all obtained from Sigma-Aldrich (St. Louis, MO, USA).

데이터처리

Pearson’s correlation was administered to test for correlation between bioactive compounds and antioxidant capacity.
Pearson’s correlation was used to test for correlation between variables, and correlations were considered significant at p < 0.05.
The differences between the means were analyzed by Duncan’s test of One-Way Analysis of variance (ANOVA).

성능/효과

In conclusion, multiple linear regression analysis on the relationship between bioactive compounds and antioxidant capacity in C. chinensis Franch shows that TPC contributes the DPPH⋅ scavenging activity, while both TAC and TFC contribute to FRAP.
Taking solvent water content as the independent variable, the highest antioxidant power was obtained using 25% water in the solvent (414.26 ± 2.44 mg FSE/g extract powder), followed by 0% water (405.75 ± 244 mg FSE/g extract powder), 50% water (402.83 ± 2.44 mg FSE/g extract powder), 75% water (387.90 ± 2.44 mg FSE/g extract powder) and 100% water (337.51 ± 2.44 mg FSE/g extract powder), indicating that 25% water content in the solvent was good for the extracts’ ferric reducing antioxidant power.
The highest average DPPH⋅ scavenging ability was obtained with 25% solvent water content (131.29 ± 2.73 mg α-TE/g extract powder), followed by 50% water (112.23 ± 2.73 mg α-TE/g extract powder), 100% water (103.81 ± 2.73 mg α-TE/g extract powder), 75% water (101.32 ± 2.73 mg α-TE/g extract powder) and 0% water (92.34 ± 2.73 mg α-TE/g extract powder).
The highest average value of TPC was obtained with 25% water (64.99 ± 0.52 mg GAE/g extract powder), followed by 50% water (63.84 ± 0.52 mg GAE/g extract powder), 75% water (63.67 ± 0.52 mg GAE/g extract powder), 0% water (59.95 ± 0.52 mg GAE/g extract powder) and the least efficient extraction condition was with 100% water (58.58 ± 0.52 mg GAE/g extract powder).
The highest average value of alkaloids extracted was obtained with 25% water content (476.88 ± 3.81 mg BCE/g extract powder), followed by 0% water (462.08 ± 3.81 mg BCE/g extract powder), 50% water (451.91 ± 3.81 mg BCE/g extract powder), 75% water (438.69 ± 3.81 mg BCE/g extract powder) and 100% water (367.68 ± 3.81 mg BCE/g extract powder).
The highest average value of flavonoids extracted was obtained with 0% water (99.12 ± 0.77 mg QE/g extract powder), followed by 25% water (98.78 ± 0.77 mg QE/g extract powder), 50% water (96.29 ± 0.77 mg QE/g extract powder), 75% water (93.16 ± 0.77 mg QE/g extract powder) and the worst extraction condition was with 100% water (82.66 ± 0.77 mg QE/g extract powder).
The results show that the highest values of TAC, TPC, and DPPH⋅ scavenging activity were obtained under conditions of 25% water/25 ℃/120 min, 25% water/25 ℃/30 min and 25% water/35 ℃/120 min, respectively.
The total DPPH⋅ scavenging ability was based on a standard curve of different concentrations (0-100 µg/mL) of α-tocopherol as a reference material (R2 = 0.9802) and the results were calculated and expressed as mg of α-tocopherol equivalent (α-TE) per g dry plant or per g extract powder of C. chinensis.
The total phenolic contents of the samples were determined by preparing a standard curve using different concentrations (0-62.5 µg/mL) of gallic acid as a reference material (R2 = 0.9977) and the results were calculated and expressed as mg of gallic acid equivalent (GAE) per g dry plant or per g extract powder of C. chinensis.
To establish the best model that could represent the influence of TAC, TFC and TPC on the antioxidant activity of C. chinensis, the stepwise elimination of variables (if p > 0.05) and stepwise insertion of variables (if p < 0.10) were performed.
When only taking solvent into consideration, we found that the highest average value of TFC and FRAP were obtained with 0% water (99.12 ± 0.77 mg QE/g extract powder), followed by 25% water (98.78 ± 0.77 mg QE/g extract powder), respectively.

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