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Natural Product Research: Formerly Natural Product Letters Publication details, including instructions for authors and subscription information: http://www.tandfonline.com/loi/gnpl20
Phenolic extracts from Rhus verniciflua Stokes bark by decompressing inner ebullition and their antioxidant activities ab
abc
Hongxia Chen , Chengzhang Wang
ab
abc
, Jianzhong Ye , Hao Zhou
ab
& Jiaojiao Yuan a
Institute of Chemical Industry of Forest Products, CAF, Nanjing 210042, P.R. China b
Click for updates
Key and Open Laboratory on Forest Chemical Engineering, SFA, Nanjing 210042, P.R. China c
Institute of New Technology of Forestry, CAF, Beijing 100091, P.R. China Published online: 17 Jan 2014.
To cite this article: Hongxia Chen, Chengzhang Wang, Jianzhong Ye, Hao Zhou & Jiaojiao Yuan (2014) Phenolic extracts from Rhus verniciflua Stokes bark by decompressing inner ebullition and their antioxidant activities, Natural Product Research: Formerly Natural Product Letters, 28:7, 496-499, DOI: 10.1080/14786419.2013.872105 To link to this article: http://dx.doi.org/10.1080/14786419.2013.872105
PLEASE SCROLL DOWN FOR ARTICLE Taylor & Francis makes every effort to ensure the accuracy of all the information (the “Content”) contained in the publications on our platform. However, Taylor & Francis, our agents, and our licensors make no representations or warranties whatsoever as to the accuracy, completeness, or suitability for any purpose of the Content. Any opinions and views expressed in this publication are the opinions and views of the authors, and are not the views of or endorsed by Taylor & Francis. The accuracy of the Content should not be relied upon and should be independently verified with primary sources of information. Taylor and Francis shall not be liable for any losses, actions, claims, proceedings, demands, costs, expenses, damages, and other liabilities whatsoever or howsoever caused arising directly or indirectly in connection with, in relation to or arising out of the use of the Content.
Downloaded by [Florida Atlantic University] at 16:08 26 January 2015
This article may be used for research, teaching, and private study purposes. Any substantial or systematic reproduction, redistribution, reselling, loan, sub-licensing, systematic supply, or distribution in any form to anyone is expressly forbidden. Terms & Conditions of access and use can be found at http://www.tandfonline.com/page/termsand-conditions
Natural Product Research, 2014 Vol. 28, No. 7, 496–499, http://dx.doi.org/10.1080/14786419.2013.872105
SHORT COMMUNICATION Phenolic extracts from Rhus verniciflua Stokes bark by decompressing inner ebullition and their antioxidant activities Hongxia Chenab, Chengzhang Wangabc*, Jianzhong Yeab, Hao Zhouabc and Jiaojiao Yuanab Institute of Chemical Industry of Forest Products, CAF, Nanjing 210042, P.R. China; bKey and Open Laboratory on Forest Chemical Engineering, SFA, Nanjing 210042, P.R. China; cInstitute of New Technology of Forestry, CAF, Beijing 100091, P.R. China
Downloaded by [Florida Atlantic University] at 16:08 26 January 2015
a
(Received 11 September 2013; final version received 1 December 2013) Decompressing inner ebullition (DIE) can reduce the extraction liquid boiling point and polyphenols oxidation in the extraction process. The aim of this study is to optimise the phenolic extraction process by DIE and to examine the antioxidant activities. The extraction process parameters were observed by central composite design. The antioxidant activity was measured using 1,1-diphenyl-2-picrylhydrazyl (DPPH) and ferric reducing power assays. The results showed that the optimal extraction condition is extract time of 90 min, temperature of 458C, solid – liquid ratio of 1:20 g/mL, vacuum degree of 2 0.08 MPa, ethanol concentration of 60%, while the phenolic content was 5.4%. The phenolic extracts from Rhus verniciflua Stokes bark had better antioxidant activities; the antioxidant activity (IC50) of the DIE was 20 mg/mL by the DPPH method. The reducing power of the phenolic extracts was significantly related to their total phenolic content (R ¼ 0.9903). The results presented show that the DIE method is an effective method for polyphenols extraction. Keywords: Rhus verniciflua Stokes; total phenolic contents; decompressing inner ebullition; antioxidant activity
1. Introduction Rhus verniciflua Stokes is a deciduous forest tree belonging to the family Anacardiaceae (Zheng & Min 1980). The species of this tree (Rhus vernicifera) grows in Japan, China and Korea. Recently, it has been found that the phenolic extracts of R. verniciflua Stokes bark have marked antioxidant and anti-microbial effects and anti-tumour properties, and it was found that the main bioactive compounds were polyphenols, such as fisetin, fusin, butein and sulphuretin (Kim et al. 2006; Kim et al. 2010). Decompressing inner ebullition (DIE) is a new extraction method, which was established by Wei et al. (2005), using the decompressing condition. DIE can reduce the boiling point of the extraction liquidand reduce the oxidation of polyphenols in the extraction process and the macromolecular substances (such as starch and polysaccharides) of dissolution. In recent years, many studies have been carried out on the activity of the R. verniciflua Stokes extracts; some have reported on the different organic solvent extractions and their antioxidant activity (Kim et al. 2010). However, little information is available concerning the extraction variables on the phenolic yield of the R. verniciflua Stokes bark. In particular, extract condition is the important parameter. In this study, the influence of some critical extraction variables on the phenolic yield is researched by the DIE method. Central composite design (CCD) was used to evaluate the effect of three factors, such as time, vacuum degree and
*Corresponding author. Email: [email protected] q 2014 Taylor & Francis
Natural Product Research
497
Downloaded by [Florida Atlantic University] at 16:08 26 January 2015
temperature which were studied for the optimum extraction condition. The aim of this study was also to examine the antioxidant activities in the optimum extracts. 2. Results and discussion CCD was used to optimise several major extract parameters, which are vacuum degree (x1), time (x2) and temperature (x3). The experimental matrix for three factors consists of 20 experiments, as shown in Table 1; the solid –liquid ratio was 1:20 g/mL, the ethanol concentration was 60%, extracted two times. The experimental calculations were performed by using Design-expert software version 7.1.3. A classical second-degree model was obtained as follows. According to the regression model equations, the fitting coefficient of three variables show 0.18 . 0.15 . 0.11, implying that time and temperature are the main variables that the phenolic content depends on. Y ¼ 5:24 þ 0:11x1 þ 0:18x2 þ 0:15x3 þ 0:055x1 x2 2 0:18x1 x3 2 0:11x2 x3 2 0:061x21 2 0:15x22 2 0:13x23 ; where Y is the total phenolic content and is expressed as gallic acid equivalents; xi is the coded level of the three variables. The statistical parameters obtained from ANOVA for the reduced models are given in Table 2. For the models, P , 0.001, implying that the models are significant; the models can predict the real experimental data. For pure error, P . 0.05, implying that the calculated values can fit with the experimental values. The x1, x2 and x3 variables have a significant effect on the polyphenolic content. For x1x3, P , 0.01, implying that the vacuum degree is closely related to temperature on the polyphenolic content.
Table 1. Experimental design and results of CCD. Coded variables No.
x1
x2
x3
Total phenolic content
1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20
21 21 21 21 1 1 1 1 21.682 1.682 0 0 0 0 0 0 0 0 0 0
21 21 1 1 21 21 1 1 0 0 21.682 1.682 0 0 0 0 0 0 0 0
21 1 21 1 21 1 21 1 0 0 0 0 21.682 1.682 0 0 0 0 0 0
4.066 4.712 4.617 5.301 5.149 4.883 5.073 5.206 4.987 5.206 4.541 5.149 4.788 5.035 5.187 5.358 5.206 5.206 5.149 5.301
498
H. Chen et al.
Table 2. Variance analysis of items in regression equation.
Downloaded by [Florida Atlantic University] at 16:08 26 January 2015
Sources of variation
Sum of squares
df
Mean square
F value
P value
1.82 0.18 0.43 0.30 0.024 0.27 0.1 0.054 0.32 0.23 0.14 0.11 0.031 1.96
9 1 1 1 1 1 1 1 1 1 10 5 5 19
0.2 0.18 0.43 0.30 0.024 0.27 0.1 0.054 0.32 0.23 0.014 0.021 6.173E 2 003
14.84 13.15 31.17 22.13 1.75 19.62 7.31 3.93 23.75 16.89
,0.001 0.0046 ,0.001 ,0.001 0.2153 0.0013 0.0222 0.0755 ,0.001 0.0021
Model x1 x2 x3 x1x2 x1x3 x2x3 x21 x22 x23 Residual Lack of fit Pure error Total
3.42 0.1017
After the CCD of the DIE method, the optimal extraction condition with an extraction time of 90 min, temperature of 458C, solid –liquid ratio of 1:20 g/mL, vacuum degree of 2 0.08 MPa, ethanol concentration of 60% was predicted using the desirability function. Comparing the extraction conditions of DIE and reflux extraction, the reflux extraction is at the same conditions of the DIE method with no vaccum degree. The total phenolic content of the DIE method is 5.4% and that of the reflux extraction is 4.12%. Park (2012) used water at 958C to extract the active material, the phenolic content was 3%; by this method the antioxidant activity (IC50) was 58.9 mg/mL by the 1,1-diphenyl-2-picrylhydrazyl (DPPH) method. We used the DPPH method to study the antioxidant activity. The antioxidant activity (IC50) of the DIE and reflux extraction methods was 20 and 30 mg/mL, respectively, which means that the DIE method had better phenolic content and antioxidant activity. It also proved that the high temperature extraction could improve the degradation of phenols. The reducing power of the phenolic extracts was significantly related to their total phenolic content (R ¼ 0.9903), while the concentration of the phenolic extracts is 40 – 50 mg/ mL and the absorbance at 593 nm is 2.5. The FRAP assay also confirmed that the phenolic extracts of R. verniciflua Stokes had stronger antioxidant activity. Supplementary material Experimental details relating to this article are available online. Acknowledgements The authors thank Dr Wang for his support and guidance with this study. The authors are grateful to the China Basic Research Foundation of National Commonweal Research Institute (CAFINT2013C03), International Cooperation Project (S2012GR0138) and “948” Technology Introduction Project (2012-4-05) for financial support.
References Kim JH, Kim HP, Jung CH. 2006. Inhibition of cell cycle progression via p27Kip1 upregulation and apoptosis induction by an ethanol extract of Rhus verniciflua Stokes in AGS gastric cancer cells. Int J Mol Med. 18:201–208. Kim JS, Kwon YS, Chun WJ. 2010. Rhus verniciflua Stokes flavonoid extracts have anti-oxidant, anti-microbial and aglucosidase inhibitory effect. Food Chem. 120:539–543.
Natural Product Research
499
Downloaded by [Florida Atlantic University] at 16:08 26 January 2015
Park SJ, Kim KH, Shin MJ. 2012. Rhus verniciflua Stokes extract having increased content of active flavonoid compound and method for preparing same. United States patent, US2012/0283322 A1[P]. Wei TY, Tong ZF, Zhao ZX. Extraction the active components of plants by decompressing inner ebullition. China Patent, 2005100328021 2005. Zheng M, Min TL. 1980. Flora of China 45(1)-Anacardiaceae. Beijing: Science press.
Natural Product Research: Formerly Natural Product Letters Publication details, including instructions for authors and subscription information: http://www.tandfonline.com/loi/gnpl20
Phenolic extracts from Rhus verniciflua Stokes bark by decompressing inner ebullition and their antioxidant activities ab
abc
Hongxia Chen , Chengzhang Wang
ab
abc
, Jianzhong Ye , Hao Zhou
ab
& Jiaojiao Yuan a
Institute of Chemical Industry of Forest Products, CAF, Nanjing 210042, P.R. China b
Click for updates
Key and Open Laboratory on Forest Chemical Engineering, SFA, Nanjing 210042, P.R. China c
Institute of New Technology of Forestry, CAF, Beijing 100091, P.R. China Published online: 17 Jan 2014.
To cite this article: Hongxia Chen, Chengzhang Wang, Jianzhong Ye, Hao Zhou & Jiaojiao Yuan (2014) Phenolic extracts from Rhus verniciflua Stokes bark by decompressing inner ebullition and their antioxidant activities, Natural Product Research: Formerly Natural Product Letters, 28:7, 496-499, DOI: 10.1080/14786419.2013.872105 To link to this article: http://dx.doi.org/10.1080/14786419.2013.872105
PLEASE SCROLL DOWN FOR ARTICLE Taylor & Francis makes every effort to ensure the accuracy of all the information (the “Content”) contained in the publications on our platform. However, Taylor & Francis, our agents, and our licensors make no representations or warranties whatsoever as to the accuracy, completeness, or suitability for any purpose of the Content. Any opinions and views expressed in this publication are the opinions and views of the authors, and are not the views of or endorsed by Taylor & Francis. The accuracy of the Content should not be relied upon and should be independently verified with primary sources of information. Taylor and Francis shall not be liable for any losses, actions, claims, proceedings, demands, costs, expenses, damages, and other liabilities whatsoever or howsoever caused arising directly or indirectly in connection with, in relation to or arising out of the use of the Content.
Downloaded by [Florida Atlantic University] at 16:08 26 January 2015
This article may be used for research, teaching, and private study purposes. Any substantial or systematic reproduction, redistribution, reselling, loan, sub-licensing, systematic supply, or distribution in any form to anyone is expressly forbidden. Terms & Conditions of access and use can be found at http://www.tandfonline.com/page/termsand-conditions
Natural Product Research, 2014 Vol. 28, No. 7, 496–499, http://dx.doi.org/10.1080/14786419.2013.872105
SHORT COMMUNICATION Phenolic extracts from Rhus verniciflua Stokes bark by decompressing inner ebullition and their antioxidant activities Hongxia Chenab, Chengzhang Wangabc*, Jianzhong Yeab, Hao Zhouabc and Jiaojiao Yuanab Institute of Chemical Industry of Forest Products, CAF, Nanjing 210042, P.R. China; bKey and Open Laboratory on Forest Chemical Engineering, SFA, Nanjing 210042, P.R. China; cInstitute of New Technology of Forestry, CAF, Beijing 100091, P.R. China
Downloaded by [Florida Atlantic University] at 16:08 26 January 2015
a
(Received 11 September 2013; final version received 1 December 2013) Decompressing inner ebullition (DIE) can reduce the extraction liquid boiling point and polyphenols oxidation in the extraction process. The aim of this study is to optimise the phenolic extraction process by DIE and to examine the antioxidant activities. The extraction process parameters were observed by central composite design. The antioxidant activity was measured using 1,1-diphenyl-2-picrylhydrazyl (DPPH) and ferric reducing power assays. The results showed that the optimal extraction condition is extract time of 90 min, temperature of 458C, solid – liquid ratio of 1:20 g/mL, vacuum degree of 2 0.08 MPa, ethanol concentration of 60%, while the phenolic content was 5.4%. The phenolic extracts from Rhus verniciflua Stokes bark had better antioxidant activities; the antioxidant activity (IC50) of the DIE was 20 mg/mL by the DPPH method. The reducing power of the phenolic extracts was significantly related to their total phenolic content (R ¼ 0.9903). The results presented show that the DIE method is an effective method for polyphenols extraction. Keywords: Rhus verniciflua Stokes; total phenolic contents; decompressing inner ebullition; antioxidant activity
1. Introduction Rhus verniciflua Stokes is a deciduous forest tree belonging to the family Anacardiaceae (Zheng & Min 1980). The species of this tree (Rhus vernicifera) grows in Japan, China and Korea. Recently, it has been found that the phenolic extracts of R. verniciflua Stokes bark have marked antioxidant and anti-microbial effects and anti-tumour properties, and it was found that the main bioactive compounds were polyphenols, such as fisetin, fusin, butein and sulphuretin (Kim et al. 2006; Kim et al. 2010). Decompressing inner ebullition (DIE) is a new extraction method, which was established by Wei et al. (2005), using the decompressing condition. DIE can reduce the boiling point of the extraction liquidand reduce the oxidation of polyphenols in the extraction process and the macromolecular substances (such as starch and polysaccharides) of dissolution. In recent years, many studies have been carried out on the activity of the R. verniciflua Stokes extracts; some have reported on the different organic solvent extractions and their antioxidant activity (Kim et al. 2010). However, little information is available concerning the extraction variables on the phenolic yield of the R. verniciflua Stokes bark. In particular, extract condition is the important parameter. In this study, the influence of some critical extraction variables on the phenolic yield is researched by the DIE method. Central composite design (CCD) was used to evaluate the effect of three factors, such as time, vacuum degree and
*Corresponding author. Email: [email protected] q 2014 Taylor & Francis
Natural Product Research
497
Downloaded by [Florida Atlantic University] at 16:08 26 January 2015
temperature which were studied for the optimum extraction condition. The aim of this study was also to examine the antioxidant activities in the optimum extracts. 2. Results and discussion CCD was used to optimise several major extract parameters, which are vacuum degree (x1), time (x2) and temperature (x3). The experimental matrix for three factors consists of 20 experiments, as shown in Table 1; the solid –liquid ratio was 1:20 g/mL, the ethanol concentration was 60%, extracted two times. The experimental calculations were performed by using Design-expert software version 7.1.3. A classical second-degree model was obtained as follows. According to the regression model equations, the fitting coefficient of three variables show 0.18 . 0.15 . 0.11, implying that time and temperature are the main variables that the phenolic content depends on. Y ¼ 5:24 þ 0:11x1 þ 0:18x2 þ 0:15x3 þ 0:055x1 x2 2 0:18x1 x3 2 0:11x2 x3 2 0:061x21 2 0:15x22 2 0:13x23 ; where Y is the total phenolic content and is expressed as gallic acid equivalents; xi is the coded level of the three variables. The statistical parameters obtained from ANOVA for the reduced models are given in Table 2. For the models, P , 0.001, implying that the models are significant; the models can predict the real experimental data. For pure error, P . 0.05, implying that the calculated values can fit with the experimental values. The x1, x2 and x3 variables have a significant effect on the polyphenolic content. For x1x3, P , 0.01, implying that the vacuum degree is closely related to temperature on the polyphenolic content.
Table 1. Experimental design and results of CCD. Coded variables No.
x1
x2
x3
Total phenolic content
1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20
21 21 21 21 1 1 1 1 21.682 1.682 0 0 0 0 0 0 0 0 0 0
21 21 1 1 21 21 1 1 0 0 21.682 1.682 0 0 0 0 0 0 0 0
21 1 21 1 21 1 21 1 0 0 0 0 21.682 1.682 0 0 0 0 0 0
4.066 4.712 4.617 5.301 5.149 4.883 5.073 5.206 4.987 5.206 4.541 5.149 4.788 5.035 5.187 5.358 5.206 5.206 5.149 5.301
498
H. Chen et al.
Table 2. Variance analysis of items in regression equation.
Downloaded by [Florida Atlantic University] at 16:08 26 January 2015
Sources of variation
Sum of squares
df
Mean square
F value
P value
1.82 0.18 0.43 0.30 0.024 0.27 0.1 0.054 0.32 0.23 0.14 0.11 0.031 1.96
9 1 1 1 1 1 1 1 1 1 10 5 5 19
0.2 0.18 0.43 0.30 0.024 0.27 0.1 0.054 0.32 0.23 0.014 0.021 6.173E 2 003
14.84 13.15 31.17 22.13 1.75 19.62 7.31 3.93 23.75 16.89
,0.001 0.0046 ,0.001 ,0.001 0.2153 0.0013 0.0222 0.0755 ,0.001 0.0021
Model x1 x2 x3 x1x2 x1x3 x2x3 x21 x22 x23 Residual Lack of fit Pure error Total
3.42 0.1017
After the CCD of the DIE method, the optimal extraction condition with an extraction time of 90 min, temperature of 458C, solid –liquid ratio of 1:20 g/mL, vacuum degree of 2 0.08 MPa, ethanol concentration of 60% was predicted using the desirability function. Comparing the extraction conditions of DIE and reflux extraction, the reflux extraction is at the same conditions of the DIE method with no vaccum degree. The total phenolic content of the DIE method is 5.4% and that of the reflux extraction is 4.12%. Park (2012) used water at 958C to extract the active material, the phenolic content was 3%; by this method the antioxidant activity (IC50) was 58.9 mg/mL by the 1,1-diphenyl-2-picrylhydrazyl (DPPH) method. We used the DPPH method to study the antioxidant activity. The antioxidant activity (IC50) of the DIE and reflux extraction methods was 20 and 30 mg/mL, respectively, which means that the DIE method had better phenolic content and antioxidant activity. It also proved that the high temperature extraction could improve the degradation of phenols. The reducing power of the phenolic extracts was significantly related to their total phenolic content (R ¼ 0.9903), while the concentration of the phenolic extracts is 40 – 50 mg/ mL and the absorbance at 593 nm is 2.5. The FRAP assay also confirmed that the phenolic extracts of R. verniciflua Stokes had stronger antioxidant activity. Supplementary material Experimental details relating to this article are available online. Acknowledgements The authors thank Dr Wang for his support and guidance with this study. The authors are grateful to the China Basic Research Foundation of National Commonweal Research Institute (CAFINT2013C03), International Cooperation Project (S2012GR0138) and “948” Technology Introduction Project (2012-4-05) for financial support.
References Kim JH, Kim HP, Jung CH. 2006. Inhibition of cell cycle progression via p27Kip1 upregulation and apoptosis induction by an ethanol extract of Rhus verniciflua Stokes in AGS gastric cancer cells. Int J Mol Med. 18:201–208. Kim JS, Kwon YS, Chun WJ. 2010. Rhus verniciflua Stokes flavonoid extracts have anti-oxidant, anti-microbial and aglucosidase inhibitory effect. Food Chem. 120:539–543.
Natural Product Research
499
Downloaded by [Florida Atlantic University] at 16:08 26 January 2015
Park SJ, Kim KH, Shin MJ. 2012. Rhus verniciflua Stokes extract having increased content of active flavonoid compound and method for preparing same. United States patent, US2012/0283322 A1[P]. Wei TY, Tong ZF, Zhao ZX. Extraction the active components of plants by decompressing inner ebullition. China Patent, 2005100328021 2005. Zheng M, Min TL. 1980. Flora of China 45(1)-Anacardiaceae. Beijing: Science press.
