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Innovative Food Science and Emerging Technologies 12 (2011) 282–287

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Innovative Food Science and Emerging Technologies
j o u r n a l h o m e p a g e : w w w. e l s ev i e r. c o m / l o c a t e / i f s e t

Optimized microwave-assisted extraction of total phenolics (TP) from Ipomoea batatas leaves and its antioxidant activity
Jiangfeng Song, Dajing Li, Chunquan Liu ⁎, Ying Zhang
Institute of Farm Product Processing, Jiangsu Academy of Agricultural Sciences, Nanjing 210014, PR China Engineering Research Center for Agricultural Products Processing, National Agricultural Science and Technology Innovation Center in East China, Nanjing 210014, PR China

a r t i c l e

i n f o

a b s t r a c t
An efficient microwave-assisted extraction (MAE) technique was employed to extract total phenolics (TP) from sweetpotato [Ipomoea batatas (L.) Lam.] leaves (TPSL). The optimal conditions for microwave-assisted extraction of TPSL were determined by response surface methodology. A face-centered cubic design (FCD) was applied to evaluate the effects of three independent variables (microwave power, extraction time and ethanol proportion) on the recovery of TPSL. The correlation analysis of the mathematical-regression model indicated that quadratic polynomial model could be employed to optimize the microwave extraction of TPSL. From response surface plots, microwave power, extraction time and ethanol proportion exhibited independent and interactive effects on the extraction of TPSL. The optimal conditions to obtain the highest recovery of TPSL were as follows: microwave power, 302 W; extraction time, 123 s; ethanol proportion, 53% (v/v). Under these optimal conditions, the experimental values agreed with the predicted ones by analysis of variance. It indicated high fitness of the model used and the success of response surface methodology for optimizing TPSL extraction. After method development, the antioxidant activity of the MAE extract was preliminarily evaluated. MAE showed obvious advantages in terms of high extraction efficiency and antioxidant activity of extract with in shortest extraction time. Industrial Relevance: Sweet potato leaves have been neglected except for a partial use as livestock feed in China. This study describes the response surface optimization of microwave-assisted extraction (MAE) process for the enhanced recovery of total phenolics from sweetpotato leaves (TPSL). MAE showed obvious advantages in terms of high extraction efficiency and antioxidant activity of extract within shortest extraction time. TPSL obtained using MAE showed potential for use in health-care food and pharmaceutical industry. Crown Copyright © 2011 Published by Elsevier Ltd. All rights reserved.

Article history: Received 16 July 2010 Accepted 5 March 2011 Editor Proof Receive Date 6 April 2011 Keywords: Sweetpotato leaves TP Microwave-assisted extraction DPPH radical scavenging activity

1. Introduction Recent studies indicate that phenolics from numerous vegetables exert several health-promoting functions, including reducing the risks of cancer and heart and neurodegenerative diseases (Joseph, Fisher, & Bielinski, 2005; Vita, 2005). Although there is still uncertainty about the relationship between polyphenols and ailments, the healthpromoting potential of these foods may be attributed to the phytochemical compounds present in the plants (Rangkadilok, Worasuttayangkurn, Bennett, & Satayavivad, 2005). Sweetpotato [Ipomoea batatas (L.) Lam.] leaves are consumed as a leafy vegetable in many parts of the world and can be harvested many times during a season, which are considered to be rich sources of phenolic compounds (Villareal, Tsou, Lo, & Chiu, 1982; S. Islam, 2006). Five major phenolic acids were identified in sweetpotato leaf tissue such as chlorogenic acid (ChlA); 3,5-dicaffeoylquinic acid (3,5⁎ Corresponding author. Tel.:+86 25 84391255; fax:+86 25 84391570. E-mail addresses: songjiangfeng102@163.com (J. Song), liuchunquan2009@163.com (C. Liu).

diCQA); 3,4-dicaffeoylquinic acid (3,4-diCQA); 4,5-dicaffeoylquinic acid (4,5-diCQA) and 3, 4, 5-tri-O-caffeoylquinic acid (3,4,5-triCQA) (Islam, Yoshimoto, Yahara, & Yamakawa, 2002; Truong, Mcfeeters, Thompson, Dean, & Shpfran, 2007; Yoshimoto et al., 2006). Previous reports indicated that the sweetpotato leaves had a higher antioxidant potential than root tissue (Islam, Yoshimoto, Ishiguro, Okuno, & Yamakawa, 2003). However, sweetpotato leaves have been neglected except for a partial use as livestock feed in China. In recent years, the use of microwave-assisted extraction (MAE) of constituents from plant material has shown tremendous research interest and potential. Compared with the traditional methods, MAE has many advantages, such as shorter time, less solvent, higher extraction rate and better products with lower cost because microwave treatment has mechanical effects that heats the solvent mixture directly, and interacts directly with the free water molecules present in the glands and vascular systems, which results in rupture of the plant tissue and release of the active constituents into the solvent (Al-Harahsheh & Kingman, 2004). So far, microwave treatment has been widely employed to extract total phenolics from different plant materials (Sutivisedsak et al., 2009; Beejmohun et al., 2007; Kalia,

1466-8564/$ – see front matter. Crown Copyright © 2011 Published by Elsevier Ltd. All rights reserved. doi:10.1016/j.ifset.2011.03.001

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Sharma, Singh, & Singh, 2008). Up to our best knowledge, usage of MAE has not been previously reported in total phenolics in sweetpotato leaves. Response surface methodology (RSM) is a statistical method that uses quantitative data from an appropriate experimental design to determine or simultaneously solve multivariate equation (Yang, Zhao, Shi, Yang, & Jiang, 2008). Unlike the conventional empirical method, RSM can generate a mathematical model, and take into account the possible interrelationship among the test variables while minimizing the number of experiments (Silva, Rogez, & Larondelle, 2007). The availability of phenolics in sweetpotato [Ipomoea batatas (L.) Lam.] leaves (TPSL) as antioxidant source is ensured. However, the economical feasibility of an industrial process also requires working in such a way that high extraction efficiency is attained. In this study, microwave technique was employed to extract TPSL. Response surface methodology was used to evaluate the effects of microwave power, extraction time and ethanol proportion on the recovery of TPSL to obtain the optimal extraction conditions, furthermore on basis of DPPH radical scavenging assay, antioxidant activity of TPSL with MAE was preliminarily evaluated. 2. Materials and methods 2.1. Materials For the Ningshu 43-8 sweetpotato leaves, the samples were harvested after 105 days of transplanting the cuttings to the fields from experimental plots at Jiangsu Academy of Agricultural Sciences, Nanjing, Jiangsu Province, China. Harvested leaves were immediately placed in plastic bags and transported in an ice-cooled container to the laboratory within a day. The leaves were wiped with wet paper towels to remove dirt and then air-dried at room temperature for 15 to 30 min before sampling, then the sweetpotato leaves were crushed onto small particle sizes using a high speed-crushing machine (FW100, 24000 r/min, Tianjin Taisite Instrument Co., Ltd.) and were passed through a 40-mesh sieve. The resultant crushed powder was subjected to the following analysis. 2.2. Chemicals DPPH (1,1′-diphenyl-2-picrylhydrazyl) was purchased from Sigma chemical company (St. Louis, MO, USA). Gallic acid were obtained

from China Medicinal Group Shanghai Chemical Reagent Co. (Shanghai, China). Deionized water used for the preparation of all the solutions was purified by Milli-Q purification system (Millipore) (Massachusetts, USA). All other chemicals used were of analytical grade. 2.3. Extraction and quantification of TPSL The process of total phenolics extraction from dried sweetpotato leaves by microwave treatment was performed using a microwave solvent extraction system (LWMC-205, 2450 MHz, Lingjiang Development Co., Ltd., Nanjing, China) with adjustable power settings and with full-power level of 900 W (Fig. 1). 1.0 g of dried sweetpotato leaves was immersed into a certain ethanol solution. The extraction process was performed at different microwave power, extraction time and ethanol proportion. The extract was filtered through a Whatman No. 1 filter paper and the filtrate was then concentrated to 15 ml with a rotary evaporator at 40 °C under vacuum. The supernatant was obtained by centrifugation for 10 min. The addition of hexane to the supernatant from ethanol-extracted samples was tested to remove polar lipids and other interfering compounds. The clear supernatant was diluted to the final volume of 50 ml and was used for TP analysis. The content of total phenolics was determined by the Folin-Ciocalteu colorimetric method (Silavin, Cheng, Luther, Kenworthy, & Yu, 2009). Gallic acid was used to construct a standard curve. The recovery of TPSL was expressed as mg of gallic acid equivalents (GAE) per gram of sweetpotato leaves on dry weight (DW) basis. 2.4. Face-centered cubic design On the basis of single-factor experiment for the recovery of TPSL, proper ranges of microwave power, extraction time, ethanol proportion and ratio of liquid to solid were preliminarily determined. A facecentered cubic design (FCD) was used to evaluate the main and interaction effects of factors: microwave power (X1), extraction time (X2) and ethanol proportion (X3) on the recovery of TPSL obtained from microwave extraction process. Based on the investigations on single-factor experiment, the variables considered were microwave power, extraction time and ethanol proportion in this experimental design. For statistical calculation, the variables were coded according to xi = Xi −X0 ΔX ð1Þ

where xi was a coded value of the variable; Xi was the actual value of variable; X0 was the actual value of the Xi on the center point; and ∆X was the step change value. The range of independent variables and their levels is presented in Table 1, which was based on the results of preliminary experiments. The recovery of TPSL was the dependent variable. All experimental data obtained are shown in Table 2, the whole design consisted of 20 experimental points carried out in random order. The software Design Expert (Trial Version 7.0.3, StatEase Inc., Minneapolis, MN, USA) was employed for experimental design, data analysis and model building. Experiments were randomized to maximize the effects of unexplained variability in the observed responses due to extraneous factors. A full quadratic equation or the

Table 1 Factors and levels tested for the designed experiment. Factor Low level (− 1) Medium level (0) High level (+ 1) 350 90 70 450 110 80

Fig. 1. Schematic diagram of microwave equipment.

Microwave power (X1, W) 250 Microwave time (X2, s) 70 Ethanol proportion (X3, %) 60

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Table 2 Face-centered cubic design and the response for the recovery of TPSL. Experiments Coded levels X1 Microwave power (W) 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 1 (450) 1 (450) 1 (450) 1 (450) 1 (450) −1 (250) −1 (250) −1 (250) −1 (250) −1 (250) 0 (350) 0 (350) 0 (350) 0 (350) 0 (350) 0 (350) 0 (350) 0 (350) 0 (350) 0 (350) X2 Microwave time (s) 1 1 −1 −1 0 −1 0 1 1 −1 0 1 0 −1 0 0 0 0 0 0 (110) (110) (70) (70) (90) (70) (90) (110) (110) (70) (90) (110) (90) (70) (90) (90) (90) (90) (90) (90) X3 Ethanol proportion (%) −1 (60) 1 (80) 1 (80) −1 (60) 0 (70) 1 (80) 0 (70) 1 (80) −1 (60) −1 (60) −1 (60) 0 (70) 1 (80) 0 (70) 0 (70) 0 (70) 0 (70) 0 (70) 0 (70) 0 (70) 57.97 57.56 53.29 53.87 58.03 53.87 58.20 56.87 61.14 53.64 58.78 60.45 59.47 56.70 60.39 59.01 60.04 60.78 59.94 61.07 Recovery of TPSL (mgGAE/gDW)

Recovery of TPSL (mg GAE/g DW)

54 53 52 51 50 49 48 47 150 250 350 450 550 650

Microwave power (W)
Fig. 3. Effect of microwave power on the extraction of TPSL (extraction time 90 s, ethanol proportion 70%, ratio of liquid to solid 30 ml/g).

2.6. Statistical analysis All experimental results were expressed as mean ± SD of three parallel measurements and all calculations were carried out with the help of the statistical package for the social science SPSS 11.5. Analysis of variance (ANOVA) was performed by “Design expert 7.1.3” software. P values b 0.05 were regarded as signi ficant and P values b 0.01 as very significant. 3. Results and discussion 3.1. The effect of different ethanol proportion on the extraction of TPSL Different ethanol proportion was set at 40, 50, 60, 70 and 80% (v/v) to investigate the influence of ethanol proportion on the recovery of TPSL when the other reaction conditions were set as follows: microwave power 350 W, extraction time 90 s, ratio of liquid to solid 30 ml/g. Fig. 2 shows that the extraction of TPSL was greatly influenced by the ethanol concentration in water. When the ethanol volume percentage in the solvent was lower than 70% (v/v), the extraction recovery of TPSL was increased with the increase of ethanol concentration. When the ethanol volume percentage in the solvent was higher than 70% (v/v), the extraction was slowly decreased with the further increase of ethanol concentration. Therefore, 60–80% (v/v) ethanol concentration in water was considered to be optimal in the following experiments. 3.2. The effect of different microwave power on the extraction of TPSL Different microwave power was set at 150, 250, 350, 450, and 550 W to investigate the influence of microwave power on the recovery of TPSL when the other reaction conditions were set as follows: extraction time 90 s, ethanol proportion 70%, ratio of liquid to solid 30 ml/g. Fig. 3 shows that the extraction of TPSL was greatly influenced by the microwave
Recovery of TPSL (mg GAE/g DW)
55 54 53 52 51 50 49 48 47 30 50 70 90 110 130 150 170 190

diminished form of this equation, shown as follows, was used for this model. Y = β0 + ∑ βi Xi + ∑ βii Xi + ∑ ∑ βij Xi Xj
i=1 i=1
i =1 j = 2 ibj

3

3

2

2

3

ð2Þ

where β0, βi, βii, and βij are the regression coefficients for intercept, linear, quadratic and interaction terms, respectively, and Xi, and Xj are the independent variables. 2.5. Assay of DPPH radical scavenging activity The DPPH radical scavenging assay was used to determine the antioxidant activity of the extract. The DPPH radical scavenging activity was measured according to the method of Peng et al. (Peng, Chen, Lin, & Lin, 2000). Two milliliters of 0.2 mM DPPH in ethanol was added to 2 ml of the TPSL ethanolic dilute solution. The absorbance was measured at 517 nm after 20 min of incubation at 25 °C. Distilled water was used as the control. The scavenging activity of DPPH radicals by the sample was calculated according to the following equation: DPPH radical scavenging activity (%) = (1 − absorbance of sample/absorbance of control) × 100.
46.5 46 45.5 45 44.5 44 43.5 43 42.5 40 50 60 70 80 90

Recovery of TPSL (mg GAE/g DW)

Ethanol proportion (%)
Fig. 2. Effect of ethanol concentration in water on the extraction of TPSL (microwave power 350 W, extraction time 90 s, ratio of liquid to solid 30 ml/g).

Extraction time (s)
Fig. 4. Effect of extraction time on the extraction of TPSL (microwave power 350 W, ethanol proportion 70%, ratio of liquid to solid 30 ml/g).

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Recovery of TPSL (mg GAE/g DW)

60 50 40 30 20 10 0 10

Table 4 Test of significance for regression coefficient. Model term Recovery of TPSL (Y) Coefficient estimate 60.11 −0.3 2.26 −0.43 −0.27 0.38 −0.54 −1.86 −1.40 −0.85 DF 1 1 1 1 1 1 1 1 1 1 Standard error 0.299 0.275 0.275 0.275 0.307 0.307 0.307 0.524 0.524 0.524 95% CI low 59.450 −0.912 1.650 −1.046 −0.950 −0.303 −1.225 −3.031 −2.571 −2.021 95% CI high 60.780 0.312 2.874 0.178 0.418 1.0653 0.143 −0.697 −0.237 0.313 Prob N F

15

20

25

30

35

40

Ratio of liquid to solid (ml/g)
Fig. 5. Effect of ratio of liquid to solid on the extraction of TPSL (microwave power 350 W, extraction time 90 s, ethanol proportion 70%).

Intercept X1 X2 X3 X1 X2 X1 X3 X2 X3 X2 1 X2 2 X2 3

0.3002 b 0.0001 0.1451 0.4061 0.2426 0.1084 0.0052 0.0230 0.1339

power. When microwave power was lower than 350 W, the extraction increased with the increase of microwave power. When microwave power was higher than 350 W, the extraction decreased with the further increase of microwave power. These data suggest that applying a higher microwave power for a short time maybe the most effective way to extract phenolic compounds from sweetpotato leaves and other plant materials using MAE. However, a higher microwave power may lead to thermal degradation of the phenols. 3.3. The effect of different extraction time on the extraction of TPSL The recovery of TPSL affected by different extraction time (30, 60, 90, 120, 150, and 180 s) is shown in Fig. 4, when other three factors (microwave power, ethanol proportion and ratio of liquid to solid) were fixed at 350 W, 70% (v/v) and 30 ml/g. The results indicate that the recovery of TPSL increased with the increase of MAE time in the beginning of extraction. The recovery could reach its maximum 53.97 ±0.15 mg GAE/g DW in 90 s during the MAE process. If the extraction time was more than 90 s, the extraction recovery of TPSL decreased with the increase of extraction time, herein the extraction time of 70–110 s was investigated in the present work. 3.4. The effect of different ratio of liquid to solid on the extraction of TPSL The recovery of TPSL affected by different ratio of liquid to solid (10, 15, 20, 25, 30 and 35 ml/g) can be seen in Fig. 5, when the other three factors (microwave power, extraction time, and ethanol proportion) were fixed at 350 W, 90 s, and 70% (v/v). The result implied the recovery of TPSL was enhanced to the critical value (48.51 ± 0.24%) at the ratio of 25, and then it maintained a mild slope when the ratio of liquid to solid increased from 25 to 30 ml/g. As there was no significant difference between the recovery at 25 and that at 30 ml/g, in order to save the solvent consumption, 25 ml/g was chosen as the ratio of liquid to solid through all the extraction optimization experiments. 3.5. Optimization of MAE conditions of TPSL Optimization of MAE conditions has been reported in many studies (Eskilsson & Björklund, 2000; Wang et al., 2010; Zheng et al., 2009).

Many researchers have used single factors, factorial and orthogonal array designs to find optimal conditions. RSM optimization is more advantageous than the traditional single parameter optimization in that it saves time, space and raw material. There were a total of 20 runs for optimizing the three individual parameters in the FCD. Table 2 shows the experimental conditions and the results of recovery of TPSL according to the FCD. Maximum recovery of TPSL (61.14 mg GAE/gDW) was recorded under the experimental conditions of microwave power 250 W, extraction time 110 s and ethanol proportion 60% (v/v). By applying multiple regression analysis on the experimental data, the mathematical model representing the recovery of TPSL as a function of the independent variables within the region under investigation was expressed by the following equation: Y = 60:11−0:30X1 + 2:26X2 −0:43X3 −1:86X1 −1:40X2 −0:85X3 −0:27X1 X2 + 0:38X1 X3 −0:54X2 X3 ð3Þ where Y is the recovery of TPSL, whereas X1, X2 and X3 are the coded variables for microwave power, extraction time and ethanol proportion, respectively. In general, exploration and optimization of a fitted response surface may produce poor or misleading results unless the model exhibits a good fit, which checks the model adequacy essential (Liyana-Pathirana & Shahidi, 2005). The P-values of the model for the recovery of TPSL were b 0.0001 (Table 3), which indicated that the fitness of the model was extremely significant. However, the fit value of the model exhibited 0.2568, without significant difference.
Recovery of TPSL (mg GAE/g DW)
2 2 2

61.1 59.45 57.8 56.15 54.5

Table 3 Analysis of variance for the response surface quadratic model for the recovery of TPSL. Source Model Lack of fit Pure error Total Degrees of freedom Sum of squares Mean square 13.49 0.98 0.53 F-value 17.84 1.86 P-value b 0.0001 0.2568

110.00 100.00 90.00 350.00 80.00 70.00 250.00 300.00 400.00

450.00

9 121.4 5 4.91 5 2.65 19 128.96 R-squared = 0.9415

Extraction time (s)

Microwave power (W)

Adj R-squared = 0.8889

Fig. 6. Response surface plots showing effects of microwave power and extraction time on the recovery of TPSL and their interaction. The ethanol proportion was constant at 70% (v/v).

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J. Song et al. / Innovative Food Science and Emerging Technologies 12 (2011) 282–287 Table 5 Effect of different extraction methods on recovery of TPSL. Extraction method MAE TE Extraction Ethanol Ratio of liquid to Recovery of TPSL time (min) proportion (%) solid (ml/g) (mg GAE/g DW) 2 120 53 60 25 30 61.15 ± 0.08 59.69 ± 0.11

Recovery of TPSL (mg GAE/g DW)

60.2 59.35 58.5 57.65 56.8

80.00 75.00 70.00 350.00 65.00 60.00 250.00 300.00 400.00

450.00

Ethanol proportion (%)

Microwave power (W)

Fig. 7. Response surface plots showing effects of microwave power and ethanol proportion on the recovery of TPSL and their interaction. The extraction time was constant for 90 s.

Coefficient (R2) of determination is defined as the ratio of the explained variation to the total variation and is a measurement of the degree of fitness (Nath & Chattopadhyay, 2007). The small value of R2 indicates the poor relevance of the dependent variables in the model. The model can fit well with the actual data when R2 approaches unity (Sin, Yusof, Hamid, & Rahman, 2006). Analysis of variance, the R2 value of the model for the recovery of TPSL was determined to be 0.9415, only 5.85% of that total variations was not explained by the model, which showed that the regression model defined well the true behavior of the system. The regression coefficient of Eq. (3) calculated and tested for their significance using Design Expert and are listed in Table 4. The P-value was used as a tool to check the significance of each coefficient between the variables. The smaller the value of P, the more significant the corresponding coefficient. It can be seen from the table that the linear coefficient (X2) and quadratic term coefficient (X2 1) were extremely significant, with very small P-value (P b 0.01), the quadratic term coefficient (X2 2) was significant, the P-value being very small. The other term coefficients were not significant (P N 0.05). The full model filled Eq. (3) was made the 3-D response surface plot to predict the relationships between the independent and dependent variables. The effects of microwave power, extraction time and ethanol proportion on the recovery of TPSL, as well as their interactions, are shown in Figs. 6–8. The principle of heating during microwave extraction is based on the direct effect of microwaves on molecules by ionic conduction and dipole rotation. Ionic conduction is the electrophoretic migration when an electromagnetic field is applied. The resistance of the solution to this flow of ions will result in friction, which will heat the solution (Song, Li, & Liu, 2009). The 3-D response surface plot based on independent variables microwave
Recovery of TPSL (mg GAE/g DW)

power and extraction time are shown in Fig. 6, while ethanol proportion was kept at 0 level, an increase of the recovery of TPSL could be significantly achieved with the increases of extraction time. It was obvious that the recovery of TPSL was increased with the increasing extraction time from 70 to 100 s, meaning that further increases of extraction time would not increase the recovery of TPSL any longer. The extending microwave time could result in a higher extraction recovery. However, the recovery of TPSL decreased with the extension of microwave time when a high microwave power was used, which was possibly due to the degradation of total phenolics by microwave wave. In Fig. 7, when the 3-D response surface plot was developed for the recovery of TPSL with varying microwave power and ethanol proportion at fixed extraction time (0 level), it can be seen that maximum recovery of TPSL can be achieved when microwave power and ethanol proportion were 338.6 W and 67.1% (v/v), respectively. The recovery of TPSL affected by different ethanol proportion and extraction time is seen in Fig. 8, when microwave power was fixed at 0 level. It is clear that with an increase of ethanol proportion from 65 to 80% (v/v), the recovery of TPSL was gradually decreased. Solubility of phenolics could be enhanced using an aqueous ethanol over a limited compositional range. In general, it was found that ethanol solutions that ranged from 40 to 80% had greater efficiency in the extraction of phenolics compared to pure ethanol (Hayouni, Abedrabba, Bouix, & Hamdi, 2007). From the industrialization point of view, low ethanol concentration with shorter extraction time would be more adequate as long extraction period rendered the extraction procedure time consuming and uneconomical. By prediction of computing program, the optimal conditions to obtain the highest recovery of TPSL were determined as follows: microwave power, 302 W; extraction time, 123 s; ethanol proportion, 53% (v/v). After extraction of TPSL under these optimal conditions, the recovery of TPSL were 61.26 ± 0.16 mg GAE/g DW, but they were not significantly different to predicted value 61.38 mg GAE/g DW within 95% confidence interval. 3.6. Comparison of MAE with traditional extraction method (TE) Table 5 shows that the optimized MAE has the advantage of reduction in extraction time, saving energy and increased extraction yield as compared with traditional extraction. MAE for about 2 min
100.00

61.3 59.825 90.00

Scavenging effect (%)

80.00 70.00 60.00 50.00 40.00 30.00 20.00 10.00 0.00 0 10 20 30 40 50 60 70 80 90
TPSL BHT

58.35 56.875 55.4 80.00 75.00 70.00 90.00 80.00 60.00 70.00 100.00

110.00

Ethanol proportion (%) 65.00

Extraction time (s)

Concentration (µg/ml)
Fig. 8. Response surface plots showing effects of extraction time and ethanol proportion on the recovery of TPSL and their interaction. The microwave power was constant at 350 W. Fig. 9. DPPH radical scavenging activity of TPSL and BHT. Values are means ± SD of three determinations.

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gave the same recovery of TPSL as that of TE for 120 min. MAE can greatly reduce the extraction time for the same level of extraction, the quantity of solvent is less and the processing time is shorter. TE is a time-consuming process based on heating and mixing to increase the mass transfer rate. The high efficiency of MAE found in this work is suggested to be because the cells of sweetpotato leaves were broken by the microwave radiation, so that total phenolics dissolved more easily in the solvent. 3.7. Scavenging activity of TPSL on DPPH radical Radical scavenging activities of MAE extract were tested using DPPH as the free radical. DPPH is a free radical compound and has been used to test free radical scavenging ability of various samples widely in present studies. In the DPPH test, the antioxidants reduce the DPPH radical to a yellow-colored compound, diphenylpicrylhydrazine, and the extent of the reaction will depend on the hydrogen donating ability of the antioxidants (Ardestani & Yazdanparast, 2007). The TPSL demonstrated a concentration-dependent scavenging activity by quenching DPPH radicals (Fig. 9). As shown in Fig. 9, TPSL was found to have a higher scavenging effect on DPPH radical at concentrations from 10 to 80 μg/ml, and even exceed the potency of BHT which has been documented to reduce and decolorize DPPH by their hydrogen donating capabilities (Li, Huang, Gong, & Ma, 2008). The scavenging effects of TPSL increased in a dose-dependent manner. This result proved that TPSL with MAE had a significant effect on scavenging of DPPH radicals. 4. Conclusions The high correlation of the mathematical model indicated that quadratic polynomial model could be employed to optimize microwave extraction process of TPSL. From response surface plots, three factors (microwave power, time and ethanol proportion) significantly influenced the extraction efficiency of TPSL, independently and interactively. The optimal conditions to obtain the highest recovery of TPSL were determined to be 302 W, 123 s and 53%. Under the optimal conditions, the experimental values agreed with the predicted values by analysis of variance. Thus, this methodology could provide a basis for the model to search for non-linear nature between independent variables and response in a short term experiment. In addition, TPSL obtained using MAE showed antioxidant potential for use in health-care food. However, further studies concerning the nutritional and health benefits are required before a large scale utilization of the sweetpotato leaves can be recommended. Acknowledgements The financial support provided by Agricultural Science and Technology & Self-innovation Foundation of Jiangsu Province (Grant No. CX (09) 628), and National Agricultural Comprehensive Development Industrialization Foundation (No. 2009KJ-16) was appreciated. References
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