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International Journal of Food Sciences and Nutrition, February 2011; 62(1): 9196

Optimization conditions for anthocyanin and phenolic content extraction form purple sweet potato using response surface methodology
MARUF AHMED1,2, MST. SORIFA AKTER1, & JONG-BANG EUN1
Department of Food Science and Technology and Institute of Biotechnology, Chonnam National University, Gwangju, South Korea, and 2Department of Food Processing and Preservation, Hajee Mohammad Danesh Science and Technology University, Dinajpur, Bangladesh
1

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Abstract Purple sweet potato our could be used to enhance the bioactive components such as phenolic compounds and anthocyanin content that might be used as nutraceutical ingredients for formulated foods. Optimization of anthocyanin and phenolic contents of purple sweet potato were investigated using response surface methodology. A face-centered cube design was used to investigate the effects of three independent variables: namely, drying temperature 55 658C, citric acid concentration 1 3% w/v and soaking time 1 3 min. The optimal conditions for anthocyanin and phenolic contents were 62.918C, 1.38%, 2.53 min and 60.948C, 1.04% and 2.24 min, respectively. However, optimal conditions of anthocyanin content were not apparent. The experimental value of anthocyanin content was 19.78 mg/100 g and total phenolic content was 61.55 mg/g. These data showed that the experimental responses were reasonably close to the predicted responses. Therefore, the results showed that treated ours could be used to enhance the antioxidant activities of functional foods.

Keywords: Purple sweet potato, response surface methodology, phenolic compounds, anthocyanin content

Introduction Purple-eshed sweet potatoes have an intense purple color in the storage roots due to the accumulation of anthocyanins (Terahara et al. 2004).The anthocyanins in purple sweet potato are mono-acylated or di-acylated forms of cyanidin and peonidin (Yang and Gadi 2008). Sweet potatoes had intermediate antioxidant activity among 43 vegetables (Huang et al. 2006). Recently natural antioxidants have attracted considerable attention due to their positive health benet (Huang et al. 2006). Rumbaboa et al. (2009) reported that anthocyanin from purple sweet potato has better radical scavenging activity than that of red cabbage, grape skin, elderberry and purple corn. Anthocyanins from purple sweet potatoes have many biological functions, such as scavenging free radicals, anti-mutagenicity, anti-carcinogen activity and antihypertensive effect (Oki et al. 2002). Several extraction methods have been used to obtain extracts rich in anthocyanin and phenolic content based on different solvents such as methanol, ethanol, acetone, water or mixture (Pathirana and Shahidi 2005, Huang et al. 2006). The stability of anthocyanin and phenolic content were inuenced by several factors (Jiang 2000). Among them, polyphenol oxidase plays an important role in the degradation of anthocyanin and phenolic content. Citric acid has been used extensively for the inhibitory activity on polyphenol oxidase and the anti-browning activity in minimally processed fruits and vegetables. Citric acid extracts have a double inhibitory effect by chelating copper at lower pH (Altunkaya and Gokmen 2009). Sweet potatoes can be processed into our, which are less bulky and more stable than the highly perishable fresh root. Flour can be used as a thickener in soup, gravy, fabricated snacks and bakery products. It could be used to enhance food products through color, avor, natural sweetness and nutrients. Sing et al. (2003) used potassium metabisulphite, citric acid and sodium chloride to improve the quality of chips from

Correspondence: Jong-Bang Eun, Department of Food Science & Technology, Chonnam National University, 77 Yongbong-ro Buk-gu, Gwangju 500-757, South Korea. Tel: 82 62 530 0255. Fax: 82 62 530 2149. E-mail: jbeun@jnu.ac.kr
ISSN 0963-7486 print/ISSN 1465-3478 online q 2011 Informa UK, Ltd. DOI: 10.3109/09637486.2010.511167

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M. Ahmed et al. 6 7%) was obtained by milling the dried slices using a blender (FM-681C; Hanil, Gwangju, Korea), and sieved through an 80-mesh (Chung gye sang gongsa, Seoul, South Korea) screen. Experimental design for RSM analysis A three-factor (X1, X2 and X3) and three-level ( 1, 0 and 1) face-centered cube design were employed in this study, and 15 individual run points were taken for analysis (Wanasundara and Shahidi 1999). The actual and corresponding values are presented in Table I. The multiple regression equation was used to t the second-order polynomial equation based on the experimental data as follows: Y b0 b1 X1 b2 X2 b3 X3 b11 X1 X1 b22 X2 X2 b33 X3 X3 b12 X1 X2 b13 X1 X3 b23 X2 X3 where Y is the response variable, b0 is the intercept, b1, b2, b3, b11, b22, b33 and b12, b13, b23 are linear, quadratic and interaction coefcients respectively, and X1, X2 and X3 are the coded independent variables. Verication of model RSM was used to optimize anthocyanin and phenolic contents from purple sweet potato. The experimental and predicted values were compared to conrm the validity of the model. Analysis of anthocyanin contents The content of anthocyanin was determined following the procedures of Proctor (1974) and Huang et al. (2006) The sweet potato our (1 g) was treated with 15 ml HCl methanol (0.15% HCl:methanol 15:85) for 4 h. The extract was ltered and its absorbance was determined at 530 nm. The anthocyanin content was calculated on the basis of the

sweet potatoes. Response surface methodology (RSM) has been successfully used to optimize biochemical and biotechnological process related to food systems (Cacace and Maza 2003, Pathirana and Shahidi 2005). Therefore, the goal of the present study was to optimize different pretreatments such as drying temperature, citric acid concentration and soaking time for production of sweet potato our with high retention of anthocyanin and phenolic content using RSM. Materials and methods Raw materials
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Sweet potato (Ipomoea batatas L. Lam variety, Sinjami) was purchased from a local farm. Roots were washed with tap water to remove dirt and soil and allowed to dry at ambient temperature (, 208C). The washed sweet potatoes were stored at 148C for 15 days without curing. Sample preparation and treatment Sweet potatoes were peeled with a hand peeler (Han Sung 27 stainless; Namdong Industry Park, Incheon, South Korea). Then peeled samples were cut into slices (1 mm thickness) using a slicing machine (HFS 350G; Hankook fujee Industries Co. Ltd. Suwon-si, Gyeonggi-do, Fujee, South Korea). Various levels of citric acid concentration (1 3% w/v) were solubilized in deionized water at room temperature (20 ^ 18C). After that, peeled slices were dipped in aqueous citric acid solutions (1 3% w/v) for different soaking times (1 3 min) at room temperature. Preparation of sweet potato our The slices were dried using a convection drying oven (Dasol Scientic Co. Ltd, Seoul, South Korea) at different temperatures 558C, 608C, and 658C for 7 8 h. The sweet potato our (moisture content

Table I. Three-factor, three-level, face-centered cube design for RSM. Factor X1 Assay number 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 Drying temperature (8C) 55 (2 1) 55(2 1) 55(2 1) 55(2 1) 60(0) 60(0) 60(0) 60(0) 65( 1) 65(2 1) 65( 1) 65( 1) 60(0) 60(0) 60(0) Factor X2 Concentration (%) 2(0) 1(2 1) 2(0) 3( 1) 3( 1) 1(2 1) 3( 1) 1(2 1) 2(0) 1(2 1) 2(0) 3( 1) 2(0) 2(0) 2(0) Factor X3 Soaking time (min) 3( 1) 2(0) 1(2 1) 2(0) 3( 1) 1(2 1) 1(2 1) 3( 1) 3( 1) 2(0) 1(2 1) 2(0) 2(0) 2(0) 2(0) Variable Anthocyanin (mg/100 g) 40.32 ^ 1.32 39.56 ^ 1.03 40.79 ^ 0.60 37.40 ^ 2.08 34.17 ^ 4.72 32.53 ^ 2.73 24.45 ^ 4.96 24.51 ^ 0.37 29.16 ^ 3.64 26.49 ^ 1.83 24.55 ^ 0.29 22.45 ^ 0.28 23.94 ^ 0.57 20.02 ^ 1.18 21.63 ^ 0.17 Total phenolics (mg/g) 56.85 ^ 8.28 47.77 ^ 1.53 47.18 ^ 5.73 47.52 ^ 4.02 44.64 ^ 0.34 51.69 ^ 0.19 56.38 ^ 0.39 63.38 ^ 1.02 46.46 ^ 1.07 51.65 ^ 2.37 51.65 ^ 2.37 52.03 ^ 2.43 45.70 ^ 0.41 49.39 ^ 0.81 58.91 ^ 1.18

Optimizing anthocyanin and phenolic content of purple sweet potato our following equation: Anthocyanin content A MW DF 100=e W where A is the absorbance, MW is the molecular weight of cyanidin-3-glucoside (MW 449.2), DF is the dilution factor, 1 is the molar absorptivity (34,300), and W is the sample weight (g). Analysis of total phenolic contents The total phenolic content was determined using Folin Ciocalteau reagent according to a slightly modied method (Swain and Hills 1959). The sample (0.1 g) was extracted three times with 20 ml of 75% methanol and was ltered through Whatman No. 2 lter paper. Extracts were combined and concentrated in a rotary vacuum evaporator (Rikakikai Co. Ltd, Tokyo, Japan) at 408C; the volume was adjusted to 20 ml with 75% methanol. One milliliter of extract, 5 ml distilled water and 2 ml of 10% Folin Ciocalteau reagent were added into a Falcon tube. After 3 min at room temperature, 2 ml of 7.5% Na2CO3 solution was added and the sample was diluted to 20 ml with distilled water. Each sample was allowed to stand for
49.22 Anthocyanins (mg/100 g) (a)

93

39.39

29.55

3.00
n(

2.33 19.72 65.00 61.67 Tempe ra 58.33 ture (C )


en

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1.67 1.00 55.00

(b)

49.22 Anthocyanins (mg/100 g)

Table II. Regression coefcients of predicted quadratic polynomial for the response anthocyanin and phenolic contents. Coefcient b0 Linear b1 b2 b3 Quadratic b11 b22 b33 Cross-product b12 b13 b23 R2 Anthocyanin content 1240.40*** 2 36.84*** 2 24.72 2 47.49*** 0.28*** 3.12*** 3.92*** 0.04 0.39 4.43*** 0.97 Phenolic content 2 176.82 6.19 2 1.65 37.69 2 0.04 2 4.76 2 1.95 2 0.16 2 0.30 2 5.97** 0.80

39.39

29.55

Co

nc

3.00

tra

Tempe ra

58.33 ture (C )

1.00 55.00

Figure 1. Response surface plots of the anthocyanin content of purple sweet potato our as affected by temperature, citric acid concentration and soaking time. (a) Temperature and concentration. (b) Temperature and soaking time.

R 2, coefcient of multiple determination. ***Signicant at P # 0.01. **Signicant at P # 0.05. Table III. Analysis of variance for the response surface quadratic model for anthocyanin and phenolic contents. Degree of freedom Sum of squares

Source

Mean square

F value

1 h at room temperature and absorbance was measured at 760 nm (UV-1201; Shimadzu, Kyoto, Japan). The total phenolic content was calculated on the basis of standard curves of gallic acid, and expressed as milligrams of gallic acid equivalents per gram of sample on a wet weight basis. Statistical analysis All determinations were carried out in triplicate and the experimental results are expressed as means ^ standard deviation. Statistical analysis of the verication results was carried out by analysis of variance and Duncans multiple-range tests using SAS (version

For anthocyanin Lack of t Pure error Total error For total phenolic Lack of t Pure error Total error

3 2 5 3 2 5

7.50 7.76 15.26 18.75 92.91 111.67

2.50 3.88 3.05 6.25 46.45 22.33

0.64

0.93

So

61.67

ak

65.00

ing

19.72

1.67

tim

e(

2.33

mi

n)

tio

%)

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(a)

(a) 65.0

Temperature (C)

62.5 64.46 60.0 Phenolic content (mg/g) 56.82

57.5

55.0 1.0

1.5

2.0 2.5 Concentration (%)


21.20 38.90 25.62 43.32 30.05 47.75

3.0

49.17

3.00
Co nc en tra tio n(

Anthocyanins (mg/100 g)

34.47

2.33

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(b) 65.0 Temperature (C) 62.5 60.0 57.5 (b)

41.53 65.00 61.67

1.67

Tempera tu

58.33 re (C)

1.00 55.00

64.46 1.5 2.0 2.5 Soaking time (min)


21.20 38.90 25.62 43.32 30.05 47.75

3.0

Phenolic content (mg/g)

55.0 1.0

56.82

Anthocyanins (mg/100 g)

34.47

9.1). The optimal conditions were estimated through three-dimensional response surface analyses of the three independent variables and each dependent variable.

Tempera tu

re (C)

Figure 3. Response surface plots of the phenolic contents of purple sweet potato our as affected by temperature, citric acid concentration and soaking time. (a) Temperature and concentration. (b) Temperature and soaking time.

Results and discussion Fitting the models Analysis of variance using SAS was performed to determine the signicance of the linear, quadratic, cross-product (Table II) and the lack of t (Table III) of the independent variables on the anthocyanin and phenolic contents. The lack-of-t test is a measure of the failure of a model to represent data in the experimental domain at the points that were not included in the regression (Montgomery 1984). However, the R 2 value of the dependent variables was approximately 0.80, indicating that a high proportion of variability was explained by the data (Varnalis et al. 2004). Therefore, the results showed that the experimental model was adequate due to no signicant lack of t and satisfactory levels of R 2. Effect of pretreatment on anthocyanin and phenolic contents The anthocyanin content of sweet potato ours ranged from 20.02 to 40.79 mg/100 g wet weight basis (Table I). The contents of anthocyanin were much higher than those of sweet potato puree (Steed and Truong 2008) and of steamed or kneaded ours (Huang et al. 2006). The results of multiple regression analysis showed that the anthocyanin contents were signicantly (P # 0.001) affected by the linear term of temperature and soaking time, the quadratic of all terms and the interaction term of concentration and soaking time (Table II). The predicted model obtained

So

58.33

1.00 55.00

ak

61.67

ing

41.53 65.00

1.67

tim

e(

Figure 2. Contour plots showing the effects of temperature, citric acid concentration and soaking time on anthocyanin content of purple sweet potato our. (a) Temperature and citric acid concentration. (b) Temperature and soaking time.

49.17

3.00 2.33
mi n)

%)

Optimizing anthocyanin and phenolic content of purple sweet potato our


(a) 65.0 Temperature (C) 62.5 60.0 57.5 55.0 1.0

95

1.5

2.0 Concentration (%)

2.5

3.0

the anthocyanin structure could be varying with pH (Cevallos-Casala and Cisneros-Zevallos 2004). The total phenolic content of sweet potato ours ranged from 44.64 to 64.32 mg/g wet weight basis (Table I). These results were much higher than those reported in literature for fresh and steamed sweet potato ours (Yang and Gadi 2008). The results of multiple regression showed that the total phenolic content was signicantly affected by the interaction term of concentration and soaking time (X2, X3, P # 0.05). The nal predictive model for phenolic content is given below: Y 2176:82 2 5:97X2 X3 The response surface plots in Figures 3 and 4 show the relationship between the phenolic content and drying temperatures, citric acid concentrations and soaking times. The total phenolic content increased with increasing drying temperatures (Figure 3a,b). This might release more bound phenolic compounds from the breakdown of cellular constituents. Huang et al. (2006) found that steaming treatment increased the total phenolic content of purple sweet potato our. Dewanto et al. (2002a) also found that the free phenolic content of sweet corn increased with increasing heating temperature and time. However, thermal processing had no effect on the phenolic content of tomato (Dewanto et al. 2002b). On the other hand, total phenolic contents decreased with increasing the concentration and soaking time (Figure 4a,b). This was probably because some phenolic compounds were more hydrolyzed or oxidized because dispersions were prepared in the presence of ambient oxygen. Optimization of pretreatments and verication of models The predicted and experimental results are presented in Table IV. For the phenolic content, the predicted response surface of the stationary point was a saddle point. Thus the estimated surface did not have a unique optimum. However, for the anthocyanin content, the predicted response surface of the stationary point was a minimum. Therefore, different optimum conditions were obtained for both responses. The optimal conditions for anthocyanin and phenolic contents were 62.918C, 1.38%, 2.53 min, whereas for total phenolic contents they were drying temperature 60.948C, citric acid concentration 1.04% and soaking time 2.24 min. However, optimal conditions of the anthocyanin content were not apparent. This is due to

Phenolic content (mg/g)

42.68 56.43

46.12 59.87

49.56 63.31

53.00

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(b) 65.0 Temperature (C) 62.5 60.0 57.5 55.0 1.0

1.5

2.0 Soaking time (min)

2.5

3.0

Phenolic content (mg/g)

42.68 56.43

46.12 59.87

49.56 63.31

53.00

Figure 4. Contour plots showing the effects of temperature, citric acid concentration and soaking time on phenolic contents of purple sweet potato our. (a) Temperature and citric acid concentration. (b) Temperature and soaking time.

for Y is given below: Y 1240:40 2 36:84X1 2 47:49X3 0:28X2 1


2 3:12X2 2 3:92X3 4:43X2 X3

Figures 1 and 2 show the response surface plots of the relationship between anthocyanin content and drying temperatures, citric acid concentrations and soaking times. Anthocyanin contents decreased with increasing drying temperatures (Figure 1a,b). This was as expected because heating opened the structure of anthocyanin to form chalcones, which was degraded further to form brown products (Delgado-Vargas et al. 2000). However, the anthocyanin content increased with increasing soaking time and concentration (Figure 2a,b). This might be due to interaction between citric acid and anthocyanin. In acidic media,

Table IV. Comparison of predicted and experimental values for the response of anthocyanin and phenolic contents. Optimum conditions Response variable Anthocyanin Total phenolics Stationary point Minimum Saddle Temperature (8C) 62.91 60.64 Soaking time (min) 2.53 1.04 Soaking concentration (%) 1.38 2.24 Values Experimental 19.78 ^ 0.97 61.55 ^ 2.9 Predicted 19.71 52.89

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Fan G, Han Y, Gu Z, Chen D. 2008. Optimization conditions for anthocyanins extraction from purple sweet potato using response surface methodology (RSM). LWT Food Sci Technol 41: 155 160. Huang YC, Chang YH, Shao YY. 2006. Effects of genotype and treatment on the antioxidant activity of sweet potato in Taiwan. Food Chem 98:529538. Jiang Y. 2000. Role of anthocyanins, polyphenoloxidase and phenols in lychee pericarp browning. J Sci Food Agric 80:305310. Montgomery DC. 1984. Design and analysis of experiments. 2nd ed. New York: John Wiley and Sons. Oki T, Masuda M, Furuta S, Nishiba Y, Terahara N, Suda I. 2002. Involvement of anthocyanins and other phenolic compounds in radical-scavenging activity of purple-eshed sweet potato cultivars. J Food Sci 67:17521756. Pathirana LC, Shahidi F. 2005. Optimization of extraction of phenolic compounds from wheat using response surface methodology. Food Chem 93:47 56. Proctor JTA. 1974. Colour stimulation in attached apples with supplementary light. Can J Plant Sci 54:499503. Rumbaboa RGO, Cornago DF, Geronimo IM. 2009. Phenolic content and antioxidant capacity of Philippine sweet potato (Ipomoea batatas) varieties. Food Chem 113:11331138. Sing S, Raina CS, Bawa AS, Saxena DC. 2003. Optimization of processing variables in the preparation of sweet potato chips using response surface methodology. Euro Food Res Technol 217:374381. Steed LE, Truong VD. 2008. Anthocyanin content, antioxidant activity and selected physical properties of owable purpleeshed sweet potato purees. J Food Sci 73:215221. Swain T, Hillis WE. 1959. The phenolic constituents of prunus domestica. I. The quantitative analysis of phenolic constituents. J Sci Food Agric 10:63 68. Terahara N, Konczak I, Ono H, Yoshimoto M, Yamakewa O. 2004. Characterization of acylated anthocyanins in callus induced from storage root of purple-eshed sweet potato, Ipomoea batatas L. J Biomed Biotechnol 5:279286. Varnalis AI, Brenan JG, Macdougall DB, Gilmour SG. 2004. Optimization of high temperature pufng of potato cubes using response surface methodology. J Food Eng 61:153163. Wanasundara UN, Shahidi F. 1999. Concentration of omega polyunsaturated fatty acids of seal blubber oil by urea complexation: Optimization of reaction conditions. Food Chem 65:4149. Yang J, Gadi RL. 2008. Effects of steaming and dehydration on anthocyanins, antioxidant activity, total phenols and color characteristics of purple-eshed sweet potatoes (Ipomoea batatas). Am J Food Technol 3:224 234.

the fact that the optimization point was a minimum. The optimal value of anthocyanin content was lower than expected values. This might be related to the anthocyanin extraction conditions by secondorder polynomials (Fan et al. 2008). The corresponding experimental responses of anthocyanin and total phenolic contents were 19.78 mg/100 g and 61.55 mg/g, respectively. These data showed that the experimental responses were reasonably close to the predicted responses. Conclusion The results of anthocyanin and phenolic contents were higher than the previous reported values for raw, steam and kneaded sweet potato ours. Therefore, treated ours could be used to make the higher quality products that would be more attractive to product developers and consumers. Declaration of interest: The authors report no conicts of interest. The authors alone are responsible for the content and writing of the paper. References
Altunkaya A, Gokmen V. 2009. Effect of various anti-browning agents on phenolic compounds prole of fresh lettuce (L. Sativa). Food Chem 117:122126. Cacace JE, Mazza G. 2003. Optimization of extraction of anthocyanins from black currants with aqueous ethanol. J Sci Food Agric 68:240248. Cevallos-Casala BA, Cisneros-Zevallos L. 2004. Stability of anthocyanin-based aqueous extracts of Andean purple corn and red-eshed sweet potato to synthetic and natural colorants. Food Chem 86:6977. Delgado-Vargas F, Jimenez AR, Paredes-Lopez O. 2000. Natural pigments: Carotenoids, anthocyanins, and betalainscharacteristics, biosynthesis, processing and stability. Crit Rev Food Sci Nutr 40:173289. Dewanto V, Xianzhong WU, Adom KK, Liu RH. 2002a. Thermal processing enhances the nutritional value of tomatoes by increasing total antioxidant activity. J Sci Food Agric 50: 30103014. Dewanto V, Xianzhong WU, Adom KK, Liu RH. 2002b. Processed sweet corn has higher antioxidant activity. J Sci Food Agric 50: 49594964.

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