food and bioproducts processing 8 8 ( 2 0 1 0 ) 209214

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Food and Bioproducts Processing

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Short communication

Effect of extraction conditions on yield and degree of esterication of durian rind pectin: An experimental design
Wong Weng Wai, Abbas F.M. Alkarkhi, Azhar Mat Easa
School of Industrial Technology, 11800, Universiti Sains Malaysia, Minden, Penang, Malaysia

a b s t r a c t
The effect of time (1 and 4 h), pH (2.0 and 2.5) and temperature (80 and 90 C) on yield and degree of esterication (DE) of durian rind pectin was investigated. The yield and DE of the extracted pectin ranged from 2.1 to 10.3% (w/w, based on dry weight of durian rind) and 45.664.8% respectively. Yield was signicantly affected by time, temperature and pH, and interactions between temperature and pH, and heating time and pH. DE was signicantly affected by heating time and pH, and interactive effects of temperature and pH, and heating time and pH. The extraction yield was not related to DE. By considering pectin yield and DE, the acid extraction of durian rind pectin at 80 C, for 4 h and at pH 2.5 could be suitable. 2010 The Institution of Chemical Engineers. Published by Elsevier B.V. All rights reserved. Keywords: Durian rind pectin; Yield; Degree of esterication; Experimental design; Regression model

1.

Introduction

Durian (Durio zibethinus), a tropical fruit native to Southeast Asia, is one of the most highly valued and desired fruits among Southeast Asians due to its distinct avour and unique taste. The fruit is ovoid or ovoid-oblong to nearly round shaped with an average size weighing between 2 and 4.5 kg depending on their varieties (Hokputsa et al., 2004). The rind which usually weighs more than half of the total fruit weight is green to yellowish brown, thick and semi-woody with sharply pointed pyramidal thorns. During the season of durian, the amounts of rind that disposed as waste could lead to environmental problems. This agricultural waste could be further utilized as a source of valuable materials of commercial importance; such as particleboard component of construction panels for energy conservation in building (Khedari et al., 2003), tablet binder, tablet disintegrator and gelling agent (Pongsamart and Panmaung, 1998; Umprayn et al., 1990a,b). Of interest is watersoluble polysaccharides extracted from durian rind containing high amount pectin (Hokputsa et al., 2004). Pectin is a complex family of heterogeneous branched polysaccharides that arise from the primary cell walls and

intercellular regions of higher plants, consisting mainly of dgalacturonic acid and neutral sugars, such as l-rhamnose, l-arabinose, and d-galactose. The term pectin or pectic substance describes a group of polysaccharides in which the presence of partly methyl-esteried galacturonic acid and rhamnose is a distinctive feature (Voragen et al., 1995). Pectin with degree of esterication (DE; the percentage of carboxyl groups esteried with methanol) higher than 50%, named high methoxyl pectin (HMP) forms gel after heating in sugar solutions at concentration higher than 55% and pH lower than 3.5. On the other hand, formation of gel with a low methoxyl pectin (LMP; DE < 50%) requires the presence of calcium ions, extending the use of this gelling agent to a broader range of foods. LMP may be used as a gelling agent in low sugar products, such as low-calorie jams and jellies, confectionery jelly products, and other foods applications. The heat reversibility of LMP gels can be utilized in bakery jams and jellies for glazing, retorting, microwaving, baking, and sterilizing or pasteurizing. Durian agro-waste could be used to produce pectin. As far as we know, there is no published work on the study of the effect of extraction conditions on the yield and DE of durian rind pectin. Extraction conditions have been reported to inu-

Corresponding author. Fax: +6 04 6573678. E-mail address: azhar@usm.my (A.M. Easa). Received 23 October 2008; Accepted 25 January 2010 0960-3085/$ see front matter 2010 The Institution of Chemical Engineers. Published by Elsevier B.V. All rights reserved. doi:10.1016/j.fbp.2010.01.010

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ence physico-chemical properties of pectin extracted from other plant sources such as beet (Levigne et al., 2002). Therefore the objective of this paper was to investigate the effect of extraction conditions (temperature, pH and time) on yield and DE of durian rind pectin.

titrated with 0.5 M sodium hydroxide to a faint pink color that persisted after vigorous shaking (end-point). This volume of titration was recorded as the saponication titer (the nal titer). The DE was calculated from the following formula: % DE = the nal titer 100 the initial titer + the nal titer

2.
2.1.

Materials and methods

Durian rind 2.4.

Statistical analysis

Durians fruit (Durio zibethinus) cultivar D24 used in this study were obtained from a fruit orchard in Relau, Penang, Malaysia in mid-July 2007. Ripened fruit that dropped naturally were collected and transported within 3 h on the same morning (at 30 2 C). Fruits were selected for uniformity of size (with the average weight 2.0 0.5 kg per fruit) and color and freedom of defects. A total of 25 fruits were selected for the study. Durians were cut open with a sharp knife. Durian aril was separated from the durian rind and seed manually. The rinds were cut as to eliminate the hard and thorny parts. Later, the rinds were cut into small pieces (about 0.5 cm thick) to facilitate the drying and grinding process. Pieces of durian rind were placed on aluminum foils and dried in a hot air oven (AFOS Oven, Hull England) at 70 C for 24 h. The dried durian rind were ground into powder by using a grinder (Micro Universal Bench Top Grinder, Retsch ZM 100, Germany) and packed into a sealable plastic bag and stored in desiccators.

Many experiments involve the study of the effect of two or more factors, factorial designs are more efcient for this type of experiment (Montgomery, 2005). This type of designs is very important to study the effect of several factors, since each replication of the experiment contains all possible combinations of the levels of the factors. The effect of three factors; temperature (X1 ), heating time (X2 ) and pH (X3 ). Two responses in the form of different components of the extract were evaluated: 1- Y1 = Yield % 2- Y2 = DE % Factorial design of type 23 was carried out to study t he effect of three factors temperature (8090 C), heating time (14 h) and pH (2.02.5) on the two responses mentioned above. This experiment was carried out in random order to minimize the effects of unexpected variability in the observed responses. Twenty-four runs were required to cover all possible combinations of factors levels with three replicates. Experimental data were analyzed to t the following regression model with interaction terms
3

2.2.

Pectin extraction

The dried durian rind (solidliquid ratio; 1:9, w/v) was stirred in a mild acid aqueous solution adjusted to desired pHs (2.0 or 2.5) with 1N HCl. Then, the solution was extracted at 80 or 90 C for 1 or 4 h. The resulting slurries were ltered through cheese cloth and allowed to cool to room temperature (25 C). Acidied ethanol (4% HCl in 95% EtOH) was added in the ratio 1:4 (v/v) and kept for 1 h. The mixture was centrifuged at 1710 g for 15 min in a Bench Top Centrifuge (Kubota 5100, Fujioka, Japan). The gel-like precipitate was collected and re-suspended in distilled water with the ratio 1:4 (w/v). Then, the solution was rewashed twice with 95% ethanol (1:2, v/v) and centrifuged for 15 min (1710 g). Precipitate was collected and dried in a vacuum oven at 25 C for 8 h. The pectin was ground and sieved (mesh no.60) for further experiments. Yield of pectin obtained was calculated as: % Yield = mass of extracted pectin 100 mass of air-dried durian rind

Y = 0 +
i=

i Xi +
i<j

ij Xi Xj +
i<j<k

ijk Xi Xj Xk (1)

where 0 , i , ij and ijk are regression coefcients.

3.
3.1.

Results and discussion

General

2.3.

Determination of degree of esterication (DE)

The DE of pectin was determined by the titrimetric method of Food Chemical Codex (FCC, 1981) and USP 26 NF 21 (2003) with slight modication. Dried sample (500 mg) was moistened with 2 ml ethanol and dissolved in 100 ml carbon dioxide-free water. After the sample was completely dissolved, ve drops of phenolphthalein were added, the sample was titrated with 0.5 M sodium hydroxide and the result was recorded as the initial titer. Then, 10 ml of 0.5 M sodium hydroxide were added, the sample was shaken vigorously, and allowed to stand for 15 min; 10 ml of 0.5 M hydrochloric acid were added and the sample was shaken until the pink color disappeared. Phenolphthalein (ve drops) was added and the solution was

The total extraction yield reected pectin yield even though some impurities such as degraded pectins and starch might be present. Table 1 summarizes the results of 24 runs to determine the yield and DE of pectin produced using different conditions. The acid-extracted pectin yield ranged from 2.13 to 10.25% of the dry weight of the durian rind. Compared to literature data, these values were generally lower than pectins extracted from sugar beet and citrus (Yapo et al., 2007; Micard and Thibault, 1999; Mesbahi et al., 2005) and banana peel (Suhaila and Zahariah, 1995; Happi Emaga et al., 2008). The highest yield was obtained when the durian rind was extracted at pH 2.0, for 4 h, at 90 C. The effects of pH and time seemed to be the most inuential on the pectin yield. At constant pH and temperature, the yields of pectin obtained for 1 h of extraction were lower than those of 4 h. On the other hand, the pectin yields from various extractions at pH 2.0 were higher than those at pH 2.5. The yield of pectin has been reported to increase with decreasing pH (or increasing acid strength) of the extraction (Levigne et al., 2002). A similar result of yield was shown at constant pH and temperature, i.e. increasing the extraction time had a marked effect on yield. However, when the temperature was

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Table 1 The results of 24-run with experimental and predicted values. Input factors X1
80 80 80 90 90 90 80 80 80 90 90 90 80 80 80 90 90 90 80 80 80 90 90 90

Actual values X2
1 1 1 1 1 1 4 4 4 4 4 4 1 1 1 1 1 1 4 4 4 4 4 4

Predicted values Y2 Y1
5.22 5.22 5.22 5.92 5.92 5.92 8.08 8.08 8.08 9.37 9.37 9.37 2.26 2.26 2.26 4.36 4.36 4.36 6.87 6.87 6.87 8.46 8.46 8.46

X3
2 2 2 2 2 2 2 2 2 2 2 2 2.5 2.5 2.5 2.5 2.5 2.5 2.5 2.5 2.5 2.5 2.5 2.5

Y1
5.14 5.04 5.48 5.67 6.20 5.89 8.06 8.01 8.16 8.74 9.13 10.25 2.24 2.13 2.41 4.34 4.32 4.43 6.88 7.10 6.64 7.91 8.90 8.57

Y2
48.87 48.87 48.87 54.97 54.97 54.97 47.52 47.52 47.52 51.98 51.98 51.98 63.91 63.91 63.91 61.80 61.80 61.80 58.43 58.43 58.43 50.64 50.64 50.64

50.39 46.21 50.00 55.43 54.50 54.99 47.84 45.57 49.16 53.72 51.18 51.03 64.05 62.90 64.78 64.46 59.48 61.47 57.73 57.93 59.64 47.39 51.61 52.93

increased while pH and time remained constant, the pectin yield increased slightly. This indicated that yield was higher when pectin was extracted at pH 2.0 for 4 h irrespective of the temperature.

els for yield and DE are given by Eqs. (1) and (2) respectively: Y1 = 71.22 0.66 X1 14.66 X2 35.48 X3 + 0.17 X1 X2 + 0.35 X1 X3 + 7.11 X2 X3 0.07 X1 X2 X3 (2)

3.2.

Yield and DE

Y2 = 289.47 + 3.41 X1 33.51 X2 + 142.76 X3 + 0.48 X1 X2 1.37 X1 X3 + 18.74 X2 X3 0.27 X1 X2 X3 (3)

Factorial design of type 23 was used to evaluate the effect of temperature X1 , heating time X2 and pH X3 on the percentage of yield (Y1 ) and DE (Y2 ). The results of 24-run including actual and predicted values are given in Table 1. The regression mod-

The regression models obtained for yield and DE are satisfactory since the value of the coefcient of determination

Table 2 The results of analysis of variance (ANOVA) for yield and DE. Source
Yield Model X1 X2 X3 X1 X2 X1 X3 X2 X3 X1 X2 X3 Error Total DE Model X1 X2 X3 X1 X2 X1 X3 X2 X3 X1 X2 X3 Error Total

Sum of squares
116.93 12.13 84.60 16.50 0.002 1.08 2.16 0.46 2.15 119.08

DF
7 1 1 1 1 1 1 1 16 23

Mean square
16.70 12.13 84.60 16.50 0.002 1.08 2.16 0.46 0.13

F-Value
124.34 90.27 629.74 122.82 0.018 8 16.08 3.46

P-Value
<0.0001 <0.0001 <0.0001 <0.0001 <0.8953 <0.0121 <0.0010 <0.0814

775.99 0.17 165.01 370.91 20.19 156.93 56.70 6.09 55.58 831.57

7 1 1 1 1 1 1 1 16 23

110.86 0.17 165.01 370.91 20.19 156.93 56.70 6.09 3.47

31.92 0.048 47.51 106.79 5.81 45.18 16.32 1.75

<0.0001 <0.8302 <0.0001 <0.0001 <0.0283 <0.0001 <0.0009 <0.2041

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(R2 ) was high and close to 1. The values of R2 for yield and DE were 0.98, 0.93, respectively. This indicates that 0.930.98 of the total variation was explained by the models and only (0.020.05) of the total variation is unexplained. The relative contribution of each factor to each response (yield and DE) was directly measured by the regression coefcient in the tted model. A positive sign for the regression coefcient in the tted model indicates that the ability of the factor to increase the response, whilst the negative sign indicates the ability of a factor to decrease the response. The results of analysis of variance (ANOVA) for each dependent variable, yield and DE are given in Table 2. The analysis of variance reveals that the models adequately tted the experimental data for yield and DE. It can be seen that the main effect

for temperature (X1 ), heating time (X2 ), pH (X3 ) and interaction terms between temperature and pH (X1 X3 ), heating time and pH (X2 X3 ) exhibited strong signicant effect on yield, whilst the interaction between temperature and heating time (X1 X2 ), and the interaction between temperature, heating time and pH did not show signicant effect on yield. It is interesting to note that pH and time were the most signicant interactive effect on pectin yield, which is similar to that reported by Happi Emaga et al. (2008) who studied extraction conditions on pectin yield from banana peel. The results of analysis for DE showed that the main effect of temperature and the interaction between temperature, heating time and pH did not exhibited signicant effect on DE, while heating time, pH, interaction between temperature and

Fig. 1 Three-dimensional response surface plots for yield as a function of (a) temperature and heating time, (b) temperature and pH and (c) pH and heating time of extraction.

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213

heating time, interaction between temperature and pH, and interaction between heating time and pH exhibited a strong signicant effect on DE. Signicant interaction indicates that the factors work independently, whilst presence of an interaction indicates that the difference in response (yield and DE) at different levels of a factor is not the same at all levels of another factor. These three parameters have been recognized to pose interactive effects on experimental responses during the extraction sugar beet pulp pectin (Yapo et al., 2007). It is to note that temperature and pH, as well as pH and time were the most signicant interactive effect on DE. Pectins that are classed as high methoxyl (HM) have DEs values higher than 50%. HM pectin requires a minimum amount of soluble solids and a pH within a narrow range, around 2.03.5, in

order to form gels (Yapo et al., 2007). Most of durian rind pectin extracted in this study showed DE value higher than 50%. It is to note that the extraction yield was not related to DE (Table 1). DE values of durian rind pectin produced in this study were inuenced by the extraction conditions, and the highest DE was obtained at extraction pH of 2.5, heating time of 1 h and temperature of 80 C, even though the yield was quite low for these conditions. The said conditions were probably the least harsh as compared to other conditions used, thus supporting the suggestion that harsh conditions of temperature and pH could increase deesterication of polygalacturonic chain (Mort et al., 1993). In this study, the highest yield of pectin was achieved by using the harshest extraction conditions (90 C, 4 h and pH 2.0), and this pectin had DE of 51% (Table 1).

Fig. 2 Three-dimensional response surface plots for DE as a function of (a) temperature and heating time, (b) temperature and pH and (c) pH and heating time of extraction.

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Experimental data were analyzed by factorial design to t the regression model mentioned in Eq. (1). The threedimensional response surface plots for yield and DE using Eqs. (2) and (3) are given in Figs. 1 and 2 showing the effect of factors on yield and DE respectively. It can be seen that different effect on yield and DE was exhibited by different combination of factors. Fig. 1(a) depicts the effect of heating time and temperature on yield. It is clear that increasing temperature and time maximizes the yield, whilst decreasing both or either one will result in a decrease in the yield. On the other hand, Fig. 1(b) and (c) exhibited different behavior on yield and can be explained in the same way. Fig. 2 shows the behavior of heating time, pH and temperature on DE. It is clear that increasing temperature and decreasing heating time increases the DE ((Fig. 2(a)). Fig. 2(b) indicates that high DE pectin could be obtained at high pHs and low temperatures combinations, while Fig. 2(c) shows a high DE pectin could be extracted at high pHs and short heating time combinations. These threedimensional response surface plots offer a clear picture of the behavior of the factors on responses of the study.

4.

Conclusion

The effect of time (1 and 4 h), pH (2 and 2.5) and temperature (80 and 90 C) on yield and DE of durian rind pectin was investigated. In general, durian rind pectin can be classied as high methoxyl type with DE higher than 50%. Yield was signicantly affected by time, temperature and pH, and interactions between temperature and pH, and heating time and pH. DE was signicantly affected by heating time and pH, and interactive effects of temperature and pH, and heating time and pH. By considering pectin yield and DE, the acid extraction of durian rind pectin at 80 C, for 4 h and at pH 2.5 could be suitable. Optimum conditions for isolation of pectin from the rind with desired rheological and functional properties are yet to be determined.

Acknowledgement
Short term grant from USM (304/PTEKIND/638069), Penang is gratefully acknowledged.

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