Optimization of maltodextrin production from avocado seed starch by response

surface methodology
Thanh Viet Nguyen, Tuyen-Hoang Nguyen Ma, Tha Thi Nguyen, Vinh-Nghi Kim Ho, and Hau Tan Vo

Citation: AIP Conference Proceedings 1954, 040004 (2018); doi: 10.1063/1.5033404

View online: https://doi.org/10.1063/1.5033404
View Table of Contents: http://aip.scitation.org/toc/apc/1954/1
Published by the American Institute of Physics

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 Optimization of Maltodextrin Production from Avocado
Seed Starch by Response Surface Methodology
Thanh Viet Nguyen1, Tuyen-Hoang Nguyen Ma1, Tha Thi Nguyen2,
Vinh-Nghi Kim Ho2 and Hau Tan Vo2,a
1
Faculty of Applied Sciences, Ton Duc Thang University, 19 nguyen Huu Tho street, Tan Phong ward, District
7, Ho Chi Minh City, Vietnam
2
Department of Food & Agro-materials Preservation and Processing, Food Industry Research Institute,
Branch in Ho Chi Minh City, Vietnam
a)
Corresponding author: hautanvo@gmail.com

Abstract. A process for maltodextrin production from avocado seed starch was reported in this study. Response
surface methodology was used to investigate the effects of three independent variables for hydrolysis of the starch
using a commercial food-grade α-amylase, Termamyl SC. These variables included enzyme concentration (0.05 –
0.15% starch), pH (5.0 – 6.0) and hydrolysis time (1.0 – 3.0 h), while the temperature fixed at 95oC. The result
showed that the optimum conditions were using enzyme concentration at 0.12%, pH at 5.5 and 2.75 h of the
incubation time. Under the optimum conditions, the recovered starch yield was 79.8% and the maltodextrin powder
had 15.8 of dextrose equivalent.

INTRODUCTION
Avocado (Persia americana Miller) is an evergreen tree native to Central America that now widely
cultivated in most the tropical and subtropical regions of the world for edible fruits, which are very rich in oil
[1]. In 2014, world avocado production was approximately 5.5 million metric tones [2]. The avocado was first
introduced into the Lam Dong province of Vietnam in 1940 by the French [3]. Although no statistical figures are
available on the area and production of avocado, the tree is widely grown in the upland parts of Vietnam such as
Dong Nai, Ba Ria-Vung Tau, Lam Dong, DakLak and Phu Tho with various local names given to them
according to their fruit shape and quality.
Avocado is an important commercial tropical fruit. The edible part of the fruit is rich in unsaturated fatty
acids, vitamins B, C and E, and other nutrients [4]. Avocado is mainly consumed as a fresh fruit but now many
value added products have manufactured as guacamole, avocado pulp and avocado oil [5]. Industrial processing
of avocado fruit generates a large amount of peels and seeds as waste. These by-products can cause
environmental problems. The avocado seed presents up 16% of the total weight of the fruit and contains starch
which is quite high, about 29% (wet basis) [6-7]. Therefore it may be considered an alternative starch source for
other applications. The research results about functional and rheological properties of avocado seed starch
(ASS) revealed that it has potential applications in products such as baby food, sauces, bread products, jellies,
candies and sausages [8-10]. Other possible uses are as a vehicle in pharmaceutical products and in
biodegradable polymers for food packaging. However, the production of maltodextrin from ASS using
commercial food-grade α-amylase has not been reported.
Commercial starch hydrolysates are classified on the basis of dextrose equivalent (DE), which is defined as
the percentage of reducing sugar in a syrup calculated as dextrose on a dry weight basis [11]. Maltodextrin (be
also known as dextrin) is the dried products or purified aqueous solution of saccharides obtaibed from edible
starch having a DE of less than 20. It may be manufactured either by acid or by acid-enzyme processes.
However, the maltodextrin produced by acid conversion of starch may slowly reassociate into insoluble
compounds causing haze in certain applications [12].
Nowadays, the maltodextrin product have wide applications in other industries [13]. As a food additive, it
performs multi-functions in food system, including bulking, caking resistance, texture and body improvement,
film formation, binding of flavour and fat and increasing of soluble solids. It have also been proven useful to

Proceedings of the 2nd International Conference on Applied Sciences (ICAS-2)

AIP Conf. Proc. 1954, 040004-1–040004-5; https://doi.org/10.1063/1.5033404
Published by AIP Publishing. 978-0-7354-1653-6/$30.00

040004-1
reduce Maillard reactions and is used in microencapsulation of food components such as vitamins, polyphenols
and colourants [14].
Response surface methodology (RSM) consists of a group of mathematical and statistical techniques that are
based on the fit of empirical models to the experimental data obtained in relation to experimental design [15]. It
is the one of most effective tools for optimizing the process when many factors and interactions affect the
desired response. RSM usually uses an experimental design such as Box-Behnken or central composite to fit a
second order polynomial [16-17].
The purpose of this study was to optimize the production process parameters of maltodextrin from ASS by a
commercial food-grade α-amylase, Termamyl SC using RSM.

MATERIALS AND METHODS

Materials: Avocado fruits (Persia americana, named Bo Sap in Vietnamese) were purchased from the local
market in Ho Chi Minh, Vietnam and kept at room temperature until they reached ready-to-eat ripeness.
Commercial food-grade α-amylase, Termamyl SC (Novo Nordisk, Denmark) which labeling enzyme activity
was 120 KNU/g. Other chemicals either HPLC reagent grades or the highest purity were available.
Starch isolation: Avocado seeds were manually seperated from the flesh and cleaned. Starch was extracted
from the fresh avocado seeds by wet fractionation technique [10]. One kilogram quantity of the avocado seed
were chopped into small pieces and soaked in a 0.075% (w/v) sodium metabisulphite solution for 24 h (the
sodium metabisulphite used as a preservative to prevent bacteria and an inhibitor of enzymatic browning). The
hydrated pieces of the avocado seed were milled into a homogenous fine pulp using a Vitamix machine (Type-
5300, America). The mash was dispersed in 10 L of the sodium metabisulphite solution (0.075%, w/v) and
filtered through a sieve of fine muslin cloth. The suspension obtained was allowed to stand for 12 h for complete
sedimentation of the starch granules before the supernatant was decanted. The starch layer was repeatedly
washed with distilled water. The resulting starch cake was dried in oven at 50oC. The dry starch was then
pulverrised to a fine powder and stored in a dry container until use.
The proximate composition of ASS were determined according to method AOAC [18]. The protein content
was calculated using the 5.95 conversion factor. The results were composed of 7.3% moisture, 88.2% total
carbohydrate (77.5% starch), 2.36% protein, 0.53% lipid and 0.72% ash.
Preparation of maltodextrin from ASS: One hundred grams of ASS (77.5% starch) were mixed with 387.5
ml distilled water (20% starch suspension) and stirred continuously at 450 rpm using a magnetic stirrer. After
the pH of the suspension was adjusted (5.0 to 6.0) by 1N HCl, Termamyl SC (0.05% - 0.15% of starch, w/w)
was added. The suspension containing enzyme was incubated in a water bath at 95 oC for a selected period of
time (1.0 - 3.0 h). At the end of the hydrolysis process, enzyme was inactivated by addition of 1N HCl to pH =
4.3. Supernatant was recovered by centrifugation at 4000 rpm for 15 min. The enzymatic maltodextrin
preparation variables are presented in TABLE 1. The recovered starch yield was expressed as:

Starch in supernatant
Starch yield (%) = x100 (1)
Starch in ASS

DE determination was made on hydrolysates or final maltodextrin powder product [19]. The reducing sugars
were determined with Lane and Eynon method and DE was computed using the formula:

% Reducing sugar
DE = x100 (2)
% Dry matter
Experimental design
In this study, RSM was used to predict the optimum hydrolysis conditions of ASS by using Design-Expert
software (version 9.0, Stat-Ease Inc., Minneapolis, MN, USA). The Box-Behnken design (BBD) with a
quadratic model was selected to investigate the combined effects of three independent variables while
hydrolysis temperature and starch concentration were fixed at constant rate of 95 oC and 20%, respectively
(determinated after several preliminary experiments, data not shown). The independent variables were
Termamyl SC enzyme concentration (0.05 – 0.15%; X1), pH (5.0 – 6.0; X2) and hydrolysis time (1.0 – 3.0 h; X3)
as in TABLE 1.

040004-2
 Statistical analysis
Experimental data were fitted to a quadratic polynomial model and regression coefficients obtained. The
computer-generated quadratic model used in the response surface was as follows:

Y=β0 + σ3i=1 βi Xi + σ3i=1 βii X2i + σ2i=1 σ3j=i+1 βij Xi Xj (3)

where Y denotes the dependent variable (recovered starch yield). The coefficients of the polynomial
equation were represented by β0 (intercept), βi (linear effects), βii (quadratic effects), and βij (cross product
effects). Design-Expert software was used to estimate the response of each set of experimental design and
optimized conditions. The quality of the fitted model was expressed by the coefficient of determination R 2, the
adjusted determination coefficient R2adj as well as the predicted determination coefficient R2pred and statistical
significance of the model was determined by F-test.
All treatments were done in triplicate and the results were expressed as a mean (± SD) for each treatment.
The significant difference between treatments reported at p ≤ 0.05.
TABLE 1. Factors and levels for RSM and BBD with the observed values for the response variable (Y)

Coded variable levels Uncoded variable levels Response (%)

Run
x1 x2 x3 X1 X2 X3 Y
1 0 1 1 0.10 6.0 3.0 79.3
2 1 -1 0 0.15 5.0 2.0 77.3
3 -1 -1 0 0.05 5.0 2.0 68.5
4 1 1 0 0.15 6.0 2.0 77.9
5 -1 0 1 0.05 5.5 3.0 70.7
6 1 0 -1 0.15 5.5 1.0 70.8
7 1 0 1 0.15 5.5 3.0 79.7
8 0 -1 1 0.10 5.0 3.0 77.8
9 -1 0 -1 0.05 5.5 1.0 56.7
10 0 0 0 0.10 5.5 2.0 76.3
11 0 1 -1 0.10 6.0 1.0 66.3
12 0 -1 -1 0.10 5.0 1.0 68.2
13 0 0 0 0.10 5.5 2.0 75.1
14 -1 1 0 0.05 6.0 2.0 66.2
15 0 0 0 0.10 5.5 2.0 77.0

RESULTS AND DISCUSSION

Fitting the model

The study used RMS to develop a prediction model for optimizing conditions of starch hydrolysis from ASS.
The experimental conditions and the corresponding response value from the BBD were presented in TABLE 1.
By performing multiple regression analysis on the experimental data, the model for the response variable Y
(recovered starch yield) could be expressed in form of coded values by the quadratic polynomial equation as
following:
Y=76.1 + 5.4X1 -0.3X2 +5.7X3 +0.7X1 X2 -1.3X1 X3 +0.9X2 X3 -3.5X21 -0.1X22 -3.1X33 (4)

To test the significance and adequacy of the model, the analysis of variance (ANOVA) for the quadratic
model was performed. The lack of fit test determines whether the selected model is adequate to explain the
experimental data or another model should be reselected. As shown in TABLE 2, the lack of fit test was not
significant relative to the pure error (p > 0.1). The model F-value of 583.22 implied the model was significant.
There was only a 0.01% chance that an this large F-value could occur due to noise.
The results of ANOVA also gave the determination coefficients for the model as shown in TABLE 2. For
the fitted model, the coefficient of determination (R2), which is a measure of degree of fit was 0.9949. This
indicates that only 0.51% of the total variations were not explained by the model. Lundstedt [20] suggested that,
R2 should be at least 0.80 and the probability (p) value of the regression model significance was less than 0.001
for a good fit of a model. Moreover, the adjusted determination coefficient (R 2adj = 0.9858) was close to R2 and
the predicted determination coefficient (R2pred = 0.9641) was in reasonable agreement with R2adj. Therefore, the

040004-3
model is adequate for prediction in the range of experimental variables and can be used to navigate the design
space.
TABLE 2. The analysis of variance (ANOVA) table for response surface quadratic model

Soure Sum of squares DF Mean square F-value p-value (Prob > F) Significant
Model 583.22 9 64.80 108.64 < 0.0001 **
X1 236.75 1 236.75 396.92 < 0.0001 **
X2 0.56 1 0.56 0.93 0.3784
X3 257.99 1 257.99 432.52 < 0.0001 **
X 1X 2 2.03 1 2.03 3.40 0.1243
X 1X 3 6.53 1 6.53 10.94 0.0213 *
X 2X 3 2.89 1 2.89 4.85 0.0790
X12 45.86 1 45.86 76.88 0.0003 **
X22 0.03 1 0.03 0.05 0.8286
X32 35.87 1 35.87 60.13 0.0006 **
Residual 2.98 5 0.60
Lack of fit 1.04 3 0.35 0.36 0.7928 Insignificant
Pure error 1.94 2 0.97
Cor. Total 586.20 14
*Significant at p < 0.05; **Significant at p < 0.001
R2 = 0.9949; R2adj = 0.9858; R2prep = 0.9641; C.V% = 1.07

Effects of independent variables on response

The significance of each coefficient was also determined using F-value and p-value (TABLE 2). It could be
seen that the effects of enzyme concentration and incubation time (X1, and X3; p < 0.05) were the major
contributing fators to the recovered starch yield (response variable Y) while pH had no significant effect (p >
0.05) within the experimantal range. In addition, it was evident that coefficients (X 1X3, X12 and X32) were
significant at the level of p < 0.05, whereas the other coefficients were insignificant (p > 0.05). The relationship
between independent and dependent variable was graphically represented by 3D response surfaces generated by
the model as in FIG 1.

FIGURE 1. Response surface plots for the effects of enzyme concentration and pH; enzyme concentration and incubation
time; pH and incubation time on starch recovery yield

Optimum condition and model verification

From the model, the optimum conditions for production of maltodextrin from ASS obtained by using
Design-Expert software were presented in TABLE 3. Under optimum conditions, recovered yield of 79.8%
starch was predicted. The suitability of the model equation for predicting the optimum response value was tested
by additional independent experiments (triplicate) using the recommended optimum conditions (TABLE 3). The
result showed the experimental starch yield (79.1%) was not significantly different from the predicted value
(79.8%).

040004-4
 TABLE 3. Optimum conditions for production of maltodextrin from ASS, predicted and experimental values from RMS

Fixed condition Optimum condition Recovered starch yield (%)

Starch
Temperature Stirring speed Enzyme Time Predicted Experimental
suspension
(oC) (rpm) (%)
pH (h) value value
(%)
20 95 500 1.12 5.5 2.75 79.8 79.1 ± 1.1

The maltodextrin liquid gained after separation solid section by centrifuge was dried by spray drier. Drying
was performed with mini spray dryer (B-290, Büchi). Inlet temperature and outlet temperature were 170 oC and
115oC, respectively. The colour of spray-dried matodextrin powder was milk white. The powder was easy
soluble in water. Total carbohydrate content of final product was 95.75% with DE = 15.8 and water content was
2.9%.

CONCLUSIONS
ASS can be used as a raw material to production value added product as maltodextrin. The RMS based on
the BBD was successfully used to optimize production parameters for maltodextrin from ASS by Termamyl SC.
The optimum conditions in the manufacture of maltodextrin from ASS were using enzyme concentration at
0.12%, pH at 5.5 and 2.75 h of the incubation time with 79.8% recovered starch. The maltodextrin powder
obtained from the conditions had 15.8 of DE.

ACKNOWLEDGMENTS
The authors gratefully acknowledge the financial support from the Vietnam Ministry of Industry and Trade.

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