VARIABLE SCREENING FOR THE PRODUCTION OF JACKFRUIT BEVERAGE WITH

CRAB CHITOSAN USING PLACKETT BURMAN DESIGN

INTRODUCTION
Chitosan is obtained by the deacetylation of chitin, a natural nondigestible
oligosaccharide which is the main component of the cell walls of fungi, and the exoskeletons of
arthropods and insects, being one of the most abundant organic materials, second only to
cellulose (Riccardo & Muzzarelli, 1977 as cited by Barry-Ryan et al., 2009). Chitin and its
deacetylated product chitosan, as well as their derivatives, have found varied applications in
agriculture, food processing, biotechnology, chemistry, cosmetics, dentistry, medicine, textiles,
veterinary medicine, and environmental sciences. Their food uses cover a wide range of
applications, including control of microbial deterioration, inhibition of lipid oxidation,
emulsification, thickening, stabilization of color, and as dietary supplements (Venugopal, 2011).
Figure 1 shows the chemical structure of chitosan.

Figure 1. Chemical structure of chitosan

Potent sources of natural food additive compounds have been found in several types of
natural material. One of these is chitosan (Mahae et al., 2011). Antioxidant activity is one of the
well-known functions of chitosan. Many studies have shown that chitosan inhibits the reactive
oxygen species (ROS) and prevent the lipid oxidation in food and biological systems. Several
mechanisms about the antioxidant action of chitosan have been proposed (Kim and Thomas,
2007 as cited by Rajalakshmi et. al., 2013).
Good experimental design is important in many studies of analytical and other chemical
processes. Complete factorial designs, which study all the factors (experimental variables)
affecting the system response, using at least two levels (values) for each factor, can give rise to
an unacceptably large number of trial experiments because even simple processes may be
affected by a large number of factors. An economical and popular approach that gives
information only on the effects of single factors, but not on interactions, is the PlackettBurman
(PB) method. This method is well suited to ruggedness testing, i.e. establishing whether the
outcome of an analytical procedure is affected by changes in each relevant factor (Analytical
Methods Committee, AMCTB No 55, 2013).
Since synthetic preservatives are suspected of being responsible for some severe toxic
effects (Roller, 2001 as cited by Toan et al., 2013) chitosan becomes a promising biopolymer.
This study aimed to screen and identify the significant variables that affect the sensory
acceptability of jackfruit beverage with chitosan in terms of color, aroma, taste, flavor and
general acceptability using Plackett-Burman design.

MATERIALS AND METHODS

Extraction of Chitin and Chitosan

Chitin and chitosan was extracted from the crab carapace according to the methods of
Abazinge et al., 2007 with some modifications. The crab exoskeletons specifically the carapace
was obtained from the processing plant of Eastern Visayas Fresh Seafood Incorporated located in
Brgy. Silanga, Catbalogan City, Samar. It was cleaned by removing the remaining meat and other
undesirable components. The cleaned carapaces were sun dried for 3 to 5 days. The dried
carapaces were crushed into smaller pieces using a mortar and pestle and then ground into
powdered form. The powdered carapace was demineralised using 5% (v/v) acetic acid in 1:10
solid/liquid ratio, deproteinized with 2% NaOH for 1h and deacetylated with 50% NaOH by
boiling the mixture at 1000C for 2h.
Preparation of jackfruit beverage with chitosan
Ripe jackfruits regardless of variety were obtained from the farmers of Inopacan, Leyte.
All other needed materials were purchased from the Baybay City Market and in Cebu City. All
materials used in this study was sanitized in chlorinated water (300ppm) using commercial
bleach containing 5.25% by weight of sodium hypochlorite for 15 minutes and air-dried.
Sterilization of bottles was done by boiling it in water for 15 minutes. Fresh ripe jackfruit was
washed with running tap water, brushed to remove the unwanted dirt and rinsed with chlorinated
water. The fruit was then cut into halves. The rags and seeds were removed from the fruit and
fruit pulp was collected and kept in the freezer until used. One hundred grams (100g) of jackfruit
pulp was placed in a blender and added with six hundred milliliters (600 ml) of water and
homogenized for one to two minutes. The homogenized pulp was then strained using a cheese
cloth and pressed to extract the juice. In a liter of juice, one hundred fifty grams (150g) of sugar,
two grams (2g) chitosan in 5% citric acid solution was added into the mixture, and the pH was

adjusted by adding citric acid. The mixture was then pasteurized at 800C for six minutes and hot
filled in a sterilized bottle.
Sensory evaluation and Plackett-BurmanVariable Screening
The medium components of the variables used in variables screening is shown in Table 1.
To trim down these variables and eliminate those that do not have significant effect on the final
product, screening of variables was conducted using Plackett-Burman Design (Table 2) with
seven (7) input variables for eight (8) runs. The seven input variables include (1) jackfruit
pulp:water ratio, (2) type of sugar, (3) level of sugar, (4) pH level, (5) concentration of chitosan,
(6) pasteurization temperature, and (7) pasteurization time. A 9-pt Hedonic scale rating was
carried out for the sensory evaluation to determine the products acceptability in terms of color,
aroma, taste, flavor and general acceptability. The sensory panel was composed of 60 third and
fourth year Food Technology majors of the Department of Food Science and Technology,
Visayas State University.
Data Analysis
All results from the sensory evaluation were tabulated in Microsoft Excel 2007 and were
analyzed using Statistica 8.0 software. Pareto charts at p<0.05 were used to show the
components that significantly affect most on the process formulation of the product.

Table 1. Medium components in variable screening using a PB design

Variables
High (+)
Low (-)
V1-Jackfruit pulp:water ratio (wt/vol)

1:2

1:4

V2-Type of sugar

white

Brown

V3-Level of sugar (g)

120

60

V4-pH level

4.2

3.0

V5-Conc. of chitosan (g/l soln)

2.0

V6-Pasteurization temperature (0C)

90

80

V7-Pasteurization time (min.)

15

10

Table 2. Plackett-Burman design matrix in 8-runs for 7 factors

Run
Pattern
V1
V2
V3
V4
V5

V6

V7

+++-+--

-+++-+-

--+++-+

+--+++-

-+--+++

+-+--++

++-+--+

-------

RESULTS AND DISCUSSION

In this study, the variables affecting and have influenced on the sensory quality
attributes of jackfruit beverage were screened using the Plackett-Burman Design.
Appendix 1a-1e and Table 7 shows the summary for the result of the screening
experiment. The variable screening revealed that jackfruit and water ratio, type of
sugar, quantity of sugar, pH level, and pasteurization time have significant effects
on one or more of the sensory attributes of the product. The concentration of
chitosan and pasteurization temperature did not significantly affect the sensory
acceptability of the product thus; both are held constant during the process
optimization. The three variables considered for the optimization process using CCD
are the quantity or amount of sugar since it affects most of the attributes; jackfruit
and water ratio for it affects both color and aroma, and lastly, pasteurization time as
it has a negative effect on the color acceptability of the product. Other variables
were held constant during the optimization process.

Table 3. Summary of the effect estimates of Plackett-Burman Design of variables affecting the
sensory attributes of jackfruit beverage with chitosan
General
Color
Aroma
Taste
Flavor
Acceptability
Mean/Interc.

7.041667**

7.295833**

6.787500**

6.879167**

6.891667**

JF:water

0.333333*

-0.308333*

-0.108333ns

-0.158333ns

-0.066667ns

type of sugar

-0.866667**

0.008333ns

0.358333*

0.375000*

0.216667ns

qty of sugar

-0.083333ns

0.341667*

0.775000**

0.741667**

0.683333**

pHlevel

0.200000ns

-0.008333ns

0.808333**

0.675000**

0.666667**

amt of CSN

-0.283333ns

0.025000ns

0.141667ns

0.108333ns

0.100000ns

past. temp

-0.283333ns

0.058333ns

-0.275000ns

-0.158333ns

-0.200000ns

past. time

-0.400000**

0.058333ns

-0.008333ns

0.025000ns

-0.050000ns

* =significant at 5%

**=significant at 1%

ns=not significant