Integration of physical and chemical treatment on the extraction of starch from Canna edulis Ker. rhizome

doi:10.4236/as.2013.49B009

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Vol.4, No.9B, 51-55 (2013) Agricultural Sci ences

http://dx.doi.org/10.4236/as.2013.49B009

Integratio n of p hy s ic al a nd chemical t re a tm e nt on th e

extraction of starch from Canna edulis Ker. rhizome

Judy R. B. Witono*, Herry Santoso, Y. I. P. Arry Miryanti, Daniel Tan

Chemical Engineering Department, Parahyangan Catholic University, Bandung, Indonesia;

*Corresponding Author: [email protected]

Received August 2013

ABSTRACT

The extraction of Canna edulis Ker. starch from

its rhizome was performed using 2 different

types of press (hy draulic press and scre w press)

and with the addition of Na-metabisulphite and

NaOH (in the range of concentration 100 - 5000

ppm each). The optimum condition for this

process was determined by Central Composite

Design of experiment and the statistical calcula-

tion was solved by Design-Expert 7.0.0. The

targets of the observed responses were high

starch yield, low ash, low fiber, and high carbo-

hydrate content. The results showed that the

starch yield and the reduction of fiber were only

influenced by the physical treatment whereas

ash content in the product was influenced by

both the NaOH concentration and physical treat-

ment. The carbohydrate content in the extrac-

tion product was affected by NaOH, by the inte-

raction bet ween the concentrations of NaOH and

Na2S2O5 and also by the physical treatment. The

hydraulic press gives much better responses

compared to the screw press. But in the se-

lected range of additives concentrations, the

screw press gives a higher starch yield (30% -

52%).

Keywords: Canna edulis Ker.; Central Composite

Design of Experim ent ; Hydraulic Press; Screw

Press; Starch Extraction

1. INTRODUCTION

One of the tropical starch resources which have not

been utilized for industrial application is Canna edulis

Ker. rhizome, partly because of the difficulties in the

extraction processing [1]. The high content of fiber and

other trace elements are a major constraint in producing

pure starch. A sample of 100 g of Canna edulis rhizome

contains 125 mg phosphorus (P), 84 mg calcium (Ca) and

1.5 mg iron (Fe) mineral [2]. The presence of those ele-

ments in food products can be categorized as a nutritional

value but it will be a disadvantage for a chemical starch-

based pr od uc t.

Salt solutions like NaCl, Na-bisulphite, and Na-meta-

bisulphite are commonly used during extraction of starch

from its natural source, to inhibit microbial growth and

deactivate plant enzyme (amylase). The salt solution can

dissolve the surface starch granule protein as well, but

for the breakdown of the integral starch granule protein,

stronger solutions are required, e.g. sodium dodecyl sul-

phate (SDS) [3 ,4] or alkaline solutions [5].

Lim et al. [6] have investigated that the use of 0.2% of

NaOH as an extraction solution for rice starch could re-

duce more than 80% of the flour protein. This was con-

firmed also by Radosavljevic et al. [7] on the extraction

of Amaranth starch; Mistry et al. [8] on the extraction of

corn flour using 0.1% and 0.4% of NaOH. It was inves-

tigated also that sodium hydroxide (NaOH) can remove

phosphorus up to 70% - 90% fr om wheat starch [9].

Since the properties of starch are, to some extent, dif-

ferent from the fiber (cellulose), a relatively low cost

method for isolation of the starch is physical treatment.

The goal of the research reported here is to determine the

optimum condition of the integration of physical and

chemical treatments in producing pure starch. For this

purpose, a Central Composite Experimental Design me-

thod (CCD) was applied, statistical calculations were made

using Design expert 7.0.0 soft wa re.

2. MATERIALS & METHODS

2.1. Materials

Freshly harvested Canna edulis Ker. rhizome (locally

known as Ganyong) was supplied by the farmers union

“Mekar Sari” at the Kulon Progo region (Central Java)

Indonesia . Analytical grade of sodium metabisulphite

and NaOH were purchased from Sigma Aldrich. α-amy-

lase was supplied by N o voz yme and Anthrone rea gent by

Merck.

J. R. B. Witono et al. / A gricultural Sciences 4 (2013) 51-55

2.2. Design of the Experiment

To determine the optimum condition from 2 numerical

factors (the Na-metabisulphite and NaOH concentrations)

and 1 categorial factor (type of mechanical treatment or

type of press instrument), the Central Composite Design

was applied. This results in 26 runs with variations as

shown in Table 1.

2.3. Starch Extraction

Fresh Canna edulis was peeled and washed, then milled

with a cross beater mill. The pulp was mixed with water

at a weight ratio 1:10. Half of mixture was filtered with

the hydraulic press and the rest with the screw press, to

separate the crude fiber.

The free crude fiber mixture which consists of the fil-

trate from each press was divided into 13 portions that

were mixed with different concentrations of Na-metabi-

sulphite and NaOH solutions as stated Tab le 1. After 12

hours the precipitated starch was separated from the liq-

uor and dried in a tray drier for about 12 hours at 45˚C

until constant weight. The dried starch was kept in closed

containers to be analyzed further.

Table 1. Overview of experimental runs.

Factor 1 Factor 2 Factor 3

Run [Na2S2O5] [NaOH] Type of treatment*

ppm ppm

1 2550 2550 Level 2

2 2550 2550 Level 2

3 100 100 Level 2

4 2550 2550 Level 1

5 2550 2550 Level 2

6 5000 100 Level 2

7 100 5000 Level 1

8 2550 2550 Level 2

9 5000 5000 Level 2

10 5000 5000 Level 1

11 2550 2550 Level 1

12 100 2550 Level 1

13 100 2550 Level 2

14 2550 2550 Level 1

15 5000 2550 Level 1

16 5000 2550 Level 2

17 2550 2550 Level 1

18 2550 2550 Level 1

19 2550 5000 Level 2

20 2550 2550 Level 2

21 100 5000 Level 2

22 100 100 Level 1

23 2550 100 Level 1

24 2550 5000 Level 1

25 2550 100 Level 2

26 5000 100 Level 1

*Level 1—hydraulic press and level 2—screw press.

2.4. Fiber Content

The fiber content was measured by heating a mixture

of 5 g of starch and 50 mL of water until 90˚C. Then 4

mL of α-amylase enzyme was added and the total was

kept at 90˚C for a further 30 minutes. After that 50 mL of

water was added and the sample was cooled to room

temperature.

This method was based on the characteristic property

of the α-amylase enzyme which degrades carbohydrates

to produce shorter chain molecules such as glucose, which

is soluble in water. The cooled mixture was filtered using

Whatman 42 filter paper under vacuum conditions to

separate the fine fiber. The Fiber content (FC) was cal-

culated using Eq.1.

( )

FC %=100%

w×

(1 )

Where w1 is the weight of fine fiber and w2 is the

weight of starch sample (dry basis).

2.5. Ash Content

Ash content measurement was also based on a gravi-

metric technique. 1 g of star ch sample was combusted in

a furnace at 600˚C for 1 hour then cooled in desiccator

and weight. This step was repeated until the weight was

constant. Ash content (AC) was calculated using Eq.2.

( )

AC %=100

w×

(2)

Where w3 is the weight of ash and w4 is the weight of

starch sample (dry basis).

2.6. Carbohydrate Content

The carbohydrate content was analyzed by anthrone

reagent [10]. The anthrone method was started by mak-

ing a standard graph which was made by diluting 10 mg

of glucose into 100 mL of water. The standards solution

was taken 0, 0.2, 0.4, 0.6, 0.8 and 1 mL (0 serves as

blank) and made up to 1 mL with distilled water. Then 4

mL of anthrone reagent was added.

Anthrone reagent was made by diluting 200 mg anth-

rone in 100 mL of ice-cold 95% H2SO4. All of the stan-

dard solutions were heated for eight minutes in a boiling

water bath and then cooled rapidly. Carbohydrate content

was measured with a spectrophotometer at 595 nm wa-

velength. A calibration graph was made by plotting con-

centrations of the standard solution on the X-axis against

and the absorbance on the Y-axis.

Starch must be hydrolyzed prior to the treatment with

anthrone solution. Therefore, 100 mg of a sample of

starch with 5 mL of 2.5 N HCl added, was boiled in a

water bath for three hours. When sample temperature

J. R. B. Witono et al. / A gricultural Sciences 4 (2013) 51-55

was back to room temperature, it was neutralized with

solid sodium carbonate (Na2CO3) until the effervescence

ceases. The solution was then made up to 100 mL and

centrifuged and 0.1 mL of the supernatant was used for

analysis. The rest of the procedure is the same for the

standard solution. The amount of carbohydrate (CC) in

the sample tube was then calculated from the absorbance

in the spectrophotometer and the calibration graph by

Eq.3.

( )

[ ]

0.9C 1000

CC %=100

×× ×

(3)

where [C] is the glucose concentration measured from

the calibration graph whereas w5 is the weight of starch

sample (dry basis).

2.7. Starch Yi eld

Starch yield was measured by comparing the weight of

obtained starch (dry basis.) with the weight of dry matter

sample (Canna edulis rhizome). The starch has been free

from dirt 100. Starch Yield (SY) was determined by

Eq.4.

( )

SY %=100

w×

(4)

where w6 is the weight starch (d.b.) whereas w7 is the

weight of the dry matter original sample.

3. RESULTS AND DISCUSSION

The analysis results from every run can be seen in Ta-

ble 2.

3.1. Starch Yi eld

Based on the ANOVA of the values obtained for the

starch yield (Table 3) it can be seen that the additives

concentrations, both Na2S2O5 and NaOH do not affect

the yield of starch. This is also proved by the probability

value (P value) from both additives which are above

0.005. The yield obtained is apparently more determined

by the method of physical treatment.

Between the two types of press used, the higher starch

yield was found when the screw press was used for se-

paration (30% - 52%) compared to the hydraulic press

(25% - 41%). However, this fraction contains 79% ± 6%

carbohydrate which is slightly lower than the product

from the hydraulic press 82% ± 8%).

3.2. Carbohydrate Content

Based on the ANOVA of carbohydrate content (Table

4) it can be seen that the NaOH concentration and the

interaction between the concentrations of Na2S2O5-

NaOH affect the amount of carbohydrate obtained. Since

Table 2. Results of the experimental runs.

Run Fiber Ash Carbohydrate Yield

% % % %

1 3.7 4.0 76.0 14.1

2 2.4 1.9 76.4 33.7

3 2.9 0.7 76.9 49.7

4 1.6 1.0 82.2 35.0

5 3.3 1.5 82.0 40.7

6 2.7 1.5 74.5 44.0

7 2.0 2.7 88.9 31.7

8 2.6 1.6 82.1 40.9

9 3.8 3.8 92.0 64.9

10 2.4 3.5 88.7 47.0

11 1.9 1.3 94.0 31.5

12 2.5 1.3 81.3 35.1

13 3.2 2.0 82.3 43.2

14 2.3 2.9 76.7 25.2

15 2.8 1.7 73.8 33.9

16 3.2 2.3 77.0 57.6

17 2.5 1.2 74.2 35.0

18 2.9 3.0 81.1 11.1

19 2.7 4.3 66.5 30.0

20 2.7 2.7 81.8 34.6

21 3.2 3.6 75.4 45.7

22 1.4 0.4 82.7 39.0

23 2.6 0.5 78.7 37.4

24 2.4 2.6 93.6 30.4

25 3.2 1.0 77.7 39.2

26 1.4 0.7 65.1 38.7

Table 3. ANOVA for starch yield response.

Factor SS DF MS F Value p-Value

[Na2S2O5] 144.47 1 144.47 3.38 0.18

[NaOH] 0.24 1 0.24 2 0.95

Treatment 442.65 1 442.65 3.37E-003 0.02

Tabl e 4. ANOVA for carbohydrate content response.

Factor SS DF MS F Value p-Value

A:[Na2S2O5] 22.39 1 22.39 0.69 0.42

B:[NaOH] 204.67 1 204.67 6.28 0.02

Treatment 62.86 1 62.86 1.93 0.18

AB 165.91 1 165.91 5.09 0.04

BC 132.74 1 132.74 4.07 0.06

the p-value of BC ≈ 0.05, so it can be predicted that there

is a relation between the physical treatment and the addi-

tive (NaOH).

From Figure 1, it can be seen that at the high concen-

tration of Na2S2O5 and the high concentration of NaOH,

within the selected ranges of this research the carbohy-

drate content reached the maximum value.

However, if the 3D surface plot is observed for each

physical treatment (Figures 2 and 3), the effect of con-

centration of additives on the carbohydrate content gives

a different profile. The separation process using the screw

J. R. B. Witono et al. / A gricultural Sciences 4 (2013) 51-55

Figure 1. 3D plot of carbohydrate content.

Figure 2. 3D surface of carbohydrate content using the hydrau-

lic press.

Figure 3. 3D surface of carbohydrate content using the screw

press.

press is slower than the hydraulic press. This results in

the need for the higher concentration of Na-metabisul-

phite (5000 ppm) to achieve high carbohydrate content

as well as to prevent microbial growth in the slurry

which can destroy the carbohydrate. This is a different

result compared to the hydraulic press in which it is

enough to use 100 ppm to gain a high carbohydrate yield.

The models resulting from the statistical methods ill u-

strate the relation between the carbohydrate content and

the significant factors as followed:

For the hydraulic press

[ ]

-3 -3

22 5

-7

22 5

CC = 83.23.710NaSO+1.110NaOH

+7.610NaS ONaOH

−× ×

(5)

For the screw press

[ ]

[]

-3 -3

22 5

-7 22 5

CC=81.11.310NaSO+1.610 NaOH

+7.610NaSONaOH

−× ×

(6)

3.3. Fiber Content

Based on the ANOVA calculations (see Table 5), the

additives concentration, both Na2S2O5 and NaOH do not

affect the reduction of fiber content. This is also proved

by the P values for both additives (>0.05). So, the physi-

cal treatment determines the amount of fiber in starch

after extraction. This result is the same as for the starch

yield.

3.4. Ash Content

Ash content shows the presence of inorganic compo-

nent in the starch. These can originate from the rhizome,

but also from the chemicals added during processing.

From the ANOVA calculations (see Table 6) it can be

seen that the concentration of NaOH and the physical

treatment affect the amount of inorganic material left in

the starch. This seems due to the solubility level of

Na2S2O5 in water which is hi gher than Na OH.

On Figure 4, it is shown that although Na2S2O5 con-

centration does not affect the ash content significantly,

there is a tendency that the higher concentrations of ad-

ditives, both Na2S2O5 and NaOH, result in a higher ash

content in the starch.

4. CONCLUSIONS

This study showe d that the integration of physical and

chemical is a promising technology for the extraction of

starch from Canna edulis. It is a relatively simple and

low cost pr oc ess and it produces a good quality st a rc h.

Tabl e 5. ANOVA for fiber content response.

Factor SS DF MS F Value p-Value

[Na2S2O5] 0.089 1 0.089 0.41 0.5261

[NaOH] 0.43 1 0.43 2.00 0.1717

treatment 4.54 1 4.54 21.18 0.0001

Tabl e 6. ANOVA for ash content response.

Factor SS DF MS F Value p-Value

[Na2S2O5] 0.63 1 0.63 1.54 0.2284

[NaOH] 20.18 1 20.18 48.80 <0.0001

Treatment 2.55 1 2.55 6.18 0.0210

J. R. B. Witono et al. / A gricultural Sciences 4 (2013) 51-55

Figure 4. 3D surface of ash content.

The use of the screw press in the separation process of

the fibers after the chemical extraction produce gives a

higher starch yield compared to the use of hydraulic press,

but the purity is lower. Therefore, the hydraulic press is

perhaps the most suitable method for preparing starch

that has to be a feed material for chemical modification

processing.

5. ACKNOWLEDGEMENTS

The authors would like to express their thanks to the Indonesian Di-

rectorate General of Higher Education—Ministry of Education which

finances this research and also the Chemical Engineering fresh gra-

duates of Parahyangan Cath olic University: Ricky Gunadi, Pamela and

David who assisted the authors in the preparation of this research.

Special thanks are addressed to the leader of “Mekar Sari” farmers

union Bapak Kemin, who facilitated the collection of the Canna edulis

Ker. Rhizome, and Indonesian Institute of Sciences (LIPI, Subang)

which allowed us to use their equipment for extracting the starch.

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