The Influence of Solvent Type and Ratio of Material With Solvent on Antioksidant Activity of Tamarillo Extract(Chyphomandra betacea Sendth.).
The Influence of Solvent Type and Ratio of Material With Solvent on
Antioxidant Activity of Tamarillo Extract (Chyphomandra betacea
Sendth).
1)
NI M. YUSA , G.A.K. DIAH PUSPAWATI 1) and I W. DIRGA 1)
1)
Faculty of Agriculture Technology Udayana University, Bukit Jimbaran,
Badung, Bali, Telp.Fax : (0361) 701801.
Correspondence Author : E-mail: nimadeyusa@yahoo.co.id
Abstract
This research was aimed to determine the influence of the solvent type and
ratio of materials with solvent on antioxidant activity of tamarillo extract and to
determine the solvent type and ratio of materials with solvent that would produce
pulp tamarillo extract with the highest antioxidant activity. This research used
Completely Randomized Design by factorial pattern with two factor. The first
factor was solvent type that contained three levels such as methanol, ethanol, and
water. The second factor was the comparison between the materials and the
solvent that contained three levels such as 1 : 4, 1 : 6, and 1 : 8. The treatments
were repeated twice and resulted in eighteen units of experiments. The observed
variables were yield, phenol total, and antioxidant activity. The research showed
that the interaction influenced the yield but did not influence the phenol total and
antioxidant activity. The solvent type influenced the yield, phenol total, and the
antioxidant activity. The ratio of the materials with solvent influenced the yield
but did not influence the phenol total and the antioxidant activity. The best results
were obtained from the ethanol solvent on a ratio of materials with solvent 1 : 4
with a yield of 3.34 %, phenol total of 7.75 % and antioxidant activity of 1,83 %
Keyword : Pulp tamarillo extract, methanol, ethanol,, and antioxidant activity.
INTRODUCTION
Tamarrilo is a kind of plant with oval shapped fruits that goes sharper on
its ends, the fruit color becomes brownish red once it is rippen. This fruit is local
to Bali island and is specially grown in Bangli regency and Bedugul area. The
usage of tamarrilo is stil limited, mostly are sold fresh and used as an ingredient of
rujak. Tamarrilo is rich with vitamin A, vitamin C, minerals and important fibers
to help keeping a healthy body (Gordon et.al, 2007). The brownish red on its
color and flesh once it is ripen indicates the rich content of anthocyanin.
Anthocyanin is a water-soluble pigment that appears naturally in various plants.
Vitamin C is also a non-enzymatic antioxidant which solves in water. Extraction
process that uses solvent is influenced by solvent types and the amount of solvent
used (Utami, 2009). This researched is aimed to determine the influence of
solvent type and the ratio of solvent material on the activity of antioxidant within
the tamarillo pulp extract.
MATERIALS AND METHODS
The research was conducted at Food Analyst Laboratory, Nutrition and
Biochemical Laboratory, and Food Processing Laboratory of Agriculture
Technology Faculty, University of Udayana, in March 2011 until May 2011. The
1
material used is tamarillo type red with optimum ripeness, which is showed by its
red color going even all over the fruit surface.
The research used factorial pattern of Completely Randomized Design which
consisted on two factors which were the treatment of solvent type and the ratio of
solvent material:
1. First factor is solvent (P), consist of :(P1 = Methanol; P2 = Ethanol dan
P3 = water solvent ).
2. Second factor is the ratio of solvent (A) which were: A1= ratio of material
and solvent 1:4, A2= ratio of material and solvent 1:6, and A3= ratio of
material and solvent1:8).
The treatment is repeated twice and resulted in 18 units of experiments.
Data obtained is analyzed by variance analysis and when the treatement showed
any real influence, it would the be continued with DMRT test to observed the
difference between the combination of treatments (Steel dan Torrie, 1993).
1. Execution of the research
1.1. The making of tamarillo pulp extract
The making of tamarillo pulp extract followed the maceration method. A
fine juice of tamarillo is weighed at 50 g then inserted into Erlenmeyer 500 ml
(wrapped in aluminum foil), then added HCl 1 % until the pH turned 2.5. Then it
was continued by maceration method by mixing material with solvent in the
shaker at 130 rpm for 24 hours at room temperature.
Tamarillo
Fruit is cut
Skin and
The flesh is kept
Crushed until becomes juice
At 50 g of weight inserted into erlenmeyer
Added by solvent and ratio of material and solvent following
the treatement and added by HCl until pH 2.5
Extracted by using shaker at 130 rpm for 24 hour at room
tempertaure
Filtered by filter paper
Dried by vacuum oven at 30 C
Antioxidant extract
2
Figure 1. Diagram of tamarillo extraction process (Saati, 2002) modified
Afterward the result was filtered by regular filter paper so then a tamarrilo extract
obtained, still mixed with the solvent. The solvent then vaporized by a vacuum
oven at ± 30 oC. Diagram of the tamarillo extraction process can be seen at Figure
1.
2. Observed variable
Variables observed covered rendemen (Sudarmadji, et al., 1989), phenol
total by using modified method of Folin-Ciocalteu phenol (Ramamoorthy and
Bono 2007), and antioxidant activity by using modified method of DPPH (Yun,
2001).
2.1 Rendemen
Tamarilo extract rendemen was obtained by a calculation (Sudarmadji, et
al., 1989). As following :
x 100 %
Rendemen (%) =
2.2 Total phenol determination
Total phenol analysis was using phenol Folin-Cioccalteu reactor. The
solvent absorbance was measured by spectrophotometer at 760 nm. The number
obtained then determined as the phenol total using gallat acid as the standard with
concentration of 0, 40, 60, 80, and 100 ppm.
2.3 Antioxidant activity
Absorbance was read at 517 nm. The determination of ability to capture
DPPH free radical used vitamin C standar curve with concentration 0, 40, 60, 80,
and 100 ppm. The antioxidant activity was measured by the following formula:
Antioxidant activity (%) =
x 100%
where :
X: sample concentration (mg/l)
TV: solvent volume total (l); FP: dilution factor; W: sample weight (mg)
RESULTS AND DISCUSSION
1. Rendemen
Average values of tamarillo rendemen can be seen Table 1.
Table 1. showed that average values of tamarrilo rendemen range at 2.24
% - 3. 34%. Treatment of solvent type that resulted in the highest rendemen was
ethanol at 3.34%, meanwhile the solvent type that resulted in the lowest
rendemen was water at 2.2.4 %.
3
Table 1. Average values of tamarillo rendemen (%)
Solvent type
Ratio between material and solvent
average
1:4
1:6
1:8
Methanol
3,64
2,49
2,32
2,82 ab
Ethanol
3,91
3,17
2,93
3,34 a
water
2,59
2,31
1,83
2,24 b
average
3,38 a
2,66 ab
2,36 b
Remark: average values that were followed by similar letter at the same column or
row showed unreal difference (P
Antioxidant Activity of Tamarillo Extract (Chyphomandra betacea
Sendth).
1)
NI M. YUSA , G.A.K. DIAH PUSPAWATI 1) and I W. DIRGA 1)
1)
Faculty of Agriculture Technology Udayana University, Bukit Jimbaran,
Badung, Bali, Telp.Fax : (0361) 701801.
Correspondence Author : E-mail: nimadeyusa@yahoo.co.id
Abstract
This research was aimed to determine the influence of the solvent type and
ratio of materials with solvent on antioxidant activity of tamarillo extract and to
determine the solvent type and ratio of materials with solvent that would produce
pulp tamarillo extract with the highest antioxidant activity. This research used
Completely Randomized Design by factorial pattern with two factor. The first
factor was solvent type that contained three levels such as methanol, ethanol, and
water. The second factor was the comparison between the materials and the
solvent that contained three levels such as 1 : 4, 1 : 6, and 1 : 8. The treatments
were repeated twice and resulted in eighteen units of experiments. The observed
variables were yield, phenol total, and antioxidant activity. The research showed
that the interaction influenced the yield but did not influence the phenol total and
antioxidant activity. The solvent type influenced the yield, phenol total, and the
antioxidant activity. The ratio of the materials with solvent influenced the yield
but did not influence the phenol total and the antioxidant activity. The best results
were obtained from the ethanol solvent on a ratio of materials with solvent 1 : 4
with a yield of 3.34 %, phenol total of 7.75 % and antioxidant activity of 1,83 %
Keyword : Pulp tamarillo extract, methanol, ethanol,, and antioxidant activity.
INTRODUCTION
Tamarrilo is a kind of plant with oval shapped fruits that goes sharper on
its ends, the fruit color becomes brownish red once it is rippen. This fruit is local
to Bali island and is specially grown in Bangli regency and Bedugul area. The
usage of tamarrilo is stil limited, mostly are sold fresh and used as an ingredient of
rujak. Tamarrilo is rich with vitamin A, vitamin C, minerals and important fibers
to help keeping a healthy body (Gordon et.al, 2007). The brownish red on its
color and flesh once it is ripen indicates the rich content of anthocyanin.
Anthocyanin is a water-soluble pigment that appears naturally in various plants.
Vitamin C is also a non-enzymatic antioxidant which solves in water. Extraction
process that uses solvent is influenced by solvent types and the amount of solvent
used (Utami, 2009). This researched is aimed to determine the influence of
solvent type and the ratio of solvent material on the activity of antioxidant within
the tamarillo pulp extract.
MATERIALS AND METHODS
The research was conducted at Food Analyst Laboratory, Nutrition and
Biochemical Laboratory, and Food Processing Laboratory of Agriculture
Technology Faculty, University of Udayana, in March 2011 until May 2011. The
1
material used is tamarillo type red with optimum ripeness, which is showed by its
red color going even all over the fruit surface.
The research used factorial pattern of Completely Randomized Design which
consisted on two factors which were the treatment of solvent type and the ratio of
solvent material:
1. First factor is solvent (P), consist of :(P1 = Methanol; P2 = Ethanol dan
P3 = water solvent ).
2. Second factor is the ratio of solvent (A) which were: A1= ratio of material
and solvent 1:4, A2= ratio of material and solvent 1:6, and A3= ratio of
material and solvent1:8).
The treatment is repeated twice and resulted in 18 units of experiments.
Data obtained is analyzed by variance analysis and when the treatement showed
any real influence, it would the be continued with DMRT test to observed the
difference between the combination of treatments (Steel dan Torrie, 1993).
1. Execution of the research
1.1. The making of tamarillo pulp extract
The making of tamarillo pulp extract followed the maceration method. A
fine juice of tamarillo is weighed at 50 g then inserted into Erlenmeyer 500 ml
(wrapped in aluminum foil), then added HCl 1 % until the pH turned 2.5. Then it
was continued by maceration method by mixing material with solvent in the
shaker at 130 rpm for 24 hours at room temperature.
Tamarillo
Fruit is cut
Skin and
The flesh is kept
Crushed until becomes juice
At 50 g of weight inserted into erlenmeyer
Added by solvent and ratio of material and solvent following
the treatement and added by HCl until pH 2.5
Extracted by using shaker at 130 rpm for 24 hour at room
tempertaure
Filtered by filter paper
Dried by vacuum oven at 30 C
Antioxidant extract
2
Figure 1. Diagram of tamarillo extraction process (Saati, 2002) modified
Afterward the result was filtered by regular filter paper so then a tamarrilo extract
obtained, still mixed with the solvent. The solvent then vaporized by a vacuum
oven at ± 30 oC. Diagram of the tamarillo extraction process can be seen at Figure
1.
2. Observed variable
Variables observed covered rendemen (Sudarmadji, et al., 1989), phenol
total by using modified method of Folin-Ciocalteu phenol (Ramamoorthy and
Bono 2007), and antioxidant activity by using modified method of DPPH (Yun,
2001).
2.1 Rendemen
Tamarilo extract rendemen was obtained by a calculation (Sudarmadji, et
al., 1989). As following :
x 100 %
Rendemen (%) =
2.2 Total phenol determination
Total phenol analysis was using phenol Folin-Cioccalteu reactor. The
solvent absorbance was measured by spectrophotometer at 760 nm. The number
obtained then determined as the phenol total using gallat acid as the standard with
concentration of 0, 40, 60, 80, and 100 ppm.
2.3 Antioxidant activity
Absorbance was read at 517 nm. The determination of ability to capture
DPPH free radical used vitamin C standar curve with concentration 0, 40, 60, 80,
and 100 ppm. The antioxidant activity was measured by the following formula:
Antioxidant activity (%) =
x 100%
where :
X: sample concentration (mg/l)
TV: solvent volume total (l); FP: dilution factor; W: sample weight (mg)
RESULTS AND DISCUSSION
1. Rendemen
Average values of tamarillo rendemen can be seen Table 1.
Table 1. showed that average values of tamarrilo rendemen range at 2.24
% - 3. 34%. Treatment of solvent type that resulted in the highest rendemen was
ethanol at 3.34%, meanwhile the solvent type that resulted in the lowest
rendemen was water at 2.2.4 %.
3
Table 1. Average values of tamarillo rendemen (%)
Solvent type
Ratio between material and solvent
average
1:4
1:6
1:8
Methanol
3,64
2,49
2,32
2,82 ab
Ethanol
3,91
3,17
2,93
3,34 a
water
2,59
2,31
1,83
2,24 b
average
3,38 a
2,66 ab
2,36 b
Remark: average values that were followed by similar letter at the same column or
row showed unreal difference (P