The Influence of Solvent Type and Ratio of Material With Solvent on Antioxidant Activity of Tamarillo Extract (Chyphomandra betacea
Sendth).
NI M. YUSA 1), 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
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.
Extracted by using shaker at 130 rpm for 24 hour at room tempertaure
Crushed until becomes juice
At 50 g of weight inserted into erlenmeyer
Dried by vacuum oven at 30 C Filtered by filter paper
Fruit is cut
The flesh is kept
Added by solvent and ratio of material and solvent following the treatement and added by HCl until pH 2.5
Tamarillo
Skin and
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 ± 30oC. 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 :
Rendemen (%) = x 100 %
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. Average values of tamarillo rendemen (%)
Remark: average values that were followed by similar letter at the same column or row showed unreal difference (P<0.05)
Table 1. also showed that the average values of tamarillo extract rendemen with the treatment of ratio material with solvent range from 2.36% to 3.38 %. The treatment of ratio between material and solvent that was able to produced the highest rendemen was ratio of 1:4 wich was at 3.38%, meanwhile the lowest rendemen is at the ratio of 1:8 at 2.36%
2. Phenol total
The experiment result on phenol total can be seen at Table 2. The illustration of interaction of solvent type and ratio between material and solvent can be seen at Figure 2.
Table 2. Total phenol average (%) of tamarillo extract in accordance with gallat acid standard treatment.
Remark: different letters at the columns and lines interaction, average of solvent, and average showed significant difference (P<0.01)
Solvent type Ratio between material and solvent average
1:4 1:6 1:8
Methanol Ethanol water 3,64 3,91 2,59 2,49 3,17 2,31 2,32 2,93 1,83 2,82 ab 3,34 a 2,24 b
average 3,38 a 2,66 ab 2,36 b
Solvent type Ratio between material and solvent average
1:4 1:6 1:8
Methanol Ethanol Water 9,68 a 7,75 bcd 9,35 ab 6,80 bcd 8,37 abc 9,66 a 6,22 de 5,48 e 10,34 a 7,56 b 7,20 b 9,78 a
Figure 2. Graphic of average value of total phenol
Table 2. showed that the interaction between solvent type and ratio of material and solvent over the phenol total of tamarillo extract range from 5.48 % to 10.34 %.
Figure 2 explained that the highest phenol total was found at the interaction of water solvent with the ratio 1:8 was 10.34 %, meanwhile the lowest phenol total was found at the interaction of ethanol solvent with ratio 1:8.
Table 2 also explained that average value of phenol total of tamarillo extract with treatment of ratio between material and solvent ranged between 7.35 % to 8.93 %.
3. Antioxidant activity
Data of the experiment on the antioxidant activity can be seen at Table 3. The illustration between the solvent type with ratio between material and solvent can be seen at Figure 3.
Table 3. Average value of antioxidant activity (%) of tamarillo extract based on vitamin C standard.
Solvent type Ratio material and solvent Average
1:4 1:6 1:8
Methanol
Ethanol
Water
0,68 b
1,83 a
0,62 b
0,62 b
1,07 b
0.77 b
0,73 b
0,76 b
0,73 b
0,67 b
1,22 a
0,70 b
Average 1,04 a 0,82 a 0,74 a
Methanol
Ethanol
Remark: different letters on the columns and lines of interaction, average of solvent type, average of ratio material and solvent indicated significant differences. (P<0.01)
Figure 3. Graphic of antioxidant activities.
Table 3. Showed that the interaction between the solvent type with the ratio between material and solvent on tamarillo extract antioxidant activity range between 0.62 % to 1.83 %. On the table 3 also was showed that the average values of antioxidant activities of tamarillo extract on the treatment of solvent type range between 0.67 % to 1.22 %
Figure 3 showed that the highest antioxidant activities was found on interaction with ethanol solvent with ratio of 1:4 at 1.83%, meanwhile the lowest antioxidant was found on the interaction with methanol solvent and water which was respectively 1:6 and 1:4 at 0.62 %
From this experiment can be concluded that ratio between material and solvent influenced significantly on rendemen and phenol total but did not influence tamarillo extract antioxidant activities. The best results was obtained from the treatment of ethanol solvent on ratio between material and solvent 1:4 with rendemen results 3.34%, phenol total 7.75% and antioxidant activities at 1.83%
Further research is needed on the length of time and the temperature of extraction of tamarillo extract in order to find out the optimum ability of capturing free radicals.
REFERENCES
Gordon A E, Rodrigues RB, Marx F, and Papagiannopoulos. 2007. Antioxidant Capacity of Tamarillo Fruit (Cyphomandra betacea) [Abstrak]. Germany, Institute of Nutrion and Food Sciences, Bonn University.
Methanol
Ethanol
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