iU

S?2nd International Conference on Biosciences and Biotechnology "Pave the Way to A Better Life"
TAMARIND LEAF EXTRACTION (Tamarindus indica L.)
ETIIANOL-DEXTRIN ENCAPSULATION: STUDY OF ANTIRADICAL
AND ANTIOXIDANT
Sri Mulyani and Lutfi Suhendra
Department of Industrial Agriculture Technology
Faculty of Agricultural Technology, Udayana University

ABSTRACT

Tamarind leaf has a flavonoid as an antiradical with antioxidant activity. Tamarind leaf
isextracted through ethanol concentration (10%, 30%, 50%, 70% dan 90%) with a ratio of 1 : 2. It
is filtrated and added dextrin concentration (5%, 7,5%, and 10%) and then dried. Tamarind
leafextract is tested by total phenolic content and diphenylpicrilhydrazyl (DPPH). Antioxidant
activity on soy beanoil istested with peroxide number (ferric thiocyanate) and malonaldehyde (TBA).
BHT synthetic antioxidant was used as the control. This research is designed using a multiple
randomized design. The results showed the following optimal levels: extraction 70% ethanol
dextrin 5%, diphenylpicrilhydrazyl and total phenolic content: 0.18% with 3.24% gallic acid
equivalent. Tamarind leaf extract in concentration 200 ppm, is not able to inhibit peroxide.
However, it does inhibit malonaldehyde on oxidized soy bean oil. Tamarind leaf extract and
BHT have a similar capability as an antioxidant in oxidized soy bean oil.
Keywords: Tamarindus indica L., dextrin, antioxidant, antiradical and soybean oil

t*

INTRODUCTION

Antioxidant compounds called phytochemicals (Pratt, 1992) that come from

natural materials are safer for humans. The derivative fiavonoids are as important a
phytochemical as an antioxidant (Johnson, 2001). Phenolic compounds in plants have
been widely studied and found to shew antioxidant activity (Pokorny, 2001V Natural
antioxidant compounds are expected fo-ehange synthetic antioxidants such as butylated
hidroksianisol (BHA) and butylate'd hidroksitoluene (BHT).
Research has been conducted showing Tamarind (Tamarindus indica L.) to have
antioxidant activity (Suwariani and Suhendra, 2008; Chanwitheesuk et al., 2005;
Maisuthisakul et al., 2008; Siddhuraju, 2007). Tamarind has shown potential as an
antidiabetic and anti-hiperlipidemik (Maiti et al, 2005; Maiti et al., 2004), by inhibiting
nitric oxide production in celis (Choi and Hwang, 2005). The antioxidant compound in
tamarind is 2-hydroxy acid-30, 40-dihydroxy aseto fenon, 3.4-dihydroxy methyl
benzoate, 3.4-dihydroxy phenyl acetic acid, epicatechin and oligomerik proanthocianidin
(Tsudaeia/., 1994).
Tamarind (Tamarindus indica L.) contains xyloglucan. The xyloglucans (XGs)
are a group of storage or structural heteropolysaccnarides from plants, and their structure
is composed of a 1,4-linked P-D-glucan main chain that is partially substituted with a-DXyl side-chains at the 0-6 atoms. Depending on the source, the side-chains can be p-DGal-l,2-a-D-Xyl or a-L-l-Fuc-1,2- p-D-Gal-1,2- a-D-Xyl (Carpita & Gibeaut, 1993; Fry,
1989; Hayashi, 1989; McNeil, Darvill, & Fry, 1984; Varner & Lin, 1989) or more
complex chains (Hantus, Pauly, Darvill, Albersheim, & York, 1997; Freitas et al., 2005;
Jia, Qin, Darvill, & York, 2003; York, Kumar-Kolli, Orlando, Albersheim, & Darvill,
1996). The XGs are water-soluble, but the individual macromolecules typically do not
fully hydrate, and consequently, aggregated species remain present even in very dilute
solutions. For encapsulation, dextrin is one of the compounds required to make tamarind
leaf extract.
Cyclodextrins (CDs) are cyclic oligomers of a-D-glucopyranose that can be
produced due to the transformation of starch by certain bacteria such as Bacillus

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International Conference on Biosciences and Biotechnology "Pave the Way to A Better Life"

(Jeang, Lin, & Hsieh, 2005; Qi, Mokhtar, & Zimmermann, 2007; Qi &
Zimmermann, 2005; Rimphanitchayakit, Tonuzuka, & Sakano, 2005). The torus-like
;yclodextrin molecules have an outer polar surface and an inner non-polar surface. The
small hydrophobic cavities within the cyclodextrins have diameters of 0.50, 0.62, and
0.79 nm respectively, for the a, |3, and y dextrins. Cavities are potential binding sites for a
wide variety of both organic and inorganic molecules. Cyclodextrins can be used to
"encapsulate" food additives, and their complexes may be useful in the separation of
enantiomers, of drugs.
In recent years, several published reviews have described the use of CDs in food
and flavour applications (see Cravotto et al., 2006; Hedges & McBride, 1999; Hedges,
Shieh, & Sikorski, 1995; Qi & Hedges, 1995; Samant & Pai, 1991; Szente & Szejtli,
2004). CDs have been recommended for applications in food processing as well as for
food additives with a variety of reason. One reason is to protect lipophilic food
components that are sensitive to oxygen and due to light- or heat-induced degradation.
The aim of the research is to find out whether tamarind leaf extract ( Tamarindus
indica L) with an ethanol-dextrin encapsulation method can act as anantiradical and

macerans

antioxidant.
MATERIALS AND METHODS

Materials and Equipment

The research is focused on local varieties the tamarind leaf (Tamarindus indica L.)
collected from Kertalangu Kesiman Village, Denpasar - Bali. The chemical were obtained
from Merck and included : Soybean oil (linoleic acid), tiobarbituric acid, phosphate
buffer, sodium carbonate, ferry and tiobarbituric thiosianat acid, and Folin Ciocalteu
phenol, ; The chemical were obtained from Brathaco Chemical included : ethanol. BHT
while Sigma provided : gallat acid, DPPH radical (-2.2-1-picryldhydrazyl dhiphenii
radical). The equipment used in this research included : Vacuum Rotary Evaporator,
spectrophotometer (Turner SP-870), Centrifuge (EC HN-S II 0000-9000 rpm), Vortex
(Thermolyne), Oven (Blue.M), and an incubator (Memmert, Model 500 ).

Experimental Design
Experiments were conducted using a completely randomized factorial design
(CRD)Factor 1 focused on : the concentration of ethanol as a solvent : Kl: 10%, K2:
50%, K3: 50%, Q4: 70% and K5: 90%. The second factor looked at : the concentration of
dextrin adding solution, (w/w), which consisteds of three levels : Rl: 5%, R2: 7.5%, and
R3: 10%. The experiment was repeated twice.

Research and Analysis

Fresh tamarind leaves were washed, drained, and then ground to make powder. The
mixture was than added to the ethanol ratio (2:1) and extracted after one hour then filtered
and leaf extracts added with dextrin as above. The compound was then dried in an oven at
a temperature of 50 C. The dried tamarind leaf extract was then ground to make a powder
and seived through a 60 mesh.

Analysis: Stage 1 involved determining DPPH and the components cf phenolic

compounds. Stage 2 looked at the antioxidant activity using TBA (Kikuzaki and
Nakatami, 1993) modified. Stage 3 focused on antioxidant activity Ferry thiosianat
(Kikuzaki and Nakatami, 1993) modified.

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*$S$J2n International Conference on Biosciences and Biotechnology "Pave the Way to A Better Life"

RESULTS AND DISCUSSION

Total Phenol Tamarind Leaf Extract

The total phenolic quantity in the tamarind leaf extract had a high solubility in 90%
ethanol. The addition of 7.5% dextrin caused a higher polyphenol binding ability than
alternative levels (Table 1). Polyphenol solubiliti differs depending upon the structure of
the compounds. Dextrin as an encapsulant has the ability to bind the polyphenol
compounds.
The binding ability of polyphenol compounds on dextrins is due to cyclodextrin
molecules that are hydrophobic. Binding capacity of polyphenol compounds depends on
the type of cyclodextrin ( a, |3 or y). The hydrophobic ability of cyclodextrin has a similar
tendency to polyphenol in that when sufficiently polarized it becomes soluble in an
ethanol. Optimum concentration of dextrin for binding to polyphenols was found to be
7.5%.

Table 1. The average of total phenolic of tamarind leaf extracts in ethanol

concentration and dextrin
Dextrin (%)
Ethanol Solvent (%)
10

30

50

70

90

1.183J

1.835'

2.599h

3.235fg

7,5

3.410ef
3.295f

6.45

2.952s

4.593c

2.624h

3.620de

3.472ef

3.797d
5.690b
5 .4 10h

10

Antiradical Activities of Tamarind Leaf Extract (DPFH test)

Extraction of ethanol with concentration 70% displayed the highest antiradical
activity (Table 2). Tamarind antioxidant compound is 2-hydroxy-30, 40-dihydroxy aseto
fenon, 3.4-dihydroxy methyl benzoate, 3.4-dihydroxy phenyl acetic acid, epicatechin and
oligomeric proanthocianidin (Tsuda et al, 1994), an has tendency to become soluble in
ethanol 70% concentration. Research using ethanol concentration of 70% for extraction
tamarind fruit also done by Martinello et al. (2006) and Ramos et al. (2003).
Addition to dextrin concentration did not influence radical activities (Table 2).
Dextrin with a concentration of 5% showed the ability to bind phenolic compounds and
protect from heat damage. Dextrin has the ability to separate molecules XGs, so that the
tamarind leaf extract forms an anhydride compound.

Table 2. Average antioxidant activity (DPPH) of tamarind leaf extract at vaiious

concentrations of ethanol and dextrin

Ethanol Solvent (%)

Dextrin (%)
10

30

50

70

0.0597'

0.0946f

0.1066d

0.1177b

7,5

0.043 0*

0.0440*

0.0980e

0.1 143

10

0.0425j

0.0372k

0.0849h

0.1232a

90

0.0857s"
0.1042d
0.1064d

Antioxidant Activity of tamarind leaf extract using the FTC method.

Measurement of antioxidant activity conducted by the FTC method based on the
formation of peroxide which is oxidation of linoleic acid from soybean oil. Peroxide will
oxidize ferrous ions to ferri, and then ferrithiosianat form when measured at h = 500 nm.
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2nd International Conference on Biosciences and Biotechnology "Pave the Way to A Better Life"
Figure la, lb, 1c: The antioxidant activity of tamarind leaf extract at addition of 5%,
7 5% & 10% dextrin with FTC test

The FTC test showed that polyphenol compounds of tamarind leaf extract haves no
ability to inhibit peroxides.
Peroxide is the primary products of oxidation which could not be inhibited / prevented by
the polyphenol compound of tamarind leaf extract (Fig. la, lb.lc).

Antioxidant Activity of Tamarind Leaf Extract with TBA Method

To determine the ability of antioxidants to inhibit the fonnation of the malonaldehid
reaction rate oxidation process on lipids using the thiobarburic acid test (TBA). In leaf
extract at addition of 5%, 7.5% and 10% dextrin with test TBA

MDA is a secondary product from the oxidation of soybean oil. Tamarind leaf extract
varied for ethanol and dextrin concentration showed a high ability to prevent / inhibit the
formation of MDA (Figure 2a, 2b. 2c). Martinello et al. (2006) examined the tamarind
fruit showed the ability to prevent / inhibit the fonnation of MDA.
CONCLUSION

f-

1. Dextrin has a high ability to isolate xylogiucans (XGs) on the encapsulation of

tamarind leaf extract
2. Tamarind leaf extract has low affect on antiradicals and has no ability to prevent
fonnation of peroxides.
3. Tamarind leaf extract has the ability to inhibit or even / prevent the formation of
MDA, similar to BHT.

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