Ultrasonic Extraction Optimization (Ultrasound-Assisted

Extraction) Saponin From Lerak (Sapindus rarak DC)

Using The Response Surface Methodology (RSM)

Nita Aryanti1, a), Dyah Rosita Heny2, b)

1
Department of Chemical Engineering, Faculty of Engineering Undip, Semarang
Jl. Prof. Sudarto Tembalang Semarang

a)
Corresponding author: nita.aryanti@gmail.com
b)
rositahenydyah@gmail.com

Abstract. Lerak belongs to the family of Sapindaceae which grows well at a height of 450 to 1500 cm above sea level. This
plant in Java grows wild, the height of the plant can reach 42 cm and has a trunk diameter of 1 m. This plant has a different
name on each region, such as in Palembang called Lamuran, in Java Lerak and in West Java is often called Rerek. The wood
is very lightly used as a cast board, matchsticks and wooden crafts. The main content of the Lerak fruit is saponin, Saponin
is a glycocide that may be present in many kinds of plants. Saponin is present in all plants with high concentrations in certain
parts, and is influenced by crop varieties and growth stages. Saponin as a surfactant derived from non-ionic plants has
excellent performance and a wide presence in nature. Due to its excellent molecular structure and the hydrophilic backbone
of the glycosides and the lipophilic triterpen derivatives, saponins have excellent soluble power to the pollutant compounds.

This research aims to optimize the extraction process of saponins from Lerak (Sapindus Rarak) using ultrasound-Assisted
Extaction (UAE). The variables are used the concentration of ethanol solvent (X 1), the ratio of the Lerak: ethanol (X2) and
the extraction time (X2). The response surface method with the Central Composite Design (CCD) design was used to acquire
a mathematical model describing the relationship between Saponins's yield on the variable that influenced . The optimum
condition of saponins extraction in Lerak using ultrasound-Assisted Extraction is best in the composition of the ratio of the
solvent to the solvents 1:22.26180 GG-1, the concentration of ethanol (X1) 83.16278%, the extraction time (X3) of 21.92890
minutes with a yield 47.15952%. The color test on the saponins extract indicates that the contained saponins is a triterpene.
The color test done with the addition of the reagent Liberman Bouchardat (LB) will produce a brown-purple ring that
indicates the presence of Triterpene saponins. The results of identification with UV-VIS spectrophotometry have the
absorbantion value of saponins compounds of 4281.45 at a maximum wavelength of 320. The identified saponins
compounds contain the O-H, C-H, C = C, C-O-C,-CHO, = CH, and pironose sugars. Lerak Saponins Extract has a saponins
content it has properties can be low the surface tension with the CMC value at a concentration of 2%.

INTRODUCTION

Lerak belongs to the family of Sapindaceae which grows well at a height of 450 to 1500 m above sea level. In Java
This plant grows wild, the height of the plant can reach 42 m and has a trunk diameter of 1 meter. This plant has a
different name on each region, such as in Palembang called lamuran, in Java Lerak and in West Java is often called
Rerek, Jambi call it Kalikea., Minang people call it Canikia.The wood is very lightly used as a cast board, matchsticks
and wooden crafts[1].
 The seeds of this plant are stretched and brownish yellow, the fruit of the Lerak consists of 75% meat and 27% of
the seeds [2]. In this piece of flesh contains saponins compounds that are quite strong poison [3]. Sapindus Mukorossi
often called the fruit of the Lerak, containing saponins about 38%, in addition to containing strong toxins, Lerak fruit
also contains a type of oil that is not easy to dry that is about 26% consisting of glycerides, acids Palmitate and stearic
acid [2].

Saponin is a high molecular weight glycosid, composed of sugars associated with the triterpene or Aglikon steroid
[4]. The classical definition of saponins is based on its surface activity like detergent properties, giving a stable foam
in water, has a bitter taste and a strong poison for fish [4]. Saponin has a molecular weight of 414.6231 grams/mol and
molecular formula C27H42O3. Saponin has a sufficiently high boiling point, up to 158ºC and density 0.5 g/cm 3 at a
temperature of 20ºC. Saponin can dissolve in various solvents such as water, ethanol and also methanol. Some may
also dissolve in ether, chloroform, benzene, ethyl acetate or acetic acid [4].

Saponin forms a colloidal solution in water and forms a steady foam if shaken and not lost with the addition of
acids[5]. Saponin active compounds strong surface and cause foam when shaken with water. Some saponins work as
anti-microbial. Saponin can be used in various fields including fisheries, textiles, cosmetics and health. The field of
saponins fisheries is used as shrimp pest repellent [6], the textile industry as a detergent, the field of cosmetics used as
a foam-forming on shampoo [7,8] and the field of health can be used as a inhibitory cancer cell growth [9].

Saponin can lower surface tension, thereby replacing the role of synthetic surfactants. As a surfactant, the ability
of saponins for foam is caused by a combination of non-polar sapogenic and water-soluble side chains. With the
complete Hydrolisa will be produced Sapogenin (Aglikon) and carbohydrates (hexose, pentose and saccharic acid).
Based on its chemical properties, saponins can be divided into two steroid groups with 27 C atoms and the triterpenoid
with 30 C atoms. Glycosides have the structure of the surfactant molecular glycocoprint consisting of two ends, the
Glikon and Aglycones (Sapogenin). Predominantly, the Glikon structure consists of one, two or three sugar chains
attached to the Aglikon. Each term monodesmosida, Bidesmosida, or Tridesmosida (from Greek desmos = chain).
Aglikon, also called Sapogenin, is a nonpolar part of the molecule. Based on the Aglikonnya backbone, Saponins is
also divided into two namely triterpenoid and steroid glycosides. Its structural characterization varies with the number
of sugar units installed in different positions [4].

The hydrolysis of saponins produces an aglycones called Sapogenin, sapogenin forms compounds that are easily
chewed with acetylated, toxic saponins are often called sapotoxin. According to the aglycones or sapogenin structures,
saponins can be distinguished into two types namely Steroida and Triterpenoida. Both of these compounds have
glycolic relationships on C-3 and the same biogenetic origins as Metilglutarilkoenzim and isoprenoid acids [10].
Saponins have a relatively large molecular weight and high polarity, hence the isolation of saponins raises its own
challenges. The Saponins insulation is done in 2 ways by extraction and chromatography [4]. Research using ultrasonic
extraction has been conducted by Wu et. al. [11]. This research aims to optimize the most influential variables such as
ethanol solvent concentrations, the ratio of lerak to ethanol solvent and the duration of extraction.
 RESEARCH METHODS

Materials and methods

Materials for extraction include Lerak fruit (Sapindus Rarak DC) obtained from Gede market Solo, ethanol
solvents 90%, 80%, 70%, 60% and 50%. Aquadest, Whatman filter paper, Standard saponins (Sigma Aldrich).
The tools used in the manufacture baking sheet, sieve 80 mesh, thermometer, digital analytical balance of
Lerak powder include a dry lender (Philip), baking sheet, sieve 80 mesh, thermometer, analytical digital balance
(Sartorius), ultrasonic bath extraction (Krisbow), vacuum rotary pump, Centrifuge, electric Oven (Miyako),
Desicator. The tools used in Saponins extract analysis include electric oven (Memmert U. 30), UV-vis
spectrophotometer and Cuvette, FTIR.

Experimental procedures
Before extracting saponins from the Lerak is done by preparing ready-made Lerak from Sapindus Rarak DC
in the form of dry powder. In this initial processing phase, several analyses were also conducted to ensure saponins
contained in the Sapindus Rarak DC.
The fruits of the Sapindus are washed clean with water flowing to remove dirt. The seeds and skin of the
Lerak are separated. The outside of the lapped fruit is dried in the oven at a temperature of 50 oC for 2 hours[12].
The dry skin is milled and filtered through a 80-sieve mesh to obtain a homogeneous fine powder of the Sapindus
Rarak. These ready-made powders are stored in closed vacuum storage at 7oC temperature and add silica gel to keep
the powder dry.

Phytochemical Saponin Analysis

Saponin inspection

Preliminary analysis was conducted to evaluate the chemical structure and the presence of saponins in
Sapindus Rarak. Before quantifying the total saponins from the source of the plant, it is advisable to perform a
simple procedure to test the presence of saponins in a sample of the extracted ingredients. A total of 2-5 grams of
plant material was inserted into the reaction tube filled with distilled water and shaken hard for 2 minutes. The stable
and persistent foam appearance of the liquid surface for 15 minutes indicates the presence of saponins.

Test the color

Samples of approximately 500 mg were inserted into the test tube which had been filled with 10 ml of
chloroform, heated for 5 minutes with a water bath while shaken. Then added several Liberman Bouchardat (LB)
reagents. If formed brown or violet rings then indicates the presence of Triterpene saponins, while the green or blue
color indicates the presence of steroidal saponins [13]
The FT-IR transmittance spectrum of Sapindus Rarak samples was noted and compared to the transmittance
spectrum of FT-IR pure saponins with wavelengths of 4000-400 cm-1 [14]
 Identification of Saponin compounds with UV-Vis spectrophotometry

Ultrasonic extraction (UAE) is carried out in an ultrasonic bath with a constant power of 280 W, at a
frequency of 40 kHz. Ultrasonic Water Bath comes with a time of digital sonikation and temperature control system.
Sapindus powder Dried Rarak in the range of 10 grams placed in a sealed glass tube and mixed with ethanol 90%,
80%, 70%, 60% and 50% as solvents. The tubes with sonicated suspension for 10, 15, 20, 25 30, 35, 40, 45, 50, 55
minutes in an ultrasonic device containing 4 L of water at a temperature of 40ºC. All the samples to be extracted are
placed in the middle of an ultrasonic tub with a depth of fifteen centimeters and subsequent each batch of samples
to be sonicated are stored at the same position. After the extraction process is completed, the extraction result is
filtered with Whatman filter paper on the rotary vacuum pump, so that the extract is obtained Lerak. These lerak
extracts are then centrifuged to separate solids and extractions. Saponins extract was tested using UV-vis spectro to
analyze its saponrates. The Absorbansi obtained is used to calculate the yield, each treatment is repeated 2 times.
Observations conducted at 200-800 nm wavelength [13]

Experimental design

Central composite Design (CCD) Three factors, to see the optimum condition of treatment effect (the ratio
of the lerak to the ethanol solvent, the concentration of ethanol, extraction time) to the yield. The best condition that
previous research gained was the basis for the determination of the treatment on the surface response method used.
The yield percentage of saponins on the Lerak, calculated by the equation below:

𝑀𝑎𝑠𝑠 𝑆𝑎𝑝𝑜𝑛𝑖𝑛 𝑒𝑥𝑡𝑟𝑎𝑐𝑡𝑖𝑜𝑛 𝑅𝑒𝑠𝑢𝑙𝑡

% Yield extraction =
𝑆𝑜𝑙𝑣𝑒𝑛𝑡 𝑀𝑎𝑠𝑠
X100% ……...(1)

The treatment and treatment codes are shown in table 1.

TABLE 1. The limitations and variable levels change/variables are free.

Variabel Level limits

-1 0 +1

Ratio lerak : etanol ,X1 (gg-1) 1: 15 1 : 20 1: 25

Solvent concentrastion etanol, X2( %) 70 % 80 % 90 %

Extraction Time (menit) 15 20 25

At this stage there will be a mathematical equation with the second order polynomial model which is the
following quadratic functions:

Y = β0 + β1X1+ β2X2+ β3X3+ β11X12+ β22X22+ β33X32+ β12X1X2+ β13X1X3+ β23X2X3 ……...(2)
 TABLE 1. Second order polynomial Model

Solvent concentration Ratio lerak : Extraction Time

Run ethanol (X2) ethanol (X1) (X3) Yield

1 70 (-1) 15 (-1) 15 (-1) 10.6798

2 70 (-1) 15 (-1) 25 (+1) 115.7198

3 70 (-1) 25 (+1) 15 (-1) 19.0098

4 70 (-1) 25 (+1) 25 (+1) 18.5198

5 90 (+1) 15 (-1) 15 (-1) 10.9698

6 90 (+1) 15 (-1) 25 (+1) 20.3198

7 90 (+1) 25 (+1) 15 (-1) 10.0198

8 90(+1) 25 (+1) 25 (+1) 33.31

9 63.18207 (-α) 20 (0) 20 (0) 10.2198

10 96.81793 (+α) 20 (0) 20 (0) 38.9198

11 80 (0) 11.59104 (-α) 20 (0) 23.8998

12 80 (0) 28.40896 (+α) 20 (0) 46.0198

13 80 (0) 20(0) 1111.59104 (-α) 15.6198

14 80 (0) 20(0) 28.40896 (+α) 35.3198

15(C) 80 (0) 20(0) 20 (0) 43.0098

16(C) 80 (0) 20(0) 20 (0) 44.0098

Where Y is the lowling saponins of the Lerak, β0 is Intersep/constant, β1, β2, β3, is a linear coefficient, β11,
β22, β33 is a quantism coefficient, β2, β13, β23 is the coefficient of treatment interaction. X1 is a ratio of saponins to
ethanol solvent (g g-1), X2 is the concentration of ethanol (percentage), X3 is the extraction time (minutes).

Analysis procedure
Moisture analysis

The petri dish is inserted into the oven (105ºC) for 24 hours after it was entered into the Desicator for 15
minutes then weighed. Samples that have been mashed, weighed 2 grams in containers that have been known for
weight and then in the oven at a temperature of 100-105ºC for 5 hours. After that it is cooled in 15 minutes dive
desicator and weighed in weight. Then heated again in the oven 30 minutes, then cooled in a desicator and weighed.
This treatment is repeated until the constant weight is reached.

𝐼𝑛𝑖𝑡𝑖𝑎𝑙 𝑚𝑎𝑠𝑠−𝑓𝑖𝑛𝑎𝑙 𝑚𝑎𝑠𝑠

% Moisture content = x100% .......... (3)
𝑠𝑎𝑚𝑝𝑙𝑒 𝑀𝑎𝑠𝑠
 Analyze the yield powder Simplisia Lerak

Concentrated Lerak extract after extracted then evaporated will be separated between pure Lerak extract
with ethanol solvent, the result is weighed in a container that has been known to weigh, then calculated the weight
of the extract is concentrated compared to the initial weight Powder.
𝑓𝑖𝑛𝑎𝑙 𝑚𝑎𝑠𝑠 𝑠𝑎𝑚𝑝𝑙𝑒
% Yield Simplisia = x100%... ….(4)
𝑖𝑛𝑖𝑡𝑖𝑖𝑠𝑙 𝑚𝑎𝑠𝑠 𝑠𝑎𝑚𝑝𝑙𝑒

FTIR Analysis

Evaluate the chemical structure of the saponins extract from the Sapindus Rarak with the Fourier transform
Infra red spectroscopy (FT-IR) method. The analysis is performed using the Shimadzu FT-IR spectrometer with a
total, weakened reflection method. Before analysis, 20 mL of samples were evaporated using an IKA RV 10 Rotary
Vacuum Evaporator to reduce moisture content. Samples of liquid saponins extract are used as samples, analysis is
also performed on standard saponins (Sigma Aldrich) as a reference.Analisis Saponins levels with UV-VIS Spectro
This method is performed based on a specific group of saponins detected by UV rays of spectrophotometers.
Using a pure saponins from Sigma, Aldrich as the standard saponins reference to create a calibration curve. The
maximum absorption wavelength is determined by the UV-VIS Spectrophotometer, the result of absorbansi of 312
nm was found as the maximum wavelength.
The calibration curve preparation is done by analyzing the absorbance of pure saponins solution, varying
concentrations (5000-100 ppm). Spectrophotometry analysis is done by measuring the absorbance at the maximum
absorption wavelength using an empty reagent as a reference. The regression equation of the standard curve is based
on the concentration (X) versus the absorbancy (Y) value created and should have a linear correlation of more than
0.95. Total analysis of saponins on Rarak Sapindus extract obtained in different extraction conditions diluted to fit
the spectrophotometer equipment. Then the absorbance (A) value is measured at the appropriate wavelength. So the
concentration of samples in the solution is calculated by the equation obtained from the standard calibration curve
of pure saponins, and it is the equivalent of pure saponins.

Surface tension measurement

The surface tension test of the Lerak extract is carried out with the Noüy ring method using the Surface
Tensiomat tool. The extract measured the surface tension dissolved with the Akua Demineralisata so that the
obtained concentration of extract solution 0%, 0.005%, 0.01%, 0.02%, 0.03%, 0.05% 0.08%, 0.1%, 0.2% 0.4%,
0.6%, 0.8%, 1% 2%, 3% 4%, 5%, 6%, 8%, 14%, Tool is the surface tension value of the extract solution.
 RESULTS AND DISCUSSION

characterisation of the Lerak

TABLE 1. Result of Lerak characterisation

Parameter result Pure saponin

Colour Brown Brown

Appearance Powder Powder

Odor Normal Normal

Moisture content 5% 8%

Liberman Bouchardat Test (LB) Brown rings Brown rings

Foam test + +

Examination of saponins compounds

At the time the pollen is inserted into the test tube filled with distilled water and used hard for 2 minutes. A
stable and persistent foam appearance of the liquid surface for 15 minutes indicates the presence of saponins.

FIGURE 1 Determination of Saponins

Moisture content and Yield

The determination of the moisture content of simppsia powder is 5%. This results in accordance with the
requirements that the water content in Simplisia should not be more than 10%. [15]. Determination of water content
aims to give a minimum or range limit about the amount of water content in the material. It is related to the purity
and presence of contaminants in Simplisia. The low Simplisia powder yield of 8.64%.

Color test
Repeated color tests on the Lerak ethanol extract show that the saponins contained is a triterpene. In the color
test carried out resulted in a brown ring after the Simplisia dissolved in chloroform and heated for 5 minutes while
shaken, added the reagent LB (Liberman Bouchardat). Saponins compounds that state that samples after added LB
reagent will produce a brown-violet ring that indicates the presence of Triterpene saponins.

FIGURE 2 Saponins color test showing the presence of triterpene compounds

Analysis of surface responsiveness of Lerak

Saponin

The results showed that the ratio of saponins to ethanol solvent, the concentration of ethanol, the extraction
length had an influence on the yield of saponins from the resulting. The experiment's value and the model prediction
result for the yield as shown in table 3. According to table 2, it is known that the value of saponins yield obtained
through experimentation and through predictive models has good compatibility. The difference in the yield value
of the experiment results and the average prediction result of 24,023%. The highest Yield obtained 46.0198% is in
the extraction condition with a comparison of the powder Lerak against the ethanol solvent 1:28 GG-1, the
concentration of ethanol 80% and the extraction time 20 minutes. Meanwhile, the lowest yield value is obtained in
the extraction condition with the ratio of the powder to the ethanol solvent 1:25 GG-1, the concentration of the Lerak
at 90% and the duration of the extraction of 15 minutes, where in this condition resulted in a yield of 10.0198%. In
this case the amount of solvent used can not dissolve the lerak perfectly because it clumpy so that a little extract is
produced.
TABLE 2. Analysis of regression variant for the yield of lerak

Faktor SS df MS F p

(Conc etanol (L) 254.525 1 254.52 2.5738 0.1597

Conci Etanol (Q) 758.356 1 758.3561 7.6687 0.0324

ratio (L) 266.878 1 266.8779 2.698 0.1515

ratio (Q) 267.567 1 267.566 2.705 0.1510

xtraction Time (L) 362.098 1 362.0977 3.6616 0.1041

Extraction time (Q) 705.952 1 705.9525 7.1388 0.0369

1 L by 2 L 0.104 1 0.1036 0.0010 0.9752

1 L by 3 L 98.632 1 98.6324 0.9974 0.3564

2 L by 3L 8.841 1 8.8414 0.089 0.7750

Error 593.336 6 98.8894

 Total SS 2615.520 15

R2 0.77315

The results of variant analysis for Saponin yield showed a value of R2 of 0.77315. This indicates that a fixed
variable (X1, X2, and X3) affects the model. P value regression of 0.0369 is smaller than the degree of α significance
= 5%. This means that the research variables give a meaningful influence in the model, both linear and quadratic
models. The initial hypothesis (H0) will be rejected if the P value is less than α, otherwise the initial hypothesis will
fail if the P value exceeds α. Then there is no reason to reject the initial hypothesis (H 0) that says there is no lack of
fit, meaning the model that has been made in accordance with Data. The mathematical Model obtained to predict the
value of Saponins is as follows:
Y = 44.4838 + 4.31743X1+4.42095X2+5.14953X3-9.04776X12-5.37434X22-8.72956X32+ 0.1137X1X2+
3.51067X1X3+ 1.05067 X2X3 ….(5)

(a) (b)

FIGURE 3 (a) Plot Contour (b) surface relation yield Lerak (Y1) with a comparison of the concentration of ethanol
(X1) with extraction time (X3)

Fig. 3a and 3b indicate that the greater the concentration ratio of ethanol (X1) is used, the yield gained is also
increasing. At extraction time less than 20 minutes, the yield of saponins Lerak reaches 46.0198%. Thus, the
extraction time of less than 25 minutes of the saponins of the bark contained in the material has been completely
extracted.

(a) (b)

FIGURE 4 (a) Plot contour (b) yield surface link Lerak (Y1) By comparison of the ratio of the Lerak: Ethanol (X2)
with extraction time (X3)

In Fig. 4a and 4b, it is noted that increasing the volume of ethanol is not able to increase the resulting yield of
saponins lerak. The ratio of the Lerak with ethanol 1:25 GG-1, and the extraction time varies, indicating that the
extraction less than 25 minutes, will be obtained high yield saponins is 33.31%. However, extraction time is less than
15 minutes and the ratio of the Lerak: Ethanol 15 GG-1, the yield of the saponins Lerak is less than 10.67%. However,
at extraction time longer than 15 to 25 minutes, the ratio of the lerak to the constant ethanol solvent is at 1:25 GG-1,
the yield of the saponins Lerak can decline 1 again from 19.0098% to 18.5198%. This occurs because the ratio of the
Lerak: ethanol and the extraction time also affects the resulting yield of saponins. Although the effect is not so great,
when compared With a comparison of the amount of Lerak powder against the ethanol solvent used. Moreover, the
ratio of a lerak to the ethanol solvent used in the 1:25 GG-1 ratio is not yet able to absorb all saponins content in the
material, so the yield saponins is not maximal.
The highest yield optimization of the extraction experiment was obtained by 47.15952% with the ratio of the
Lerak to ethanol 1:22.2618 GG-1, the concentration of ethanol 83.162785 and the extraction time of 21.92 minutes
can be seen in table 5
TABLE 5. Extraction Experiment Optimization Results

Factor Observed Min Critical values Observed

Maximum

conc. ethanol 63.18207 83.16278 96.81793

Ratio lerak:ethanol 11.59104 22.26180 28.40896

Extraction time 11.59104 21.92890 28.40896

Yield 47.17592
 (a) (b)

FIGURE 5. (a) Plot Contour and (b) surface relationship Yield (Y1) with a comparison of the concentration of
ethanol (X1) Lerak ratio: ethanol (X2)

In the Fig. 5a and 5b shows that the greater the ratio of ethanol concentrations to the ratio of the Lerak: ethanol
used, the ratios are also increasing. At the ratio of the Lerak: Ethanol 15 g/g and the concentration of ethanol from
70% increased to 90%, increased yield from 10.6798-10.9698%, but increased concentration of ethanol tends to
increase the yield of saponins but not significant. This is because it has reached the saturation point, then the addition
of solvent concentrations will not increase the yield in accordance with the expected.

Identification of a cluster of isolates function with infrared spectrophotometry FTIR Saponin

of Lerak (Sapindus Rarak)

The peak of Sapoin extract is similar to the apex of pure saponins, almost all of the relative peaks show the
pure FTIR saponins also shown in Saponins extract, Both of samples show infrared. The absorption characteristic of
the hydroxyl group (OH) 3393.94 cm-1 in pure saponins and 3422.13 cm-1in saponins extract. Carbon-Hydrogen (C-
H) absorption at 2930.16 Pure saponins and 2935.14 cm-in saponins exstracts . Absorbance C = C observed in 1609.73
cm-1and 1638.54 cm-1on pure saponins and saponins extract. The apex of oligosaccharides indicating the presence of
Sapogenic (Aglikon), C-O-C, existed at 1137.15 cm-1 in pure saponins and at 1007.43 cm-1in saponins extract. When
the peak at 1392.58 cm-1in pure saponins and 1382.07 cm-1in saponins Extract Group – CHO from a structure of
oleanane. At the weak peak at 1,243.9 cm-1in pure saponins and 1,261.12 cm-1in The saponins extract indicates
aromatic = C-H. And the wide peaks at 617.71 cm-1and 655.48 cm-1in the saponins extract and the standard saponins
present the pyanose sugar in the Glikon structure.
Characterization of saponins using the infrared absorption functional group has been performed on Lerak,
Quilaja Saponaria [16], Basellasaponin [17] and steroidal saponins [18]. Previous studies reported that existence –
OH, C-H, C = C, and C-O-C were identified as saponins of Triterpenoid Oleanane [19].
The study also stated that the group C = O was to exhibit bidesmosidic saponins [20]. However, in Fig. 6
There is no infrared absorption to confirm the existence of the group C = O. Therefore, saponins may be a triterpenoid
oleanane monodesmosidik saponins. The top Group C = O also cannot be found on the standard saponins. Shows that
the Lerak saponins used in this study had compatibility with saponins from the standard saponins. The molecular
structure of the saponins of the Rarak Sapindus is also provided by Morikawa et al. [21]
 90

80

70

Transmitance (%)
60
C=C

50
Pyranosa sugar
40

C=C C-O-C
30 C-H -CHO
Saponin standard
20 -OH
Lerak extract

10
4000 3500 3000 2500 2000 1500 1000 500
Wave of Length (cm-1)

FIGURE 6. Spectral FTIR Saponin Standard and Lerak Extract

Surface tension test Compound Saponin Lerak

The Surface tension Drop chart overview shown in Fig. 7 The graph shows the Critical value Micelle
Concentration (CMC) through the intersection of the surface tension reduction line Concentration of extracts where
the surface tension value is relatively similar, Then extrapolated so that it is known at that concentration occurs CMC.
CMC can occur with the addition of surfactants in the solution will cause a decrease in the surface tension of the
solution, after achieving a certain concentration, the surface tension will be constant even though surfactant
concentrations are increased. When surfactants are added over this concentration the Surfactants aggregate form the
Misel. The concentration of the formation of this misel is called Critical Micelle Concentration (CMC). The surface
tension will decrease until CMC is reached. Once the CMC is reached, the surface tension will be constant indicating
that the interface becomes saturated and forms the misel that is in balance.
Fig. 7. obtained the surface voltage measurement of the A1 extract, obtained equation of the line: y =-49.333
x + 48,829 and y =-0.5705 x + 24769 so that after calculating the results obtained x = 2.11 Thus the value of CMC for
the extract of Lerak at 2% concentration.
 FIGURE 7. Surface Tension of Saponin

CONCLUSION

The color test on the saponins extract indicates that the contained saponins is a triterpene. The color test done with
the addition of the reagent Liberman Bouchardat (LB) will produce a brown-purple ring that indicates the presence of
Triterpene saponins.

The optimum condition of saponins extraction in Lerak using ultrasound-Assisted Extraction is best in. the
composition of the ratio of the solvent to the solvents 1:22.26180 GG-1, the concentration of ethanol (X1) 83.16278%,
the extraction time (X3) of 21.92890 minutes with a yield 47.15952%. The results of identification with UV-VIS
spectrophotometry have the absorbantion value of saponins compounds of 4281.45 at a maximum wavelength of 320.
From isolates of identified saponins compounds containing the O-H, C-H, C=C, C-O-c, -CHO, =CH, dan pironosa sugar.

Lerak Saponins Extract has a saponins content that has properties can lower the surface tension with the CMC value
at a concentration of 2.11%.

REFERENCES

1. U. Laba, (Lerak (Sapindus rarak) Soap Substitute Industrial Plant) Center for Research and Development of
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