Jurnal Teknologi Industri Pertanian Ike Sitoresmi M Purbowati, Khaswar Syamsu,

ISSNEndang Warsiki,
0216-3160 Herastuti
EISSN Sri R
2252-3901
26(1):23-30 (2016) Terakreditasi DIKTI No 56/DIKTI/Kep/2012

OPTIMIZATION OF PHENOLS EXTRACTION FROM ROSELLE (Hibiscus sabdariffa) BY

MICROWAVE ASSISTED EXTRACTION AS ANTIBACTERIAL AND ANTIOXIDANT AGENTS

OPTIMASI EKSTRAKSI FENOLIK DARI ROSELA (Hibiscus sabdariffa) BERBANTU GELOMBANG

MIKRO SEBAGAI BAHAN ANTIBAKTERI DAN ANTIOKSIDAN
Ike Sitoresmi M Purbowati1)*, Khaswar Syamsu2), Endang Warsiki2), Herastuti Sri R1)
1)
Departement of Agricultural Technology, Faculty of Agriculture, Jenderal Soedirman University
Dr. Suparno Street, Karangwangkal, Purwokerto53123, Indonesia
E-mail: [email protected]
2)
Department of Agroindustrial Technology, Faculty of Agricultural Engineering and Technology, Bogor Agricultural University
PO BOX 220 16602, Bogor, Indonesia

Paper: Received 1 January 2015; Revised 23 April 2015; Accepted 14 March 2015

ABSTRAK

Kelopak bunga rosela merupakan sumber potensial senyawa phenolik. Untuk mendapatkan senyawa
bioaktif tersebut digunakan metode ekstraksi berbantu gelombang mikro yaitu ekstraksi menggunakan alat
microwave skala rumah tangga. Response Surface Method digunakan untuk menentukan kondisi optimum
variabel ekstraksi, yaitu daya gelombang mikro, konsentrasi etanol dan lama waktu ekstraksi. Kondisi optimum
proses ekstraksi yaitu pada daya gelombang mikro 250 Watt, konsentrasi etanol 78,36% dengan lama waktu
4,91 menit. Hasil verifikasi penelitian total phenol yang diperoleh pada kondisi optimum adalah 23,77 + 0,25
mg/g, lebih rendah dari estimasi optimasi proses, yaitu 24,61 mg/g. Kandungan anthosianin, vitamin C dan
rendemen MAE berturut turut 14,80 + 0,08; 10,74 + 0,14 mg/g dan 22,09 + 3,3%. Aktivitas antioksidan
ditunjukkan dengan zona hambat yang terbentuk 12,6 + 0,6 mm terhadap E. coli dan 11,6 + 0,3 mm terhadap S.
aureus. Aktivitas antioksidan ditunjukkan dengan nilai IC50 202,47 µL/mL
Keywords: microwave assisted extraction, Phenols, rosella, response surface method

ABSTRACT

Roselle calyx is rich with phenolic substances. In this study, microwave-assisted extraction was used to
extract the phenolic compounds from Roselle calyx. By using response surface methodology, the effects of
microwave output power, ethanol concentration and extraction time on total phenols yield were investigated and
the optimal conditions were determined as follows: microwave output power 250 W, ethanol concentration
78.36% and extraction time 4.91 min. The estimated values for total phenols yield was 24.61 mg/g obtained at
these conditions. The yield of a verification experiment at the optimum condition was 23.77 + 0.25 mg/g total
phenols higher than those of conventional method 19.84 + 0.46 mg/g was obtained. The detailed yield consist of
anthocyanin, vitamin C and yield of microwave assisted extraction were 14.80 + 0.08; 10.74 + 0.14 mg/g and
22.09 + 3.3% which were higher and significantly different than conventional extraction , which were 9.28 +
0.04; 9.99 + 0.16 mg/g and 16.18 + 1.9%, respectively. Inhibitory zone was 12.6 + 0.6 mm against E. coli and
11.6 + 0.3 mm against S. aureus. IC50 value of the extract was 202.47 µL/mL. Compared with conventional
method, with inhibitory zone 10. 2 + 0.4 mm against E. coli and 9.8 + 0.6 mm against S. aureus. Antioxidant
activity expressed with IC50 value 293.09 µL/mL.
Keywords: microwave assisted extraction, Phenols, roselle calyx, response surface method
INTRODUCTION growth. These substances that inhibit pathogen and
have little toxicity to cell are considered prospective
Roselle (Hibiscus sabdariffa) is also known candidates for developing new antimicrobial drug.
as rosella in Indonesia. Initially, it is cultivated in According to Pietta (2000), dried calyces contain the
this region for its fiber, but now also for leaf, fleshly flavonoid gossypetin, sabdaretin, hibiscetin and
calyx and seed according to their respective anthocyanin.
properties. The thick red and fleshly cup shaped Mourtzinos et al. (2008) stated that the
calyx of the flower are consumed worldwide as a calyx of Roselle was rich in phenolic compounds
cold beverage and as warm tea. This plant is also including anthocyanin. It was reported that those
used in traditional medicine against many compounds could be considered as a great source of
complaints that including high blood pressure, liver natural antioxidants. As anthocyanin are derivatives
diseases and fever (Tsai et al., 2004). of the basic flavylium cation structure, which has an
Roselle as medicinal plant are known to electron deficient nucleus, they generally are highly
produce bioactive compounds which react with other reactive. This reactivity shows the capabilities of
organism in environment, inhibiting bacterial anthocyanin as antioxidant. In the other hand, this

Jurnal Teknologi
*Correspondence Industri Pertanian 26 (1): 23-30
Author 23
Optimization of Phenols Extraction from Roselle …………

reaction usually involves decolorization on the (50%, 60%, 70%, 80%, and 90%), microwave power
pigment. The rate of anthocyanin destruction extraction (100, 175, 250, 325, and 400 W) and time
depends on many factors such as pH, temperature, of extraction (1, 3,5,7, 9, and 11 minutes) were
intermolecular copigmentation, ascorbic acid, conducted for each parameter using non factorial
oxygen, etc. design. Microwave used for extraction was
Usually, a conventional techniques as Electrolux EMM 2007X. The extraction was done
heating, boiling or refluxing can be used to extract in pH 2, the ratio dried roselle and solvent 1:10 w/v
the phenols. However the disadvantages are the loss (wet weight), 10 g dried roselle powder, in 100 mL
of phenols due to oxidation, hydrolysis and solvent. The slurry was radiated in microwave oven
ionization during extraction as long as the long at regular intervals (one minute radiation and two
extraction time. Other techniques such as minute off) to keep the temperature not rise above
Supercritical Carbondioxide Extraction, Subcritical the boiling point (Li et al., 2009). Roselle extract
Water Extraction, Ultrasonic Assisted Extraction and was filtered and concentrated with vacuum
Microwave Assisted Extraction (MAE) have become evaporator at 70oC, 44 cmHg and blowing with N2 to
interesting alternatives for the conventional methods. ensure the solvent totally evaporate. The main
Among these, MAE is the most economical and response parameters that analyzed were total
simplest technique for extraction of many plant phenols. The best combination was also analyzed
derived compound (Li et al., 2009; Wang et al., vitamin C, (AOAC, 2000) and total anthocyanin
2010). Product recovery by microwave is generally content (Fuleki and Francis, 1968).
attributed to its heating effect, which occurs due to
the dipole rotation of the solvent in the microwave Determination of Total Phenols
field (Zhang et al., 2005; Hemwimon et al., 2007). Total phenols were determined by the
Nevertheless, no reports on MAE of phenol Folin-Ciocalteu method of Chew et al. (2009).
compounds from roselle have been published, thus Briefly, 1.5 mL Folin-Ciocalteu reagent (10% v/v)
this experiemen using MAE technique should be was mixed with 1.5 mL 7.5% (w/v) Na2CO3
found. solution, then 0.4 mL sample solution was added.
Based on the dependent characteristic of After a 90-min incubation at room temperature in
phenols, and the need to find the most efficienct dark, the absorbance was measured at 765 nm using
extraction have emphasized the need of optimizing Spectrofotometer UV Vis 1800 Shimadzu. Gallic
the extraction process. The Response Surface acid was used as a standard compound for the
Method (RSM) is a collection of mathematical and standard curves. The results were presented in mg
statistical technique for the investigation and Gallic acid equivalent (GAE)/g extract. All the
modeling of complex problem processes whose experiments were carried out in four replicates.
response of interest is influenced by several
variables and objective to optimize this response Screening of Antibacterial Activity (Doughari,
(Montgomery, 2001). RSM takes interactions into 2006).
consideration and optimizes the process parameters Screening of antibacterial activity of the
to reasonable range, with the advantage of less the plant extract was performed by disc diffusion
number of replicates and the total time required to technique which is highly effective for rapidly
perform the experiments (Lee et al., 2006). RSM growing microorganism. The 20 µL test
uses an experimental design such as the central microorganism were inoculated on to the respective
composite design (CCD) to fit a model by least NA medium by pour plate method with 24 hours
squares technique. There has been no report on incubation. After solidification the filter paper disc
optimizing MAE of phenols from roselle using RSM (6 mm diameter) impregnated with 10 µL crude
technique. The objectives of the work were to extract sample, standard antibiotic (Kloramfenicol)
establish an optimised condition of MAE for roselle and a blank disc impregnated with 10 µL respective
calyx phenols, and to characterize of the extract of solvent had included. This screening was tested
the optimum condition. against gram positive bacteria Staphylococcus
aureus and gram negative bacteria Escherichia coli.
MATERIAL AND METHODS The inoculated plates were incubated for 24 hours at
30oC. The antibacterial activities of the extracts
Material were determined by measuring the clearance zone
Roselle calyxes were bought from surrounding the disc.
Beringharjo Market, Yogyakarta Indonesia. Solvent
used in this experience was ethanol (p.a, 101986, Minimum Inhibitory Concentration and
Merckmillipore, Germany). Minimum Killed Concentration (Doughari, 2006).
Ethanol extract with varying concentration
Extraction were added to 1 mL NB containing of test bacteria.
The dried roselle calyxes were ground for 1 Each of the solution was incubated at 37oC, 24 hrs.
minute using grinder and sieved in 60 mesh. The The lowest concentration of an antimicrobial that
extraction with various concentrations of ethanol will inhibit the visible growth of microoaganism is

24 Jurnal Teknologi Industri Pertanian 26 (1): 23-30

 Ike Sitoresmi M Purbowati, Khaswar Syamsu, Endang Warsiki, Herastuti Sri R

expressed as the MIC value. This clearance solution Ethanol was used in this research because it nontoxic
was mixed with NA medium and poured to the plate and relative commonly used. Methanol was not
incubated at 37oC for 24 hours. The Minimum Killed used in this investigation although having higher
Concentration was determined by the least dissipation factor. It could absorb the microwave
concentration extract without any bacteria growth. power and change to heat energy better, but the
toxicity was higher made it not food grade
Antioxidant Activity (Al Hashimi, 2012) (Hemwimon et al., 2007). For this reason, additions
0,6 mL of sample was dissolved in 0.12 mL ethanol with water have to be done. Water was
of 98% ethanol and 2.88 mL of a 2.51% linoleic acid added to reach the concentration as follows: 40, 50,
and 9 mL of a 40 mM phosphate buffer (pH 7.0) 60, 70, 80 and 90%. In these experiment, variables
were added. The mixture was incubating at 40⁰C in were used 250 Watt of microwave power and 5
a test tube in the dark for 3 days (72 hours). After minutes extraction time.
incubation, a 0.1 mL was taken from the mixture and The result of the research is shown in
diluted with 9.7 mL of 75% ethanol, followed by the Figure 1. Ethanol concentration influenced the
addition of 0.1 mL of 30% ammonium thiocyanate. yield, of phenols. The yield of phenol was
Precisely three minutes after adding the 0.1 mL of increasing with the increasing of ethanol
20 mM ferrous chloride in 3.5% hydrocloride acid, concentration. However the increase of ethanol
the absorbance of the red color was measured at 500 concentration above 80% made the yield of phenol
nm, using Spectrofotometer UV Vis 1800 Shimadzu. decreased. In extraction process, the polarity of
The level of lipid peroxidation inhibition by each solvent and the substances play an important role
fraction was calculated from the absorbance ratio to
that of a blank without any sample. A half lipid 14
peroxidation inhibition expressed with IC50.
Total Phenols (mg/g)
12
10
Antioxidant activity (%) = 8
Control absorbance - sample absorbance x 100% 6
control absorbance 4
2
0
Experimental Design 0 40 50 60 70 80 90
One response was measured: Total phenolic
yield (Y), defined as the ratio of total phenols in the Ethanol Concentration (%)
extract to total amount of raw material expressed as
GAE milligrams per gram of raw material (wet Figure 1. The effect of ethanol concentration to the
weight). Each of variables to be optimized was yield of total phenols
coded at 3 levels: -1, 0, and 1. A Central Composite
Design (CCD), was arranged to allow for fitting of a The polarity of solution is determined by its
second-order model. The model proposed for the dielectric constant. Markom (2007) stated that
response (Y) was: dielectric constant of water and ethanol respectively
were 80.20; 24.30 and according to Durst and
Y =β0 + βixi+ βiixi2 + βij xi xj + € Worldstad (2005) in roselle almost 80-90% phenols
content was anthocyanin. The polarity of
Where 0 was the value of the fitted response at the anthocyanin was classified as semi polar which is
center point of the design, which is point (0, 0, 0). 0, around 30-40 (Richter et al., 2006), mean that the
i, ii, and ij were the constant, linear, quadratic and polarity of phenols as a result of secondary
cross-product regression terms, respectively. metabolism on plant was fitted with the polarity of
ethanol 80%. It was decided to use the ethanol 80%
RESULT AND DISCUSSION for the next experiment.

The Effect of Ethanol Concentration on Total The Effect of Extraction Time to The Yield of
Phenol Extraction Phenols
The influencing factors on MAE were Extraction time is the factor which must
ethanol concentration, microwave power and time of have to be investigated to increase the effectiveness
extraction. To determine the range for each variable, of extraction. The research was done in varying
and degree polynomial should be used, the time: 1, 3, 5, 7, 9 and 11 minutes and 250 Watt of
influencing factors must be investigated separately microwave power. Figure 2 shows that the yield of
by doing the single experiment for every variable (Li total phenols is increasing until 5 minutes of
et al., 2009; Wang et al., 2010). extraction time. The extraction time longer than 5
Solvent selection, which is fitted with the minutes decreases the yield of phenols. The reason
method of extraction used, was the first step to of this condition is that the deterioration of phenols
develop the method of extraction included MAE. is depending on many factors including temperature.

Jurnal Teknologi Industri Pertanian 26 (1): 23-30 25

Optimization of Phenols Extraction from Roselle …………

The longer time this substance irradiated with

microwave power, it would increase the contact Optimization of MAE Condition of Phenol
duration between the bioactive compound and heat, This research evaluated 3 parameters,
causing the loss of phenols, although the temperature which are: (A) microwave power, (B) ethanol
was stable at 50⁰C. Chumsri et al. (2008) stated that concentration, and (C) extraction time. Based on
the rate of anthocyanin destruction depend on many the previous study, there were three level for each
factors such as pH, temperature, intermolecular parameter and second order model used, shown in
copigmentation ascorbic acid, and oxygen. The Table 1.
purpose of determining the extraction time was to
know the time needed for the absorption process on Table 1. Code and variable of experimental design
extraction with minimum loss of total phenols. In Variable Code -1 0 1
this case no need to do the extraction experiment Microwave power
more than 5 minutes. (Watt) A 175 250 325
25.00 Ethanol
Total phenols (mg/g)

Concentration %) B 70% 80% 90%

20.00
Time of extraction
15.00 (min) C 3 5 7
10.00
5.00 In order to reduce the number of
0.00 experiment, a Central Composite Design was used
0 1 3 5 7 9 11 (Table 2). In this way, only 20 experiment were
Time extraction (minutes)
necessary. Multiple regression analysis is following
second-order polynomial stepwise equation:
Figure 2. The effect of extraction time to the yield of
total phenols Y = 24.50 + 1.76A - 2.32B - 0.63C - 0.89AB +
0.48BC - 2.05A2 - 3.03B2 -1.39C2 + 2.00A2B +
The Effect of Microwave Power to The Yield of 0.93A2C
Phenols
Microwave power is the important The result of ANOVA is shown in Table 3.
parameter in microwave assisted extraction. The The model F-value of 145.86 implies that the model
varying microwave power used were 100, 175, 250, is significant. The coefficient of determination (R-
325 and 400 W, 80% ethanol for 5 minutes Squared) is the proportion of variability in the data
extraction time. Figure 3 shows that microwave explained or accounted for by the model. The “R-
power of 250 W was enough to the next experiment. Squared” of 0.9979 is desirable, mean that the model
was fitted for this case.
20
Total Phenols (mg/g)

It is shown from Table 3 that the calculated

15 of probability value (p) is smaller than 0.01.
Consequently, the model is statistically significant
10
for microwave power and ethanol concentration
5 factor. The interaction between microwave power
and ethanol concentration also significantly different
0
0 100 175 250 325 400
within 99% of confidence level but not for the
interaction of all factors. A quadratic change of
Microwave Power (W)
factor would significantly affect the response (Y).
Figure 3. The effect of microwave power to the yield Figures 4 shows three dimensional response
of total phenols surface was presented for the independent variables
(microwave output power, and ethanol
The extraction effectiveness improved concentration) which were obtained by keeping
when the microwave power increased from 100 to another variable (extraction time) constant. The
250 W. This condition can be explained that the figure indicate the changes in total phenols yield
higher microwave power resulted the higher effect of under different MAE conditions. Phenol substances
microwave energy on biomolecules by ionic just like anthocyanins could solute in polar solution
conduction and dipole rotation which results in ethanol, aceton and water. But from polarity level,
power dissipated inside the solvent and plant between phenol as salute and ethanol as solvent are
material and then generate molecular movement and disbalance.
heating (Chen et al., 2008). Microwave power
above 250 W decreased the yield of phenols.
Phenols are very sensitive to the heat according to
Gao et al. (2006) in higher microwave power (400-
1200W).

26 Jurnal Teknologi Industri Pertanian 26 (1): 23-30

 Ike Sitoresmi M Purbowati, Khaswar Syamsu, Endang Warsiki, Herastuti Sri R

Table 2. Matrix of central composite design

Unit Type A B C Total phenol
(Watt) (%) (Minute) (mg/g)
1 center 250 80 5 24.49
2 center 250 80 5 24.49
3 Fact 175 70 3 17.59
4 Fact 175 90 7 18.98
5 Fact 325 90 3 16.46
6 Fact 325 70 7 19.84
7 Fact 325 90 7 17.59
8 center 250 80 5 24.61
9 Fact 325 70 3 19.04
10 Fact 175 70 7 16.11
11 Fact 175 90 3 16.98
12 center 250 80 5 24.61
13 Axial 250 80 1.64 22.00
14 Axial 376.13 80 5 22.02
15 center 250 80 5 24.38
16 Axial 123.87 80 5 16.11
17 center 250 80 5 24.38
18 Axial 250 63.18 5 20.20
19 Axial 250 80 8.36 19.89
20 Axial 250 96.82 5 12.38

Table 3. ANOVA for the complete codified mode

Sum of Mean F p-value
Source df
Squares Square Value Prob>F
Block 0.76 2 0.38
Model 251.39 13 19.34 145.86 0.0001**
A-Power 17.46 1 17.46 131.72 0.0003**
B-Concent 30.58 1 30.58 230.62 0.0001**
C-Time 2.23 1 2.23 16.79 0.0149*
AB 6.28 1 6.28 47.39 0.0023**
AC 0.25 1 0.25 1.87 0.2428
BC 1.81 1 1.81 13.69 0.0208*
A2 60.56 1 60.56 456.79 <0.0001
B2 132.37 1 132.37 998.40 <0.0001
C2 27.67 1 27.67 208.67 0.0001
ABC 0.55 1 0.55 4.14 0.1116
A2B 13.30 1 13.30 100.34 0.0006
A2C 2.89 1 2.89 21.78 0.0095
AB2 6.02 1 6.02 45.44 0.0025
Ket: **significantly difference in 1% confidence level range
Design-Expert® Software

Total fenol
* significantly difference in 5% confidence level range
Design points above predicted value
Design points below predicted value
24.61

12.38

X1 = A: Daya 24.8
T o ta l P h e n o l ( m g /g )

X2 = B: Konsentrasi

Actual Factor 23.125

C: Waktu = 5.00

21.45

19.775

18.1

90.00 325.00
85.00 287.50
80.00 250.00

Ethanol Concentration (%) 75.00 212.50 Microwave Output Power (W)

70.00 175.00

(A) (B)
Figure 4. The Interaction between microwave power and ethanol concentration to the yield of total phenols (A)
Contour 3D, (B) Plotting Contour

Jurnal Teknologi Industri Pertanian 26 (1): 23-30 27

Optimization of Phenols Extraction from Roselle …………

The polarity of ethanol according Richter et 25

MAE
al. (2006) is 24, and the polarity of the phenol
substances and anthocyanins around 30-40. So, it 20 Conventional
needs addition of water to increase the extraction
efficiency. In the other side King et al. (2009) stated 15

mg/g
that the increasing of temperature will decrease
water polarity. Based from these reasons made the 10
influenced cause by the interaction between
microwave power and ethanol concentration is 5
significant different.
0
Optimization of Microwave-Assisted Extraction Phenols Anthocyanin Vit. C
The optimum conditions were obtained by
running the program of central composite design.
The optimum conditions for independent variables Figure 5. Phytochemical characterization
and the predicted values of the responses also are
presented as follows: 250 W microwave output 90
80

Antioxidant activity (%)

power, 78.36% ethanol concentration and extraction MAE
70
time 4.91 min. The estimated values for total
60
phenols yield, 24.61 mg/g was obtained at those
50 Conven
conditions. A verification experiment at the 40 tional
optimum condition, consisting of 4 runs, was 30
performed and the experimental yield of 23.77 + 20
0.25 mg/g total phenols higher than using 10
conventional method 19.84 + 0.46 mg/g was 0
obtained. 0 300 450 600
In this research, microwave assisted
Extract dose (µl/L)
extraction was compared with conventional
extraction techique using electricity boiling at 50°C
(temperature of MAE), 5 minutes extraction time, Figure 6. Antioxidant activity
and 78.36% of alcohol concentration. Those
variables are the optimum variables using MAE. In So that, the MIC and MKC from optimum
this research, the heat generated by the microwave at extraction, lower than the conventional ones (Table
the optimum condition are equal to 50°C based from 4). The results of this study clearly indicated that the
preliminary research. roselle extract inhibit the growth of tested
From the Figure 5, compared with microorganism, however the effectiveness are
conventional method, total phenol, anthocyanin, and varied. The antibacterial activity can be attributed to
vitamin C and yield of microwave assisted extraction the action of phytochemical. Phenols substance
were 23.77 + 0.25, 14.80 + 0.08, 10.74 + 0,14 mg/g known to be synthesized by plants in response to
and 22.09 + 3.3% respectively which are higher and microbial infection and it should not be surprising
significantly different within 99% confidence level that they have been found in vitro to be effective
than conventional extraction, which were 19.84 + antimicrobial substances against microorganism.
0.46; 9.28 + 0.04; 9.99 + 0.16 mg/g and 16.18 + Their activity is probably due to their ability to
1.9% respectively. complex with extracellular and soluble proteins and
Compared to the conventional method of to complex with bacteria cell walls. This results
extraction, MAE extraction produces a higher total agree with the previous (Al-Hashimi, 2012; Alonso
phenols, anthocyanin, total solid and vitamin C. The et al., 2006) who found that plant polyphenols have
fact that the increased extraction of phenols by the been demonstrated as potential antibacterial.
microwave power is related to the direct effects of
microwave energy on biomolecules by ionic Microscopic Characterization
conduction and dipole rotation which result in power In order to study cell damages during the
dissipated inside the plant material and the solvent convensional extraction (boiling in 50oC at the same
which generate the molecules movement and heating time and the same ethanol concentration) and MAE,
(Chen et al., 2008). roselle calyx samples were examined by mini SEM
The different of total phenol, anthocyanin (Scanning Electron Microscopy) with 7000x of
and vitamin C also made the different of antioxidant magnification. The result shows that there are
activity between the two extract, shows in Figure 6. structural difference between MAE and
Extract using MAE has IC50 value 202.47 µL/mL convensional extraction. Figure 7 shows that the
lower than conventional method which has IC50 surface of the sample is found to be greatly
293.09 µL/mL. destroyed after MAE.

28 Jurnal Teknologi Industri Pertanian 26 (1): 23-30

 Ike Sitoresmi M Purbowati, Khaswar Syamsu, Endang Warsiki, Herastuti Sri R

Table 4. Antibacterial activity of roselle extract

E. coli S. aureus
Method Clear zone MIC MKC Clear zone MIC MKC
(mm) (ppm) (ppm) (mm) (ppm) (ppm)
MAE 12.6 + 0,6 5500 6000 11.6 + 0.3 5500 6000
Conventional 10. 2 + 0.4 7000 7500 9.8 + 0.6 7000 7500

(a) Conventional extraction (b) MAE extraction

Figure 7. Cell of roselle powder after the extraction in 7000x of magnification

This observation suggests that the REFERENCES

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30 Jurnal Teknologi Industri Pertanian 26 (1): 23-30