Artikel Penelitian

Instant Powder Formulation for Anti Anemia and Optimazation

Exctraction Condition of Moringa pterygosperma Gaertn Leaves Using MAE

Yosita Anggraeni, Mahdi Jufri, dan Abdul Mun’in

ABSTRACT: Iron supplement containing ferrous sulfate is commonly used for anaemia.
Unfortunately, it has bad taste, can cause nausea, and made adverse effects if taken in large
doses for long periods. It is necessary to ind an alternative source of raw materials, including
those from plants. Moringa pterigospera Gaertn leaves was selected because it contains iron
and other nutritions. The purpose of this work was to make instant powder formula for anti
anaemia using Moringa leaves extract as an alternative for ferous suphate iron suplementation.
The extraction was performed by Microwave Assisted Extraction method. Optimization of
extraction condition was performed by creating some variations in solvent composition (0-
70% ethanol), microwave power (450 to 900 watts) and extraction time (3 to10 min). Iron
content was determined by Atomic Absorption Spectrophotometer at wave length of 248 nm.
Instant powder formula was made in 3 concentrations of sodium CMC as suspending agent.
Results of the study showed that the most optimal extraction condition which resulted the
highest iron content (2.4 mg/g extracts) achieved with 900 watts microwave power, 10 min
extraction time and aqua demineralisata. According to 30 panelists of hedonic test, formula
which used 5% w/w of sodium CMC got the highest scores. Its powder form had 2.31% of loss
on drying and 7.74 g/sec of low rate for powder form and pH of 5.78, viscosity of 15.98 cps for
reconstituted form.

Keyword: anti anemia, instant powder, iron, microwave assisted extraction, Moringa
pterygosperma Gaertn

ABSTRAK: Suplemen besi yang mengandung ferrous sulfat umum digunakan untuk anti
anemia. Sayangnya sediaan ini memiliki rasa tidak enak, menyebabkan mual dan jika
digunakan dengan dosis besar dan jangka waktu lama dapat menyebabkan efek samping.
Perlu dicari alternative sumber lain, termasuk dari tanaman. Daun Moringa pterigospera
Gaertn dipilih karena mengandung besi dan suplemen lain. Penelitian ini bertujuan untuk
membuat formulasi serbuk instan untuk anti anemia sebagai alternatif suplementasi
zat besi selain dari ferous sulfat, dengan menggunakan ekstrak dari daun kelor (Moringa
pterygosperma Gaertn). Ekstrak diperoleh dengan metode Microwave Assisted Extraction.
Optimasi kondisi dibuat dengan memvariasikan pelarut etanol (0-70%), daya listrik
microwave (450-900 watt) dan waktu ekstraksi 3-10 menit. Analisa kadar besi dilakukan
menggunakan Spektrofotometer Serapan Atom pada panjang gelombang 248 nm. Formula
serbuk instan dibuat dengan 3 konsentrasi natrium CMC sebagai bahan pensuspensi.
Penelitian menunjukkan bahwa kondisi optimal ekstraksi yang menghasilkan kandungan
besi paling besar (2,4 mg/g ekstrak) dicapai dengan daya listrik 900 watt, waktu ekstraksi
10 menit dan pelarut air suling. Berdasarkan uji hedonis dengan 30 panelis, formula dengan
5% natrium CMC paling disukai. Bentuk serbuk memiliki kadar air 2,31%, laju alir 7,74 g/
detik dan bentuk rekonstitusinya memiliki pH 5,78, dan viskositas 15,98 cps.

Fakultas Farmasi Kata Kunci: anti anemia, serbuk instan, besi, Microwave Assisted Extraction, Moringa
Universitas Indonesia pterygosperma Gaertn
Korespondensi:
Yosita anggraeni
Email: yosita.anggraeni@gmail.com

136 Jurnal Farmasi Indonesia ■ Vol. 7 No. 3 ■ Januari 2015

 Yosita Anggraeni, Mahdi Jufri, dan Abdul Mun’in

INTRODUCTION percolation and soxhlet [3], Supercritical Fluid

Extraction [4], re lux [5]. The method requires a
Iron supplements -currently available as anti- complicated process, a long time and required a
anemia- is commonly used raw materials ferrous lot of solvents, therefor, today was developed a
sulfate which has a bad taste, causing nausea, microwave extraction method. The advantages of
and if taken in large doses for long periods can using microwaves for extraction is the shorter ex-
cause heart failure [1]. Therefore, it is necessary traction and cooling time and use less solvent [6].
to ind an alternative of raw materials includ- To optimize the extraction process, some ex-
ing source from plants contain high iron, such perimental design optimization of extraction con-
as Moringa leaves. Moringa leaves is selected for ditions are made. The experiments are designed-
antianemia therapy because it contains iron and by modifying multiple extraction conditions, such
nutrients, such as protein [2] and abundant in as electric power of microwave, extraction time
Indonesia. The formula of instant powder made and solvent composition in order to obtain the
from Moringa leaves should instantly soluble in maximum iron content. Optimization of the in-
water, practical, easy to carry, easy to use and stant powder formula of Moringa leaves are also
“customer friendly”. Anti-anemia preparations conducted in order to obtain a formula that most
should contain suf icient iron content, therefore, consumers preferred. Some physical parameter-
it is necessary to ind the optimal extraction con- swere conducted to evaluate the instant powders
dition in order to obtain high iron content that including loss on drying, low rate and angle of
could be extracted from Moringa leaves. Several repose and for the reconstituted form, the tests
previous studies conducted for extracting the were pH, viscosity and hedonic test (colour, aro-
leaves of Moringa include decoction, maceration, ma and taste).

Figure 1 : Result of Residual test performed with GC-FID

Jurnal Farmasi Indonesia ■ Vol. 7 No. 3 ■ Januari 2015 137

Instant Powder Formulation from Moringa pterygosperma Gaertn Leaves

Figure 2 : Physicochemical test for instant powder

MATERIALS, INSTRUMENT AND METHOD Methods:

Materials: Preparation
Dried Moringa leaves obtained from Sukabu- Dried Moringa leaves were cleaned then blend-
miand determined by the Indonesian Institute ed to become a powder. The amount of 1,430 g
of Sciences, Bogor Botanical Gardens; n-hexane, of powdered Moringa leaves were then macerated
ethanol, and aquademineralisata (Brataco, Indone- with 4 liters of hexane for 24 hours. The process is
sia), iron standard solution (Merck, Germany), ni- repeated 7 times. The macerated powder was dried
tric acid (HNO3) (Merck, Germany), HClO4 (Merck, in drying cabinet.
Germany), manitol (Merck, Jerman), maltodextrin
(Zhucheng Dongxiao Biotechnology Ltd, China), Microwave Assistance Extraction Process
aerosil (Cabot Blue Star, China), D-mannitol (Merck, Before extraction process, the macerated powder
Jerman), CMC sodium (Dai-ichi, Kagyo), potassium was stirred using a magnetic stirrer for 10 min-
sorbate (local, Indonesia), lavor and coloring Grape utes until obtained a homogeneous mixture, then
(local, Indonesia). extracted by MAE method. The optimization was
applied by varying the type of solvent as follow: dis-
Instruments: tilled water, ethanol 30% , 50% and 70% ; varying
MAE - Microwave Assistance Extraction (Mode- the electrical power; and varying the time of extrac-
na MV Series) which had been modi ied by installing tion. The parameters were analyzed using d–opti-
additional double reverse condensor, Rotary Vac- mum design from Design Expert. As the result there
uum Evaporator (Buchi), destilation lask, Atomic were 29 types of extraction condition that will be
Absorption Spectrofotometer (Shimadzu AA-6300 made for the optimization with modi ication.
PC), GC-FID, Whatman # 42 ilter paper, waterbath,
magnetic stirer, thermometer, spatula, viscometer Testing for extract
Brook ield, analytical balance and moisture balance. Los on drying test using oven : The amount of 1

138 Jurnal Farmasi Indonesia ■ Vol. 7 No. 3 ■ Januari 2015

 Yosita Anggraeni, Mahdi Jufri, dan Abdul Mun’in

Table 1. Formula of Moringa Extract Granule

Ingredients Formula (gram) Function

Moringa leaves extract 1.375 Active Ingredient
Maltodextrin 3.321 Filler
Aerosil 0.704 Adsorbence
Total 5.400

Table 2. Instant Powder Formula

Formula (gram)
Ingredients
I II III
Moringa’s leaf extract dried granul 5.400 5.400 5.400
Mannitol 8.745 8.370 7.650
Sodium CMC 0.375 0.750 1.275
Grape’s lavour and colouring paste 0.450 0.450 0.450
Potassium sorbate 0.030 0.030 0.030
Total 15.00 15.00 15.00

gram sample were weighed and placed in a plati- mg single dose. The amount of 1.375 g of Mori-
num crucible, then heated in oven at 105 ° C for 3-5 nga extract was required to make a 3,33 mg single
hours. dose of iron (assuming that the content of Iron in
Total ash: The amount of approximately 1 gram the extract is 2.4 mg/g).
of sample was put in a platinum crucible and then Dried granules was prepared by absorbing
ignite slowly until the charcoal is depleted then Moringa leaves extract with aerosil then mixed
cooled and weighed . with maltodextrin until obtained a homogeneous
Acid insoluble ash : After determination of the mass. Then, dried granule in drying oven at 40°C
ash content, the sample was added 25 mL of H2SO4 for 6 hours, then crushed and sieved with mesh
10 % then heated 10 minutes to boil. the liquid is #40 .
iltered with Whatman ilter paper in a beaker glass. Instant powder made by adding sodium CMC
Whatman’s paper was then placed into the porce- as suspending agent, pasta lavor and colorants,
lain plate and put into the oven for ± 2 hours at a preservatives then stirred homogeneously. The
temperature of ± 600°C. The plate was allowed to mixture was then dried at a temperature of 40°C
stand in a desiccator for ± 1 hour. The porcelain for 2x6 hours, then crushed and sieved with a
plate is weighed until obtain the ix weight . mesh # 40. The powder was illed into alu-foil
Hexane residual test: using GC - FID instrument. sachet and stored in a tight container
Organoleptic test: visual observed to the appear-
ance, color , and taste. Evaluation of granules :
Identi ication of iron (qualitative) : react the il- Moisture content measured with a moisture
trate with 3 drops of NaOH, potassium hexacyano- balance, low rate and angle of repose, measured
ferrate (II), potassium hexacyanoferrate (III) by lowmeter

Instant Powder Preparation Evaluation of Reconstituted powder :

The recommended daily dose of iron is in the Reconstitution time, pH and viscosity and or-
range of 10 mg which coud be divided into a 3.33 ganoleptic test aroma, color, taste).

Jurnal Farmasi Indonesia ■ Vol. 7 No. 3 ■ Januari 2015 139

Instant Powder Formulation from Moringa pterygosperma Gaertn Leaves

Table 3. Iron Content Obtained Form The Optimization of Extraction Condition

Solvent Eletrical Power Extraction Time (minutes) Iron content

No
composition (Watt) Teoritic Actual (mg)
1 0 450 4.38 4.30 0.9865
2 0 450 5.97 6 0.9836
3 0 450 9.56 9.30 0.8824
4 0 630 3.55 3.30 0.8564
5 0 630 8.02 8 0.8733
6 0 630 10.00 10 0.9633
7 0 900 3.00 3 1.4406
8 0 900 6.23 6 0.9422
9 0 900 9.78 10 2.4046
10 30 450 5.34 5.30 2.0774
11 30 450 9.17 9 1.2672
12 30 630 3.99 4 0.9958
13 30 630 5.56 5.30 1.4650
14 30 630 9.56 9.30 2.3741
15 30 900 4.93 5 1.3211
16 30 900 6.48 6.30 1.5422
17 30 900 10.00 10 1.6070
18 50 450 5.21 5 1.3275
19 50 630 4.73 4.30 1.5342
20 50 630 4.73 4.30 1.3165
21 50 900 6.35 6.30 1.4952
22 50 900 7.26 7.30 1.2985
23 50 900 7.26 7.30 1.1058
24 70 450 7.05 7 0.8152
25 70 450 7.05 7 0.8227
26 70 450 7.05 7 0.8877
27 70 630 6.77 7 0.7403
28 70 630 6.77 7 0.7812
29 70 900 3.86 4 1.3160

RESULTS AND DISCUSSION stored in the refrigerator until a few months [7].
Furthermore, the ine moringa leaves powder
Dried Moringa leaves obtained from Suka- were macerated with hexane, to extract non-
bumi determined in the Indonesian Institute of polar compounds that may interfere the assay
Sciences, Bogor Botanical Gardens Conservation of the active substance (Fe) which are polar and
Center. The results of the determination states to eliminate bad odors of Moringa leaves. The
that the plant is Moringa pterygosperma Gaertn. essential oils contained in the leaves of Mor-
The value of nutritional contents in Moringa leaf inga were 1, 2-Benzenedicarboxylic acid, mono
powder is reported not change even if it is not (2-ethylhexyl) ester, nonacosane, heptacosane,

140 Jurnal Farmasi Indonesia ■ Vol. 7 No. 3 ■ Januari 2015

 Yosita Anggraeni, Mahdi Jufri, dan Abdul Mun’in

Table 4. Total Score of Hedonic Test

Parameter Formula 1 Formula 2 Formula 3

Texture 108 105 109
Colour 99 100 111
Aroma 103 112 107
Taste 93 107 68
Total 403 424 395

β-Amyrin [4]. Maceration process was repeated der Moringa leaves is higher than the amount of
for 7 times, since the solvent of the 7th process 16.42 mg/100 g dried Moringa leaves which was
began clear indicating that most of non-polar extracted used MAE method but without macer-
substances had been extracted. The rendemen ated with hexane [8]. However, it was lower than
of this hexane-maceration was 79.47 % . 28.2 mg/100 g of dry powder Moringa leaves
The regression liniear equation obtained obtained by Fuglie [2]. This is possibly due to
from several measurement of iron (Fe) reference the difference of extraction method as well as
standard solution at the concentration of 2, 4, the difference of geographical location where
6 and 8 ppm was y = 0,043755x - 0.0318 with r the plants grow.
= 0.9971. The visual test of the viscous extract Visual observation of the powder prepara-
showed a brown, hygroscopic with a distinctive tion showed that the formula 1, 2 and 3 formed
aroma and lavor of Moringa. The viscous extract ine granules, colored purple with the aromas of
was then dried in a water bath.The physical tests grape. After dissolving in water, the dark purple-
were conducted to the dried extract and result- slight grape aroma solution was formed. Physi-
ed loss on drying of 1.06 %, total ash of 12.76 %, cal low rate test of formula 1 and 2 showed a
acid insoluble ash of 1.69 %, the protein content good low ability between 4-10 g/sec, while the
of 0.18 % and the residual hexane of below 100 formula 3 showed a poor low ability between
ppm using GC-FID, which was far below the limit 1.6 to 4 g/sec. The angle of repose test show
290 ppm. that the formulas 1, 2 and 3 showed a good
Qualitative test identi icatioin with NaOH angle of repose which is between 25-300. The
produces a reddish brown color (positive Fe), reconstitution time test (with 200 mL water) of
using potassium hexacyanoferrate (III) pro- all formulas are below 1 minute, but with a large
duces a dark blue color (positive Fe3+), and us- standard deviation. It is probably in luenced by
ing potassium hexacyanoferrate (II) produced the manual reconstitution method which vary in
a light blue color (positive Fe2+).The presence of the stirring speed and strength.
iron in Fe2+and or Fe3+ form did not affect the The results of pH test of the reconstitution
anti-anemia function, as both can be absorbed form, showed a pH of 5.76± 0.01 of formula 1;
either in the body . 5.78 ± 0.02 of formula 2 and 5.81 ± 0.02 of for-
The data showed that the maximum iron con- mula 3. This difference were mainly caused by
tent was obtained under the extraction condi- the different amount of sodium CMC added into
tion as follow: Aqua demineralisata as solvent, the formula. The higher the amount of sodium
900 watt microwave power and 10 minutes ex- CMC, the higher value of pHwill increase. All for-
traction time which resulted of 2.4 mg iron/g ex- mulas have a pH near to 6 so it did not irritate
tract or 25.4 mg iron/100 g of dry powder Mo- the stomach. Viscosity test was performed us-
ringa leaves. Comparing to the previous study, ing Brook ield viscometer. The result is 9.61 cP
the iron content of 25.4 mg/100 g of dry pow- of formula 1; 15.98 cP of formula 2 and 80.03 cP

Jurnal Farmasi Indonesia ■ Vol. 7 No. 3 ■ Januari 2015 141

Instant Powder Formulation from Moringa pterygosperma Gaertn Leaves

of formula 3. CONCLUSION
A hedonic test was performed to the prepara-
tions before and after reconstitution. The param- The optimum condition for extraction of iron
eters tested were texture (before reconstituion), molecule from Moringa leaves using MAE is
color, aroma and taste (after reconstitution). 30 obtained by using aqua demineralisata solvent,
panelists were randomly selected, which aimed to 900 watt (P 100%) of microwave power and
reduce variables that may interfere the results [9]. 10 minutes of extraction time. This condition
In this study, the total of 30 people with vary in gen- resulted the maximum amount of Iron of 2.4 mg/
der and age as follow: female (80 %) and male (20 gram of extract. The formula 2 wich applying the
%), age of 16-20 years (36.7 %), 21-25 years (56.7 amount of 5% w/w of sodium CMC as suspending
%) and 26-30 years (6.7 %). The collected data was agentwas the most prefered formula selected by
analysed using a numerical scale (scoring ) ranged panelist with total score of 424. The formula has
from 1 to 5 The meaning of the value was as fol- a physical properties of 2.31% loss on drying,
lows: 1 = not like, 2 = less like, 3 = normal, 4 = like 7.74 g/sec of low rate, 5.78 of pH, and 15.98 cP
and 5 = very like [10,11]. All data were analyzed us- of viscosity.
ing SPSS version 17.

REFERENCES 6. Jain, Tripti., V Jain., R Pandey., A Vyas, and SS Shuk-

la. (2009). Microwave assisted extraction for phy-
1. Smolin, L. A. dan Mary, B. G. (2002). Nutrition from toconstituents–An overview. Asian Journal of Re-
science to life. Philadelphia: Harcourt College Pub- search in Chemistry. 2(1): Jan-March.
lishers, 440. 7. Verma, Arti R., Vijayakumar, M., Mathela, Chandra
2. Fuglie, LJ. (2001). The Miracle Tree: Moringa oleifera: S., Rao, Chandana V. (2000). In vitro and in vivo
natural nutrition for the tropics. Church World Ser- antiox-sidant properties of different fractions of
vice, Dakar. 68 pp; revised in 2001 and published as Moringa oleiferaleaves. Food Chemical Toxicology
The Miracle Tree: The Multiple Attributes of Moringa, 47 (2009) 2196-2201.
172 pp.http://www.echotech.org/bookstore/ad- 8. Triandita, Gita. (2013). Optimasi kondisi micro-
vanced_search_result.php?keywords=Miracle+tree. wave assisted extraction (MAE) terhadap kadar kal-
3. Boonyadist V., Pongtip S., Supachoke M., Suchitra T., siumdan zat besi dari daun kelor (Moringa oleifera
Yuvadee W., Wandee,G. (2013). Maximizing total phe- L). skripsi. Depok : Fakultas Farmasi UI.
nolics, total lavonoids contents and antioxidant ac- 9. Meilgaard, Morten C., Carr, B. Thomas., Civille, Gail
tivity of Moringa oleifera leaf extract by the appropri- Vance. (2007). Sensory Evaluation Techniques 4th
ate extraction method, Industrial Crops and Products Ed, USA : Chapman & Hall/CRC Taylor & Francis
44 : 566– 571 Group
4. Zhao, Suweiand Zhang, Dongke., Supercritical luid 10. Peryam, D.R. and Pilgrim, F.J. (1957). Hedonic scale
extraction and characterisation of essential oil from method of measuring food preferences. Food Tech-
Moringa oleifera leaves. Crawley WA : The University nology 9-14.
of Western Australia [diakses pada 19 April 2013 pk 11. Lim, Juyun (2011). Hedonic Scaling : A review of
13.11]. methods and theory. Food Quality and Preference
5. Siddhuraju, Perumaldan Becker, Klaus. (2003). Anti- 22 (2011) 733–747
oxidant properties of various solvent extracts of total 12. Rowe, R.C., P.J. Sheskey, dan S.C. Owen. (2009).
fenol constituents from three different agroclimatic Handbook of Pharmaceutical Excipients 6th Ed.
origins of drumstick tree (Moringa oleifera Lam) London : Pharmaceutical & American Pharmacist
leaves. Journal of Agri Food Chemistry 51: 2144-2155. Association.

142 Jurnal Farmasi Indonesia ■ Vol. 7 No. 3 ■ Januari 2015