Formulation and evaluation of serum from

red, brown and green algae extract for anti-

aging base material
Cite as: AIP Conference Proceedings 2175, 020078 (2019); https://doi.org/10.1063/1.5134642
Published Online: 20 November 2019

Melati Septiyanti, Lilis Liana, Sutriningsih, Bayu Kumayanjati, and Yenny Meliana

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© 2019 Author(s).
Formulation and Evaluation of Serum from Red, Brown and
Green Algae Extract for Anti-aging Base Material

Melati Septiyanti1,a), Lilis Liana2,b), Sutriningsih2,c), Bayu Kumayanjati3,d), and

Yenny Meliana1,e)

1
Research Center for Chemistry, Indonesian Institute of Sciences, Kawasan Puspiptek Serpong, South Tangerang,
Banten, Indonesia 15314.
2
Faculty of Pharmacy, Universitas 17 Agustus 1945, Sunter Agung, North Jakarta, DKI Jakarta, Indonesia 14350
3
Research Unit for Natural Product Technology, Indonesian Institute of Sciences, Jln. Yogya - Wonosari km 31,5
Gading, Playen, Gunungkidul, Yogyakarta, Indonesia
.
a)
Corresponding author: mela003@lipi.go.id
b)
lilis.liana98@gmail.com
c)
vinnelaras@yahoo.co.id
d)
bayu011@lipi.go.id
e)
yenn001@lipi.go.id

Abstract. Red, brown and Green algae contain antioxidant and have the potential as anti-aging but still rarely developed
for cosmetic ingredients. The purpose of this study was to investigate the chemical constituent, total phenol and flavonoid
content, antioxidant activity and toxicity of algae extract and formulated into serum preparation. Padina Australis, Ulva
Reticulata, Sargassum Oligocystum, dan Euchema Cottonii contain chemical constituent hexadecanoic acid, methyl ester,
fatty acid and phytol which possess antioxidant activity. P. australis has the highest total phenol and flavonoid content
that is 106 mgGAE/100 gr and 30.516 mgQE/g respectively. Antioxidant activity of P. australis, U. reticulata, E. cottoni
and S. oligocystum is stated in IC50 value which are 2693.33, 2679.16, 4407.57 and 1700 μg/ml respectively wherein it
still considered as low antioxidant activity. Algae extract in this study also proven to have low toxicity with LC50 higher
than 1000 ppm. Algae crude extract was formulated into anti-aging serum formulation and evaluated for one month by
accelerated stability test method. This stability test observed physical properties pH, viscosity, refractive index, and
specific gravity and chemical properties. Based on the stability test results on each anti-aging serum formulas, formula
with E. cottoni extract showed the stable formulation for serum preparation with no significant chemical and physical
change due to temperature alteration during accelerated stability test.

INTRODUCTION
Recently, aging become a concern and many research to prevent premature aging has been developed in order to
improve the quality of life [1]. Aging process is different on each individual based on many factors which can affect
and accelerate aging [2]. Aging is a biological process that can be affected by internal factors such as genetic, cell
metabolism and hormones. The external factors are expose of sunlight, pollution, radiation, chemicals and toxin.
These factors cause structural and physiological change which visible in skin appearance. Main target of anti-aging
research is to prevent oxidative damage and collagen metabolism. UV-B expose cause free radical formation of

Proceedings of the 5th International Symposium on Applied Chemistry 2019

AIP Conf. Proc. 2175, 020078-1–020078-11; https://doi.org/10.1063/1.5134642
Published by AIP Publishing. 978-0-7354-1922-3/$30.00

020078-1
Radical Oxygen Species (ROS) which is an unstable molecule and can be formed in all epidermis layer and even
deeper tissue [3]. ROS can damage cell component and cause premature aging on skin marked with dry skin,
wrinkle and dull. To prevent those effect, it is needed cosmetic preparation contains antioxidant to neutralized ROS
[4].
Phaeophyceae (P. australis and S. oligocystum) showed highest antioxidant activity among Rhodophyceae and
Chlorophyceae. Phenolic compound and its derivatives are the main antioxidant component in Phaeophyceae [5].
Phenolic compound contained in brown algae is phlorotanin for about 0.74%-5.06%. This compound which able to
capture free radical caused by UV radiation [6]. E. cottoni is one of red algae that contain protein, lipid,
carbohydrate, α tocopherol, mineral, vitamin C and vitamin E [7]. It is also known contain phenolic compound and
flavonoid such as catechin, flavonols, flavonol glycosides, caffeic acid, hesperidin, myricetin which act as
antioxidant [8].
Green algae commonly contain chlorophyll A and B and carotene compound which act as antioxidant. U.
reticulata forsskal is rich of vitamin A, B1, C and fatty acid. Commonly green algae produce terpenoid compound
and aromatic compound as anti-inflammation, anti-microbe, anti-virus, anti-mutagen and insecticide [9]. In previous
study, antioxidant activity of P. australis had IC50 value of 87.082 ppm and E. cottoni 106.021 ppm [10]. While S.
oligocystum has IC50 value of 219.68 ppm and U. reticulata 365.95 ppm [9].
Indonesia is rich for marine biota such as algae that has not much used as medicines and cosmetics [11]. Thus, in
this study red, brown, and green algae which are E. cottoni, P. australis, U. reticulata, dan S. oligocystum was
extracted and formulated into serum preparation as anti-aging cosmetics raw material. Serum is liquid preparation
with low amount of solvent and high bioactive agent [12]. This formula is easy to apply into skin and has comfort
sensation. It has high concentration thus it can give effective effect because it contain high concentrate active agent
and easy to absorb by skin [13].
This study investigated the extraction of red, brown and green algae and its formulation into serum preparation.
Algae extract was characterized with total phenol and flavonoid, toxicity, antioxidant activity and chemical
constituent using Gas chromatography-mass spectrometry (GCMS). The stability of algae serum was tested using
accelerated stability test method in temperature 5oC and 40oC alternately for 1 month. The parameter tested were
Foutier Fourier Transform Infra Red (FTIR), Viscosity, pH, density, refractive index and spreadability.

METHODOLOGY/EXPERIMENTAL

Material

The materials used in this study were macro algae P. australis, U. reticulata, S. oligocystum, dan E. cottonii
from Tual Coast, Maluku, Indonesia, ethanol for analysis, natrium bicarbonate from Merck, Germany, carbomer,
glycerine, triethanolamine (TEA) technical grade from Brataco, Indonesia, euxyl® PE 9010 from Schülke,
Germany, nanosilver with size 24 nm and deionized water.

Experimental Procedure
Simplicia of P. australis, U. reticulata, E. cottonii, S. oligocystum was from Dulah, Tual Coast, Maluku,
Indonesia. The sample was dried in oven at 50oC and then grinded into small particle. The algae extraction was done
by maceration method. 100 g of prepared algae sample was immersed in 600 ml of ethanol pro analysis while stirred
150 rpm for 24 hours this method was done twice. Algae extract solution was filtered using filer paper and then
concentrated using vacuum rotary evaporator at 40oC. From the extraction process obtained crude extract with
physical appearance S. oligocystum dark brown, P. australis green, E. cottoni light green, U. reticulata green. Yield
result of algae extraction P. australis, U. reticulata, E. cottonii, S. oligocystum respectively 1.18%, 1.3%, 1.12% and
1.7%.
Algae crude extract was tested with parameters antioxidant activity, total phenol, flavonoid, toxicity and GCMS
from Agilent, USA, to analyze the chemical constituent. Antioxidant activity was done using 2,2-Diphenyl-1-
picrylhydrazyl (DPPH) method. 1 ml DPPH with concentration 100 μg/ml was added by 3 ml methanol. The extract
samples were diluted in methanol with concentration 1000 ppm, 750 ppm, 500 ppm and 250 ppm, then all samples

020078-2
incubated at 37oC for 30 minutes. Each sample was measured its absorbance using spectrophotometer UV-Vis from
Agilent, USA, with wavelength 517 nm. Antioxidant parameter was expressed with IC50 value [14].
Total phenol content was done by using follin ciocalteu method. Gallic acid as standard was diluted with
methanol with concentration 0, 10, 20, 30, 40, 50 ppm. 25 mg algae extract sample was diluted with methanol 25 ml.
The absorbance is measured with wavelength 765 nm. Total flavonoid content was done using quercetin as standard
with methanol as solvent with concentration 0, 10, 20, 30, 40, 50 ppm. 25 mg algae extract sample was diluted with
methanol 25 ml. The test is using the mixture of NaNO2, AlCl3 and NaOH. The sample was measured using
spectrophotometer UV-Vis with wavelength 510 nm [15].
The toxicity test used Brine Shrimp Lethality Test (BSLT) method. 2.5 mg Artemisia salina leach eggs was
aerated for 48 hours. The samples were diluted with concentration 2000 ppm, 1000 ppm, 200 ppm, 100 ppm dan 0
and put into the well, then 10 larvae were inserted into the well. Plate well was incubated for 24 hours and the dead
larva was counted. The toxicity of the sample was expressed using LC50 value [16].
The algae crude extract was formulated into serum preparation. The composition of algae serum is shown in
Table 1. Algae extract serum was made by dispersing carbomer in warm water with speed 150 rpm using
mechanical stirrer until gel phase formed. Natrium bicarbonate, nanosilver and etanol 96% was mixed and then
added into gel phase. Euxyl® and glycerin was added into compound until homogen. Algae extract was diluted with
ethanol 96% then added into serum base and stirred for 30 minutes, later TEA was added gradually with speed 500-
700 rpm. The stability of the algae serum stability was tested by accelerated stability test method with temperature
alteration 5oC and 40oC every 24 hours for 4 weeks then the parameter tested was chemical properties using FTIR
Prestige-21 Shimadzu, Japan, viscosity using DV-E viscometer Brookfield USA, pH using pH meter Eutech 700,
USA, density using picnometer, refractive index using refractometer Atago RX5000α, Indonesia and spreadability
[17], [18].

TABLE 1. Algae Extract Serum Formulation

Formula (wt.%)
Components
I II III IV
P. australis extract 0.2 - - -
U. reticulata extract - 0.2 - -
E. cottoni extract - - 0.2 -
S. oligocystum extract - - - 0.2
Natrium bicarbonate 0.02 0.02 0.02 0.02
Carbomer 1.5 1.5 1.5 1.5
Glycerine 10 10 10 10
Euxyl® 0.5 0.5 0.5 0.5
Nanosilver 0.3 0.3 0.3 0.3
TEA 0.3 0.3 0.3 0.3

RESULTS AND DISCUSSION

Chemical Constituent of Algae Extract

The chemical constituent of algae extract was analysed by using GCMS. GCMS chromatogram for each algae
extract is shown in Figure 1 and the compound is listed in Table 2. It is seen on the table the main constituent of P.
australis is n-Hexadecanoic acid, U. reticulata is 2-Hexanol Butane, E. cottoni is Hexadecanoic acid, methyl ester
and S. oligocystum is 2-Hexanol Butane. Fatty acid group is a major compound in algae, fatty acid can be act as
antibacterial because its properties which is similar like surfactant in bacteria cell membrane thus inhibit the protein
sintesis. Fatty acid also acts as antioxidant because easily to oxidize. Ghavam et al (2015) reported that fatty acid
group such as hexadecanoic acid can be used as sunscreen, emollient, moisturizer and cleanser [19]. Hexadecanoic

020078-3
acid, methyl ester that contained in P. australis and E. cottoni has bioactivity as antioxidant, hypocolesterolemic and
5-alpha reductase inhibitor [20]. Methyl stearate, cyclodecasiloxane, eicosamethyl, n-hexadecanoic acid, ethyl oleate
which contained in U. reticulata, E. cottoni and S. olygocystum, included in fatty acid group which have antioxidant
and antibacterial activity. Phytol in P. australis is diterpenoid group which has antioxidant, antibacterial, anti-
inflammation activity [21].

(A) (B)

(C) (D)

FIGURE 1. GCMS Chromatogram Algae Extract (A) P. australis (B) U. reticulata (C) E. cottoni
(D) S. olygocystum

Total Phenol, Flavonoid Content and Antioxidant Activity of Algae Extract

Based on the GCMS analysis, algae possess chemical constituent which has antioxidant activity. Further to
quantify the activity of the algae extract, the total phenol, flavonoid and antioxidant test was established, and the
result is shown in Table 3. Total phenol was tested using follin-ciocalteu method. Gallic acid used as standard in this
test because gallic acid is the derivative form hidroxibenzoic acid which is classified as simple phenol and has been
discovered to be equivalent to phenolics content in a substance [22]. Maximum wavelength for total phenol using
follin-ciocalteu is 765 nm. Standard curve equation from gallic acid was y=0.0876x + 0.023 with coefficient

020078-4
determination R2 0.9987. Further this standard curve was used in total phenol calculation represent in Gallic Acid
Equivalent (GAE) mgGAE/100g as shown in table below.

TABLE 2. Algae Extract Chemical Compound from GCMS Analysis

Name of Compound Retention Time Area %

1-methoxy-2-hexanol 4.499 11.07
Acetamide 9.485 1.73
n-Hexadecanoic acid 19.832 16.54
P. australis Hexadecanoic acid, ethyl ester 20.159 6.05
Phytol 21.282 2.40
Ethyl oleate 21.801 8.18
Cyclotrisiloxane 26.580 4.79
2-Hexanol Butane 4.495 15.02
1,2-Bis(trimethylsilyl) benzene 4.658 8.95
U. reticulata Cyclopentene 6.158 3.59
Sarcosine, N-(cyclohexylcarbonyl)-
8.389 2.43
heptyl ester
Hexamethyl Cyclotrisiloxane 26.480 3.42
1-methoxy-2-methyl-2-hexanol 4.482 4.77
Heptadecane 17.098 5,16
E. cottoni Octadecamethyl cyclononasiloxane 19.391 3.29
Hexadecanoic acid, methyl ester 19.492 6.33
Methyl stearat 20.200 3.55
2-Hexanol Butane 4.495 18.15
cyclopentene 6.158 3.48
S. olygocystum Cyclooctasiloxane, hexadecamethyl 16.178 1.67
Cyclononasiloxane, octadecamethyl 17.879 1.88
Cyclodecasiloxane, eicosamethyl 19.391 1.52

TABLE 3. Total Phenol, Flavonoid and IC50 of Algae Extract

Total Phenol Content Total Flavonoid Content

Sample IC50 (ppm)
(mgGAE/100g) (mgQE/g)
P. australis 106 30,516 2693,33
U. reticulata 91 24,008 2679,16
E. cottoni 40 2,433 4407,57
S. oligocystum 78 12,333 1700

It is seen that the highest total phenol content is in P. australis. P. australis which is brown algae contain higher
total phenol content compared that red algae [23]. Phenolic content depends on its chemical structure. Phenolic
group which has many hydroxyls functional group produce high total phenol content. This substance works as
antioxidant through termination of radical reaction chain and donate hydrogen atom of which the stable radical
formed [24]. Phenolic content in algae varies based on season, cultivation, geographic location, and species type
[25].

020078-5
 Total Flavonoid test principle is the reaction between flavonoid and AlCl3 with the addition of NaOH forms a
complex which its absorbance can be measured in wavelength 510 nm [26]. Quercetin is used as standard in total
flavonoid test and stated in QE (Quercetin equivalent). Regression of quercetine standard curve equation is y =
0,0012x + 0,0342 with coefficient of determination R2 0,9876. The standard curve was used to calculate total
flavonoids content as shown in the table above.
The highest flavonoid content is in P. australis which is 30,516 mg QE/g. Flavonoid is secondary metabolite
which is found in brown, red and green algae. Antioxidant compound correlated with phenolic content in algae.
Flavonoid act as antioxidant through donation of hydrogen atom [27]. Plant with flavonoid content has antioxidant,
antibacterial, anti-inflammation activity [28].
Antioxidant was investigated on each alga extract with DPPH method. The sample concentration was varied 250
ppm, 500 ppm, 750 ppm and 1000 ppm. Quercetine concentration as positive control is 12.5 ppm, 25 ppm, 50 ppm,
75 ppm. All sample was measured with wavelength 515 nm. The result of absorbance, % inhibition and IC50 is
shown in the table.
IC50 Quercetin as positive control is 34.33 ppm. A substance is said to be very active antioxidant if the IC50 value
is less than 50 ppm [29]. The result in the table showed the antioxidant activity of ethanolic algae extract was range
between 1700-4000 ppm. The highest IC50 value is S. oligocystum compared other algae extract which is 1700 ppm.
Based on Molyneux, IC50 value of 200-1000 ppm was stated less active yet still has potential antioxidant activity
[30]. Overall result, ethanolic algae extract contained phenolic and flavonoid substances yet has relatively low
antioxidant activity. Ethanol as polar solvent attract flavonoid components that attached to glycine which absorbed
in polar solvent. Phenolic also extracted into polar solvent due to the presence of hydroxyl group. Antioxidant will
potentially extracted using semi-polar and non-polar solvent [31].

Toxicity of Algae Extract

To make sure that the algae extract is not toxic and safe for human use, the toxicity test was conducted. Toxicity
test was done using BSLT method, the toxicity level was identified from mortality of shrimp larva with sample
concentration 20 ppm, 200 ppm, 1000 ppm and 2000 ppm. LC50 value of P. australis, U. reticulate, E. cottoni, S.
oligocystum, and respectively 2511886,43 ppm, 10000 ppm, 5011,87 ppm and 15848,93 ppm. Based on previous
study by Sharo et al (2013) the LC50 of E. cottoni was 58.0128 ppm which is toxic [32]. This is due to the difference
Dimethyl Sulfoxide (DMSO) solvent amount added into the well. DMSO can be toxic to A. salina leach if the
amount of DMS is more than 0.75%. Generally, the value of LC50 is higher than 1000 ppm and it is considerate that
the algae extract is not toxic. The plant extract is said to be toxic when the mortality of Artemia salina leach larva is
50% in sample concentration less than 1000 ppm. The extract with low toxicity can be developed into food,
supplement and cosmetic raw material [9]. Further, this algae extract was formulated in serum preparation for anti-
aging use.

Stability of Algae Serum Formulation

The formulation of serum used each algae extract. The composition is shown in Table 1. Serum composition
contained carbomer which belong to acrylate acid compound with hydrophilic and stable properties. This base was
chosen because it has good compatibility with all the ingredient contained in the formulation and it is good to be
cosmetic ingredient because it doesn’t leave trace and crack and also give comfort when it applied [33]. The stability
test was done by accelerated stability test method where the serum of algae extract was stored at alternating
temperature 5oC and 40oC every 24 hours for 1 month and the parameters were observed every week. The stability
test is correlated with the durability of algae extract serum through storage time. The parameter evaluated in this
stability test are physical properties such as viscosity, pH, density, spreadability, refractive index and the chemical
properties by IR spectra analysis. The result of stability test for 1 month observation from physical properties
parameter is shown in Table 4. The chemical groups observed before and after stability test is shown in Figure 2.

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 TABLE 4. Algae Extract Accelerated Stability Test Observation for 4 Weeks

F1 pH Viscosity (cPs) Spreadability (cm) Refractive Index Density (g/ml)

1 5.83 874 5.5 1.33844 10,118
2 6.31 982 5.2 1.33848 10,127
3 5.58 1138 5 1.33878 10,081
4 5.74 986 5.7 1.33901 10,110
St Dev 0.31 109 0.3 0.00027 19.916
Mean 5.87 995 5.4 1.33868 10,109
RSD (%) 5.36 10.91 5.81 0.02 0.20

F2 pH Viscosity (cPs) Spreadability (cm) Refractive Index Density (g/ml)

1 5.54 1274 3.7 1.34318 10,200
2 5.79 1299 3.7 1.34318 10,209
3 5.44 1338 4.2 1.34383 10,203
4 5.36 1318 4.8 1.34378 10,194
St Dev 0.19 27 0.5 0.00036 6.245
Mean 5.53 1307.25 4.1 1.34349 10,202
RSD (%) 3.38 2.09 12.75 0.03 0.06

F3 pH Viscosity (cPs) Spreadability (cm) Refractive Index Density (g/ml)

1 4.9 832 5.3 1.34555 10,267
2 5.62 890 5 1.34601 10,271
3 5.42 979 5 1.34618 10,256
4 4.7 966 5.9 1.34638 10,282
St Dev 0.43 69 0.4 0.00035 10.739
Mean 5.16 916.75 5.3 1.34603 10,269
RSD (%) 8.36 7.50 8.00 0.03 0.10

F4 pH Viscosity (cPs) Spreadability (cm) Refractive Index Density (g/ml)

1 5.26 346 5.9 1.34473 10,267
2 5.42 384 5.8 1.34501 10,271
3 5.18 451 5.4 1.34544 10,256
4 5.2 438 6 1.3456 10,282
St Dev 0.11 49 0.3 0.00040 10.739
Mean 5.27 404.75 5.8 1.34520 10,269
RSD (%) 2.07 12.04 4.55 0.03 0.10

The pH analysis aimed to investigate the safety of serum preparation when it is applied thus the serum
preparation doesn’t irritate the skin [34]. Standard pH range for topical preparation is 4.5-6.5 [35]. The value of pH
for all algae extract serum in week-1 is suitable with skin pH but when it comes to week-4 the tendency of pH is
lower than the first week. It is because the release of hydrogen ion or ion contamination in serum preparation
through the extreme temperature alteration between 5oC and 40oC. The change of pH can be affected by several
factors like temperature, the improper storage and active agent instability due to oxidation [35]. The preparation

020078-7
must have certain pH value because if the pH is too low it cause skin irritation meanwhile if it is too high it cause
the skin dry and scaly [36].
The viscosity of the preparation affected by several factors such as mixing process, thickener agent selection and
particle size [37]. Each of algae extract gave different consistency. After the accelerated stability test the viscosity
become higher than the initial condition. High viscosity of the preparation caused by the shear force from the mixing
process. Shear force can change polymer structure become tenuous; thus it has low viscosity after mixing. After
storage, the polymer structure change and back to the stable structure and it became more viscous [38]. After test
condition on week 4 the viscosity of all preparation tend to decrease. Viscosity value of F1, F2 and F3 in range of -
900-1150 cPs while formula F4 has the lowest viscosity. Refractive index result support viscosity parameter because
refractive index directly proportional with viscosity. As seen on the table, refractive index increases as the viscosity
increase. The density also corresponds to viscosity, as seen on the table there is no significant change before and
after accelerated stability test.
Spreadability is the ability of preparation to spread while applied in skin. Spreadability inversely proportional
with viscosity. The good formulation has spreadability diameter 5-7 cm. The widest spreadability, the more active
agent can give the effect due to broad contact with skin [39]. From the cycle test the spreadability didn’t change
significantly. The decrease of spreadability tendency caused by the increase of molecule unit because it absorb the
solvent and increase the resistance to spread and flow [40]. From the Table 4 it is seen that formula F2 shows poor
spreadability due to high viscosity. In week-4 the spreadability improved due to storage time [39].

(A) (B)

(D)
(C)

FIGURE 2. FTIR Spectra Before and After Accelerated Stability Test of Serum (A) P. australis (B) U. reticulata (C) E.
cottoni (D) S. olygocystum

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 TABLE 5. FTIR Functional Group of Algae Extract Serum Before and After Accelerated Stability Test

Functional P. australis U. reticulata E. cottoni S. oligocystum

Group Before After Before After Before After Before After
-OH 3626.33 3603.18 3554.96 3492.27 3612.83 3477.80 3634.71 3544.35
C=C 1624.13 1624.13 1634.74 1634.74 1664.64 1637.67 1624.42 1633.78
C-O 1041.61 1041.61 1042.57 1041.61 1041.61 1043.53 1040.64 1042.57
C-H 734.91 698.26 746.48 726.23 742.63 639.43 712.73 731.05

The chemical groups of the serum preparation before and after accelerated stability test is shown in Figure 2 and
the list of functional groups is shown in Table 5. On P. australis extract serum before and after the cycle test it is
seen OH stretch with wide peak in wavenumber 3600 cm-1 and it is clarified by the presence of high intensity in
wavenumber 1041 cm-1 which shows C-O bending. This two absorbance indicate OH alcohol presence which bond
to carbon atom. Wavenumber 1600 cm-1 indicate C=C aromatic functional group, this corresponds to absorption in
wavenumber between 600-700 cm-1 which indicate C-H aromatic functional group. From the IR spectra it is
represent that serum from P. australis extract contain phenolic and flavonoid compound. This is marked by
functional group O-H and C=C.
In Serum U. reticulata it is seen OH stretch in wavenumber 3500 cm-1, correspond with C-O bending in
wavenumber 1040 cm-1 with high intensity. The wavenumber around 1600 cm-1 denote typical functional group
C=C aromatic along with low intensity of C-H aromatic in wavenumber 740 cm-1. Similar with P. australis, U.
reticulata serum shows phenolic and flavonoid functional group marker OH group and several aromatic group with
C=C functional group.
E. cottoni serum also has broad peak of OH stretch in 3600 cm-1 and C-O alcohol bending in wavenumber 1040
cm-1 with high intensity. C=C aromatic bond also appeared by absorption in wavelength 1600 cm-1. In S.
oligocystum also shown similar IR spectra profile where there is a presence of OH stretch in wavenumber 3600 cm-1
and also C-O alcohol bending at wavenumber 1040 cm-1. It is also seen absorbance of C-H aliphatic at wavenumber
730 cm-1. C=C stretch of alkene also seen at wavenumber 1600 cm-1. This also correspond to the presence of
phenolic and flavonoid compound with typical OH functional group and C=C aromatic group.
It can be concluded that the chemical groups in serum algae wasn’t affected by temperature alteration during
accelerated stability test for 1 weeks. From the stabilization observation the formula with the smallest value of
relative standard deviation (RSD) is Formula 3, serum preparation with E. cottoni extract.

CONCLUSION
An evaluation of red, brown and Green algae extract in serum formulation has been done. The major constituent
of P. australis, U. reticulata, S. oligocystum, dan E. cottonii contain hexadecanoic acid, methyl ester, fatty acid and
phytol which possess antioxidant activity. It is also found P. australis has the highest total phenol and flavonoid
content that is 106 mgGAE/100 gr and 30.516 mgQE/g respectively. The highest antioxidant activity is S.
oligocystum with IC50 value 1700 μg/ml which still considered as low activity but still has potential. All algae
extract in this study also proven to have low toxicity with LC50 higher than 1000 ppm. The most stable algae serum
is formulation with E. cottoni from parameter evaluation pH, viscosity, refractive index, and density and chemical
properties with no significant chemical and physical change due to temperature alteration during accelerated
stability test proven by the smallest RSD value.

ACKNOWLEDGMENTS

The authors would like to thank Research Center for Chemistry, Indonesian Institute of Sciences (LIPI) for
supporting this research in laboratory practice, testing and analysis. Faculty of Pharmacy, Universitas 17 Agustus
1945 for laboratory practice support. Conservation Unit for Tual Marine Life, Indonesian Institute of Sciences for
providing algae raw material.

020078-9
 REFERENCES

1. V. M. Monnier and D. R. Sell, “ Prevention and Repair of Protein Damage by the Maillard Reaction In Vivo ,”
Rejuvenation Res., vol. 9, no. 2, pp. 264–273, 2006.
2. W. Cunningham, “Aging and Photo-aging,” in Textbook of cosmetic dermatology, 5th ed., R. Baran and H. I.
Maibach, Eds. CRC Press, 2017.
3. K. M. Hanson and R. M. Clegg, “Observation and Quantification of Ultraviolet-induced Reactive Oxygen
Species in Ex Vivo Human Skin,” Photochem. Photobiol., vol. 76, no. 1, pp. 57–63, 2002.
4. M. Wilkinson and R. Harry, Harry’s Cosmeticology, 7th ed. New York: Chemical PUblishing Company, 1982.
5. R. Putranti, “Skrining fitokimia dan aktivitas antioksidan ekstrak rumput laut sargassum duplicatum dan
turbinaria ornata dari Jepara,” Universitas Diponegoro, 2013.
6. K. Chojnacka, “Biologically Active Compounds in Seaweed Extracts - the Prospects for the Application,”
Open Conf. Proc. J., vol. 3, no. 1, pp. 20–28, 2012.
7. B. D. Wandansari, L. Agustina, and N. S. Mulyani, “Fermentasi rumput laut Euchema cottonnii oleh
Lactobacillus plantarum,” Chem Info, vol. 1, no. 1, pp. 64–69, 2013.
8. Y. Yumiko, H. Ya Pei, and S. Takeshi, “Distribution of flavonoids and related compounds from seaweed in
Japan,” J. Tokyo Univ. Fish., vol. 89, pp. 1–6, 2003.
9. S. Tamat, T. Wikanta, and L. Maulina, “Aktivitas antioksidan dan toksisitas senyawa bioaktif dari ekstrak
rumput laut hijau ulva reticulata forsskal,” J. Ilmu Kefarmasian Indones., vol. 5, no. 1, pp. 31–36, 2007.
10. F. Maharany, Nurjanah, R. Suwandi, E. Anwar, and T. Hidayat, “Kandungan Senyawa Bioaktif Rumput Laut
Padina australis dan Eucheuma cottonii Sebagai Bahan Baku Krim Tabir Surya,” Jphpi, vol. 20, no. 1, pp. 10–
17, 2017.
11. M. A. Gammone, G. Riccioni, and N. D’Orazio, “Carotenoids: Potential allies of cardiovascular health?,” Food
Nutr. Res., vol. 59, no. 7, pp. 1–11, 2015.
12. F. A. Simion, E. S. Abrutyn, and Z. D. Draelos, “Ability of moisturizers to reduce dry skin and irritation and to
prevent their return,” J. Cosmet. Sci., vol. 56, pp. 427–444, 2005.
13. T. Mitsui, New Cosmetic Science. Amsterdam: Elsevier, 1996.
14. A. M. Pisoschi and G. P. Negulescu, “Methods for Total Antioxidant Activity Determination : A Review,”
Biochem. Anal. Biochem., vol. 1, no. 1, pp. 1–10, 2011.
15. H. L. Wildenradt and V. L. Singleton, “The Production of Aldehydes as a Result of Oxidation of Polyphenolic
Compounds and its Relation to Wine Aging,” Am. J. Enol. Vitic., vol. 25, no. 2, pp. 119 LP – 126, Jan. 1974.
16. Z. Zuraida, “Toxicity Analysis of Forestry Plants Using Brine Shrimp Lethality Test ( BSLT ) Method,” J.
Penelit. Has. Hutan, vol. 36, no. 3, pp. 239–246, 2018.
17. W. K. Restu, Y. Sampora, Y. Meliana, and A. Haryono, “Effect of Accelerated Stability Test on
Characteristics of Emulsion Systems with Chitosan as a Stabilizer,” Procedia Chem., vol. 16, pp. 171–176,
2015.
18. Y. Meliana and M. Septiyanti, “Karakterisasi Sediaan Topikal Anti Aging Dari Kombinasi Ekstrak Pegagan
Dan Kulit Buah Manggis,” J. Sains Mater. Indones., vol. 17, no. 4, pp. 178–183, 2016.
19. M. Ghavam, “Chemical composition of the leaf essential oil of Smirnovia iranica Sabeti from Iran,” J. Herb.
Drugs, vol. 5, no. 4, pp. 175–178, 2015.
20. M. Sermakkani and V. Thangapandian, “GC-MS analysis of Cassia italica leaf methanol extract,” Asian J.
Pharm. Clin. Res., vol. 5, no. 2, pp. 90–94, 2012.
21. Sheeba Gnanadeebam and P. Viswanathan, “GC-MS Analysis of Phytocomponents in Spermacoce articularis
L . f . leaf,” Res. Pharm., vol. 4, no. 4, p. 2015, 2015.
22. V. L. Singleton, R. Orthofer, and R. M. Lamuela-raventos, “Analysis of Total Phenols and Other Oxidation
Substrates and ANtioxidants by Means of Folin-Ciocalteu Reagent,” Methods Enzymol., vol. 299, pp. 152–178,
1999.
23. P. Matanjun, S. Mohamed, N. M. Mustapha, K. Muhammad, and C. H. Ming, “Antioxidant activities and
phenolics content of eight species of seaweeds from north Borneo,” J. Appl. Phycol., vol. 20, no. 4, pp. 367–
373, 2008.
24. S. B. Nimse and D. Pal, “Free radicals, natural antioxidants, and their reaction mechanisms,” RSC Adv., vol. 5,
no. 35, pp. 27986–28006, 2015.

020078-10
25. G. Rajauria, B. Foley, and N. Abu-Ghannam, “Identification and characterization of phenolic antioxidant
compounds from brown Irish seaweed Himanthalia elongata using LC-DAD–ESI-MS/MS,” Innov. Food Sci.
Emerg. Technol., vol. 37, pp. 261–268, 2016.
26. L. Jack and A. Rohman, “Daya antioksidan ekstrak etanol Daun Kemuning ( Murraya paniculata ( L ) Jack )
secara in vitro Antioxidant potency of ethanolic extract of Kemuning,” Maj. Farm. Indones., vol. 16, no. 3, pp.
136–140, 2005.
27. M. S. and C. H. I. Cuppett, S., “Natural Antioxidant – Are They Reality,” in Natural antioxidants, chemistry,
health effect and application, F. Shahidi, Ed. Illinois: AOCS Press, 1954, pp. 12–24.
28. R. Gusnedi, “Analisis Nilai Absorbansi dalam Penentuan Kadar Flavonoid untuk Berbagai Jenis Daun
Tanaman Obat,” Pillar of Physics, vol. 2, pp. 76–83, 2013.
29. M. Marjoni and A. Zulfisa, “Antioxidant Activity of Methanol Extract / Fractions of Senggani Leaves,”
Pharm. Anal. Acta, vol. 8, no. 8, pp. 1–6, 2017.
30. P. Molyneux, “The use of the stable free radical diphenylpicryl- hydrazyl ( DPPH ) for estimating antioxidant
activity,” Songklanakarin J. Sci. Technol., vol. 26, no. 2, pp. 211–219, 2004.
31. A. Sushant, M. K. Baniya, K. Danekhu, P. Kunwar, R. Gurung, and N. Koirala, “Total Phenolic Content,
Flavonoid Content and Antioxidant Potential of Wild Vegetables from Western Nepal,” Plants, vol. 8, no. 96,
pp. 1–12, 2019.
32. N. M. Sharo, R. Ningsih, A. Nasichuddin, and A. Hanapi, “UJI TOKSISITAS DAN IDENTIFIKASI
SENYAWA EKSTRAK ALGA MERAH ( Eucheuma cottonii ) TERHADAP LARVA UDANG Artemia
salina LEACH,” Alchemy, vol. 2, no. 3, pp. 170–177, 2013.
33. M. Rabisková, M. Sedláková, M. Vitková, and M. Kuna, “Carbomers and their use in pharmaceutical
technology,” Ceska Slov. Farm., vol. 53, p. 300, 2004.
34. M. Anief, Ilmu meracik obat. Yogyakarta: Gadjah Mada UNiversity Press.
35. R. I. Tranggono and F. Latifah, Buku pegangan ilmu pengetahuan kosmetik. Jakarta: PT. Gramedia Pustaka
Utama, 2007.
36. S. Ansari, “Skin pH andskin folra,” in Handbook of cosmetics science and technology, 3rd ed., New York:
Informa Healtcare, 2009, pp. 222–223.
37. D. Ansel, Pengantar bentuk sediaan farmasi. Jakarta: UI Press, 1989.
38. A. S. Martin, Farmasi Fisik, 3rd ed. Jakarta: UI Press, 1983.
39. K. Sayuti and R. Yenrina, Antioksidan Alami dan Sintetik. Padang: Andalas University Press, 2015.
40. N. M. . Sukmawati, C. I. . Arisanti, and N. P. A. . Wijayanti, “Pengaruh Variasi Konsentrasi PVA, HPMC, dan
Gliserin terhadap Sifat Fisika Masker Wajah Gel Peel-Off Ekstrak Etanol 96% Kulit Buah Manggis (Garcinia
mangostana L.),” J. Farm. Udayana, vol. Vol. 2, no. No. 3, pp. 35–42, 2013.

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