JURNAL FARMASI SAINS DAN KOMUNITAS, month year, pp Vol. X No.

X
p-ISSN 1693-5683; e-ISSN 2527-7146
doi:

Antioxidant activity of Rosa damascene flos ethanol extracts using

hydroxyl and nitrite oxide scavenging methods
Elfina Br, T1., Chrismis, N. G2*., Linda, C2., I Nyoman, E. L2
1
Master Programsof Biomedical Science, Universitas Prima Indonesia, Medan, Indonesia
2
Faculty of Medicine, Universitas Prima Indonesia, Medan, Indonesia

* Corresponding author: Chrismis, N. G

email: [email protected]

co-Author email:
Author2: [email protected]
Author3: [email protected]
Author4: [email protected]

ABSTRACT

The purpose of this study were to determine antioxidant activity of Rosa damascene
flos ethanol extracts. The ethanol extracts were extracted from the crown of rose and rose
base by maceration using ethanol 70% solvent. Antioxidant activity was determined with
hydroxyl and nitrite oxide scavenging methods and the IC50 analyzed using SPSS 23. The
IC50 of crown rose ethanol extract (CREE) and rose base ethanol extract (RBEE) on
hydroxyl and nitrite oxide scavenging were 7.61 ± 0.38μg/mL, 17.55 ± 0.37 μg/mL and were
349,57 ± 0,35μg/mL, 54.93 ± 4.49 μg/mL.The crown of rose ethanol extract (CREE) and rose
base ethanol extract (RBEE) has activity as antioxidant.

Keywords: Antioxidant; Rosa damascene flos; hydroxyl; nitrite oxide

*Corresponding author: corresponding author’s name

Email: corresponding author’s email
 Jurnal Farmasi Sains dan Komunitas, year, vol(no), pp

INTRODUCTION
Rosa damescene of the family Rosaceae, is one of the most important commercial
flower crops with over 150 species, more than 20 000 cultivars andwith colour spectre
ranging from subtle whites, yellows and pinks to intense purple, orange and red tones. Its
flower colour is attributed to the presence of anthocyanins and carotenoids (Cai et al, 2005).
The crown of rose have been consumed as a food ingredient in teas, cakes, and flavour
extracts as well as medicinal remedies of various illnesses (Lin et al, 2014). The rose flower
is known as an astringent, stomachic, and is used traditionally as an agent for activating blood
circulation to relieve blood stasis, and counteracting toxin. Being rich in anthocyanin content,
rose petals are a good colourant and potentially a good source of antioxidants (Poonam et al,
2017).

Antioxidant activity of a plant is important because of two reasons. First the

consumption of a food rich in antioxidants has been suggested to prevent or delay oxidation
of major biomolecules within the cell by chelating metals or scavenging free radicals that are
produced as consequences of metabolism (Hatice et al, 2016). Free radicals can defined as
some free entities having one or more unpaired electrons which play a vital role in the
development of various human diseases including aging (Sanchez et al, 1999). Antioxidant
constituents can protect the human body from free radicals such hydroxyl radicals (OH) and
nitrite oxide radicals (NO) (Su et al, 2017). Secondly antioxidants are used in food
preservation to prevent the food from oxidation, and increase their shelf life by retarding the
process of lipid peroxidation, which is one of the major reasons for deterioration of food and
pharmaceutical products during processing and storage (Karuna et al, 2017).

There are several ways to test the strength of antioxidant activity of a natural products,
including scavenging OH and NO as free radicals. OH is considered as one of the most
reactive spesies, it can attack biomolecules and cause irreversible damage (Filippi et al,
2019). While NO is a free radical that plays a role in the process of vasodilation,
inflammation and the immune system. Decreasing the number of OH and NO radicals with
scavenging method can inhibit cell damage9.Several constituents are present in the R.
damascena including flavonoids, anthocyanins, terpenes and glycosides, which have useful
effects on body. The investigation confirmed flavonoids and other contents in R. damascene
has antioxidant effect (Akram et al, 2019).

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METHODS
Plant and chemicals materials
Rosa damascena floswere collected from Parsoburan Village, Toba Samosir, North
Sumatra, Indonesia. Rosa damascene floswas identified in Herbarium Medanense (MEDA)
Universityof Sumatera Utara. The chemicals materials used in this study weresodium
nitroprusside (Sigma), sulphanilamide (Sigma), phosfat acid (Merck), N-(1-naphtyl)
ethylenediamine dihydrochloride (Sigma), Ethanol (Merck), ferrous ammonium sulfate
(Merck), hydrogen peroxide (Merck), buffer phosfat, L-ascorbic acid (Sigma), deoxyribose
(Sigma), trikloroasetat acid (TCA) (Merck), tiobarbiturat acid (TBA) (Merck) and aqudest.
Preparation of CREE and RBEE
The air-dried and powdered flos of Rosa damascene(Lour,) (500 g) were repeatedly
macerated with ethanol 70% (3x3 d, 7.5 L), The filtrate was evaporated with a rotary
evaporator with a temperature of ± 40oC to give a viscous extract (Satria et al, 2019).
Scavenging OH Assay of CREE and RBEE
Enter 2 µl of the sample into well blank and well sample. Add 10 µl of FeCl 3-EDTA
to the sample well and control well. Add H2O2 of 5 µl to the sample well and control well.
Add 5 mL of L-Ascorbic Acid 1 mM to the sample well and control well. Add 10 µl of
deoxyribose to the sample well and control well. To well blank, add 120 µl buffer. To the
well control, add 70 µl buffer. To the sample well, add 69 µl buffer. Incubation plate for 30
minutes at 37°C. Add 25% 25 µl TCA solution to the sample well and control well. Add a
25% 25% TBA solution to the sample well and control well. The plate for incubated 30
minutes at 80°C. Absorbance was measured using a microplate reader at λ = 532 nm (Bhat et
al, 2019). The equation to determine scavenging activity:

Scavenging NO Assay of CREE and RBEE

Enter 10 µl of the sample into well blank and well sample. Add 40 µl SNP to the
sample well and control well. To well blank, add 140 µl ethanol. To the well control, add 10
µl ethanol. The plate incubated for 2 hours at room temperature. Add 100 µl Greiss solution
to sample well and control well. Absorbance was measured using a microplate reader at λ =
546 nm (Parul et al, 2013).The equation to determine scavenging activity:

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Statistical Analysis
The results were presented as means ± SD. The statistical analysis was carried out
byusing SPSS edition 23.

RESULTS AND DISCUSSION

Antioxidant activity of CREE and RBEE, it is begin from extraction Rosa

damascenaflos. The extraction results can be seen in Table 1. Crown rose and base rose is a
part of Rosa damascenaflos were extracted by maceration methods using ethanol 70%. CREE
and RBEE effect as antioxidant was carried out by OH and NO scavenging methods.The
comparison of antioxidant test results through NO scavenging by CREE and RBEE was
showed in Table 2. There was a statistically significant difference followed by the Post Hoc
test using the Tukey HSD method. The average percentage of NO scavenging antioxidant
activity by CREE was higher than by BREE. The results showed an increase that was in line
with the high concentration. While antioxidant activity of CREE and BREE using OH
scavenging method showed the same results. The was showed in Table 3, CREE has better
antioxidant OH scavening activity compared to BREE. Based on the data above, it can be
calculated the IC50 value of each test sample. IC50 results can be seen in Table 4.

CREE and RBEE effect as antioxidant was carried out by OH and NO scavenging
methods. The average percentage of NO scavenging antioxidant activity by CREE was higher
than by BREE. At concentrations of 2.08 μg/mL, 4.17 μg/mL, 8.33 μg/mL, 16.67 μg/mL,
33.33 μg/mL and 66.67 μg/mL CREE each has an antioxidant activity of 28.33 ± 4.30%,
32.83 ± 1.96%, 34.46 ± 2.23%, 38.59 ± 1.79%, 46.62 ± 2.90%, and 64.30 ± 0.90%. BREE at
the same concentration each had antioxidant activity of 24.10 ± 1.39%, 27.74 ± 2.32%, 28.67
± 2.84%, 31.75 ± 2.49%, 40.27 ± 0.84% and 55.45 ± 1.47%.

Antioxidant activity of CREE and BREE using OH scavenging method showed the same
results. CREE has better antioxidant OH scavening activity compared to BREE. CREE with
concentrations of 0.83 μg/mL, 1.67 μg/mL, 3.33 μg/mL, 6.67 μg/mL, 13.33 μg/mL and 26.67
μg/mL each have antioxidant antivitivity 36.32 ± 5.27%, 36.34 ± 3.80%, 42.63 ± 1.12%,
50.37 ± 1.75%, 61.73 ± 1.80%, and 85.59 ± 0.65%. BREE compounds at the same
concentration each had antioxidant activity of 19.60 ± 1.85%, 23.79 ± 1.30%, 27.34 ± 3.32%,
33.05 ± 0.97%, 44.00 ± 1.01% and 64.29 ± 1.14%.

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The content of chemical compounds in roses is a strong reason that roses are
antioxidants. Rose contains various active compounds including tannin, geraniol, nerol,
citronellol, geranic acid, terpenes, flavonoids, polyphenol pectin, vanillin, carotenoids,
stearopten, farnesol, eugenol, pheniletilakohol, and vitamin C in the fight against free
radicals. According to the results of the present study, the fresh and spent flower extracts
obtained from Rose damascena could be a good natural antioxidant source (Baydar and
Hasan, 2013). Based on the data above, it can be calculated the IC50 value of each test sample.
IC50 results can be seen in Table 4.

IC50 value was obtained through the calculation of the absorbance value using the
linear regression equation y = a + bx by comparing the extract concentration with the NO and
OHscavenging values. In the table 4, it can be seen that the IC50 value of NO scavenging
activity shows that the CREE has an IC50 value was 39.29 ± 0.47 µg/mL smaller than the
BREE which has an IC50 value was 54.93 ± 4.49 µg/mL. IC50 CREE was also smaller when
compared to BREE in OH scavenging testing. IC 50 value of CREEwas 7.61 ± 0.38 μg/mL
while BREE was 17.55± 0.37 μg/mL. IC50 values indicate the sample concentration needed to
inhibit 50% of free radical activity. The smaller the IC 50 value produced, the better the ability
of a compound in free radical scavenging activities (Widowati et al, 2014).

CONCLUSION

Based on the results we obtained ethanol extract of Rosa damascena flos had a
potentially antioxidant activity.

ACKNOWLEDGEMENT

We gratefully thank to Universitas Prima Indonesia for support in the study.

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Table 1. Extract content of CREE and RBEE

No Sample Gram

1 CREE 40.50

2 RBEE 53.67

Description: The samples were extracted by maceration method. Each samples weight used was 250 gram.

Table 2. NO scavenging activity of CREE and BREE

Concentration NO scavenging activity (%)

(ug/mL) CREE BREE

2.08 28.33±4.30a 24.10±1.39a

4.17 32.83±1.96ab 27.74±2.32ab

8.33 34.46±2.23ab 28.67±2.84ab

16.67 38.59±1.79b 31.75±2.49b

33.33 46.62±2.90c 40.27±0.84c

66.67 64.30±0.90d 55.45±1.47d

Description: The difference in superscript letters (a, ab, b, c, d) in the same column shows a significant
difference at P <0.05 (Tukey HSD post hoc test).

Table 3. OH scavenging activity of CREE and BREE

Concentration NO scavenging activity (%)

(ug/mL) CREE BREE

0.83 36.32±5.27a 19.60±1.85a

1.67 36.34±3.80a 23.79±1.30ab

3.33 42.63±1.12ab 27.34±3.32b

6.67 50.37±1.75b 33.05±0.97c

13.33 61.73±1.80c 44.00±1.01d

26.67 85.59±0.65d 64.29±1.14e

Description: The difference in superscript letters (a, ab, b, c, d) in the same column shows a significant
difference at P <0.05 (Tukey HSD post hoc test).

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Table 4. IC50 values of CREE and BREE

IC50 (µg/mL)
Sample
NO scavenging OH scavenging

CREE 39.29 ± 0.47 7.61± 0.38

BREE 54.93 ± 4.49 17.55± 0.37

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