ROSELLA FLOWERS PETAL EXTRACT (HIBISCUS SABDARIFFA,

LINN) TO IMPROVE ERYTHROCYT LEVELS OF FEMALE

ADOLESCENTS WITH ANEMIA

SRI SUMARNI 1, LULUK HANDAYANI 2, DONNY KRISTANTO 3

marninugroho@yahoo.com, lulukhandayani82@gmail.com, Donny.kristanto@yahoo.com
1
Poltekkes of Health Ministry Semarang
2
Master of Applied Midwifery, Poltekkes of Health Ministry Semarang
3
Research and Development Center of Health Magelang

ABSTRACT
Objectives : This research proved the use of providing Rosella petals, Hibiscus
Sabdariffa, Linn, to increase the erythrocyt levels for female adolescent with anemia.
Method: This true experiment with randomized pretest and posttest control group
involved 40 female adolescents as the participants. The researchers selected them based
on the inclusion criteria with randomization. The intervention group, twenty
participants, received 279 mg Rosella petal extract and Fe 60 mg tablet. Then, the
control group, consisting of 20 participants, received Fe 60 mg tablet and placebo for 1
x 1 in 14 days. The data analysis used paired t-test and independent t-test.
Results: The results showed the differences of erythrocyt levels of both groups. The
researchers found the erythrocyt level of intervention group 0.77 million/mm3 while the
control group was 0.35 million/mm3. The researchers also found significant statistics
differences between erythrocyt levels of female adolescents from both groups with
p-value < 0.05.
Conclusion : The administration of Rosella petals (Hibiscus Sabdariffa, Linn), was
effective increase the erythrocyt levels in blood for female adolescent with anemia.

Keywords : Rosella Petals, Erythrocyt Levels, Anemia

INTRODUCTION
Anemia is a nutritional problems
suffered by women, especially pregnant
women and female adolescents. Basic
Health Research (2018) found the
prevalence of anemia in Indonesia was
mostly observable on women, 27.3%.
They were from the category of 15-24
years old, 32%. From the total of 21
million female adolescents, 4.8 million of
them experienced lack of blood cell
(Kemenkes RI, 2018), (Badan Penelitian
dan Pengembangan Kesehatan, 2018). The
prevalence of anemia in Indonesia is
mostly caused by iron deficit with a
percentage of 50% (Bappenas, 2019).
Anemia on female adolescents
occurs due to menstrual bleeding so the
adolescents lose their iron for 0.4-0.5
mg/day. Besides that, the growth speed of
female adolescents is high, known as
growth spurt, that influences anemia.
Then, chronic disease, infectious disease,
and inadequate dietary pattern can also
cause anemia (Masthalina, 2015).
The provision of complementary
supplement to improve the blood has
many hindrances to manage anemia. One
of them is due to inadequate nutritional
consumption based on nutritional
adequacy rate (NAC) (Sari et al., 2020).
The nutritional consumptions, especially
inadequate iron and vitamin
consumptions, lower the effectiveness of
blood boosting tablets. Akib et al found
73.3% adolescents suffer anemia due to
inadequate vitamin C and iron
consumptions (Akib and Sumarmi, 2017).
For these reasons, the researchers
assumed Rosella petals had high iron and
vitamin C levels. The content levels of the
substances are 8.98 mg iron and 244.4 mg
vitamin C (Sembiring, Tanjung and Sabri,
2013). Rosella petal extract has higher
contents. They are 13.59 mg iron and
336.3 mg vitamin C per 100 gr Rosella
petal extract. The iron and vitamin C
contents of Rosella petal extracts are
higher than moringa leaves with iron level
of 6mg/100gr, long beans with
6.2mg/100gr, and red spinach 7mg/100gr.
The vitamin C level of the extract is
higher than leave with 32.41 mg/100 gr
and papaya with 78mg/100gr.
The bioactive compounds of th petal
is obtained from extracting process. The
researcher found the flavonoid and
anthocyanin bioactive compounds during
phytochemical screening test. The
compounds are mostly detected on
sorghum caudatum leaves to medicate
anemia (Agbangnan et al., 2012). The
anthocyanin stimulates the erythropoietin
production to influence the erythrocyt
production (Da-Costa-Rocha et al., 2014),
(Oktaviani, 2018). The administration of
Rosella petal extract for 279 mg/60 Kg of
weight, converted from the ratio of mice
and human weights, could manage
anemia. Sembiring found the
administration of 48 mg/20 Kg weight
was effective to increase the erythrocyt
levels of mice (Sembiring, Tanjung and
Sabri, 2013).
Therefore, this research aims to
prove the administration of Rosella petal
extract (Hibiscus Sabdariffa, Linn) to
improve the hematological blood of female
adolescents with anemia.
C METHODS
Research Design
This true experiment with
randomized pretest and posttest control
group design involved 40 participants.
Then, the researchers grouped them into
intervention and control groups.
The researchers used randomization
process to group the samples on each
group by preparing 40 envelops labeled A
20 and B 20. The researchers used the
envelopes randomly. All subjects that met
the criteria received the envelop with
conditions: they had to select the envelop
then the envelopes were opened by a
enumerator (Single Blind)
In this research, the label A was for
intervention while the label B was for
control group. The intervention group
received the extract with 279 mg and Fe 60
mg tablet for fourteen days. On the other
hand, the intervention group received Fe
60 mg tablet and placebo for fourteen days.
The researchers checked the erythrocyt
with hematology analyzer in IBL
Laboratory and a Laboratory in Srondol
Public Health Center.
The researchers obtained the grant
from the Health Research Ethical
Committee of Dr. Moewardi Regional
Hospital, Medical School, Universitas
Sebelas Maret, March 18, 2021, numbered:
333 /III /HREC /2021. The researchers
used informed consent to obtain the
willingness of the research subjects. This
research used parametric test, paired t-test,
for each group and independent t-test for
different groups
The Extraction Process of Rosella Petals
The researchers blended the dry
Rosella petals into simplicia forms. The
applied extraction method was UAE
macreation. It is a simplicia of 1 kg
ingredient with ethanol 96% to soak and
using ultrasonic assisted device 48 KHz +/-
30o. The researchers filtrated it to separate
the pulp and the maserate. The products,
the maserates of the petals that were
screened with bucher funnel, were steamed
with wind evaporator in 30-400
temperatures to remove the solvent
content, the ethanol.
The result was Roesella petal
extract, (Hibiscus Sabdariffa, Linn), in the
form of semi-solid materials with darker
color and condensed extract. The
researchers dried the extract until it was
condensed. After the crystallization
process, the researchers added the
simplicia residues in the condensed
extract based on the needs. Then, the
researchers mixed to make it more
homogeneous before dried with a food
dehydrator into powders. The extract, in
the form of powders, is ready to be
encapsulated. Then, capsule for the extract
is equal to 279 mg/60 Kg weight.

RESULT
Table 1
The Distribution of the Subjects’ Characteristics based on Menstruation Periods
and Food Intakes (Protein, Vitamin C, and Iron)
Characteristics Intervention Group Control group P value
Mean±SD Mean ± SD
Mentruation Periods 7 ± 0,51 6 ± 0,59 0,395b
Protein 50,2 ± 2,99 53,0 ± 2,41 0,131a
Vitamin C 66,7 ± 6,88 71,9 ± 7,64 0,637b
Iron 7,4 ± 0,99 8,3 ± 0,98 0,674a
a
Independent t-test, b Mann-Whitney
The table shows that both groups of protein intake show p > 0.05 for both
research subjects have normal menstrual groups. It indicates no differences of
categories. For 7 days in the intervention protein intakes of both groups.
group and 6 days in the control group. The Mann-Whitney test for the
The protein intakes show both subjects characteristics, in terms of
groups are normal. The vitamin C and iron menstruation period, vitamin C intake,
intakes of intervention group are not and iron intake of both group obtains a
normal while the control group’s intakes p-value > 0.05. It indicates no differences
are normal. of menstruation periods, vitamin C
The independent t-test of the intake, and iron intake of both groups.
subjects characteristics based on the
Table 2
The Normality Test Results of Erythrocyt
Level of Both Groups
Variables Intervention Group Control group
Mean±SD p-value Mean±SD p-value
Erythrocyt Pretest 3,40±0,14 0,264 3,38±0,13 0,075
(µ) Posttest 4,18±0,27 0,604 3,73±0,20 0,303
Δ 0,77±0,23 0,739 0,35±0,17 0,147
* shapiro-wilk

The normality test ensures the data before bivariate test. The researchers used
normality to examine the hypotheses
Shapiro-Wilk normality test because the groups obtains p > 0.05. It indicates the
sample was lesser than 50 members. data distribution is normal.
The normality test result with
Shapiro-Wilk on erythrocyt levels for both

Table 3.
The Level Changes of Erythrocyt levels before and after Intervention
of Intervention and Control Groups
Variables Intervention Group Control group
Mean ± SD Mean ± SD p-value
Erythrocyt level Pretest 3,40 ± 0,14 3,38 ± 0,14 0,570 b
(Million/mm3) Posttest 4,17 ± 0,27 3,73 ± 0,21 0,001 b
p-value 0,001a 0,001a
Difference 0,77 ± 0,23 0,35 ± 0,17 0,001 b
a
Paired t-test, bIndependent t-test
Erythrocyt level erythrocyt levels than the administration
Erythrocyt level of the intervention of Fe 60 mg tablet and placebo.
group increases higher than the control
group. The erythrocyt level after the DISCUSSION
administration of Rosella petal extract and Erythrocyt level
Fe tablet increases into 0.77 million/mm 3, Rosella, Hibiscus Sabdariffa, Linn,
higher than the control group with Fe and is a medicinal plant. The petal is useful to
placebo administrations,0.35 million/mm3. cure anemia in Benin. The researcher
The paired t-test of erythrocyt in the found the flavonoid and anthocyanin
intervention group obtains a bioactive compounds during
p-value < 0.05, indicating significant phytochemical screening test (Obouayeba
differences of before and after the et al., 2015). Most compounds are
intervention with 279 mg Rosella petal available sorghum caudatum leaves The
extract and Fe 60 mg tablet for 14 days. applied to medicate anemia (Agbangnan
The paired t-test of erythrocyt in the et al., 2012). The flavonoid contents in
control group obtains a p-value < 0.05, Rosella are gossypetin, hibisten and
indicating significant differences of before sabdaretin. Some metabolite compounds
and after the intervention with Fe 60 mg are also available in Rosella. They can
tablet and placebo for 14 days. increase the erythrocyt (Da-Costa-Rocha
The independent t-test obtains p- et al., 2014) (Oktaviani and Megantara,
value < 0.05 for the erythrocyt levels of 2018).The anthocyanin stimulates the
both groups. It indicates significant erythropoietin production to influence the
differences of both groups. erythrocyt production.
Thus, the administration of 279 mg Sembiring administered Rosella
Rosella petal extract per day and Fe 60 mg extract for mice, Mus Musculus, L
per day was effective to increase the suffering anemia with three variants of
doses. They were 14 mg, 28 mg. and 42
mg for 14 days. The results showed 42 mg
administration was more effective to
increase the erythrocyt than the other
doses. The average of the increasing rate
for 42 mg dose is 20.23 cells/mm 3 while
the 28 mg and 14 mg doses are 15.87
cells/mm3 and 14.50 cells/mm3. The
statistics test of both groups obtained a p
value < 0.05. It proved the extract could
increase the mice’s erythrocyt with the
increasing extract concentration on the
intervention group. In this research, the
dose was converted from the mice itno
human with more effective dose by
Sembiring. The dose was 42 mg/20Kg
weight. After the dose conversion from
mice to human, the dose of the Rose petal
extract was 279 mg (Sembiring, Tanjung
and Sabri, 2013).
Ghislain found drinking Rosella
petal extraction was significant toward the
hematological parameter with p-value <
0.05. The increased erythrocyt shows that
drinking Rosella petal extraction
containing phytochemical can trigger the
erythropoietin production (Ghislain et al.,
2014) .
A previous study by Nurjannah
administered long bean leave extract for
female adolescents in 14 days with 200
mg doses can increase the erythrocyt for
0.53 million/mm3. The increasing
percentage was 13.9%, p-value = 0.013,
and size effect 0.25 (weak) (Nurjanah,
Hadisaputro and Fatmasari, 2020).
Sari administered Rosella petal
extract on Sprague Dawley female mice
with oral dose of 850.90 mg/Kg weight. It
did not show any mortality symptoms and
toxic effects during the delayed
observation in 14 days (Sari, Nurkhasanah
and Bachri, 2016). Setianingsih found that
the administration of red Rosella ethanol
extract with 500mg dose in 30 days could
increase erythrocyt level without toxic
effect. It was found after 30 days of
observation on the consumption of
Rosella petal ethanol extraction and
during the 15 days of observation on
healthy participant after the consumption.
In this research, the female
adolescents did not experience the side
effects during the consumption. This
research was done in the evening so the
female adolescents could have meals
before consuming the extract. It was
important due to the high vitamin C
content. Thus, Rosella is relatively safe to
consume.
The current research and the
previous studies, with the same ingredient
and research period except the dose, 279
mg/60Kg weight in 14 days for female
adolescents with anemia, was effective to
increase erythrocyt levels. The average of
erythrocyte level average, 4.18
million/mm3, showed that the healthy
respondents, with normal erythrocyte
level, had 0.77 million/mm3 difference
with a percentage of 22.6% and effect size
2.0, strong effect. This research is
different with Nurjannah because the long
bean leave extract administration, 200mg,
in 14 days for female adolescents with
anemia, obtained an averages of 4.34
million/mm3, increasing rate 0.53
million/mm , increasing rate 13.9%, and
3
effect size 0.25 (weak effect). This current
research has higher increasing rate and
effect size than the previous study because
of the different vitamin C and iron
contents. It might be also caused by other
factors that could trigger the numbers of
erythrocyte. The use of Rosella petal
extract was more effective to increase the
erythrocyte than the use of brewing
method with hot water.
CONCLUSION
The administration of Rosella petal
extraction, Hibiscus Sabdariffa, Linn, 279
mg and Fe 60 mg tablet in 14 days
significantly increase the erythrocyt levels
of female adolescents with anemia. It was
higher than the administrations with Fe 60 FUNDING
mg and placebo for control group. Nol
The administration of Rosella petal
extract, Hibiscus Sabdariffa, Linn, as side AUTHORS CONTRIBUTIONS
herbal therapy program with Fe tablet All the authors have contributed equally
could improve the anemia status. CONFLICT OF INTEREST
The authors declare no conflict of interest
ACKNOWLEDGMENT
Thanks to the chief of the Integrated
Laboratory of Diponegoro University, the
IBL Laboratory, and Laboratory of
Srondol public health to support this
research.
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