Effect of Sodium Nitrite (NaNO2) to Erithrocyte and Hemoglobin Profile in White Rats (Rini Ambarwati)

EFFECT OF SODIUM NITRITE (NaNO 2 ) TO ERITHROCYTE AND HEMOGLOBIN PROFILE

IN WHITE RAT (Rattus norvegicus)

Rini Ambarwati
Nursing Department, Health Polytechnique
Ministry of Health, Surabaya

ABSTRAK

Natrium Nitrit (NaNO 2 ) merupakan bahan pengawet makanan yang cenderung disalahgunakan. Keracunan Natrium Nitrit yang
sering terjadi adalah berkurangnya kemampuan hemoglobin untuk mengikat oksigen dan terbentuknya reactive oxygen species
(ROS) yang dapat menyebabkan perubahan jumlah eritrosit dan kadar hemoglobin. Penelitian ini bersifat eksperimental laboratoris
dengan menggunakan Post Test Control Group Design untuk mengetahui efek pemberian Natrium Nitrit terhadap profil eritrosit
yang meliputi jumlah eritrosit, kadar hemoglobin pada tikus putih. Sampel 33 tikus (Rattus norvegicus) jantan berat 175 -200 gram
dibagi menjadi 3 kelompok masing-masing terdiri dari 11 tikus. Kelompok 1 adalah kelompok kontrol sedangkan kelompok 2 adalah
kelompok yang diberikan Natrium Nitrit dosis 0,5 mg/200 gram BB tikus dan kelompok 3 adalah kelompok yang diberikan Natrium
Nitrit dosis 1,5 mg/200gram BB tikus selama 30 hari. Hasil penelitian menunjukan ada perbedaan yang bermakna p=0,000 (p<0,05)
pada jumlah eritrosit dan kadar hemoglobin. Pemberian Natrium Nitrit 0,5 mg/200gram BB tikus yang merupakan dosis aman sudah
menunjukkan perubahan profil eritrosit dan dosis Natrium Nitrit 1,5 mg/200gram BB tikus menunjukkan perubahan pada jumlah
eritrosit dan kadar hemoglobin yang lebih besar. Kesimpulan dari penelitian ini adalah pemberian Natrium Nitrit dengan dosis 0,5
mg/200gram BB tikus dan 1,5 mg/200gram BB tikus dapat menurunkan jumlah eritrosit dan kadar hemoglobin tikus putih
jantan.(FMI 2012;48:1-5)

Kata kunci: Natrium Nitrit, eritrosit, hemoglobin.

ABSTRACT

Sodium Nitrite (NaNO 2 ) is a food preservative that tend to be misused. Poisoning from Sodium Nitrite that often occurs reduces
hemoglobin ability to bind oxygen and the formation reactive oxygen species (ROS) that can changes erythrocyte count and
hemoglobin level. This was an experimental laboratories study using Post Test Control Group Design to identify the effect of Sodium
Nitrite administration on the profile of erythrocytes, including erythrocytes count, and hemoglobin levels in white rat. Samples
comprised 33 male rats (Rattus norvegicus) weight 175 -200 grams, divided into 3 groups, each consisting of 11 rats. Group 1 was
the control group, group 2 received Sodium Nitrite of 0.5 mg/200 grams BW rats and group 3 received Sodium Nitrite of 1.5 mg/200
gram BW rats for 30 days. Results of research showed that there was a high difference in means p=0.000 (p<0.05) on erythrocytes
count and hemoglobin level. The Sodium Nitrite of 0.5 mg/200gram BW for white rat, which was a safe dose, has already changed
erythrocytes count, and Sodium Nitrite of 1.5 mg/200gram BW showed higher changes either in erythrocytes count and hemoglobin
level. In conclusion, Sodium Nitrite administration of 0.5 mg/200gram BW/white rat, and 1.5 mg/200gram BW rats can decrease
erythrocytes count and hemoglobin level of male white rats.(FMI 2012;48:1-5)

Keywords: Sodium Nitrite, erythrocyte, hemoglobin.

Correspondence: Rini Ambarwati, Nursing Departement, Nursing Department, Health Polytechnique

Ministry of Health, Jl. Prof Moestopo 8C, Surabaya, Indonesia. Email: jurkep@poltekkesdepkes-sby.ac.idSurabaya

INTRODUCTION "nitrosomioglobin" making it a bright red (like corneed,

Sodium Nitrite (NaNO2) is preservatives for processed
meat foods that tend to be abused because of weak
oversight and the number of people who have not or do
not know the dangers posed, so the manufacturers take
advantage of these conditions (Afrianti 2010). Nitrite is
a compound like crystal or crystal powder which is
odorless, white color, and water soluble (Cahyadi 2006).
Nitrite is an active compound of NO that formed when
Nitrite is dissolved. In the preservation of meat, NO will
join the meat pigment (called myoglobin) to form
1
sausages, burgers) these compounds are carcinogenic
(Afrianti 2010).
Williams et al (2000) has reported that the binding of
nitrite and hemoglobin cause the formation of reactive
oxygen groups (Reactive Oxygen Species/ROS). ROS
working cause oxidative stress on erythrocyte
membrane, erythrocyte consequently not able to
maintain supple and hemolysis occurs early. Although it
has been known that nitrites are carcinogenic to
hemolytic red blood cells, there is a safe dose to be
Folia Medica Indonesiana Vol. 48 No. 1 January - March 2012 : 1-5

consumed according to FAO/WHO, namely: 0.07 correlation test and Multi-analysis of variance (Manova)
mg/kgBW/day so for 70 kg body weight was 7 mg. was used for post hoc analysis, p < 0.05 was considered
Meanwhile, according to Depkes RI 0.4 mg/kg/day as statistically significant
(Department of Health 2006, Afrianti 2010).

The purpose of this study is to obtain the number of RESULTS

erythrocytes and hemoglobin levels of white rats (Rattus
norvegicus) males treated with sodium nitrite for 30 The following is the spread data based on age and the s
days. This study is an experimental laboratory that uses tandar deviation of erythrocytes and levels of
Post Test Control Group Design to determine the effect hemoglobin.
of sodium nitrite on erythrocyte profile ie the number of
erythrocytes, levels of hemoglobin, hematocrit and Table 1. Average test and standard deviation
reticulocyte in white rat. erythrocytes profile

Group
Erythrocytes Controls NaNO 2 = NaNO 2 =
MATERIALS AND METHODS Profile (N=11) 0.5 = 1.5 mg
(N=11) (N=11)
This research was experimental laboratories, using a Average, SD
Erythrocytes (x 10 6 L) 9.61, 0.54 7.70, 0.84 5.99, 0.74
random design complete with 2 treatments to adult male Hemoglobin (g/dL) 14.51, 1.23 12.27, 1.19 8.61, 1.20
white rat (Rattus norvegicus) wistar strain with heavy
175-200 gram in 3-4 month age with physically health. Table 1 shows decreasing average test number of
Total 24 Rats white kept in cage, each cage contains 8 erythrocytes and levels of hemoglobin respectively in
male rats white (total 24 lambs). Rats had aclimaticated the control group, the group treatment NaNO2 dosage
first for 1 weeks. Rats were given feeds and drink et 0.5 mg/200gram BB rats, and the treatment NaNO2
libitum and given oral solution sodium nitrite during the dosage 1.5 mg/200gram BB rats. Changes can be seen
30-day, with the dose per day for each group as follows. in the figure 1 below:
In group one, rats gift feeds and drinking water standard
3 ml/200g BB rats/day (during the 30-day / sonde)
without Sodium Nitrite. Then, group two gift Feeds
standard plus Sodium Nitrite 0.5 mg in 3ml aquades per
200g BB rats/day (for a period of 30 hr/ sonde). While
group three gift feeds standard plus Sodium Nitrite 1.5
mg in 3ml aquades/200g BB rats/hr (during the 30 hr/
sonde).

Technical data

Taking blood specimens to group 1-3 would be done on

the 30th day after having been given preferential
treatment to know the total amount erythrocytes and
hemoglobin. Taking blood specimen came from the
heart rats, where previously done anesthesia by using
Ketamin Hcl (dosage 20-40 mg/kg BW in
intraperitonial) as many as 1 ml. After taking blood
specimens then decapitation done (Kusumawati 2004).
Blood Specimens in all the groups, was included in the
tiny reactions that have been given EDTA then given
identity in accordance with the sequence specimens in
the group and the cook in Balai Besar Health
Laboratory Surabaya.
The checking procedures of erythrocytes and levels of
hemoglobin analysis methods with a computerized with
the instruments Sysmex KX- 21. Measuring result was
visualized in numbers and histogram (Bakta 2006).
Statistical analysis bothered by perfomed Pearson
2
Figure 1. Bar chart of average number of erythrocytes /
RBC (x 10 6 per L)
Results of The Analysis Multivariate Anova
(Manova)
To know the difference between mean of each groups:
number of erythrocytes and levels of hemoglobin in the
control group, the group NaNO2 dosage 0.5 mg per
200gram BB rats and NaNO2 dosage 1.5 mg per 200
grams BB rats , then used tests Manova. Test result
Manova showed the p=0.000 (p< 0.05), so it can be said
that there are huge differences between ; number of
erythrocytes as many as 72.1 (Sum of Square Squares
 Effect of Sodium Nitrite (NaNO2) to Erithrocyte and Hemoglobin Profile in White Rats (Rini Ambarwati)
Type III) and levels of hemoglobin as many as 195.0.
Whereas in Mean Square the number of erythrocytes are
36.0 and hemoglobin 97.5 in the current controls, the
group NaNO2 dosage 0.5 mg per 200gram BB rats and
NaNO2 dosage 1.5 mg per 200gram BB rats.
Figure 2. Bar Chart of average levels of hemoglobin/
HGB (g/dl)
DISCUSSION
Effect of Giving Sodium Nitrite to Number
of Erythrocytes
Nitric is chemicals that are not excreted by the human
body so that it is accumulated and can cause health
problems (Lundberg et al 2008). Number of
erythrocytes is one of the important parameters to assess
health, considering his role is very big in carrying
oxygen in the body. Blood circulation describes basic
mechanisms chemical distribution throughout the body
(Loomis 1978).
Results of research shows that there is a decline of the
average value for the number of erythrocytes
respectively: 9,61x10 6 L ; 7,7x 10 6 L ; 5,99x 10 6
L. Manova test on the number of erythrocytes shows
there is a big difference means with p= 0.000 (p<0.05)
from the sodium Nitrite dosage 0.5 mg/200 grams BB
white rat, and 1.5 mg/200 grams BB rats, as shown in
table 1 and the figure 1
The Sodium Nitrite dosage 0.5 mg/200 grams BB white
rat that is dosage conversion from safe dosage is
allowed according to Permenkes 1988 is 0.4 mg/kg
BW/day while safe dosage that nominated FAO, WHO
is 0.07 mg/kg BW/day, but on the dose of 0.5 mg/200
grams BB rats has shown that there is a decrease in the
number of erythrocytes. According to Lundberg et al
(2008), Sodium Nitrite if in consumption will continue
3
to insist on and excessive cause stock exchange toxic
substances in humans.
On the gift Sodium Nitrite dosage 1.5 mg/200 grams BB
rats, effects on a decrease in the number of erythrocytes
is greater. This is because Sodium Nitrite into the blood
circulation will bond hemoglobin produce Reactive
Oxygen species (ROS) namely anions superoxide (O2 -
). ROS cause oxidative stress to the proteins in
cytoskeleton erythrocytes and damage membrane
structures erythrocytes by ties crossing (cross linking)
proteins, near break easily and cause erythrocytes (lysis)
(May et al 2000). Hemolysis early triggers bone
marrows to speed up the formation of new erythrocyte,
up to 10 times the normal speed marked with the
increase in the number of erythroblast and reticulocyte
accelerated into blood so produce erythrocytes that has
not been mature (in the normal situation, life expectancy
erythrocytes are 120 days). The destruction of
erythrocytes faster than producing it causes a decrease
in the number of erythrocytes.
Decreasing in the number of erythrocytes, affects the
formation of the occurrence of eritropoietin and
stimulate eritropoiesis, but because erythrocytes that
there is a newly formed, then hemoglobin level that is in
erythrocytes has yet to reach the optimum level.
Number of hemoglobin that had not been stable, cause
hemoglobin ability to tie was far from optimal oxygen.
According to Derave and Taes (2009), the number of
oxygen that decreased will lead to liver secretes more
globulin, the kidneys producing more renal eritropoietin
factors. In the blood, globulin and factors eritropoietin
kidney will hold an interaction formed eritropoietin that
will stimulate the occurrence of eritropoiesis.
Speed of eritropoiesis stated in such a manner so that a
number of erythrocytes in the circulatory system there is
more or less constant. Every time the number of oxygen
transported to tissue become low usually will increase
the speed formation of erythrocytes (Murray et al 2009).
Research done by Hord et al (2009), the Sodium Nitrite
with great dosage will speed up destruction the red
blood cells, so reducing the number of erythrocytes
circulation and tends to cause anemia. Based on the
research Umbreit (2007) stated that all chemicals that
were used as preservatives is the poison, but toxicity
that is determined by the number of that is needed to
produce the effects and health problems or sick.
Effect of Giving Sodium Nitrite to Levels
of Hemoglobin
From the result of the research, that is obtained the
decreasing average hemoglobin level with the sequence:
14.51 g/dL; 12.27 g/dL; 8.61 g/dL. Based on statistic
Folia Medica Indonesiana Vol. 48 No. 1 January - March 2012 : 1-5
test Manova on the number of hemoglobin there is a
significant difference with p = 0.000 (p<0.05) from the
Sodium Nitrite dosage 0.5 mg/200 grams BB rats, and
1.5 mg/200 grams BB rats, as can be seen in table 1 and
the figure 2.
Hemoglobin is an erythrocytes component and a
conjugation protein in transport oxygen and carbon
dioxide. The determination levels of Hemoglobin often
used to know that there is an anemia (Tahono &
Wuryaningsih 2000). In this research the sodium Nitrite
dosage 0.5 mg/200 grams BB rats and 1.5 mg/200
grams BB rats showed average test levels of
hemoglobin that decreased. This condition caused Nitric
bound with hemoglobin forms methemoglobine, skin
allergic that is hemoglobin that is oxidized and is a form
of hemoglobin which bind water, not oxygen ; NaNO2
+ hbfe2+ ? NO + hbfe3+ + O2 - , formation of
methemoglobine, skin allergic cause blood hemoglobin
capacity to carry oxygen is decreased, carbonation level
so it can cause hypoxia or lack of oxygen on body
tissues (Hoffbrand et al 2005, Murray et al 2009).
The gift with a dosage of Sodium Nitrite 0.5/200gram
BB rats cause experienced haemoglobin in normal limit
(11.1-18g/dl) but cause the Sodium Nitrite with the dose
1.5 mg/200gram BB rats drop levels hemoglobin under
normal limit (<11,1g/dl), it shows that there is a rats
tendency to anemia. Sodium nitrite is chemicals that
usually used to preservative meat and fish to get a good
color and prevent the growing microbes, but for a
continual intake can cause harmful effects for the health
and can cause anemia.
Research done by Faisol (1987) apparently anemia
affected power concentration, perception and attention
elementary school children this caused by hemoglobin
is the primary protein in the human body that has a
function to carry the oxygen from the lungs to the
peripheral and carrying carbon dioxide from peripheral
tissue to the lungs, to maintain retention or the capacity
to concentrate, there is required energy that is available
in the body. Energy was obtained from foods that go
into the body through a series of processes metabolism.
Exchange substance or the process of metabolism is a
chemical reactions series in the body with the aim is to
produce energy. To be able a continuous metabolism
process in the body, it is needed oxygen as fuel that is
responsible from the process respiration. Hemoglobin
molecule is responsible for carrying oxygen and carbon
dioxide in the blood. Through this fusion oxygen
brought up from the lungs circulated throughout all
body tissues that require (Dejam et al 2005).
4
Factors that are very affected supplies oxygen to tissues
of the body is the number of the red blood cells and the
number of hemoglobin that were found in it, so that
those who are suffering from anemia, oxygen supply to
the tissue of the body will experience disruption since
the internal organs is less, automatically oxygen that is
carried will decreased. With less oxygen in the tissue so
the body metabolism process would be disturbed and
the implication is the need for the learning process
energy had problems. The more levels of Hemoglobin
in the blood, the more oxygen that can be transported to
various tissues vice versa (Ama 1987).
CONCLUSION
The sodium nitrite (NaNO2) dosage 0.5 mg per 200
gram BB in rats (Rattus norvegicus) during the 30-day
decrease the number of erythrocytes, levels of
hemoglobin, The sodium Nitrite (NaNO2) dosage 1.5
mg per 200 gram BB in rats during the 30-day higher
decrease the number of erythrocytes, levels of
hemoglobin.
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