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Life Science Archives (LSA)

ISSN: 2454-1354
Volume 1; Issue - 3; Year 2015; Page: 175 - 181

Research Article

HEAMATOLOGICAL EFFECT OF CROCIN ON MALE ALBINO RAT

K. Jayalakshmi*,
Department of Zoology, Arignar Anna Government Arts and Science College, Karaikal 609005, UT of
Puducherry, India.
Abstract
The active ingredient present in Crocin is Paracetamol (Acetaminophen). It is one of the world's
largest sold paracetamol tablets. The main aim of this study was to assess the effect of Crocin suspension on
male albino rat. The effect was assessed on the basis of the results of acute toxicity tests and on the
comparison of the results of blood haematological examinations of control and experimental group. Saline
solution for control and Crocin suspension chemical were used for treatment in the study. Following to oral
administration of crocin, male rats showed changes of blood WBC, Lymphocyte, Neutrophil, Heamoglobin
and Packed Cell Volume. Investigations carried out using male albino rats weighing about 240 - 260gms.
Crocin was administered with 96 mg/kg of body wt. for first dose at 0-hr, second dose at 6th hr, third dose at
12th hrs and fourth dose at 18th hrs. Blood was collected from optical vein following at 6 th hrs, 12th hrs and
24th hrs after the first dose time. Crocin effectively produced a decreased amount of packed cell volume,
WBC, neutrophil in blood and increased lymphocytes and hemoglobin amounts compared with control.
Changes of the blood biochemical compounds leads to adverse effect in the different physiological role in
cellular activity.
Key words: Haematology, Crocin suspension,
Packed Cell and RBC.

Article History
Received : 02.04.2015
Revised : 10.05.2015
Accepted : 15.05.2015
1. Introduction

Hematology, also spelled haematology

(Gk: haima "blood") is the study of blood, the
blood - forming organs, and blood diseases.
Hematology includes the study of etiology,
diagnosis, treatment, prognosis, and prevention of
blood diseases that affect the production of blood
and its components, such as blood cells,
haemoglobin, blood proteins, and the mechanism
of coagulation.
* Corresponding author: K. Jayalakshmi
Tel.: +91-9600916225

E-mail: jayalakshmichand26@gmail.com

Carotenoids by acting as biological

antioxidants, protecting cells and tissues from
damaging effects of free radicals and singlet
oxygen play a significant role in human health
(Rios et al., 1996). Crocin is a natural carotenoid
chemical compound that is found in the flowers
crocus and gardenia. It is the di-ester formed from
the disaccharide gentiobiose and the dicarboxylic
acid crocetin. It has a deep red color and forms
crystals with a melting point of 186C. When
dissolved in water, it forms an orange solution.
Crocin is the chemical ingredient primarily
responsible for the color of saffron.

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K. Jayalakshmi / Life Science Archives (LSA), Volume 1, Issue 3, Page 175 - 181, 2015

Presence of caffeine in crocin allows it to

works faster and better than ordinary paracetamol
tablets. Crocin is medication that comes under the
class of OTC drugs.OTC drugs or Over-thecounter medicines are drugs you can buy without a
prescription. These rugs are considered safe
enough to sell over-the-counter. Most OTC drugs
help relive muscular pain, cold, flu, tooth decay,
headaches and itches. Symptoms of overdose
include vomiting, nausea, stomach upset,
dizziness, constipation, restlessness, headaches
and collapse. If you notice dark yellow colored
urine seek immediate medical attention because an
overdose may lead to jaundice. Overdose of crocin
can lead to liver or kidney impairments. Patients
with previous history of liver or kidney
impairments must take the medication if clearly
needed or if prescribed by the doctor. If you have
taken more than 7.5 g of Crocin, seek immediate
medical attention. You may need urgent medical
attention or hospitalization.
Crocin is a di-gentiobiose ester of crocetin
and a water-soluble carotenoid of C. sativus that
has attracted scientific attentions because of the
various pharmacological effects such as
hypotensive effect (Imenshahidi et al., 2010),
antidepressant activity (Hosseinzadeh et al., 2004)
genoprotective (Hosseinzadeh et al., 2008),
improve sexual activity (Hosseinzadeh et al.,
2008), antioxidant (Hosseinzadeh et al., 2009),
inhibition of lipid peroxidation in renal
(Hosseinzadeh et al., 2005), hippocampal
(Hosseinzadeh et al., 2005) and muscle skeletal
homogenates during ischemia-reperfusion-induced
oxidative damage in rats (Hosseinzadeh et al.,
2009).
Crocin is also reported to increase the
blood flow in the retina and choroid and could be
used in treatment of ischemic retinopathy and agerelated macular degeneration. Moreover, crocin
inhibits skin tumor growth (Konoshima et al.,
1998), improves the learning behavior that has

176

been impaired by ethanol (Sugiura et al., 1995) or

hyoscine (Hosseinzadeh & Ziaei, 2006), and
prevents the inhibitory effect of ethanol on longterm potentiation in rats (Sugiura et al. 1994). In
animal study, LD50 values of intraperitoneally
injection of saffron stigma and petal extracts were
1.6 and 6 g/kg, respectively in mice. In sub-acute
study, the aqueous extract of stigma decreased
levels of hematocrit, hemoglobin and erythrocytes,
but it did not induce any significant pathological
effects on different organs (Karimi et al., 2004).
Information on toxicology and safety of
saffron is not consistent. In some reports, saffron
doses between 1.2 and 2 g showed nausea
followed by vomiting, diarrhea and bleeding
(Schmidt et al., 2007). Saffron tablets with high
doses (200 and 400 mg/day) changed some
hematological and biochemical parameters in
healthy adult volunteers. However, these
alterations were in normal ranges and they were
not clinically important (Modaghegh et al., 2008).
As crocin, showed a variety of interesting
pharmacological activities, its acute toxicity was
evaluated in rats.
Acute and sub-acute toxicity of crocin, a
constituent of Crocus sativus, were studied in
mice and rats. The acute (upto 3 g, orally and
intraperitoneally) and chronic (15, 45, 90 and 180
mg/kg, intraperitoneally) toxicity were evaluated
for 2 and 21 days, respectively. In chronic
toxicity, changes in weight and amount of food
intake as well as biochemical, hematological and
pathological tests were studied in rats after 21
days. High oral and intraperitoneal doses of crocin
(3 g/kg) did not cause death within 2 days of
study. A dose 180 g/kg of crocin in sub-acute
study increased platelets and creatinin levels. The
weight loss and a reduction in food intake were
also observed with this concentration. The lower
doses of the substance decreased albumin and
ALP, and raised the LDL level dose
independently. A decline in alveolar size in lungs

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K. Jayalakshmi / Life Science Archives (LSA), Volume 1, Issue 3, Page 175 - 181, 2015

of 180 mg/kg crocin group was also observed.

Moreover, in hearts of this group myosin light
chain atrophy and a reduction in size of cells and
nuclei was seen (Hosseinzadeh et al., 2010). The
main target of the present investigation is to
determine the effect of using crocin suspension on
male albino rat blood.
2. MATERIALS AND METHODS
Healthy male albino rats were purchased
from JIPMER animal house, Puducherry,
weighing (240 - 260 g) were selected for present
study. They were kept in polypropylene cages
measuring 45 77 15 cm, at temperature
250.5C, relative humidity 60 5% and
photoperiod of 12 hrs per day. Male albino rats
were grouped into two groups of six rats each:
Control Group A- oral administration to saline
solution 4 ml/kg and Experimental group B
exposed to 96 mg/kg bwt. Administration of
Crocin
suspension
(Acetaminophen
oral
suspension) for first dose at 0 hr, second dose after
6hr from first dose, third dose after 12 hrs from
first dose and fourth dose after 18 hrs from first
dose for each time. Similarly, control group
administered saline solution orally for above said
timings respectively. Blood samples of Control
group 'A' and Experimental group 'B' were
collected from after 6 hrs, 12 hrs and 24 hrs from
first dose 0 hrs time. Blood samples of first and
second were taken directly from optic vein, while
third sample of blood collected from ventricles of
dissected rat with the help of sterilized disposable
syringes fitted with hypodermic needles and
collected in sterilized centrifuge tubes with EDTA
for blood study.
2. Materials and Methods
Estimation of Haemoglobin
Haemoglobin was estimated by the acid
hematin method (John, 1972) employing Sahlis
hemoglobinometer. Sahlis method is a visual
comparison method (John, 1972). The principle
was based on the conversion of haemoglobin to
acid-haematin in the presence of 0.1N HCl. Acid-

177

haematin is a brown coloured compound, which is

visually matched with the brown coloured glass
standard provided in the hemoglobinometer for the
estimation of haemoglobin content. HCl (0.1N)
was taken in a graduated test tube up to the mark
20. Blood specimen (0.02 ml) was added with the
help of a blood pipette. The contents of the tube
were mixed well using a dry glass rod and
incubated for 5 minutes at room temperature. The
test solution was then diluted with the acid until it
visually matched with that of brown coloured
glass standard provided with haemoglobinometer.
The colour intensity of the test solution was noted
in comparison to the glass standard and
haemoglobin content was expressed as gram
percentage.
Blood cell counts
a) Total Leukocyte (WBC) Count
The leukocyte count was estimated by the
haemocytometer method (John, 1972). The WBC
pipette was filled to the 0.5 mark with whole
blood and diluted to the 11 mark with Turk's fluid,
resulting in a 1:20 dilution of the blood sample.
The Neubauers chamber of the haemocytometer
was then charged with diluted blood, and
leukocytes present in the four large corner squares
(mm 3) were counted. Values were expressed as
103 leukocytes per ml of blood.
b) Leukocyte differential count
The leukocyte differential count was
determined by the Leishman Staining method.
Blood smears were taken on microscopic glass
slides and left at room temperature for 3 minutes
to dry. A known volume of staining fluid was
added to the smear on the slides, and left for 3
minutes followed by addition of an equal amount
of phosphate buffer for 7 minutes. The slides were
then rinsed with tap water and dried in air. The
dried film was examined under an oil immersion
lens of a microscope. Leukocytes were classified
as lymphocytes, eosinophils, neutrophils, and

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K. Jayalakshmi / Life Science Archives (LSA), Volume 1, Issue 3, Page 175 - 181, 2015

monocytes based on their morphology and their

content was expressed as percentage of each cell
type.
Packed cell volume
The packed cell volume (PCV) was determined by
centrifuging heparinized blood in a capillary tube
at 10,000 RPM for five minutes (Marler, 1975).
This separates the blood into two layers. The
lower layer contains the total cell mass. The
volume of packed red blood cells divided by the
total volume of the blood sample; it implies the
PCV of the given sample.
2. RESULTS AND DISCUSSION

178

For treated Male albino rat, haemoglobin and

lymphocyte (%) (Table - 1 & Table - 4) were
increased, while WBC, Neutrophil and Packed cell
volume content (Table - 2, Table - 3 & Table - 5)
were decreased when compared with the control.
Percentage difference between control and treated
Male albino rat were calculated. Percentage
difference of Lymphocyte was found high and
showed up to 34% after 24 hrs of crocin
administration and hemoglobin percentage was
found 1.85% increase after 12 hrs crocin
administered male rat. Whereas, Total count of
WBC, Neutrophil and Packed cell volume content
were found decreased to 7.5% (at 12 hrs), 10.5%
(at 24 hrs) and 9.0% (at 12 hrs) respectively.

The haematological results of control and treated

rat blood were shown in Tables -1 to Table - 5.
Table - 1: Hemoglobin (g %) amount of male albino rat present after 6 hrs, 12 hrs and 24 hrs
duration from 0 hr oral administration of crocin (96 mg/kg b.wt)
Parameter
Hemoglobin
Control
Treated

Duration
6 hrs
12 hrs
24 hrs
14.021.02 13.931.13
14.110.965
14.241.2

14.190.846

14.330.881

Values were represented by Mean SEM; Mean values were calculated by six male rats.

Table - 2: WBC (103 mm-3) count of male albino rat present after 6 hrs, 12 hrs and 24 hrs
duration from 0 hr oral administration of crocin (96 mg/kg b.wt)
Parameter
WBC

6 hrs
6300.72 7.11

Duration
12 hrs
24 hrs
6424.52 10.73 6382.81 16.32

6174.2 10.72

5937.0 12.74

Control
5927.5 14.42

Treated
Values were represented by Mean SEM; Mean values were calculated by six male rats.

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K. Jayalakshmi / Life Science Archives (LSA), Volume 1, Issue 3, Page 175 - 181, 2015

179

Table - 3: Neutrophil count (%) of male albino rat present after 6 hrs, 12 hrs and 24 hrs
duration from 0 hrs oral administration of crocin (96 mg/kg b.wt)
Parameter
Duration
Neutrophil 6 hrs
12 hrs
24 hrs
78.732.34 79.741.54 80.742.62
Control
75.222.04 74.521.87 72.241.96
Treated
Values were represented by Mean SEM; Mean values were calculated by six male rats

Table - 4: Lymphocytes count (%) of male albino rat present after 6 hrs, 12 hrs and 24 hrs
duration from 0 hr oral administration of crocin (96 mg/kg bwt)
Parameter
Lymphocytes

6 hrs
17.220.857

Duration
12 hrs
16.741.94

24 hrs
18.3461.06

22.741.53

23.642.25

24.721.61

Control
Treated
Values were represented by Mean SEM; Mean values were calculated by six male rats

Table - 5: Packed cell volume (%) of Male albino rat present after 6 hrs, 12 hrs and 24 hrs
duration from 0 hr oral administration of crocin (96 mg/kg b.wt)
Parameter
Packed cell volume

6 hrs
38.31 2.63

Duration
12 hrs
24 hrs
39.61 3.11 39.72 1.42

36.71 1.15

35.78 0.75

Control
36.22 1.63

Treated
Values were represented by Mean SEM; Mean values were calculated by six male rats

Oral administration of crocin into the male

albino rat shows their increase of blood
lymphocytes
and
hemoglobin
leads
to
erythrocytosis
and
hypoxia.
Increased
lymphocytes crowd the bone marrow, and
interfere with normal blood cell production which
cause to anaemia. Crocin may have induced an
increased in the metabolic rate, with the resultant
increased in the generation of free radicals with
the attendant cellular damage. The immune system
responds to this damages caused by production of
oxidants during stressful conditions. During such

responses, free radicals are produced by the

neutrophils; the first-responds to inflammatory
cells to remove damage cells. Being highly
mobile, neutrophils quickly congregate at the
focus of infection, attracted by cytokines
expressed by activated endothelium, mast cells
and macrophages (Ear and McDonald, 2008).
Neutrophils also recruit and activate other cells of
the immune system.
The PCV was lower in crocin treated rats
compared to control group. The observed decrease

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K. Jayalakshmi / Life Science Archives (LSA), Volume 1, Issue 3, Page 175 - 181, 2015

in PCV is believed to be as a result of the

decreased RBC which is consistent with the
findings of (Eyong et al., 2004). This may
possibly explains the low values of haematological
parameters recorded in crocin treated rats in this
study; suppresses the immune system, causes
disruption or suspension of haematopoiesis
(Hodgson and Smart, 2001).
4. Conclusion
The experimental results based on hypoxia,
erythrocytosis and anaemia effects were caused
by the oral repeatable administration of crocin
suspension compound. It suppresses the immune
system, causes disruption or suspension of
haematopoiesis. So prevent the repeatable
administration of crocin suspension to in animal
models and human beings. In summary, oral
repeatable administration of crocin suspension
increased hemoglobin and lymphocyte in the male
albino rat blood and decreased WBC, Neutrophil
and Packed cell volume when comparing to the
experimental groups. Our study confirmed that
crocin seems to be toxic to rat. However, further
studies are necessary, to confirm this evidence.
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