JFS T: Toxicology and Chemical Food Safety

Mutagenicity and Safety Evaluation of Water

Extract of Coriander sativum Leaves
NGELES, AND LOURDES RODRIGUEZ-FRAGOSO
MARIANA RAMIREZ REYES, JORGE REYES-ESPARZA, OSCAR TORRES A

ABSTRACT: Coriander has been used as a spice and medicinal plant for centuries. Several studies have described its
biological properties and some reports have indicated its pharmacological actions in some human pathology. However, data on its toxicity and metabolism are limited or null, and no research has been conducted with mammalian
cells. The purpose of this study was to evaluate the mutagenicity and safety of Coriandrum sativum extract. The
mutagenic effects of C. sativum extract were evaluated by Ames test. Mutagenicity was present when the C. sativum
extract was used in high concentrations in both tested strains (Salmonella typhimurium TA97 and TA102). Our research showed that C. sativum extract reduced the cell survival of human cell lines (WRL-68 and 293Q cells) by
inducing apoptosis and necrosis in the cases where extract concentration was the highest. The C. sativum extract
altered the cell cycle; it increased the G1 phase of hepatic cells and reduced the G2+M phase in both cell lines in
a dose-response manner. These results showed correlation with a reduction in the mitotic index. The extract also
induced severe malformations during embryonic development. Exposure of chicken embryos to the C. sativum extract resulted in a dose-dependent increase of anomalies. Present results show that C. sativum extract reduced the
axial skeleton and affected the neural tube, the somites, the cardiovascular structures, and the eye. According to the
present results, the C. sativum aqueous extract cannot be considered safe. These results indicate that some significant adverse effects of C. sativum extract could be observed in vivo.
Keywords: apoptosis, Coriandrum sativum, cytotoxicity, embryotoxicity, mutagenicity

Introduction

for medicinal purposes. These include powered seeds or dry extract

(2 to 5 g/d), tea (4 to 8 g/100 mL), tinctures (1.8 g/mL), decoctions,
and infusions ( Breevort 1996; De Smet 2002). One would expect
each of these preparations to have a different proportion of each of
the previously mentioned components; the tinctures and extracts
will have more nonpolar components, while hydrophilic components will predominate in the teas and decoctions. The way these
preparations are made must also be considered (cooking time, temperature, amount of water, time of rest, amount of plant used, and
so on). Tinctures are more concentrated than infusions and decoctions, while the preparation of extracts could lead to loss of volatile
oils. Having taken all this into account, the efficacy or toxicity of C.
sativum could vary depending on the composition and proportion
of constituents in the preparations made for human use.
Recent research involving in vivo pharmacological use of extracts from this plant in experimental models has shown their high
therapeutic effectiveness (Medhin and others 1986; Gray and Flatt
1999; Kubo and others 2004; Lal and others 2004; Ramadan and
others 2003; Emamghoreishi and others 2005). But very few (if
any) modern clinical studies have been conducted on coriander.
Its approved modern therapeutic applications are based on its long
history of use in well-established systems of traditional medicine,
pharmacological studies conducted on animals, nutrient composition, dietary value studies, and phytochemical research (Burdock
and Carabin 2009).
In spite of the wide-ranging, extant research on the therapeutic effects of C. sativum, little is known about its toxicological
effects, since no extensive studies have been conducted on in
vitro and in vivo models (Burdock and Carabin 2009). An important
aspect of natural products, especially those that are readMS 20090347 Submitted 4/17/2009, Accepted 8/29/2009. Authors are with
Facultad de Farmacia, Univ. Autonoma del Estado de Morelos. Avenida ily available to the public, is safety. Many people assume natural
Univ. 1001 Col. Chamilpa 62210, Cuernavaca, Morelos. Mexico. Direct in- products are safe, but there is recent, abundant evidence involving
quiries to author Rodrguez-Fragoso (E-mail: mrodriguezf@uaem.mx).
serious adverse effects and deaths associated with the use of dietary

oriander is an annual herb native to Mediterranean Europe

and Western Asia, naturalized in North America, and now extensively cultivated in many temperate countries (Wichtl and Bisset 1994; BHP 1996; Leung and Foster 1996). Fresh leaves are used
as a flavoring agent and dried coriander seeds are used as spices
in food preparation. Both an annual and a perennial herb, coriander is rich in various food elements (Grieve 1971). It contains about
1% volatile oil, of which 55% to 74% is linalool; monoterpene hydrocarbons (a- and b-pinene and limonene), anethole, and camphor comprises 20%; oleic, petroselinic, and linolenic fatty acids
make up 26%; approximately 20% is comprises flavonoid glycosides
(quercetin, isoquercitrin, and rutin), chlorogenic and caffeic acids,
tannins, and sugars while proteins comprise 11% to 17%. The remainder (approximately 1%) contains coumarins, mucilage, and
starch (Lister and Hrhamme 1973; Hansel and others 1992; Wichtl
and Bisset 1994; Budavari 1996; Leung and Foster 1996).
Coriander essential oil has a long history in traditional medicine
(Uma and others 1993; Kiple and Ornelas 2000). Galenical preparations of coriander seed have been used in traditional Chinese, Indian, Greco-European, and Latin American indigenous medicine.
The British Herbal Pharmacopoeia (BHP 1996) and The Merck Index
also report its therapeutic qualities as a carminative and aromatic
(Budavari 1996).
In traditional medicine, Coriandrum sativum has been used to
treat a number of medical problems such as dyspepsia, loss of appetite, convulsions, insomnia, and anxiety (Breevort 1996; De Smet
2002). C. sativum is empirically used in different doses and forms

T: Toxicology &
Chemical Food Safety

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R
2009 Institute of Food Technologists
doi: 10.1111/j.1750-3841.2009.01403.x


C

Further reproduction without permission is prohibited

Toxicity of Coriandrum sativum . . .

supplements and nutriments (Marcus and Grollman 2002; Wooltorton 2002; Wooltorton and Sibbald 2002; Morrow and others 2005).
The purpose of this study was, therefore, to evaluate the toxicological effect of C. sativum extract on in vitro models and chick embryo
development.

Materials and Methods

cator to avoid water absorption until used. Hydroalcoholic extraction using 80% methanol was conducted by percolating 200 to 300 g
of the dried and powdered plant material for 5 d, which was then
filtered through Whatman filter paper nr 1. The solvent was evaporated using a Rotavapor and the extract was kept in a stoppered
sample vial at 4 C until used.

Determination of content and

essential oil composition

Preparation of C. sativum extract

Leaves of C. sativum were purchased from the Oaxaca Sierra,
Mexico, and transported to the state of Morelos (Mexico) in October
2006 (Figure 1). These were then identified by a taxonomist and a
voucher sample representing Herbarium nr FL6006 was deposited
at the Herbarium of the Autonomous Univ. of the State of Morelos.
The leaves were air dried at room temperature, ground, and kept in
amber colored bottles until processed. Aqueous extraction was performed by soaking a weighed amount of the dry powder in distilled
water and shaking it for 3 h with an electric shaker. The suspension was filtered through muslin gauze and the filtrate kept in deep
freeze for 24 h, then lyophilized. The lyophilized dry powder was
collected in a stoppered sample vial, weighed, and kept in a desic-

Coriander leaf oils were analyzed as 1% solutions in hexane,

using a PerkinElmer autosystem gas chromatograph under the
control of PerkinElmer Omega (version 5.2) software. GC analysis was performed on a Hewlett-Packard 5890 series II gas chromatograph equipped with 2 flame ionization detectors (255 C), 2
fused capillary columns of different polarities, a methyl silicone,
and a polyethylene glycol 20000 column (HP-1 and HP-Wax, 50 m
0.25 mm, film thickness 0.25 mm), which were used simultaneously, and a split injector at 255 C (split ratio) 1 : 150). The temperature was programmed from 90 to 220 C at a rate of 3 C/min.
Nitrogen was used as the carrier gas at a flow rate of 0.8 mL/min.
GC-MS was performed on a PerkinElmer Autosystem gas chromatograph Q-mass 910 quadrupole mass spectrometer, equipped
with 2 fused silica columns, a nonpolar J&W DB-1 (30 m
0.25 mm, 0.25 m film thickness) and a polar DB-Wax (30 m
0.25 mm, 0.25 m film thickness). The GC parameters were the
same as those mentioned previously, but helium was used as the
carrier gas. Mass units were monitored from 45 to 350 at 70 eV.
The oil components were identified by (1) determining their retention indexes (RI) in relation to a homologous series of fatty acids
methyl esters (C4-C18) and (2) comparing their mass spectra with
published values (Adams 1995; Baratta and others 1998; Gil and
others 2002) (Table 1).

Cell culture
Two cell lines were used for this study: 293Q cells derived from
normal epithelial cells of human fetal kidney (CRL-1573 ATCC) and
WRL-68 cells derived from epithelial cells of human fetal liver (CRL48 ATCC). Cell lines were cultured in minimal essential medium
(MEM, GIBCO BRL Inc., Grand Island, N.Y., U.S.A.), supplemented
with nonessential amino acids (GIBCO BRL Inc), 10% fetal calf
serum (GIBCO BRL Inc), L-glutamine (2 mol/L), and antibiotics.
Cells were plated in 100-mm culture dishes (106 cells/dish), and
maintained at 37 C under an atmosphere of 5% CO2 in humidified air. The medium was replaced every 2 d and the cells were
harvested and diluted 5-fold every 7 d. Subcultures were obtained
Figure 1 --- Coriander (Coriander sativum) (Source: Oaxaca by trypsinization (0.025% trypsin solution containing 0.01% N,Ndiethyldithiocarbamic acid sodium salt, EDTA).
Sierra, Mexico, 2006).
Table 1 --- Identification of C. sativum components.
Identity

1
2
3
4
5
6
7
8
9
10
11
12
13

-pinene
camphene
-pinene
myrcene
p-cymene
limonene
-terpinene
linalool
camphor
decanal
geraniol
decanol
geranyl-acetate

GC

RI

GC-MS

Component level (%)

+
+
+
+
+
+
+
+
+
+
+
+
+

+
+
+
+
+
+
+
+
+
+
+
+
+

+
+
+
+
+
+
+
+
+
+
+

4.8
0.7
0.4
0.9
0.7
2.7
4.9
75.4
5.1
0.3
2.8
0.3
3.0

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Identification method
Peak

Toxicity of Coriandrum sativum . . .

Assessment of cell viability
Cells were plated at 10000 cells/well on 96-well plates. After 24 h,
the culture medium was replaced by a fresh one supplemented with
different concentrations of C. sativum extract (0.4, 0.8, 1.6, 3.2, 4.8,
6.4, and 8 g/mL). After 24 h incubation, cells were collected and
processed. Cell viability was measured by a 3-(4, 5-dimethylthiazol2-yl)-2, 5-diphenyltetrazolium bromide (MTT) assay (Wang and
others 1996). Briefly, 20 L MTT (5 g/L) was added to each well
and incubated with the culture for an additional 4 h at 37 C, 5%
CO2 . Culture media were then discarded, followed by the addition
of 200 L DMSO with 25 L Srensens glycine buffer (glycine 0.1 M,
NaCl 0.1 M, pH 10.5) to each well. When the blue crystals were dissolved, the optical density was determined on a microplate reader
(Bio-Rad) at 450 nm.

Survival of cells and detection

of percentage of apoptosis and necrosis
Survival of cells was evaluated by a flow cytometric method
(Nicoletti and others 1991). Annexin V is a Ca2+-dependent
phospholipid-binding protein that has a high affinity for
phospholipid-like phosphatidylserine (PS) and is useful for
identifying apoptotic cells with exposed PS. Cells (105 ) were
washed twice with cold PBS and then re-suspended in 1 binding
buffer at a concentration of 1 106 cells/mL. A total of 100 L of the
solution (1 105 cells) were transferred to a 5 mL culture tube. Five
milliliters of Annexin VFITC and 10 L of PI were added. Cells were
incubated for 15 min at room temperature (20 to 25 C) in the dark
and then 400 L of 1 binding buffer were added to each tube. The
results were analyzed with a CELLQuest program in a FACSCalibur
flow cytometer (Becton Dickinson, Calif., U.S.A.). Staining cells
simultaneously with FITC-Annexin V (green fluorescence) and the
nonvital dye propidium iodide (red fluorescence) enables bivariate
analysis of the discrimination of intact cells the discrimination
of intact cells (FITC-PI-), early apoptotic (FITC+PI-), and late
apoptotic or necrotic cells (FITC+PI+).

Bactericidal toxicity test

Samples (1 to 5 mg/plate) were prepared with 0.1 mL of fresh
culture of the tester strain (approximately 108 cells/mL), 0.1 mL of
the WFTS, 0.2 mL of phosphate buffer (0.2 M, pH 7.4), and 0.5 mL
of S-9 mix (metabolic activator) or, instead, phosphate buffer. The
mixture was diluted sequentially with phosphate buffer and 1 mL
of diluted solution was mixed with 12 mL of nutrient agar. After incubation at 37 C for 48 h, the number of colonies was counted.
A bactericidal toxicity effect was confirmed if the standard plate
count of tested compound was lower than that of the control (without adding tested compound).

Salmonella mutagenicity test/Ames test

T: Toxicology &
Chemical Food Safety

The Ames test was performed as a standard plate incorporation

assay with Salmonella typhimurium strains TA97a and TA102 with
metabolic activation (Maron and Ames 1983). Strain-specific genetic markers were verified prior to use. The selection of the strains
was based on the testing and strain selection strategies of Mortelmans and Zeiger (2000). For each tested strain, a specific positive
control was always used to assess the experimental flaws, if any.
Nitro phenylenediamine and sodium azide were used as positive
controls for TA97 and TA102, respectively. To ensure sterility, the
different concentrations of the extract were exposed for 15 min
to UV-C light (TUV 30W G30T8 Philips, Holland). The absence of
contaminant growth was checked in a blank set containing the extract but without the addition of bacteria. For each concentration,
100 mL of the test solution was used per plate. Positive controls
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JOURNAL OF FOOD SCIENCEVol. 75, Nr. 1, 2010

(20 mg/plate nitrophenylenediamine for TA97a and 1.5 mg/plate

sodium azide for TA102) and negative control (distilled water) were
concurrently maintained. Samples were tested on triplicate plates
in 2 independent experiments. Following 48 h incubation at 37 C,
genotoxic activities were expressed as induction factors (induction
factors of reversions), that is, as multiples of the background levels. The interpretation of the Ames test results followed the U.S. Environmental Protection Agency (1996) guidelines for genotoxicity
testing of chemicals. According to the guidelines, a mutagenic potential is assumed if the revertant frequency is 2 or higher over the
solvent control (Mortelmans and Zeiger 2000).

Analysis of the cell cycle

Cells were scraped and washed with PBS. Cells (105) were fixed in
75% ethanol for 24 h and then washed in PBS and resuspended
in 0.1% NP40 (Nonidet P40, Biochimica Fluka, St. Louis Mo.,
U.S.A.) and 10 g/mL RNAse (Ribonuclease A, DNAse-free preparation) for 20 min at room temperature (Darzynkiewicz and others
2001). Propidium iodide (PI) was then added (final concentration
5 g/mL) for 12 h at 4 C in darkness. Samples were analyzed using a FACSCalibur flow cytometer. The results were analyzed using
CELLQuest program.

Mitotic activity
Cells were fixed in 3% paraformaldehyde with 0.25M mannitol (45.54 g/L) for 2 h, rinsed in PBS and stained with DAPI (4 ,6diamidino-2-phenylindol, 480 nm). After rinsing in PBS, the cells
were embedded in Citifluor mounting medium. The mitotic index
was counted with a fluorescent microscope (Optiphot 2 Nikon). For
each experiment, the indices were determined per 1000 cells and
with 4 replicates.

Embryotoxicity studies
Fertile White Leghorn chicken eggs were obtained from A.L.P.E.
S.A. (Puebla, Mexico) and were stored at 6 C. Total of 72 fertilized
eggs were weighed, sterilized, and divided into 9 groups. First group
served as a nontreated control. The next 6 groups received the C.
sativum extract (0.4, 0.8, 1.6, 3.2, 6.4, and 8 g/mL). The last group
received caffeine (10 mg/mL) and was considered the positive control. A teratogenicity assay was carried out as described by Jelinek
and Marthan (Jelinek and Marhan 1994). Test solutions (1 mL) were
added to the air sac under sterile conditions. Each solution was injected after drilling into the shell at the blunt end of the egg; after
injection, the holes were immediately sealed with melted paraffin
wax. The eggs were then transferred to and maintained in a forced
draft incubator at 37.5 C with a relative humidity of 55% until the
desired stage of development was reached.
To determine the concentration dependency of C. sativum extract teratogenicity, a histological analysis was carried out. Embryos in each group were fixed in buffered formal saline (pH 7.4),
dehydrated, and embedded in paraffin blocks. Paraffin tissue sections of 6 m were stained with acetocarmine for routine histological examination. The embryo was examined and staged according
to morphological criteria previously outlined by Hamburger and
Hamilton (1951). Embryonic stages at the time of the C. sativum
extract application varied from 14 to 16, which correspond approximately to developed somites numbered 22 to 28.

Statistical analysis of results

In vitro data were reported as mean SD of 3 independent
experiments conducted in quadruplicate. Mean values were compared using Students t-test or analysis of variance (ANOVA) using
SPSS 10.0 software (SPSS Inc., Chicago, Ill., U.S.A.). Significant differences were established at P < 0.001.

Toxicity of Coriandrum sativum . . .

Results and Discussion

. sativum extract caused a marked reduction in the survival of

2 human cell lines after 24 h incubation (Figure 2). Concentrations higher than 1.6 g/mL produced different effects on cell lines.
A significant 60% survival reduction was observed in WRL-68 (hepatic) cells with concentrations of 1.6 to 8 g/mL (P < 0.001); 293Q
(renal) cells showed a 30% survival reduction at 1.6 g/mL. Higher
concentrations (3.2 to 8 g/mL) reduced cell survival by 60% or

Figure 2 --- Changes in the survival of cells after treatment

with different concentrations of C. sativum extract during
a 24-h incubation period (control = 100%). The results are
presented as means SD of 3 independent experiments.

P < 0.001 in comparison with control value.

more (P < 0.001). The decreased percentage of live cells was accompanied by an increase in the amount of apoptotic and necrotic cells
(Table 2), P < 0.001. It was found that apoptotic cells outnumbered
necrotic cells, especially after treatment with the highest concentrations of the extract (1.6 to 8 g/mL). The increase in apoptotic
cells took place in a dose-dependent manner. Some reports have indicated that low concentrations of some compounds induce apoptosis, while high concentrations induce necrosis (Del Bino and
others 1991). The appearance of necrotic cells may also be a result
of incomplete apoptosis (Leist and Nicotera 1998). Recent studies
by Bakkali and others (Bakkali and others 2005) have shown that an
extract of C. sativum induced cytotoxic effects on the yeast Saccharomyces cerevisiae, and that the effects were stronger in exponential
rather than stationary phase cells. The results of the present study
showed that C. sativum is cytotoxic for both human cell lines.
The results of previous comet assays show that C. sativum does
not induce toxicity in cultured fibroblasts of rat embryo (Heibatullah and others 2008). The present study demonstrated that C.
sativum extract is mutagenic in the tested range of 1.6 to 8 g/mL
according to the Ames test. The results of mutagenicity assays, presented as mean revertants per plate, are shown in Table 3. The
mutagenic potential of the extracts showed a positive dose-related
increase in the number of revertant colonies in both strains of S.
typhimurium. The number of revertant colonies ranged between
437 16 and 7905 45 in TA97a and 420 38 and 1742 58 in

Table 2 --- Changes in the percentage of normal, apoptotic, and necrotic cells after 24 h incubation with different
concentrations of C. sativum extract.
Cell line
WRL-68

293Q

Concentration of
C. sativum (g/mL)

Normal cells (%)

0
0.4
0.8
1.6

90 6
86 4
85 3
63 6

62
8 1.9
93
28 5

41
53
63
9 2

3.2
4.8
6.4
8
0
0.4
0.8
1.6
3.2
4.8
6.4
8

48 4
42 7
40 8
42 3
91 5
86 3
83 2
70 6
58 7
40 4
38 5
37 5

35 3
39 4
42 1
47 0.5
52
11 3
82
21 4
27 3
37 5
38 4
41 3

13 5
18 6
18 4
11 2
41
3 0.5
93
9 1
15 3
23 4
24 5
22 4

Apoptotic cells (%)

Necrotic cells (%)

The results are presented as means SD of 3 independent experiments. P < 0.001 in comparison with control value.

Table 3 --- Mutagenicity of C. sativum in tester strain of S. typhimurium.

Revertant colonies (UFC/plate)e
TA97a, mean SD
T102, mean SD
Distilled watera
0.4
0.8
1.6
3.2
4.8
6.4
8
NDPb
SAb

-S9d

S9

98 3
106 15
134 28
437 16c
538 22c
610 36c
690 21c
734 34c
790 45c

89 4
188 7
296 12
990 9
1030 14
2345 17
3423 24
3567 27
3974 34

a
Negative control.
b
Positive control; NDP = nitrophenylenediamine (20g/plate) and SA = sodium azide (1.5
c
Number of revertant colonies more than twice that of corresponding control (P < 0.001).
d

-S9

S9

149 21
198 34
265 42
420 38c
680 27c
945 34c
1230 43c
1742 58c

490 6
630 14
698 24
830 14
3143 25
4423 31
4987 41
5423 34

1985 124c

6423 29

g/plate).

S9 is a metabolic activation system consisting of the postmitochondrial fraction of the livers of rats. The results are presented as means SD of 3 independent
experiments. SD = standard deviation.
e
The number represent the number of mutants/plate.

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Concentration of C. sativum (g/mL)

Toxicity of Coriandrum sativum . . .

T: Toxicology &
Chemical Food Safety

TA102, with concentrations between 1.6 and 8 g/mL of C. sativum

extract (P < 0.001). According to the 2-fold rule, the doubling of
spontaneous reversion rate at 1 or 2 test chemical concentrations
constitutes a positive response (Mortelmans and Zeiger 2000). The
mutagenic activity was associated to a metabolic activation by S9
mix. Bakkali and others (Bakkali and others 2005) demonstrated
that C. sativum induced cytoplasmic petit mutations on the yeast S.
cerevisiae. The results of the present study showed that C. sativum
extract is mutagenic toward both tester strains, so that the presence
of apoptosis in human cell lines suggests it might also induce mutations in those cells.
The results also showed differences in the effects of C. sativum
extract on the cell cycle of both cell lines after 24 h incubation
(Figure 3). A 30% increase in the number of cells in G1 phase was
observed in WRL-68 cells with all used concentrations; there
was also a 50% to 80% reduction in the G2/M phase with concentrations of 3.2 to 8 g/mL (P < 0.001). No changes in S phase were
observed with any concentration. On the other hand, 293Q cells
showed a reduction of 60 to 80% in the G2+M phase with concentrations higher that 0.8 g/mL (P < 0.001), while no changes
in G1 and S phases were observed. C. sativum extract also reduced
the percentage of cellular divisions. A reduction of 20% to 25%
with lower concentrations and 35% to 40% with concentrations
higher than 3.2 g/mL was observed in WRL-68 cells, P < 0.001
(Figure 4). A 20% to 34% reduction in the mitotic index of 293Q
cells was observed when they were treated with C. sativum extract
(P < 0.001). For many cells, G1 phase is the major period of cell
growth. During this stage, new organelles are synthesized and the
cell requires both structural and functional proteins, resulting in
active protein synthesis and, therefore, a high metabolic rate cell.
The presence of cell cycle arrest and apoptosis suggests that cells
are suffering DNA damage. Previous studies have suggested that cycle regulation-mediated apoptosis is a mechanism of cell growth
inhibition (Ahmad and others 1997; Deigner and Kinscherf 1999;
Ahmad and others 2001). The apoptosis is a physiological process
that functions as an essential mechanism of tissue homeostasis and
is regarded as the preferred way to eliminate unwanted cells. Previous studies by Cortes-Eslava and others (2004) demonstrated that
an aqueous crude coriander juice significantly decreased the mutagenicity of some metabolized aromatic amines and that the coriander juice (50 to 1000 L per coincubation flask) was neither toxic
nor mutagenic. Previous studies have also shown that C. sativum
extract possesses antioxidative properties (Chithra and Leelamma
1999; Satyanarayana and others 2004; Sreelatha and others 2008).
Because we are testing an aqueous extract (a mix of essential oils),
it is possible that some toxic constituents are inducing cell damage
and others are protecting the cell.
Hepatic and renal cells play an important role in drug
metabolism because they have drug-metabolizing enzymes. Mutagenic activity in the presence of S9 suggested that C. sativum
extract might be metabolized in vivo and cause several biological
activities. Metabolic products are often less active than the parent drug and may even be inactive. However, some biotransformation products have enhanced activity or toxic properties. Certain
herbal medicines have been identified as a cause of acute and
chronic hepatitis, cholestasis, drug-induced autoimmunity, vascular lesions, and even hepatic failure and cirrhosis. Several factors
may contribute to the hepatotoxic effects of herbal preparations.
Herbal medicines are usually a mixture of several ingredients or
plants harvested during different seasons and extracted by variable procedures, which make the identification of both pharmacologically active and toxic compounds difficult. This is the case
of C. sativum (Smallfield and others 2001; Gil and others 2002;
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Ramadan and others 2003). Also, contamination of herbal

medicines with microorganisms, fungal toxins (such as aflatoxin),
pesticides, heavy metals, and synthetic drugs could confuse analysis. Moreover, it has been reported that herbal extracts may also
exert renal toxicity through inherent properties (Wojcikowski and
others 2004). There is currently not enough information regarding
the metabolism of C. sativum in human cells. However, because
this plant contains a mixture of essential oils and the C. sativum
extract here studied induced necrosis, it is possible that C. sativum
extract could be metabolized into toxic metabolites in hepatocytes
and renal cells (Smallfield and others 2001; Gil and others 2002). It
has been reported that ingestion of coriander oil led to the incorporation of petroselinoyl (triacylglycerols of coriander) into heart,

Figure 3 --- Percentage of cells in G1, S, G2/M phases of the

cell cycle after treatment with different concentrations
of C. sativum extract during a 24-h incubation period. (A)
WRL68 cells, (B) 293Q cells. The results are presented as
means SD of 3 independent experiments. P < 0.001 in
comparison with control values.

Figure 4 --- Changes in the mitotic index of cells after incubation for a 24-h period with different concentrations of C. sativum extract. The results are
presented as means SD of 3 independent experiments.

P < 0.001 as compared with all groups; #P < 0.001 as

compared with 0.4, 0.8, and 1.6 g/mL.

Toxicity of Coriandrum sativum . . .

Figure 5 --- Morphological appearance
of chick embryos treated with
different concentrations of C.
sativum. Arrows indicate
morphological alterations in the
heart, central nervous system (CNS),
placodes (optical and otic), neural
tube, and somites.

Table 4 --- Teratogenic effects of C. sativum.

Embryonic region affected

Caffeinea 10 mg/mL

Controlb

0.4

0.8

3/9
6/9
7/9
7/9
7/9
5/9
9/9

0/9
0/9
0/9
0/9
0/9
0/9
0/9

0/9
0/9
0/9
0/9
0/9
0/9
3/9

1/9
0/9
0/9
0/9
1/9
0/9
4/9

CNS
Neural tube
Somites
Vasculature
Heart
Eye
Axial skeleton

C. sativum extract (g/mL)

1.6
3.2
4.8
3/9
3/9
2/9
3/9
3/9
1/9
7/9

5/9
5/9
4/9
5/9
5/9
2/9
8/9

7/9
7/9
5/9
7/9
7/9
4/9
9/9

6.4

8/9
8/9
8/9
8/9
8/9
6/9
9/9

9/9
9/9
9/9
9/9
9/9
8/9
9/9

a
Positive
b

control.
Negative control.
The fractions represent the number of abnormal embryos and the total examined for each developmental region of the embryo.

Conclusions

he toxicological evaluation of C. sativum aqueous extract

showed that said extract is toxic for human cell lines and
chicken embryos. Study results reveal that C. sativum extract induced cytotoxic effects on renal and liver cells in a dose-dependent
manner (P < 0.001). C. sativum extract was able to produce apoptosis, necrosis, and alterations in the cell cycle, and changes in
the mitotic index in renal and hepatic cell lines. The extract also

presents mutagenicity according to the Ames test and the results

showed it induced severe malformations during embryonic development. The C. sativum aqueous extract cannot be considered safe
and these results indicate that some significant adverse effects of C.
sativum extract could be observed in vivo.

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