Vol.2, No.5, 395-399 (201
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0) Health
Openly accessible at/HEAL H
The effect of detergent as polluting agent on the
photosynthetic activity and chlorophyll content
in bean leaves
Branislav R. Jovanić1*, Srdjan Bojović2, Bratimir Panić1, Božidar Radenković3,
Marijana Despotović3
1Institute of Physics, Belgrade University, Zemun, Serbia; *Corresponding Author: branislav.jovanic@ipb.ac.rs
2Institute for Biological Research “Siniša Stanković”, Belgrade, Serbia
3Faculty of Organization Science, Belgrade, Serbia
Received 28 October 2009; revised 10 January 2010; accepted 12 January 2010.
The paper investigates effects of detergent for
domestic use on the photosynthetic activity and
chlorophyll content in intact bean leaves. The
plants were watered for 21 days with a solution
of domestic washing powder of 0.60 g r/l. It was
established that the activity of photosynthetic
apparatus in the plant leaf PhACNorm [%] decrea-
ses exponentially with the length of plant treat-
ment/watering. At the end of the treatment (21st
day) the activity of photosynthetic apparatus in
the dosed plant leaf was no more than 45% of
that in control plant (those which were not wa-
tered with detergent solution). With increased
plant treatment duration the changed chloro-
phyll concentration ΔChlNorm [%] rose non-linearly
in plant leaves. The highest change ΔChlNorm [%]
was observed on the 21st day and amounted to
Keywords: Chlorophyll; Detergent; Plant;
Photosynthesis; Pollution; Water
High technologies and technological processes are al-
ways accompanied with products which pollute the en-
vironment to varying extents. Very few of these products
are not pollutants. This is the reason why environmental
study is becoming increasingly important for the sur-
vival of plant and animal world and ultimately of hu-
mankind itself. It should be noted that a culprit for envi-
ronmental pollution should not be sought only in out-
dated or new technologies. Sources of pollution can be,
which is often ignored, some domestic processes in ur-
ban environment, such as food preparation or personal
hygiene. The subject of this study is an investigation into
water pollution resulting from everyday domestic hygi-
enic procedures. There are hardly any households with-
out a washing machine connected with pipes to sewage
for discharge of used water with detergent. Used water is
discharged into the nearest river or a lake and together
with it detergent. Undoubtedly, with time detergent con-
centration in the river/lake goes up and the direct conse-
quence of this is a dramatic change in the biosphere.
Significant pollution in ground water was observed in
Tehran [1]. There are other numerous examples of pol-
luted rivers and lakes with industrial detergents. For
example it was found out that the Caspian Sea waters
and Volga Terek, and Sulak rivers were extremely pol-
luted with a high detergent concentration [2]. The Аsa
River in Nigeria is dramatically polluted with industrial
detergents [3]. Likewise the Coastal Zone of the Sea of
Okhotsk and Avacha Bay are polluted with detergents
[4]. Regardless whether it is river or lake water, it is used
in gardening for watering vegetables used for human
consumption. Doubtless, this water will cause changes in
vegetables which can have an adverse effect on people
eating them.
2.1. Methods
Bean (Phaseolus vulgaris L.) seeds were grown for 3
weeks. They were placed in a growth chamber adjusted
to the identical growing conditions (humidity, lighting,
temperature, nutrition of soil). The seedlings were wa-
tered daily during all investigated periods with tap water.
After this period the plants were divided in two groups:
control and stress. Growing conditions were also identi-
cal for control and stressed samples and the only differ-
ence was the presence or absence of detergent in soil.
Concentration of domestic use detergent in the water
B. R. Jovanić et al. / HEALTH 2 (2010) 395-399
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used for watering was 0.60 g r/l. The control samples
were watered with water without detergent. Therefore,
any differences between fluorescent spectra and fluores-
cence induced curve for stressed versus control plants
could only be the result of the presence of detergent.
In all experiments the plant’s leaves remain intact
(cutting is an additional stress) and we could make sev-
eral measurements on the same plant at any time. In fur-
ther text, the subscripts (S) and (C) will denote the
stressed or control (nonstressed) conditions, respectively.
Photosynthetic activities were determined using well
known Kautczhy method. Photosynthesis measurements
of light-adapted plants, non-destructive measurements of
potential quantum yield (Fv/Fm), were taken using a
photodiode connected with 14bit AD card for collecting
modulated fluorescence. In front of the photodiode was
placed interference filter 690 nm 5 nm. Excitation
source for fluorescence induction curve was high inten-
sity LED 470 nm/12 mW. The beans were transferred to
a darkened laboratory for 5 minutes for adaptation be-
fore measuring fluorescence kinetics at 690 nm [5]. Each
point in Figure 1 and Figure 2 represents mean value of
15 measurements on different leaves which completely
satisfies the demand for value measurement and calcula-
tion precision [ΔChl(a,b) and PhAcNorm] to be higher
than 1% [6].
For excitation the leaves and obtained fluorescence
spectra the leaf was irradiated by high power LED (470
nm/12 mW). The fibre inlet was placed 15 mm from the
leaf surface. The LED beam diameter on the leaves was
~10 mm. The LED light beam was always directed onto
the upper surface of the leaves at a 90 angle of the leaf
axis, and the optical fibre was set at a 90 angle to the
leaves’ surface on the same side. Fluorescence emitted
radiation from intact leaf was collected and directed
through an optical fibre (N.A. of 0.22 and 1000 µm di-
ameter) that was coupled to a portable 2048-element
CCD spectrometer (AVANTES 1000 PC). Data collec-
tion and spectrum processing were conducted in real
time with microcomputer and commercial software OOI
Base (AVANTES Inc.). The results for each groups of
bean represent an average of the measurement of ten
leaves. Fluorescence measurements took 1.5 min for
each measured leaf.
Chlorophyll content Chl(a,b) in bean leaf was deter-
mined from experimentally obtained fluorescence spec-
tra and well known relation between chlorophyll content
and fluorescence intensity ration. It is well known that
the ratio of the two chlorophyll fluorescence peaks
(F730/F690) in leaves correlates well with amount of chlo-
rophyll content in the bean plant leaves [7]. Therefore
chlorophyll content was determined using: a) Fluores-
cence bean spectra and b) relation between chlorophyll
content and the fluorescence intensity ratio FIR defined
as the ratio of the fluorescence intensity measured at
730 nm (F730) and 690 nm (F690) FIR = FIR690/FIR730.
For the bean linear correlation (r2 = 0.954) between
chlorophyll(a,b) content and FIR is Chl(a,b) = 42.93 –
12 FIR and was obtained from literature data [5]. In or-
der to eliminate errors which can appear due to differ-
ences in individual chlorophyll contents in different bean
samples we introduced a relative change of the chloro-
Chl(a, b)Chl(a, b)Chl(a, b)12FIRFIR 
FIRC and FIRUV are the fluorescence intensity ratio for
control plant which were not exposed to the UV radia-
tion and the plant which were exposed to the UV radia-
tion. This method was successful in the experiment with
a pumpkin exposed to the γ-nuclear radiation [8].
Figure 1. Change in chlorophyll content ΔChl(a,b) in the bean
leaves during irrigation with water contain detergent.
Figure 2. Change of the normalized photosynthesis activity
PhAcNorm in the bean leaves as function of time exposition to
radiation with detergent.
B. R. Jovanić et al. / HEALTH 2 (2010) 395-399
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To avoid effects of plant age on chlorophyll concentra-
tion the control and dosed plants were of the same age,
To avoid determining real chlorophyll concentration,
relative chlorophyll concentration reduction was deter-
mined: chlorophyll concentration in the dosed plants was
marked ΔChlNorm as opposed to the control samples of
the same age which were not treated with detergent. The
reduction in chlorophyll concentration in the marked
values ΔChlNorm over detergent treatment time is shown
in Figure 1. Figure 1 shows that with time relative
change of chlorophyll concentration increases quickly to
reach its peak on the seventeenth day at about 11.5%.
Obviously in this period the detergent has adverse effect
on chlorophyll by destroying it constantly. After this the
relative change of chlorophyll concentration remains
steady at the same value. This pattern can be explained
with a hypothesis that the plant adapted to the given un-
favourable conditions and developed a mechanism to
maintain the reduced chlorophyll concentration.
When determining photosynthetic activity PhAc of
photosynthetic apparatus in a plant leaf, equally when
determining reduction in chlorophyll concentration in
control and dosed plants, the plants were of the same age
to avoid the effect of plant age. To avoid determining
absolute photosynthetic activity, relative reduction of
PhAc was determined. The PhAc in detergent treated
plant was marked PhAcNorm to distinguish them from
control samples of the same age which were not treated
with detergent. The obtained changes in marked values
PhAcNorm over the detergent treatment time are shown in
Figure 2. Unlike chlorophyll concentration, photosyn-
thetic activity PhAc constantly, exponentially decreases
to reach only 45% of initial activity on the 21st day. This
pattern indicates that this water has adverse effects on
Scientific papers offer data indicating different effects of
different detergents on plants. In most cases detergents
have adverse effects on plant pigments and morphology
and inhibit metabolic processes. It was found the toxic
effect of sodium dodecyl sulfate (SDS) and the house-
hold synthetic detergents (HSDs) Kristall and Tix (0.1, 1,
and 10 mg/l) on the diatom Alga Thalassiosira pseu-
donana [9]. By the presence in water of detergents for
wool domestic washings the native enzyme lost 50% of
activity after 20 min of incubation [10]. The effect of
detergent on plants varies depending on how the plant is
exposed to it. For example, when bean was watered with
0.01% (w/v) solution the nondenaturing, zwitterionic
detergent [3-{3-cholamidopropyl}-dimethylammonio]-
1-propane-sulfonate) 0.01% (w/v) it induced root hair
deformation [11]. Also, in mungbean (Vigna radiata)
seeds synthetic detergent induced reduction in dehydro-
genase activity [12]. The leaves completely lost their
turgor pressure and displayed chlorosis when they are
treated with detergents [13]. The biophysical character-
istics of the membrane were changed after detergent
(Brij 58) treatment [14]. Detergent inhibited growth,
metabolic activity took place only for 1 to 5 days, after
which metabolic activity also ceased [15]. Also, high
concentration of detergent might cause loss of the native
configuration of β-carotene [16]. Cell growth and fission
inhibition, as well as morphological changes and block-
ing of chlorophyll a synthesis, were recorded at 10 mg/L
concentration of detergent of household synthetic ab-
stergent (HSA) [17]. When β-carotene is treated with a
high concentration of detergent [18] this might cause
loss of the native configuration [18]. Higher plant thyla-
koid membranes can be fractionated with various deter-
gents [19].
As can be seen from our data chlorophyll is sensitive
to detergent which tallies with other research results. The
plant treated with water content detergents showed high
inhibitory effect on chlorophyll content in sunflower
leaves [20]. The aggregation of chlorophyll is partly
inhibited by detergent Triton X-100 [21]. In addition to
reducing its concentration, detergents have other effects
on chlorophyll. Studies on light harvesting complexes
LHCs show that detergent-induced dissociation of LHCs
and caused decline in bonding Chl b and Chl a [22]. The
results on pigment-protein complexes of Pisum sativum
thylakoids treated with detergent Triton X-100 and
n-octyl β-D-glucopyranoside show that reversible disso-
ciation of pigment-protein complexes occur [23]. Cell
growth and fission inhibition on cryptophytic alga
Chroomonas salina (Wils.) Butch. (Cryptophyta), as well
as morphological changes and blocking of chlorophyll a
synthesis, were recorded at 10 mg/L concentration of
household synthetic abstergent (HSA) «Tix» [17] .
In addition to the above discussed effect of detergent
on chlorophyll, it was to be expected that similar possi-
bly even identical effect will be observed on photosyn-
thesis. Detergent-induced reversible denaturation of the
photosystem reaction [24]. Detergents have strong effect
on the fluorescence properties of the light-harvesting
complexes of photosystem II [25]. Detergent (Triton
X-100) has effect on relaxation dynamics of photosys-
tem II [26]. Some results have shown that low concen-
tration of nonionic detergent Triton X is sufficient for
saturation of photosynthesis in terrestrial higher plants
[27]. Even a short exposure to detergent effects causes
extensive changes in photosynthesis. For example ex-
posing for 10 min in water containing a few drops of
liquid detergent induce the increase of photosyntesis
[28]. It was concluded that detergent (Triton X-100)
causes damage of the donor part of photosystem 2 in
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isolated chloroplasts [29]. Detergent treatment of the
membranes resulted in loss of PS I activities [30].
Non-ionic detergent n-dodecyl-α, D-maltoside cause
disintegration of the potosystem II (PS II) into separated
PS II in stacked and unstacked thylakoid membranes
from spinach [31]. The addition of the detergent Triton
X-100 to the ‘chromatophore’ facilitated the photooxida-
tive destruction of the antenna BChl [32]. In addition to
inhibition of activities PHII detergent can cause mor-
phological changes of these centres. Detergent treatment
of stacked thylakoid or BBY membranes usually gives to
PS II–LHC II varying size [31].
Detergents in the water for watering plants have adverse
effect. In the bean plants which were watered with de-
tergent water, significant changes were observed. Chlo-
rophyll concentration dropped by 12%. The activity of
photosynthesis apparatus in leaves decreased by around
This work was supported by Grants No. 141007 of MNRS. Authors
want to thank Mr. P. Hiddinga on kindness in supporting equipments.
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