DETERMINATION OF NITROGEN AND SULFUR IN PLANTS AND

SOILS SAMPLED FROM THE AREA NEAR A FERTILIZER PLANT

A. CICEK
Environmental Applications and Research Center, Anadolu University, Eskişehir, Turkey
(∗ author for correspondence, e-mail: acicek@anadolu.edu.tr, Fax: +90 222 323 9501)

(Received August 2002; accepted 10 January 2003)

Abstract. The purpose of this study was to investigate NOx and SO2 -induced nitrogen and sulfur
levels accumulated over vegetation and soil in the surrounds of the fertilizer plant in Kütahya, Turkey.
Therefore, leaf samples collected from the seven locations adjacent to the plant were morphologic-
ally observed (pollution-related injuries and color deformation) and chemically analyzed. Besides,
nitrogen and sulfur contents were determined in the soil samples collected from each location. A
significant correlation was achieved between locations and different vegetation samples and soil
samples. The highest nitrogen level present in the trees was 3.07 ± 0.105% (3rd location, Robinia
pseudoacacia), with the highest sulfur level being 5.79 ± 0.085% (3rd location, Salix alba). The
highest nitrogen and sulfur levels in soils were 0.31 ± 0.045 and 0.57 ± 0.052% (3rd location), re-
spectively. Furthermore, the highest nitrogen and sulfur levels were 4.69 and 2.20%, respectively, in
agricultural plants in the surrounds of the fertilizer plant. A higher level of pollutants was observed
in the samples collected from the locations in the direction of the prevailing wind. Also, differences
between locations as well as between plant specimens were statistically evaluated.

Keywords: accumulation, fertilizer plant, nitrogen, vegetation, soil pollution, sulfur

1. Introduction

The potential impact of atmospheric deposition of pollutants on vegetation has

been the subject of many papers (Mesanza et al., 1996; Kantarcı and Müezzinoğlu,
1997; Johnson et al., 1999).
Plant injury caused by air pollution is most common near large cities and indus-
trial enterprises. Damage in isolated areas occurs when pollutants are spread over
long distances by wind currents.
The three oxides of nitrogen normally encountered in the atmosphere are N2 O,
NO and NO2 . Microbially generated nitrous oxide is relatively unreactive and
probably does not significantly influence important chemical reactions in the lower
atmosphere. However, colorless, odorless nitric oxide and pungent red-brown ni-
trogen dioxide, collectively designated NOx , are very important in polluted air.
Although extensive damage to plants is observed in areas receiving heavy exposure
to NO2 , most of this damage probably comes from secondary products of nitrogen
oxides, such as Peroxy Acetyl Nitrate formed in smog. Atmospheric sulfur dioxide
is harmful to plants. Acute exposure to high levels of the gas kills leaf tissues.

Water, Air, and Soil Pollution: Focus 3: 243–253, 2003.

© 2003 Kluwer Academic Publishers. Printed in the Netherlands.
244 A. CICEK

Chronic exposure of plants to sulfur dioxide causes chlorosis, a bleaching or yel-

lowing of the normally green portions of the leaf. Sulfur dioxide in the atmosphere
is converted into sulfuric acid, so that in areas with high levels of sulfur dioxide
pollution, plants may accordingly be damaged by sulfuric acid aerosols (Manahan,
1997).
Acid rain occurs within, and downwind of, areas of major industrial emissions
of sulfur dioxide and the oxides of nitrogen. After sulfur dioxide and oxides of
nitrogen are emitted into the atmosphere, they are transformed into sulfate or nitrate
particles and, by combining with water vapor, into mild sulfuric and nitric acids.
These acids then return to earth as dew, drizzle, fog, sleet, snow, and rain (Henry
and Heinke, 1996).
There are known to be many pollutants released from fertilizer plants, such
as SO2 , H2 S, NH3 , CO, HF and particle matter. However, this study particularly
focused on nitrogen and sulfur contents among those pollutants in vegetation and
soil samples around the fertilizer plant in Kütahya, Turkey. A plume of dark yellow
color released from the fertilizer plant, which has been in operation in Kütahya
for quite a long time, threats the people residing in this area. It is a well-known
fact that the plant has so far been unable to filter the menacing smoke it releases
into the atmosphere due to high cost. Moreover, this plant cannot discharge waste
water by law. However, no current studies into this problem are under way, either.
The aim of this study was to determine the degree to which noticeably dangerous
nitrogen oxides, released from the aforementioned fertilizer plant, had accumulated
in soil and vegetation. As was expected, there was an overwhelming accumulation
of nitrogen and sulfur in vegetation and soil in the area around the fertilizer plant. It
was also observed that there were symptoms induced by pollution. These taken into
consideration, chemical analyses were carried out in support of the presumption of
the study. The annual production of fertilizer with nitrogen of this plant amounts to
230,000 tonnes and also produces 25,000 polyethylene bags. Comparison of results
of this study and those of vegetation and soil samples analyses suggested that the
fertilizer plant continues to be a growing threat to the environment in respect of
the poisonous gases it lets out, and therefore should take precautions against this
problem.

2. Material and Methods

2.1. D ESCRIPTION OF THE RESEARCH AREA

2.1.1. Geological Properties

Kütahya is located between 38◦ 70 –39◦ 80 N and 29◦ 00 –30◦ 30 E and in West-
Middle Anatolia in Turkey. Its altitude is 950 m. To the south of the city is Yellice
Mountain (1764 m) and Gümüs Mountain (1901 m), to the north is Yesil Mountain
(1533 m) to the west Egrigöz (2181 m), to the east Türkmen Mountain (1829 m).
 AIR POLLUTION AND IMPACTS ON FORESTS 245
TABLE I
Some climatic data of the study area

Meteorological data Kütahya

Annual mean temperature (◦ C) 10.6

Annual mean precipitation (mm) 564.6
Mean relative humidity (%) 67
Monthly mean local pressure (mb) 904.7

This area consists of alluvial soils (Paleozoic, Mesozoic and Cenozoic), the oldest
schist limestone belonging Devonian period.

2.1.2. Vegetation
The surveyed area forms a transitional zone among the Mediterranean, European-
Siberian and Irano-Turanian phytogeographical regions. Forest cover on the high
plateaus and mountainous area of the Kütahya consist of Pinus nigra, Quercus
cerris, Q. infectoria, Q. libani, Juniperus excelsa, J. foetidissima, Fagus orientalis,
Platanus orientalis.

2.1.3. Climate
Meteorological data of Kütahya are given in Table I. In the research area the pre-
vailing wind blows from the NW (wind speed 27.7 m s−1 ) and N (wind speed 1.7 m
s−1 ).

2.2. S AMPLING AND CHEMICAL ANALYSES

The most widespread vegetation samples surrounding the fertilizer plant were se-
lected for the study. With the exception of Northwest of the fertilizer plant, where
there is only poor vegetation, vegetation and soil samples were collected from
seven different locations. The discrepancy between the distances of the locations,
from which samples for the study were collected, resulted from the fact that veget-
ation occurred at only those spots. During the collection process, tree leaves were
painstakingly collected from the heights of 1.5 and 2 m of the trees in considera-
tion to achieving homogeneity, while the leaves of agricultural plant leaves were
collected indiscriminately.
Following are 25 plant samples that stand for 17 plant species, with 3 differ-
ent plant samples belonging to each plant species being collected, 75 altogether;
(deciduous tree species: Platanus orientalis L., Robinia pseudoacacia L., Populus
tremula L., Malus sylvestris Miller, Salix alba L., Pyracantha coccinea Roemer.,
Vitis vinifera L., Ceratonia siliqua L., and coniferous tree species: one-year-old
leaves belonging to Pinus nigra subsp. pallasiana (Lamb.) Holmboe., Picea pun-
246 A. CICEK

Figure 1. Sampling areas.

gens Engelms, Thuja orientalis L., and agricultural plant species: Medicago sativa
L., Spinacia oleracea L., Triticum sativum L., Zea mays L., Capsicum annuum L.,
Poa sp.) and 21 soil samples were collected between February–March 2001 around
the Fertilizer Plant in Kütahya. Locations, from which vegetation and soil samples
were collected, their distance from the fertilizer plant and directions and the total
species and samples are presented in Figure 1.
The unwashed and washed (with tap water and distilled water, and then air-
dried) leave samples were dried at 60 ◦ C for 24 hr and powdered in a hammer
mill and then passed through a 0.5 mm sieve. Soil samples were collected at
depths of 0–20 cm in each plot. Soils were dried and then sifted a 0.5 mm sieve.
Nitrogen contents of the plant samples and soils are determined by Kjeldahl Diges-
tion method (Brenner, 1965; Hortwitz, 1970). Sulfur contents of the plant samples
and soils were determined by nephelometric method (Butters and Chenery, 1959;
Massoumi and Cornfield, 1963).
Statistical analysis was done separately for each location using one-way AN-
OVA test (pairwise multiple comparison) using a software programme. For the par-
ticular plant species and soil samples, the independent sample t-test was conducted
to determine whether the equality means in statistically significant.
 AIR POLLUTION AND IMPACTS ON FORESTS 247
TABLE II
Total nitrogen and sulfur contents (dry weight, % ± standard deviation) of unwashed and washed
plant samples

Locations Plant samples n Nitrogen Sulfur

Unwashed Washed Unwashed Washed

1 Platanus orie. 3 2.01±0.035 1.88±0.081 3.03±0.125 1.31±0.079

2 Pinus nigra 3 1.43±0.070 1.09±0.098 0.77±0.075 0.40±0.087
Robinia pseud. 3 2.31±0.121 2.30±0.190 0.99±0.096 0.82±0.122
Thuja orie. 3 1.79±0.105 1.07±0.160 4.08±0.325 1.25±0.096
Populus trem. 3 1.51±0.078 1.44±0.055 2.18±0.098 1.84±0.079
Platanus orie. 3 1.70±0.045 1.66±0.052 2.96±0.060 2.71±0.078
3 Pinus nigra 3 1.73±0.062 1.64±0.036 1.33±0.070 0.31±0.026
Robinia pseud. 3 3.07±0.105 2.39±0.072 3.67±0.088 1.85±0.078
Malus sylve. 3 1.24±0.070 0.97±0.075 1.87±0.036 1.35±0.062
Salix alba 3 2.14±0.101 1.91±0.036 5.79±0.085 5.66±0.072
4 Thuja orie. 3 1.29±0.098 1.17±0.062 3.66±0.075 1.34±0.173
Pyracantha coc. 3 1.44±0.079 1.37±0.043 4.56±0.122 3.36±0.170
Pinus nigra 3 1.19±0.045 1.14±0.055 1.34±0.101 1.24±0.122
Picea pungens 3 1.24±0.070 1.08±0.079 1.34±0.055 1.12±0.081
Poa sp. 3 1.66±0.081 1.37±0.131 2.75±0.166 2.05±0.052
5 Populus tremu. 3 1.53±0.060 1.20±0.141 0.99±0.095 0.64±0.127
Medicago sati. 3 2.06±0.079 1.87±0.081 1.18±0.078 1.12±0.060
Spinacia oler. 3 4.30±0.259 4.20±0.336 3.54±0.130 2.10±0.085
6 Triticum sati. 3 4.69±0.196 3.85±0.235 4.24±0.134 3.98±0.127
7 Pinus nigra 3 1.32±0.108 1.29±0.081 0.48±0.065 0.21±0.034
Thuja orie. 3 1.33±0.043 1.29±0.080 1.14±0.122 0.40±0.080
Zea mays 3 1.82±0.088 1.74±0.055 0.57±0.036 0.38±0.069
Vitis vinifera 3 1.82±0.079 1.64±0.105 1.63±0.095 0.78±0.088
Capsicum ann. 3 2.90±0.098 2.61±0.141 2.21±0.115 1.63±0.090
Ceratonia sili. 3 1.92±0.113 1.69±0.165 1.89±0.091 1.04±0.065

3. Results and Discussion

Results of nitrogen and sulfur analyses done in vegetation samples in the vicinity
of the fertilizer plant are presented in Table II. Statistical evaluations of differ-
ent nitrogen and sulfur contents in consideration to all locations are presented in
Table III.
Significant differences were determined in respect to sulfur content in washed
leaves between 1st and 4th locations on analysis of the data of deciduous leaves
 TABLE III
248

Statistical differences between nitrogen and sulfur contents in vegetation within locations, using a pairwise multiple comparison of one way ANOVA (95%
confidence interval of the mean)
Location Deciduous tree (n = 33) Coniferous tree (n = 24) Agricultural plant (n = 18)
pairs Nitrogen Sulfur Nitrogen Sulfur Nitrogen Sulfur
uw w uw w uw w uw w uw w uw w

Mean difference ± Std. Dev.

1–2 NS NS NS NS – – – – – – – –
1–3 NS NS NS NS – – – – – – – –
1–4 NS NS NS –2.05±0.98 – – – – – – – –
1–5 NS NS –2.04±0.85 NS – – – – – – – –
1–6 – – – – – – – – – – – –
1–7 NS NS NS NS – – – – – – – –
2–3 NS NS –1.73±0.49 –1.16±0.56 NS –0.56±0.06 NS NS – – – –
A. CICEK

2–4 NS NS –2.51±0.69 NS 0.37±0.66 NS NS NS – – – –

2–5 NS 0.60±0.27 NS NS – – – – – – – –
2–6 – – – – – – – – – – – –
2–7 NS NS NS NS 0.25±0.66 –0.24±0.45 1.48±0.58 NS – – – –
3–4 0.71±0.310 NS NS NS 0.49±0.84 0.51±0.05 NS NS – – – –
3–5 NS NS 2.78±0.69 2.31±0.80 – – – – – – – –
3–6 – – – – – – – – – – – –
3–7 NS NS 2.01±0.54 2.04±0.63 0.37±0.84 0.32±0.05 NS NS – – – –
4–5 NS NS 3.57±0.85 2.72±0.98 – – – – –1.52±0.58 –1.66±0.58 NS NS
4–6 – – – – – – – – –3.03±0.67 –2.48±0.68 NS –1.93±0.42
4–7 NS NS 2.80±0.73 2.45±0.85 –0.12±0.05 –0.19±0.04 1.17±0.05 NS NS NS NS 1.04±0.37
5–6 – – – – – – – – –1.51±0.58 NS –1.88±0.67 –2.37±0.37
5–7 NS NS NS NS – – – – NS NS NS NS
6–7 – – – – – – – – 2.33±0.58 NS 2.85±0.67 2.97±0.37

uw: Unwashed leaves; w: Washed leaves; Std. Dev.: Standard deviation; NS: Not significant.
 AIR POLLUTION AND IMPACTS ON FORESTS 249

in Table III. The reason could simply be that sulfur content is much higher leaves
occurring in the 4th location, which is in the direction of the prevailing wind. It is
also possible to attribute concentration of sulfur, which is present over the surface
of leaves as particle matter, to the fact that it penetrates into leaves by means of the
physiological activities of leaves.
The difference observed between 3rd and 4th locations in respect to nitrogen
in unwashed deciduous leaves could also be attributable to the anatomy of leaves.
In particular, the fact that the surface of leaves of Salix alba species are hairy may
well contribute to a high accumulation of sulfur. The high rate of accumulation
in Robinia pseudoacacia in the 3rd location could result from the fact that R.
pseudoacacia is a good accumulator (Aksoy et al., 2000). That 3rd location is
in the direction of the prevailing wind and therefore has the most exposure to the
gases released from the fertilizer plant result in statistical differences in comparison
with the other locations.
Statistically significant difference was determined between washed and
unwashed deciduous leaves collected from 3–5th, 3–7th, 4–5th and 4–7th loca-
tions, respectively. This taken into account, it was concluded that pollution in 3rd
and 4th locations was as much as it had been expected.
Deciduous tree leaves (n:33), coniferous tree leaves (n:24) and agricultural plant
leaves (n:18) used for the study produced results as follows; on analyses comparing
coniferous leaves with deciduous leaves, it was determined that nitrogen levels in
unwashed samples showed difference in comparison with 3rd and 4th locations.
Nonetheless, no significant difference was determined between the aforementioned
locations in respect to sulfur. Also, similar properties were observable between 3rd
and 7th locations. It is possible to attribute the differences in pollution levels not
just to locations themselves but also to the very properties of the leaves.
When analyzing agricultural plant leaves, it was concluded that nitrogen levels
were less in 4th location than those in 6th and 7th ones. The reason could well be
the fact that Poa sp. are continually watered.
A significant difference was determined for sulfur contents in washed and un-
washed leaves, and for nitrogen contents in only unwashed leaves in 5–6th and
6–7th locations. However, such a difference was thought to stem from the normal
levels of nitrogen and sulfur present in leaves.
There is a downside to the study that not all vegetation samples were existent
in each location, which led to a restriction. For this reason, statistical analyses of
generalized vegetation samples have been made and appear in Table IV.
Significant differences were determined for deciduous leaves and coniferous
leaves in terms of nitrogen content in both washed and unwashed samples. Similar
differences were also true for sulfur.
As a result of statistical analyses, no significant difference was determinable
between deciduous and agricultural leaves in terms of sulfur content, which could
be due to the fact that leave samples were look-alike. There was a significant differ-
ence between nitrogen contents of agricultural and deciduous leaves. The reason
250 A. CICEK

TABLE IV
Statistical evaluations of nitrogen and sulfur contents between plant species (washed
and unwashed leaves) by paired t-test, considering the global significance level of
5%

Parameter Plant species Mean difference ± Std. Dev. Significance

pairs

Nitrogen (uw) a–b 0.545±0.204 ∗

a–c –0.490±0.204 ∗

b–c –1.035±0.204 ∗∗

Nitrogen (w) a–b 0.375±0.169 ∗

a–c –0.510±0.169 ∗∗

b–c –0.885±0.169 ∗∗

Sulfur (uw) a–b 0.950±0.330 ∗

a–c 0.370±0.330 NS
b–c –0.580±0.330 ∗

Sulfur (w) a–b 0.605±0.239 ∗

a–c –0.009±0.239 NS
b–c –0.700±0.239 ∗

a. Deciduous tree leaves; b. Coniferous tree leaves; c. Agricultural plant leaves.

uw: Unwashed leaves; w: Washed leaves.
∗ p < 0.05; ∗∗ p < 0.01; Std. Dev.: Standard deviation; NS: Not significant.

TABLE V
Total sulfur and nitrogen contents in soil samples (dry
weight, % ± Standard deviation)

Locations Nitrogen Sulfur

1 0.18±0.036 0.18±0.017
2 0.26±0.026 0.18±0.026
3 0.31±0.045 0.57±0.052
4 0.31±0.026 0.32±0.036
5 0.19±0.026 0.20±0.045
6 0.16±0.026 0.16±0.036
7 0.16±0.026 0.15±0.043
 AIR POLLUTION AND IMPACTS ON FORESTS 251
TABLE VI
Statistical differences between nitrogen and sulfur contents in soils by paired t-test,
considering the global significance level of 5%, within locations (n = 3)

Location Nitrogen Sulfur

pairs Mean difference Significance Mean difference Significance
± Std. Dev. ± Std. Dev.

1–2 –0.08±0.026 ∗∗ 0.00±0.031 NS

1–3 –0.13±0.026 ∗∗ –0.39±0.031 ∗∗
1–4 –0.13±0.026 ∗∗ –0.14±0.031 ∗∗
1–5 –0.01±0.026 NS –0.02±0.031 NS
1–6 0.02±0.026 NS 0.02±0.031 NS
1–7 0.02±0.026 NS 0.03±0.031 NS
2–3 –0.05±0.026 NS –0.39±0.031 ∗
2–4 –0.05±0.026 NS –0.14±0.031 ∗∗
2–5 0.07±0.026 ∗ –0.02±0.031 NS
2–6 0.10±0.026 ∗∗ 0.02±0.031 NS
2–7 0.10±0.026 ∗∗ 0.03±0.031 NS
3–4 0.00±0.026 NS 0.25±0.031 ∗∗
3–5 0.12±0.026 ∗∗ 0.37±0.031 ∗∗
3–6 0.15±0.026 ∗∗ 0.41±0.031 ∗∗
3–7 0.15±0.026 ∗∗ 0.42±0.031 ∗∗
4–5 0.12±0.026 ∗∗ 0.12±0.031 ∗∗
4–6 0.15±0.026 ∗∗ 0.16±0.031 ∗∗
4–7 0.15±0.026 ∗∗ 0.17±0.031 ∗∗
5–6 0.03±0.026 NS 0.04±0.031 NS
5–7 0.03±0.026 NS 0.05±0.031 NS
6–7 0.00±0.026 NS 0.01±0.031 NS
∗ p < 0.05; ∗∗ p < 0.01.
NS: Not significant; Std. Dev.: Standard deviation.

was thought to result from the fact that agricultural leaves absorb nitrogen from
both air and soil.
It is vital to point out that the structure of coniferous leaves is dissimilar to that
of deciduous ones in that it is both smooth and thick. Therefore, accumulation
of pollutants occurs in a less amount in and over the surface of leaves, which
supported by statistical analyses.
Nitrogen and sulfur contents of soil samples are given in Table V. Statistical
evaluations showing parameters of the distribution of pollutants in soil are presen-
ted in Table VI.
In consideration of Tables V and VI, nitrogen and sulfur contents of soil are
252 A. CICEK

categorized from the most polluted locations to the least ones. For nitrogen: 3 ≈ 4
> 2 > 5 > 1 ≈ 6 ≈ 7; for sulfur: 3 > 4 > 1 ≈ 2 ≈ 5 ≈ 6 ≈ 7.

4. Conclusion

In this study, the chemical analyses of nitrogen and sulfur contents in the leaves
of some vegetation species scattered around Kütahya Fertilizer Plant were carried
out. The same analyses were conducted for the soil in which these plants grow.
It was observed that the NOx and SO2 released from the factory, which does not
have a stack filtering system for flue gases, have negative effects on the vegetation.
On morphological observations, the development of black and brown lesions was
observed especially on the leaves of deciduous trees, peppers (Capsicum annuum
L.), grapes (Vitis vinifera L.) and corns (Zea mays L.). The bronzing and silvering
were also observed on the leaves of acacia (Robinia pseudoacacia L.) and plane
tree (Platanus orientalis L.).
As a result of all analyses conducted in the study, it was concluded species of
leaves differed from one another in respect to accumulating nitrogen and sulfur.
Due to the differences between the anatomy and morphology deciduous and con-
iferous tree leaves, it was observed that the deciduous tree leaves absorbed more
pollutants than did coniferous ones. Furthermore, significant differences were re-
corded between agricultural plant leaves and coniferous leaves depending on their
locations. As it was also supported by statistical analyses of soil samples, pollution
concentrated mostly in the prevailing wind direction.
In the literature normal nitrogen levels are 0.5–3.5% and sulfur levels are 0.5–
1.5%, respectively. Generally, the percentage of nitrogen at the upper portion of
cultivated soil is between 0.06 and 0.5, whereas the nitrogen content of lower por-
tion is below 0.02%. The total nitrogen content of organic soil is greater than 2.5%.
Total sulfur content is about 0.002% for sandy soil and 0.06% for heavy textured
soil. Generally sulfur levels in most soils are between 0.01 and 0.05% (Kacar,
1970). As to our study, these levels were observed to be exceeded particularly in
nitrogen and sulfur levels present in soil samples.

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