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ISSN: 2573-458X

Environment Pollution and
Climate Change

Costa et al., Environ Pollut Climate Change 2022, 6:8

Research Article

Open Access

Determination of Pesticides in the Lower Sao Francisco River in Brazil
Hanna Francyelle Barbosa Costa1, Emerson Carlos Soares e Silva1, Mariane Lessa Costa1, Sandra Helena Vieira de Carvalho2, Themis Jesus
Silva1, João Inácio Soletti2 and Mozart Daltro Bispo2*
1
2

Aquaculture and Water Analysis Laboratory (LAQUA), Center for Agrarian Sciences, Federal University of Alagoas (UFAL), Brazil
Process Separation and Optimization System Laboratory (LASSOP), Federal University of Alagoas (UFAL), Brazil

Abstract
A significant proportion of the pesticides that are widely used in agriculture are highly toxic pollutants, despite their low
concentrations. They have high capacity for polluting water, soil and air, especially during their application. The difficulty
and cost of analyses to identify them and the inefficiency of conventional water treatment for human consumption,
combined with the lack of legislation on this subject, has favored their accumulation in the environment. The present
study was conducted in the lower part of the São Francisco River, in Brazil, between the Xingó hydroelectric power plant
(municipality of Piranhas, Alagoas State [AL]) and the mouth of the river (municipality of Piaçabuçu, Alagoas State [AL]).
This stretch of the river is approximately 240 km long. Water was sampled in November 2021, at transects downstream
from the municipalities of Piranhas AL, Pão de Açúcar AL, Traipu AL, São Brás AL, Propriá (Sergipe State) and Penedo
AL. The technique used for analytical monitoring was gas chromatography coupled to mass spectrometry (GC-MS).
The presence of 14 pesticides was detected, among the 31 that were evaluated. Among the compounds identified,
three are classified as extremely toxic, six are classified as highly toxic and five are classified as moderately toxic. The
presence of three compounds was identified in the municipality of Pão de Açúcar and two of these were highly toxic.
In Traipu, there were four compounds, among which one was extremely toxic and two were highly toxic. In São Brás,
there were four compounds, among which three were the same as found in Traipu. Propriá and Penedo presented the
highest diversity of pesticide contaminants, respectively nine and ten pollutants.

Keywords: Chromatography; Electric Vehicle; Emerging pollutants
Introduction
With advances in agriculture, sustainable means for combating
pests that threaten food supply have increasingly been sought [1].
Pesticides are widely used against these food crop pests but their
distribution can reach the wider environment in aqueous solution,
especially during rainfall. They percolate down through the soil and
eventually reach the groundwater [2,3]. Because of the persistence of
pesticides in the environment, great mobility, high bioaccumulation
and toxicity, they have become a source of pollution at the global level
[4,5].
Despite the importance of pesticides for protecting and ensuring
good-quality crops, they are of concern regarding human health
[6,7]. Their use and the handling practices involved give rise to high
levels of exposure and adverse health effects. Exposure of humans to
pesticides results in immunosuppression, hormonal interruption,
reduced intelligence, reproductive distortion and cancer. The impacts
of this exposure can be divided into acute problems, such as the onset
of Parkinson’s disease, and chronic problems such as vision reduction,
for example [8,9].
As a measure to protect public health, guidance levels for pesticides
in drinking water have been implemented by national governments.
There are several guidance values, and a few of them are issued by the
World Health Organization (WHO) and the United Nations Food and
Agriculture Organization (FAO), which establish acceptable levels of
pesticide residues.
Currently, in Brazil, there are almost 5,000 pesticides that have
been released for use. These contain more than 550 active ingredients
that have been authorized by the Ministry of Agriculture for use in
formulations on the market, as agricultural pesticides on crops and
for plant protection products. Registration of pesticides is regulated
through Decree No. 4,074/2002; this is a responsibility shared between
the Ministry of Agriculture, Livestock and Supply (MAPA), Ministry of
the Environment (MMA) and Ministry of Health.
Environ Pollut Climate Change, an open access journal

Because of the risks that these pollutants bring to health and the
environment, it is necessary to constantly monitor the presence of
pesticides in water, with regard to the toxicity level of each compound.
Moreover, to understand the effects of contaminants, it is essential
to quantify and monitor concentrations at the emission source, in
environmental compartments and in living organisms [10]. In Brazil,
the National Water Quality Assessment Program (PNQA, 2022) aims
to contribute to sustainable management of water resources and guide
the development of policies for the recovery of environmental quality
in inland water bodies.
Selection of these compounds for regulation is not easy to evaluate,
regarding both identification and quantification. Mass spectrometry
(MS), an analytical identification technique for compounds of
interest, has been gaining a prominent position for use in organic
environmental analyses [11]. An approach consisting of use of gas
chromatography with mass spectrometry (GC-MS) combined with
solid-phase microextraction (SPME) techniques is mentioned in
regulated methods (ISO 27108:2013 and ASTM D 6520:2000). Such an
approach is essential in order to undertake sensitive positive structural
identification of pesticides and other pollutants in the environment
[12].
SPME can be done in two ways: headspace mode (HS-SPME)
*Corresponding author: Mozart Daltro Bispo, Process Separation and
Optimization System Laboratory (LASSOP), Federal University of Alagoas
(UFAL), Brazil; E-mail: mozartdaltro@hotmail.com
Received: 04-Aug-2022, Manuscript No. EPCC-22-71220; Editor assigned: 06Aug-2022, PreQC No. EPCC-22-71220 (PQ); Reviewed: 20-Aug-2022, QC No.
EPCC-22-71220; Revised: 22-Aug-2022, Manuscript No. EPCC-22-71220 (R);
Published: 29-Aug-2022, DOI: 10.4172/2573-458X.1000293
Citation: Costa HFB, Silva ECS, Costa ML, Carvalho SHV, Silva TJ, et al. (2022)
Determination of Pesticides in the Lower Sao Francisco River in Brazil. Environ
Pollut Climate Change 6: 293.
Copyright: © 2022 Costa HFB, et al. This is an open-access article distributed
under the terms of the Creative Commons Attribution License, which permits
unrestricted use, distribution, and reproduction in any medium, provided the
original author and source are credited.

Volume 6 • Issue 8 • 1000293

Citation: Costa HFB, Silva ECS, Costa ML, Carvalho SHV, Silva TJ, et al. (2022) Determination of Pesticides in the Lower Sao Francisco River in
Brazil. Environ Pollut Climate Change 6: 293.

Page 2 of 6
or through immersion of fibers in liquid samples (direct SPME).
When complex arrays are processed, HS-SPME is preferred because
interference can clog the extractor that covers the fiber; analytes need
to be volatile enough to pass easily into the headspace [13]. In some
studies, a limitation of SPME has also been described, relating to the
nature of the hydrophobic analytes that are attached to the wall of the
sample containers, thereby reducing the accuracy of determinations. In
this context, it is believed that on-site SPME sampling is the approach
to be used in the future for resolving this problem [14].
According to FAOSTAT (2022), Brazil is considered to be the
country with the highest availability of renewable water resources in
the world. The São Francisco River, the main river in eastern South
America, is classed in Brazil as a water body of great economic, social
and ecological importance. With a length of 1,811 miles, it is the fourth
largest river system on the continent and the largest river entirely in
Brazil. It is an important source of hydroelectric energy, irrigation,
water supply, fishing, aquaculture and navigation in the east and
northeast of Brazil [15].
However, intensive anthropic use has resulted in impacts on the
river. Thus, a need to adopt public policies and means for revitalization
of this asset has arisen [16].
The present study was carried out on the lower São Francisco River,
along the stretch of the river between the downstream side of the Xingó
hydroelectric power plant, in the municipality of Piranhas, Alagoas
(AL), and the mouth of the river, in the municipalities of Piaçabuçu,
AL, and Brejo Grande, Sergipe (SE). For this, a single extraction
procedure was established, using SPME combined with use of GCMS, in the selected ion monitoring mode (SIM) and scanning mode
(SCAN), with the aim of screening for 31 pesticides in surface waters.
The methodology was applied at different sampling stations as part
of a wide-ranging water quality monitoring survey, which included
sampling downstream from six municipalities in the basin in order to
assess the risk caused by pesticides in the São Francisco River.

Materials and methods
This study was conducted in the region of the lower São Francisco
River, corresponding to the area between the municipality of Piranhas
and the mouth of the river between the states of Alagoas and Sergipe.
This stretch of the river is approximately 240 km long. Samples were
collected in November 2021, at strategic points in the river (Table 1,
Figure 1).
Initially, interviews were conducted among producers and local
farmers in the lower São Francisco River region during 2020, regarding
their main crops and use of agrochemicals and fertilizers. Through this,
it was possible to select the main products used in the study area. The
pesticide standards, totaling 31 chemical compounds, were acquired
from Supelco (USA) in 1 mL ampoules. Table 2 presents some data
on the monitored compounds, including: CAS (registered name for
Table 1: Water sample collection points downstream from each municipality.
Cities

Date of collection

Geolocation of sample collection
Latitude

Longitude

Piranhas

11/1/2021

9°38'8.82"S

37°46'36.27"O

Pão de Açúcar

11/2/2021

9°44'52.18"S

37°27'30.02"O

Traipu

11/3/2021

9°58'20.72"S

37° 0'31.10"O

São Brás

11/4/2021

10° 7'35.06"S

36°54'56.03"O

Própria

11/5/2021

10°12'17.79"S

36°50'21.44"O

Penedo

11/7/2021

10°16'21.64"S

36°35'34.89"O

Environ Pollut Climate Change, an open access journal

Figure 1: Geographical location of collection points.
Table 2: Compounds monitored in the lower San Francisco River.
Compounds

CAS No.

Chemical Formula

Molecular
Mass
(g.mol-1)

Ethylenediaminetetraacetic
acid (EDTA)

60-00-4

C10H16N2O8

292.2

Alpha-BHC

319-84-6
319-85-7

C6H6Cl6

290.8

Beta-BHC
Lindane

58-89-9

290.8

Delta-BHC

319-86-8

C6H6Cl6

Heptacloro

76-44-8

373.3

Aldrin

309-00-2

C10H5Cl7

HeptachlorepoxideIsomer B

1024-57-3

γ-Chlordane

5103-74-2

C6H6Cl6

290.8

C6H6Cl6

290.8

C12H8Cl6

364.9

C10H6Cl8

409.7

C9H6Cl6O3S

406.9

C10H5Cl7O
C10H6Cl8

389.4
409.7

α-Chlordane

5103-71-9

Endosulfan I (alpha)

959-98-8

DDE

72-55-9

C14 H8 Cl4

318

Dieldrin

60-57-1

C12H8Cl6O

380.9

C9H6Cl6O3S

406.9

C12H8Cl6O

380.9

(ClC6H4)2CHCCl3

354.4

Endrin

72-20-8

Endosulfan II (Beta Isomer)

33213-65-9

DDD

72-54-8

Endrinaldehyde

7421-93-4

Endosulfan sulfate

1031-07-8

DDT

50-29-3

Endrin Cetona

53494-70-5

Methoxychlor

72-43-5

Diquat

85-00-7

Metribuzin

21087-64-9

Glifosato

1071-83-6

Ametryn

834-12-8

Atrazine

1912-24-9

Prometon

1610-18-0

Prometryn

7287-19-6

Propazine

139-40-2

Simazine

122-34-9

Terbutryn

886-50-0

C12H8Cl6O

(ClC6H4)2CHCHCl2
C9H6Cl6O4S
C12H8Cl6O

380.9
320
422.9
380.9

C16H15Cl3O2

345.6

C8H14N4OS

214.2

C9H17N5S

227.3

C12H12N2Br2
C3H8NO5P
C8H14ClN5

C10H19N5O

344
169
215.6
225.2

C10H19N5S

241.3

C7H12ClN5

201.6

C9H16N5Cl

C10H19N5S

229.7
241.3

the chemical compound in the Chemical American Society database),
chemical formula and molecular mass.
The compounds were identified using mass spectrum data from the
NIST 14 library (NIST/EPA/NIH Mass Spectra Library, version 2.2,
USA), linear retention indexes, and data from the literature and pattern
Volume 6 • Issue 8 • 1000293

Citation: Costa HFB, Silva ECS, Costa ML, Carvalho SHV, Silva TJ, et al. (2022) Determination of Pesticides in the Lower Sao Francisco River in
Brazil. Environ Pollut Climate Change 6: 293.

Page 3 of 6
injection. Linear retention indexes (LRI) were calculated in accordance
with the Van den Dool equation in Kratz (1963), using n-alkane (C7 to
C30) standards with purity above 99.5% (Sigma-Aldrich).

SPME procedure using headspace mode combined with GCMS (HS-SPME/GC-MS)
To extract contaminants of interest, which had boiling points less
than or equal to 270ºC, solid-phase microextraction in headspace
mode at 70ºC was applied. 15 mL vials equipped with a ‘mininert’ valve
(Supelco, Bellefonte, PA, USA) were accurately filled up with about 5
mL of the water samples collected.
A divinylbenzene/carboxin/polydimethylsiloxane (DVB/CAR/
PDMS) fiber of film thickness 50/30 μm (Supelco, Bellefonte, PA, USA)
was used, housed in a manual support (Supelco, Bellefonte, PA, USA).
A gas chromatograph was used together with an ion-trap mass
spectrometer (Shimadzu, GC-MS/QP2010 Plus, Kyoto, Japan). The
conditions used were the following: injector temperature, 260ºC;
injection mode, without division; capillary column, DB-5, 60 m long,
0.25 mm internal diameter, film thickness 0.25 μm (Agilent J&W); oven
temperature, 45ºC maintained for 5 min, then increased to 80ºC at a
rate of 10ºC min−1 and to 240ºC at 2ºC min−1; helium gas at a constant
pressure of 100 kPa; transfer line temperature, 250°C; and acquisition
range, 40-500 m/z.

Validation of the HS-SPME/GC-MS method
Preparation of standard solutions
Standard stock solutions were prepared at an approximate
concentration of 1 mg.L-1 in dichloromethane or acetone, according to
the solubility of the analyte. From these solutions, a working solution
was prepared at a concentration of 10 μg.L-1 with all standards. For the
analysis by means of HS-SPME/GC-MS in SIM mode, analytical curves
were constructed at concentrations between 0.005 and 10 μg.L-1. The
curve values were determined and adjusted using the maximum value
allowed according to CONAMA Resolution No. 357 [17], shown in the
Table 3.
Calibration curve and detection and quantification limits
Linearity was determined in terms of the evolution of the
regression curves of volume versus standard concentration and was
expressed as the linear coefficient (R2). The limits of detection (LD) and
quantification (LQ) were determined based on the standard deviation
of the intersection of the analytical curve(s) and the slope of the line(s).
These are presented in Equations 1 and 2, respectively.

s
LD = 3,3
S
LQ = 10

s
S
Table 3: Limit values from the Brazilian legislation.

Analytical Parameters

Limit Values
(μg L-1)

DDT

0.002

Aldrin

0.05

HeptachlorepoxideIsomer B

0.01

All the analyses were performed in triplicate, using the SIM mode
and using the major and secondary ions. The analyses on the samples
did not present peak interferents, which thus conferred selectivity to
the method developed.

Results and Discussion
With the calibration curve, the linearity range investigated for
six analytes (n = 6) can be seen, along with the linear correlation
coefficients, LD, LQ and maximum values, as established by the
Ministry of Health through Ordinance no. 518 of 2004. The presence
of six compounds (Table 4), with R2 ranging from 0.981 to 0.996, was
confirmed. The detection limits for the method remained in the range
of 0.005 to 1.0 μg.L-1, and the quantification limits were between 0.001
and 0.003 μg.L-1, i.e. in accordance with the values established by the
legislation. The pesticides identified did not have values above (LQ),
and therefore were below the permitted concentration, as defined by
CONAMA Resolution No. 357 [17].
All the analytical monitoring was performed using gas
chromatography coupled to mass spectrometry. The chromatographic
profile in SIM mode is shown in Figure 2.
Table 5 shows the compounds identified through the SIM mode
in the six municipalities studied along the lower São Francisco River.
Table 6 shows the compounds identified in SCAN mode in the six
municipalities along the lower São Francisco River.
Using the HS-SPME/GC-MS method, in SIM mode, for the
municipality of Piranhas, none of the contaminants studied were
detected. However, in SCAN mode, through the calculation referring
to the LRI, four compounds with great toxicological potential could
be identified: carbendazim, fluoroacetamide, naphthalene and toluene.
The fungicide carbendazim is widely used on fruit and vegetable crops
in Brazil. The chromatographic profile is shown in Figure 3.
Table 7 presents the classification applied, along with the
toxicological class as defined through the Collegiate Board Resolution,
RDC 294 (2019), correlated with the occurrence downstream of the
municipalities.
Among the 14 compounds identified, three are classified as
Table 4: Analytical parameters obtained through SPME/GC/MS in SIM mode.
Analytical Parameters

R2

(μg L-1)

LD

LQ

(μg L-1)

(μg L-1)

DDT

0.005 – 0.05

0.996

0.0009

0.001

Aldrin

0.005 – 0.05

0.981

0.0008

0.002

Beta-BHC

0.05 – 0.5

0.993

0.0008

0.001

HeptachlorepoxideIsomer
B

0.005 – 0.05

0.99

0.0007

0.001

Lindane

0.05 – 0.5

0.996

0.0009

0.002

y-Chlordane

0.05 – 0.5

0.989

0.0008

0.003

Table 5: Compounds identified using HS-SPME/GC-MS, SIM mode.
Analytical Parameters Piranhas Pão de Traipu
Açúcar

São
Brás

Própria Penedo

DDT

cd.

cd.

Aldrin

cd.

cd.

Beta-BHC

cd.

cd.

cd.

cd.

cd.

cd.

HeptachlorepoxideIsomer B

Lindane

0.02

y-Chlordane

0.04

Lindane

Toluene

2

y-Chlordane

Environ Pollut Climate Change, an open access journal

Linear Range

cd.

cd.

cd.

cd.

cd.

Volume 6 • Issue 8 • 1000293

Citation: Costa HFB, Silva ECS, Costa ML, Carvalho SHV, Silva TJ, et al. (2022) Determination of Pesticides in the Lower Sao Francisco River in
Brazil. Environ Pollut Climate Change 6: 293.

Page 4 of 6

Figure 2: Chromatographic profile in SIM Mode of monitored compounds.

Figure 3: Chromatographic profile of the sample from the municipality of Piranhas.
Table 6: Compounds identified using HS-SPME/GC-MS, SCAN mode.
Analytical Parameters

Piranhas

Pão de Açúcar

Traipu

São Brás

1,4-Diclorobenzene

Própria

Penedo

cd.

cd.

2-Etil Hexanol

cd.

2,6,10,15-Tetramethylheptadecane

cd.

3-Trifluoroacetoxypentadecane
Carbendazim

cd.
cd.

Fluoroacetamide

cd.

Naphthalene

cd.

cd.

Oxima-, methoxy-phenylToluene

cd.

extremely toxic (I), six are classified as highly toxic (II) and five are
classified as moderately toxic (III).
In the municipality of Pão de Açúcar, the presence of three
compounds was identified: lindane, fluoroacetamide and toluene;
two of these were highly toxic. In Traipu, four compounds were
found: heptachlor epoxide isomer B, lindane, 2,6,10,15-tetramethyl
heptadecane and fluoroacetamide; one of these was extremely toxic
and two were highly toxic. In São Brás, four compounds were found:
heptachlor epoxide isomer B, lindane, 3-trifluoroacetoxy pentadecane
and fluoroacetamide; three of these were identical to those found
Environ Pollut Climate Change, an open access journal

cd.

cd.

cd.

cd.

cd.

cd.

cd.

cd.

cd.

in Traipu. Propriá and Penedo presented the highest diversity of
pesticide contaminants: DDT, aldrin, beta-BHC, lindane, y-chlordane,
1,4-dichlorobenzene, fluoroacetamide, oxime methoxy phenyl and
toluene. In addition to these, the pollutant 2-ethyl hexanol was also
detected in Penedo. Therefore, in Propriá and Penedo, nine and ten
pollutants out of the 31 studied were identified, respectively.
It was observed that the compound fluoroacetamide, which is an
acute and highly toxic rat poison, was present in all samples collected
in the six municipalities. This is a matter of great concern.
This survey aimed to identify the pesticides present in water
Volume 6 • Issue 8 • 1000293

Citation: Costa HFB, Silva ECS, Costa ML, Carvalho SHV, Silva TJ, et al. (2022) Determination of Pesticides in the Lower Sao Francisco River in
Brazil. Environ Pollut Climate Change 6: 293.

Page 5 of 6
Table 7: Occurrence and classification of the pesticides and chemicals identified.
Analytical Parameters

Application

Toxicological Class

Occurrence

1,4-Diclorobenzene

Insecticide

III

Propriá / Penedo
Penedo

2-Etil Hexanol

Organochlorine Pesticide

III

2,6,10,15-Tetramethylheptadecane

Hydrocarbon

II

Traipu

Aldrin

Insecticide

III

Propriá / Penedo
Propriá / Penedo

Beta-BHC

Fungicide / Insecticide

II

Carbendazim

Fungicide

1

Piranhas

DDT

Insecticide

II

Propriá / Penedo

Fluoroacetamide

Insecticide

II

All

HeptachlorepoxideIsomer B

Organochlorine Pesticide

I

Traipu / São Brás
Except Piranhas

Lindane

Insecticide

II

Naphthalene

Insecticide / PAH

I

Piranhas

Oxima-, methoxy-phenyl-

Fungicide

III

Propriá / Penedo

Toluene

BTEX

III

Except Traipu e São Brás

y-Chlordane

Fungicide / Insecticide

II

Propriá / Penedo

in municipalities along the lower São Francisco River. The results
demonstrated the extent to which this water body is contaminated
by agrochemicals. A study by Ferreira et al. (2022) portrayed how
pesticide contamination in water not only directly impacts the quality
of drinking water in local areas, but also gives rise to indirect damage
through transfer between species such as in the food chain and in the
soil [18].
Studies have proven that combined or separate exposure to
agrotoxins carries risks to human health due to the action of agents
such as carcinogens, neurotoxins, toxins and endocrine disruptors
[19]. In addition, pesticides may give rise to transient or permanent
changes to the immune and hematological systems [20, 21]. Kim et al.
(2017) also found in their experiments that exposure to agrochemicals
is associated with the incidence of some diseases in the population,
such as asthma, Parkinson’s disease and cancer [22].
It is of great importance to detect and identify the pesticides
present in rivers and other water bodies so that ways to reduce the
adverse effects that these contaminants have on human health and the
environment can be found.

Conclusion
The HS-SPME/GC-MS method developed in this study proved
to be fundamental for determining the existence of pesticides in the
lower San Francisco River, with extreme sensitivity and selectivity. It
met the requirements in the range of μg.L-1, even in the presence of
organic matter, and at different concentrations. The SIM and SCAN
modes applied in pesticide determinations showed good results, even
in the presence of organic matter. The detection and quantification
limits found were low, but sufficient to detect these substances, at a
level lower than that established by the Brazilian legislation.
This study has proven to be very important from the point of view
of alerting the population and officials responsible for public policy
and the environment, and for raising their awareness. In addition,
the occurrence of new “unregulated” contaminants requires more
constant monitoring. Therefore, this study will provide support for
implementation of new policies and specific regulations, which are
fundamental for achieving and maintaining the healthy balance of the
environment.

Acknowledgements
The following laboratories were involved in developing this study:

Environ Pollut Climate Change, an open access journal

Separation System and Process Optimization Laboratory (LASSOP),
of the Technology Center; Natural Resources Research Laboratory
(LPqRN), of the Center for Agrarian Sciences; and Aquaculture and
Water Analysis Laboratory (LAQUA), of the Center for Agrarian
Sciences.
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Citation: Costa HFB, Silva ECS, Costa ML, Carvalho SHV, Silva TJ, et al. (2022) Determination of Pesticides in the Lower Sao Francisco River in
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