ISSN 00271314, Moscow University Chemistry Bulletin, 2011, Vol. 66, No. 6, pp. 345–350. © Allerton Press, Inc.

, 2011.
Original Russian Text © E.M. Basova, M.A. Bulanova, V.M. Ivanov, 2011, published in Vestnik Moskovskogo Universiteta. Khimiya, 2011, No. 6, pp. 419–425.

Photometric Detection of Urea in Natural Waters

E. M. Basovab, M. A. Bulanovab, and V. M. Ivanova
a
Division of Analytical Chemistry, Department of Chemistry, Moscow State University, Moscow, Russia
email: mvonavi@mail.ru
bDubna International University of Nature, Society, and Humans

Received February 1, 2011

Abstract—The photometric detection of urea with the use of pdimethylaminobenzaldehyde as a reagent has
been develeped. The method allows one to reliably determine 10 mg/L of urea with the volume of an aliquot
of 10 mL. The method has been applied for the determination of urea in river water.

Keywords: urea, pdimethylaminobenzaldehyde, natural water, photometric detection.

DOI: 10.3103/S0027131411060022

The indicator method of investigation for interwell
space of an oil deposit is widely used in geophysics for
the control of displacement of oil by flood water [1].
The method is based on the injection of the required
volume of liquid labeled with the indicator into the
studied bench through the injection well, edging it by
the flood water to controllable extraction wells, and
investigation of time change of indicator concentra
tion in liquid flow outward from the bench. The results
of the indicator study are used to create geological and
hydrodynamic models of oil fields, for estimation of
oil reserves, and projecting for mining. Whereas, when
moving the indicator over the reservoir, dilution up to
104 times takes place, it is necessary to have a method
of control of its content at the level of 1 mg/L.
As indicators, fluorescent dyes, phosphate, thio
cyanate, and nitrateions are widely used [1]. Urea
meets all the requirements of the indicators in geo
physical studies [1]: very soluble in water, doesn’t
affect the processes of oil refining, environmentally
safe, cheap, and provides cost efficiency of indicator
studies. Urea is used as an indicator in the Republic of
Belarus. Simultaneously, another 3–4 indicators are
used, which request methods of selective determina
tion for each of them and there is no disturbing influ
ence of other indicators.
There are a few methods of urea determination.
Total nitrogen content in end products is controlled by
the titrimetric method after decomposition of urea by
concentrated H2SO4 and converting it into ammo
nium ion [2]. The sample mass is 1 g in the determina
+ mined solution with pdimethylaminobenzaldehyde
tion of NH 4 by the formaldehyde method and 5 g
when using the predistillation of ammonia. The titri
metric method of analysis is not suitable for determi
nation of a low content of urea.
The determination of urea in blood, urine, and
other biological fluids is based on the quantitative
transfer of it into ammonium carbonate by the enzyme
urease with subsequent spectrophotometric determi
+
nation of NH 4 ions by the Nessler reagent [2, 3]. The
method allows us to determine 5–20 g/L of urea nitro
gen [3]. Organic nitrogen in natural and waste waters
is also determined by the Nessler reagent after decom
position by the concentrated H2SO4 [3, 4].
While analyzing natural waters a voluntary volume
of the sample should be 500 mL, in addition, ammonia
is predistilled from the alkaline solution [3]. While
analyzing waste waters 100 mL of the sample is
enough, and the determination is carried out directly
(without distillation) [3]. Spectrophotometric deter
mination of ammonium ions by the Nessler reagent is
sensitive enough—from 0.05 to 4.00 mg/L in natural
and waste waters, 50 mL of the sample is required for
analysis [5]. Whereas the samples of natural waters, as
well as biological fluids, already contain ammonium
nitrogen, its concentration should be established pre
viously and then subtracted from the concentration,
and obtained after the decomposition of urea. The dis
advantage of the method is the necessity to work with
nonammonia water (as one way bidistilled water is
passed through a column with the cation exchanger
KU2).
Direct spectrophometric determination of urea is
of great interest. The appearance of yellowgreen
coloration in the process of reaction of the deter
(DMABA) in the presence of HCl is used for the
detection of urea, and the detection limit is 2 mg/L
[6]. The method of spectrophometric determination
of urea in the reservoir water using DMABA is laid

345
346 BASOVA et al.
Table 1. Dependence of the absorbency on the concentra
tion of urea in water
c, mg/L 1 5 10 15 20 25
A (400 nm) 0.005 0.016 0.033 0.046 0.068 0.080
out on the internet in the analytical forum
ANCHEM.RU and characterized by a minimum
detectable concentration (2 mg/L) [7]. Formation of
the yellow compound of urea with DMABA underlies
the method of the determination for mass fraction of
amide nitrogen in fertilizers. The range of detectable
contents is 20–46 mass % [8]. Thereby, to construct a
calibration curve, 160–320 mg of urea were added into
100mL flasks. In summary, the method [8] allows one
to analyze water samples with the concentration of
1.8–3.6 g/L, which is three orders of magnitude
greater than in method [7].
The purpose of this work is to estimate the reliabil
ity of information from the internet and development
of the method for detection of urea in natural waters by
the spectrophotometric method using DMABA.
EXPERIMENTAL
Reagents. Pure urea (EKROS, Russia), analyti
cally pure DMABA (Laboratornaya tekhnika, Rus
sia), CH3COOH, 99.5% (Panreac, Spain), and ana
lytically pure conc. HCl (EKROS, Russia) were used.
Distilled water was also used in the work.
Solutions. Standard (2.5 g/L) solution of urea was
prepared by dissolving an exact weight of specimen in
water. The process solution with the concentration of
urea 250 µg/mL was prepared by diluting the initial
standard solution with water.
To prepare the solution of DMABA 20 (or 40) g of
reagent was dissolved in 50 (or 100) ml of conc. HCl in
a 1L volumetric flask, diluted with water to the mark
and stirred. The next day the solution was filtered. A
diluted solution of hydrochloric acid was prepared by
diluting 5 mL concentrated solution in a 100mL vol
umetric flask with water.
Apparatus. The absorbency of solutions was mea
sured on photometers KFK2MP or KFK3 at 400 or
440 nm, respectively, in cells with l = 5 cm relative to
the solution of the control experiment.
Experimental technique. The solutions of urea,
DMABA, acetic acid, and HCl (if necessary) were
added sequentially into 25mL volumetric flasks,
diluted with water to the mark and stirred. The absor
bency was measured relative to the reference solution.
MOSCOW UNIVERSITY CHEMISTRY BULLETIN
RESULTS AND DISCUSSION
Reproduction of the Technique from the internet
[7]. The solutions contained 15 mL of water, 10 mL of
reagent DMABA with a concentration of 20 g/L, and
5 mL of CH3COOH; the total volume of solutions was
35 mL. Dependence of absorbency on urea concen
tration in water is shown in Table 1. It is linear,
described by the equation A = 0.00324c with a correla
tion coefficient of r = 0.998, but values of absorbency
are very low. Therefore, minimum detectable concen
tration of urea can not be 2 mg/L, as indicated in work
[7]. Objectively, surely water samples with a concen
tration of urea over 15 mg/L can be analyzed.
Selection of optimum conditions for the complex
ation of urea with DMABA. A compound of urea with
DMABA has a maximum absorption at 420 nm [8]. It
was shown that the absorption of solution on KFK2MP
is maximum when using a colorfilter with transmission
wavelength of 400 nm, and on KFK3—440 nm.
Influence of DMABA, HCl, and CH3COOH con
centration on the absorbency was studied, and the
results are presented in Table 2. It can be seen that the
introduction of CH3COOH increases the absorbency
by 1.5 times. It should be noted that in technique [8]
this acid was not added to the solution. The optimum
content of conc. CH3COOH is 5 mL. Additional add
ing of HCl is inappropriate.
Dependence of the absorbency on the concentra
tion of DMABA is complicated: one can clearly dis
tinguish two plateaus in the range of 6–8 mL and 12–
18 mL for the solution of DMABA with a concentra
tion of 20 g/L. Thereby, introduction of a large
amount of reagent leads to an increase in absorbency
of the solution that should allow us to detect lower
concentrations of urea. As the best, 7 and 12 mL of
DMABA solutions (20 g/L) were chosen. Since the
introduction of a large volume of reagent solution
decreases the volume of aqueous solution containing
urea the solution of DMABA with a concentration of
40 g/L was prepared whose optimum volume is 6 mL.
The change of the absorbency for the solution of
the complex on standing was studied. It was shown
that the absorbency of the solution is constant for
5 min after mixing the solutions for both concentra
tions of reagents, then, decreases slightly. So, in the
solution containing 7 mL of DMABA (20 g/L), the
absorbency decreased by 11% after 45 min. In the
solution containing 6 mL of DMABA (40 g/L), the
absorbency decreased by 6% after 15 min, and by 8%
after 25 min. So, the absorbency of solutions should be
measured for 5 min after preparation.
Complex formation reaction of urea with DMABA.
Stoichiometry of the components in the process of
complex formation was studied by molarratio
method [9]. For this a dependence of the absorbency
of solutions with constant concentration of DMABA,
Vol. 66 No. 6 2011
 PHOTOMETRIC DETECTION OF UREA IN NATURAL WATERS 347

Table 2. The effect of quantity of reagents on the formation of colored compound of urea with DMABA (total volume of
solution is 25 mL, 20 g/L of DMABA, l = 5 cm, 400 nm, KFK2MP)
Added, mL
Urea added, mg A
DMABA CH3COOH HCl
1.250 2.00 7.00 0 0.050
4.00 7.00 0 0.042
6.00 7.00 0 0.114
8.00 7.00 0 0.113
10.00 7.00 0 0.150
12.00 7.00 0 0.162
14.00 7.00 0 0.238
0.500 2.00 5.00 8.00 0.015
4.00 5.00 6.00 0.060
5.00 5.00 5.00 0.100
6.00 5.00 4.00 0.122
7.00 5.00 3.00 0.122
8.00 5.00 2.00 0.127
10.00 5.00 0 0.095
12.00 5.00 0 0.175
16.00 5.00 0 0.170
18.00 5.00 0 0.180
0.500 6.00 5.00 4.00 0.122
6.00 5.00 6.00 0.103
6.00 5.00 10.00 0.080
0.500 6.00 3.00 4.00 0.114
6.00 5.00 4.00 0.122
6.00 7.00 4.00 0.112
6.00 10.00 4.00 0.093
1.250 14.00 0 0 0.153
14.00 7.00 0 0.238
0.250 12.00 0 0 0.080
12.00 5.00 0 0.120

equal to 0.01072 M on the concentration, was con
structed (Table 3).
It can be seen, that the break point on the satura
tion curve is unclear, the molar ratio of DMABA and
urea, obtained by extrapolation of linear segments of
the curve to intercrossing, is 2.08 : 1.0, which is close
to 2 : 1. An unclear kink on the saturation curve indi
cates the slight stability of the complex formed.
Urea has the properties of a nucleophile and weak
base [10]. Reactions of carbonyl compounds with
bases are catalyzed by both strong and weak acids [11].
Interaction between urea and DMABA can be repre
sented by the following scheme.

O
C
NH2 H
NH C(OH) N(CH3)2
O C + O C
NH2 NH2

N(CH3)2
O
C
H NH C(OH) N(CH3)2
+ O C
NH C(OH) N(CH3)2
N(CH3)2
Scheme.

Since urea is a weak base (pKb = 13.82), pulling the necessary to increase the reactivity of the carbonyl
electrons of the carbonyl group with acid catalyst is group towards nucleophile—nitrogen atom of urea.

MOSCOW UNIVERSITY CHEMISTRY BULLETIN Vol. 66 No. 6 2011

348 BASOVA et al.
Table 3. Dependence of the absorbency of solutions with a constant concentration of DMABA, equal to 0.01072 M,
on the concentration of urea
curea, mg 1.25 2.50 5.00 7.50 10.00
A (440 nm) 0.102 0.230 0.400 0.530 0.616
Therefore, the reaction should occur only in acidic
medium. However, in strongly acidic medium prefer
ential protonation of a nucleophilic agent (urea is pro
tonated at the oxygen atom rather than nitrogen [10])
and reduction of the reactivity of a free electron pair
take place, which leads to reduction of the degree of
complexation—to a decrease in the absorbency with
increasing the concentration of HCl and CH3COOH
(Table 2). It is not possible to significantly decrease the
concentration of HCl or replace it with acetic acid,
because it is necessary to dissolve the reagent. In work
[8] for the preparation of 1 L of DMABA solution
(4 g/L) 40 mL of conc. HCl was used.
Despite the fact that the stoichiometry of the form
ing complex of urea with DMABA corresponds to the
ratio of 1 : 2, excess reagent, required to reach a con
stant value of absorbency at fixed content of urea of
500 µg is large—96 and 193fold for the first and the
second plateau, respectively (Table 2). Probably, in the
area of the first plateau complex 1 : 1 is dominated, and
in the area of the second—1 : 2. Differences in the sta
bility constants of the complexes are negligible, so
when constructing the saturation curve two points of
intersection were not obtained, but only one.
Metrological characteristics of the technique. To
construct calibration curves, the work solution of urea,
7 mL of DMABA (20 g/L) or 6 mL of DMABA (40 g/L),
and 5 mL of CH3COOH solutions are added into
25mL flasks, diluted with water to the mark, and
mixed. For 5 min the absorbency of the solutions is
measured at 400 nm (KFK2MP) or 440 nm (KFK3)
in 5 cm cells relative to the solution of the control
experiment. Calibration curves are linear in the range
up to 750 µg of urea. The equations of calibration
curves are presented in Table 4. When constructing the
curves an average of two measurements was used.
While determining 375 µg of urea the relative standard
deviation was 0.06 (n = 6, 7.00 mL of reagent solution
was added, the absorbency is 0.125–0.141), and while
determining 125 µg—0.12 (n = 6, 7 mL of reagent
solution was added, the absorbency is 0.030–0.039).
Average values of the molar absorption coefficients
for the regions of the first and second plateau, which
differ by almost two times at 440 nm, are calculated
(Table 4), which may also give evidence about the for
mation of the complex 1 : 2 at high concentrations of
reagents, because of the color of the compound caused
by the presence of DMABA structure in the molecule.
Devices KFK2MP and KFK3 can determine the
absorbency in the range of 0.0–2.0. The value of
MOSCOW UNIVERSITY CHEMISTRY BULLETIN
12.50 15.00 20.00 25.00 30.00 37.50
0.694 0.723 0.737 0.750 0.760 0.766
absorbency, which can be measured with the required
accuracy (sr < 0.33), is about 0.01 [12]. For use in the
work cells with l = 5 cm and calculated values of the
molar absorption coefficient (Table 4), detection lim
its are 30 µg (7 mL of DMABA solution with a con
centration of 20 g/L is added) or 20 µg (6 mL of
DMABA solution with a concentration of 40 g/L is
added). The recommended concentration range of urea is
100–750 µg (in a 25mL flask).
Analysis of the objects. The technique was used to
analyze a sample of river water by the method of
“introducedfound.” A sample of river water was
selected on 11/23/2010 in the Uglich reservoir, Volga
river, Kalyazin city (FGVU “Tsentrregionvodhoz”
Dubna ecoanalytical laboratory). The sample of water
contained calcium, magnesium, sulfate, chloride,
nitrate, and silicateions at the level of tens and units
mg/L; pH was 7.39.
The sample of water had the highest chroma—
46 degrees at MAC (maximum allowable concentration),
for fishing ponds—20. Color was determined photo
metrically at the wavelength of 436 nm [13]. It was
shown that the absorbency for 10 mL of the sample,
diluted with water to 25 mL, measured at 440 nm, is
0.019 (an average of two measurements). Thus, in the
process of analysis of natural samples the absorption of
the matrix at the chosen wavelength should be taken
into account.
Performing the detection. The sample of analyzing
water is passed through the filter “blue ribbon.” Ten mL
of the sample is put into 25mL flasks, 6 mL of
DMABA with a concentration of 40 mg/L (or 7 mL of
DMABA with a concentration of 20 g/L), and 5 mL of
CH3COOH are added, diluted with water to the mark,
mixed, and the absorbancy is measured in 5 min at 400
or 440 nm in the cells with l = 5 cm. Optical density,
which is determined by the color of initial sample, is
substracted from the optical density obtained. For this
purpose, 10 mL of the water analyzed is introduced in
10mL flask, 5 mL CH3COOH is added, diluted by water
to mark, stirred, and optical density is measured relative
to water at 400 and 440 nm in the cells with l = 5 cm.
Urea content in an aliquot of the sample (m, µg) is
found from the calibration curve. Concentration of
urea (c, mg/L) in the sample is calculated according to
the formula c = m/10. The results of detection are pre
sented in Table 5. The technique allows one to analyze
reliably samples with a content of urea 10 mg/L and
higher.
Vol. 66 No. 6 2011
 PHOTOMETRIC DETECTION OF UREA IN NATURAL WATERS 349

Table 4. Calibration curves for the detection of urea

Apparatus (λ, nm) VDMABA, mL murea, μg A n ε, mol–1 l cm–1 Equation r
KFK2MP (400) 7 (20) 125 0.042 6 78 A = 0.00024m 0.996
250 0.067
375 0.096
500 0.119
625 0.152
750 0.164
KFK3 (440) 6 (40) 25 0.013 9 158 A = 0.00051m 0.973
50 0.029
100 0.050
150 0.078
200 0.110
250 0.135
375 0.192
500 0.260
625 0.305
KFK3 (440) 7 (20) 125 0.041 6 97 A =0.000326m 0.998
250 0.081
375 0.123
500 0.162
625 0.204
750 0.245

Table 5. The results of urea detection in samples by the method of introducedfound (P = 0.95)
Sample
c, Found,
Matrix volume, n Found, µg c, mg/L sr D, % Conditions
mg/L µm
mL
River 10 10.0 100 3 103 ± 7 10.3 ± 0.7 0.03 +3 KFK3, 440 nm, 6 mL
water of DMABA (40 g/L)
10 20.0 200 3 191 ± 13 1.9 ± 1 0.03 –4.5
Distilled 100 1.25 125 4 135 ± 16 1.4 ± 0.2 0.07 +8 KFK3, 440 nm, 6 mL
water of DMABA (40 g/L), evaporating
100 3.75 375 3 336 ± 54 3.4 ± 0.5 0.06 –10.4
River 100 5.0 500 3 467 ± 48 4.7 ± 0.5 0.04 –6.6 KFK3, 440 nm, 7 mL
water of DMABA (20 g/L), evaporating
100 2.0 200 3 140 ± 66 1.4 ± 0.7 0.19 –30 KFK3, 440 nm, 6 mL
of DMABA (40 g/L), evaporating

Concentrating. To reduce the lower limit of the
detected content the possibility of concentrating was
studied. The easiest way to concentrate water samples
is evaporation. However, in boiling water urea hydro
lyzes with formation of ammonia and carbon dioxide,
moreover, this reaction is catalyzed by acids and bases
[10]. Evaporation of the model solution, prepared
with the use of distilled water, was studied. It was evap
orated to a volume of ~8 mL on an electric stove, care
fully watched, and left there to dry. As is seen from the
data of Table 5, error in the determination of urea in
the model solution after evaporation does not exceed
11%. After evaporation of 100 mL of river water with
the addition of urea satisfactory results were also

MOSCOW UNIVERSITY CHEMISTRY BULLETIN Vol. 66 No. 6 2011

350 BASOVA et al.
obtained (Table 5). In the latter case, two more sam
ples were evaporated in parallel in order to take into
account the contribution of the color of the initial
sample of water in the absorbency of the complex; the
average value of the absorbency, due to the color, was
0.111. It should be noted, that the pH of the analyzing
sample of water was close to neutral. If the pH of the
sample differs greatly from neutral, the sample should
be neutralized to reduce the catalytic effect of hydrox
onium or hydroxideions.
Performance of the detection with preconcentration
by evaporation. A sample of analyzed water is passed
through the filter “blue ribbon,” pH is measured,
diluted solutions of HCl or NaOH are added dropwise
to reach a value of pH 6.5–7.5, if it is necessary. One
hundred mL of the sample is placed into a 100–150mL
beaker, evaporated on a rangette to a volume of ~8 mL.
The solution is quantitatively placed into a 25mL
flask, rinsing with 5 mL of water, 6 mL of DMABA
solution (40 mg/L), and 5 mL of CH3COOH are
added, diluted with water to the mark, and mixed.
Within 5 min absorbency of solutions is measured at
400 or 440 nm in cells (l = 5 cm). From the obtained
value of absorbency, absorbency, caused by the color of
initial sample, is subtracted. For this, 100 mL of ana
lyzed water is evaporated on a hot plate to a volume of
~8 mL, placed into 25mL flask, 5 mL of CH3COOH
is added, diluted with water to the mark, mixed, and
absorbency is measured at 400 or 440 nm in 5cm cells
relative to water. Urea content in an aliquot (m, µg) is
found by the calibration curve. Concentration of urea
(c, mg/L) in the initial sample is calculated by the
equation: c = m/100.
The results are presented in Table 5. In both cases,
the found content of urea is less than the introduced
content, which may indicate the partial hydrolysis of
analyte; a degree of hydrolysis is higher in a more
dilute solution. The technique allows one to analyze
reliably samples with the urea content of 2 mg/L.
REFERENCES
1. RD 39014742823589 Metodicheskoe rukovodstvo po
tekhnologii provedeniya indikatornykh issledovanii i
interpretatsii rezul’tatov dlya regulirovaniya i kontrolya
protsessa zavodneniya neftyanykh zalezhei (Guidance
on the technology of indicator studies and interpreta
MOSCOW UNIVERSITY CHEMISTRY BULLETIN
tion of the results for adjustment and control of the
flooding of oil reservoirs), Groznyi, 1989.
2. GOST 208192 Karbamid. Tekhnicheskie usloviya
(Urea. Technical conditions). http://www.himtrade.ru/
g208192.htm.
3. Kolorimetricheskie (fotometricheskie) metody oprede
leniya nemetallov (Colorimetric (photometric) meth
ods of the determination of nonmetals), Moscow, 1963,
p. 80.
4. Babko, A.K. and Pilipenko, A.T., Fotometricheskii
analiz. Metody opredeleniya nemetallov (Photometric
Analysis. Methods for the Determination of Nonmetals),
Moscow, 1974, p. 11.
5. PND F 14.1:2.195. Metodika vypolneniya izmerenii
massovoi kontsentratsii ionov ammoniya v prirodnykh i
stochnykh vodakh fotometricheskim metodom s reak
tivom Nesslera (Technique of the measurement of the
mass concentration of ammonium ions in natural and
waste waters by photometric method with the use of
Nessler reagent), Moscow, 1995.
6. http//ru.wikipedia.Org/wild/%DO%9C%DO%BE%
D1%87%DO%B5%DO%B2%DO%B8%DO%BD%BO.
7. Internet portal khimikovanalitikov ANSHEM.RU>>
Forumy>>1. Analiticheskii forum (otvet pol’zovatelya
18.02.2005).
8. GOST 30181.594. Udobreniya mineral’nye. Metod
opredeleniya massovoi doli amidnogo azota v slozhnykh
udobreniyakh (spektrofotometricheskii metod) (Mineral
fertilizers. The method for the determination of mass
fraction of amide nitrogen in compound fertilizers
(spectrophotometric method)), 1996.
9. Bulatov, M.I. and Kalinkin, I.P., Prakticheskoe ruko
vodstvo po fotometricheskim metodam (Practical Guide
for the Photometric Methods), Leningrad, 1986,
p. 245.
10. Organicheskaya khimiya. Kn. 1 (Organic Chemistry.
Book 1), Tyukavkina, N.A., Ed., Moscow, 2008, p. 498.
11. Bekker, G., Vvedenie v elektronnuyu teoriyu organ
icheskikh reaktsii (Introduction to the Electronic The
ory of Organic Reactions), Moscow, 1977, p. 296.
12. Osnovy analiticheskoi khimii. Kn. 2 (Fundamentals of
Analytical Chemistry. Book 2), Zolotov, Yu.A., Ed.,
Moscow, 2002, p. 273.
13. RD 52.24.4972005. Metodicheskie ukazaniya. Meto
dika vypolneniya izmerenii tsvetnosti poverkhnostnykh
vod sushi fotometricheskim metodom (Guidelines. The
technique for measurement of chromaticity of surface
waters of land by photometric method), Rostovon
Don, 1995.
Vol. 66 No. 6 2011