Microchemical Journal 64 2000.

161171

A modified spectrophotometric method for the

determination of trace amounts of phenol in water

Chunli KangU , Ying Wang, Runbo Li, Yaoguo Du, Jun Li,
Bowen Zhang, Liming Zhou, Yuzhong Du
Department of En ironmental Science, Jilin Uni ersity, Changchun 130023, PR China

Received 22 July 1999; received in revised form 13 October 1999; accepted 31 October 1999

Abstract

A modified spectrophotometric method has been developed for the determination of phenol in water. The method
is based on the reaction between phenol, sodium nitroprusside and hydroxylamine hydrochloride in a buffer medium
of pH 10.611.8. The linear dynamic range is 0.055.0 mg ly1. The optimum determination wavelength is at 700 nm.
The molar absorptivity is 1.23= 10 4 mol ly1 cmy1. The relative standard deviation of the standard solution of phenol
is 1.3% n s 10, C s 0.5 mg ly1 .. The average recovery of water samples spiked with phenol of 2.0 mg ly1 is 98.8%
n s 10.. The relative error of standard sample C s 0.394" 0.003 mg ly1 . is 2.0%. F values of the statistical analysis
show that there is no significant difference between the proposed method and the 4-aminoantipyrine method. The
crystal of the product of the colored reaction was synthesized. According to its IR spectrum, Raman spectrum,

Mossbauer spectrum and elemental analysis, the composition and structure of the product have been deduced and
the mechanism of the colored reaction has also been investigated. 2000 Elsevier Science B.V. All rights reserved.

Keywords: Phenol; Spectrophotometry; Water

1. Introduction plastics, organic chemicals, steel and petroleum

industries. As protoplasm toxins, they have a
Phenolic compounds have been extensively used strong influence on the taste and odor of water
as pesticides, herbicides and fungicides and are and fish. The growing utilization and pollution of
present in the wastewater from the production of phenolic compounds makes the development of
sensitive and specific analytical methods neces-
sary.
U The conventional method for the determina-
Corresponding author.
E-mail address: duyg@mail.jli.edu.cn C. Kang. tion of phenols is based on the colored derivative

0026-265Xr00r$ - see front matter 2000 Elsevier Science B.V. All rights reserved.
PII: S 0 0 2 6 - 2 6 5 X 9 9 . 0 0 0 2 2 - 3
162 C. Kang et al. r Microchemical Journal 64 (2000) 161171
formed by its coupling with 4-aminoantipyrine
4-AAP. w1x. Severe deficiencies of this method
are as follows: the enolketo system disturbs
phenol analysis; the reaction products of some
phenolic compounds can not be extracted by
chloroform; and certain p-substituted phenols
show negligible response or do not react at all
with 4-AAP. Other disadvantages are its sensitiv-
ity to pH variation and the consequent difficulty
of finding suitable buffers for the reaction. So,
many other spectrophotometric methods w26x
have been developed in recent years.
One reported procedure w4x, worthy of more
attention, is based on the blue-colored reaction
between phenol, sodium nitroprusside and hy-
droxylamine hydrochloride in a buffer medium.
This method is comparatively rapid and sensitive.
However, in the initial report, the procedure is
adding the various reagents and water sample to
a 25-ml volumetric flask first and diluting to the
mark with water. So, the water sample was di-
luted. The lowest detectable concentration of the
method is 0.1 mg ly1 . This procedure is not
suitable for the determination of lower concen-
trations of phenol. Also, the mechanism of the
colored reaction is not well established. The pre-
sent paper describes a new spectrophotometric
determination of phenol on the basis of improve-
ments on the method established by Nagaraj et al.
w4x. In the proposed method, various reagents
were directly added into a definite volume of a
water sample and there is no dilution of the water
sample, if not considering the volume of the
reagents. Therefore, the lowest detectable con-
centration was lowered to 0.05 mg ly1 . The crystal
of the product of the colored reaction was synthe-
sized. According to the characterization of the
product, its composition and structure have been
deduced and the reaction mechanism has also
been investigated.
2. Experimental
2.1. Instrumentation
The following equipment was used: a Model
721 spectrophotometer with 2-cm glass cells
Shanghai, China.; a pHS-2C digital acidimeter
Hangzhou, China.; a Bruker IFS 66V FTIR
Spectrophotometer Germany.; a Bruker RFS 100
FT Raman Spectrophotometer Germany.; a
Model 1106 element analyzer Italy.; a Model
P-E 403 atom absorption spectrophotometer
USA.; a Model MS-500 OXFORD Mossbauer
spectroscope England.; and a Model 7300 vibrat-
ing sample magnetometer USA..
2.2. Reagents
All the solutions were prepared from analytical
reagent A.R.. grade reagents and distilled water
without phenol.
The stock standard solution 1000 mg ly1 . was
prepared by dissolving 1.00 g of phenol in 1000 ml
of distilled water. The solution should be stable
for at least 30 days and kept in a refrigerator. The
working standard solutions of 20 mg ly1 were
prepared by appropriate dilution of the stock
standard solution with distilled water and used
within 1 day.
The sodium nitroprusside solution 1.0= 10y2
mol ly1 . was prepared by dissolving 0.7450 g of
sodium nitroprusside  Na 2 wFeCN.5 NOx 2H 2 O4
in 250 ml of distilled water.
The hydroxylamine hydrochloride solution 4.0
= 10y2 mol ly1 . was prepared by dissolving 0.6950
g of hydroxylamine hydrochloride in 250 ml of
distilled water.
The buffer solution pH 12. was prepared by
adding 500 ml of NaH 2 PO4 solution 0.1 mol ly1 .
and 17.5 ml of NaOH solution 6 mol ly1 . to a
1000-ml volumetric flask and diluting it to the
mark with distilled water.
2.3. Procedure
2.3.1. Standard solution
The appropriate volumes of the working stan-
dard solution 0.25, 0.5, 1.0, 2.0, 2.5, 5.0, 7.5 ml.
were, respectively, put into a series of 50-ml color
tubes with matching corks. and diluted to the
mark with distilled water. The series of the solu-
tions contained phenol in the range of 0.055.0
mg ly1 , to which 1.0 ml of 1.0= 10y2 mol ly1
sodium nitroprusside solution, 1.0 ml of 4.0= 10y2
 C. Kang et al. r Microchemical Journal 64 (2000) 161171
mol ly1 hydroxylamine hydrochloride solution,
and 3.0 ml of buffer solution were added, respec-
tively. After mixing well, they were allowed to
stand for 15 min. The absorbance at 700 nm in
2-cm cells was measured against a distilled water
blank.
2.3.2. Sample analysis
For water samples, a pre-distilled step w1x was
adopted before analysis. A 50-ml sample of the
distillate was transferred into a 50-ml color tube
with matching cork. and analyzed as described
above.
3. Results and discussion
3.1. Choice of the determination wa elength
The absorption spectra of the reaction system
are shown in Fig. 1. It can be seen that the
Fig. 1. Absorption spectra of the reaction system: . phenol; and B. blank.
163
maximum absorbance is at 700 nm when there is
phenol in the reaction system, and the blank in
which there is no phenol displays a very weak
absorbance at 700 nm. So, the optimum determi-
nation wavelength was selected at 700 nm.
3.2. Effects of the amounts of the reagents
Fig. 2 shows the effects of the amounts of
sodium nitroprusside, hydroxylamine hydrochlo-
ride and buffer solution. Evidently, 1.0 ml of
sodium nitroprusside, 1.0 ml hydroxylamine hy-
drochloride and 3.0 ml of buffer solution can
produce the maximum absorbance. It is worth
noting that the addition of excess hydroxylamine
hydrochloride could make the sensitivity drop.
3.3. Reaction time and stability
The color reaction goes very fast and it can
occur almost completely within 5 min see Fig. 3..
164 C. Kang et al. r Microchemical Journal 64 (2000) 161171

Fig. 2. Effects of the amounts of the reagents: B. hydroxylamine hydrochloride; . sodium nitroprusside; and '. buffer.

The maximum absorbance can be obtained within
15 min and it can remain stable without any
obvious change within 5 h.
3.4. Effects of other conditions
The results of the tests showed that phenol
must be added first and the order of addition of
other reagents had no effect on determination
results. At the range of 060C, temperature
hardly showed any effect on the analysis, but the
absorbance dropped rapidly at a temperature
higher than 60C.
Of the reagents used in this test, only the
working standard solution of phenol must be pre-
pared on the same day; the solutions of hydroxyl-

Fig. 3. Effect of the reaction time. The concentrations of phenol are: 3.00 mg ly1 '.; 1.50 mg ly1 B.; and 0.50 mg ly1 .,
respectively.
 C. Kang et al. r Microchemical Journal 64 (2000) 161171 165

Table 1
The reaction features of various phenolic compounds

Compounds Color ma x nm. max mol 1y1 cmy1 .

Phenol Greenblue 700 1.23= 104

o-Cresol Greenblue 700 1.62= 104
m-Cresol Greenblue 700 1.35= 104
p-Cresol Greenyellow 700 41.9
2,4-Dimethylphenol Light-green 700 1.67= 102
1-Naphthol Greenblue 680 5.46= 103
4-Chlorophenol Greenyellow 700 93.1
4-Chloro-3-methyl phenol Greenblue 720 2.93= 102
2,4-Dichlorophenol Greenblue 700 1.01= 102
4-Aminophenol Greenblue 700 1.38= 103

amine hydrochloride and buffer can be kept for a pre-distilled step is adopted, the effects of the
long time and the solution of sodium nitroprus- above-mentioned ions could be eliminated. EDTA
side can also be kept for a few months in the also interfered with the analysis. Thus, it cannot
dark. at room temperature. be used as a masking agent.

3.5. Calibration cur e and precision 3.7. Responses of other phenolic compounds

Under the above optimum conditions, the lin-
ear regression equation is A s 0.2669C y 0.01516
n s 7, r s 0.9996, C s 0.055.0 mg ly1 ..
For a 0.5-mg ly1 standard solution of phenol,
R.S.D.%s 1.3, which was obtained by the parallel
determinations of two samples each day within 5
consecutive days.
3.6. Effects of the ions
For phenol solutions 3.0 mg ly1 ., if the rela-
tive error of 5% is permitted, 100 mg ly1 of Kq,
Naq, Ba2q, Cly, SO42y, CH 3 COOy did not dis-
turb the analysis, and 100 mg ly1 of Mg 2q, Cu2q,
Fe 2q, Fe 3q, NHq 4 , Al
3q
, PO43y, HCOy y
3 , HSO 3 ,
2y
S did disturb the analysis seriously. As the
Under the experimental conditions, many
phenolic compounds can give the color reaction,
but they display different colors, max and molar
absorptivity see Table 1.. Some compounds not
included in the table, such as 1,6-dihydroxyl ben-
zene can also exhibit a brownred color, but its
max is in the ultraviolet region. Nitrophenols
including 2-nitrophenol, 3-nitrophenol, 4-
nitrophenol and 2,4-dinitrophenol all exhibit a
light-yellow color, but they almost have no absor-
bance at the range of 500800 nm.
It is clear from Table 1 that the color intensi-
ties of o-cresol and m-cresol can match that of
phenol and they are also common phenolic com-
pounds in aquatic environments. Under the ex-
perimental conditions, the linear regression equa-

Table 2
Determination of phenol in gas wastewater

Sample 4-AAP method The proposed method Relative error

mg ly1 . mg ly1 . %.

1 0.240 0.244 1.7

2 0.201 0.208 3.5
3 0.282 0.275 y2.5
4 0.320 0.323 0.94
5 0.242 0.245 1.2
166 C. Kang et al. r Microchemical Journal 64 (2000) 161171
tions of o-cresol and m-cresol are: A s 0.3425C
q 0.006441 n s 5, r s 0.9993. and A s 0.2934C
y 0.01814 n s 5, r s 0.9997., respectively.
3.8. Precision of sample analysis
For the 0.394" 0.003-mg ly1 standard sample
of phenol no. 69509202, the Environmental Mon-
itoring Central Station of Jilin Province, China.,
the R.E.s 2.0%.
For South Lake water samples which were
spiked with 2.0 mg ly1 phenol, the R.S.D.s 2.6%
and average recovery was 98.8% n s 10..
3.9. Comparison between the proposed and 4-AAP
methods
Five samples were collected from a gas waste
water. The analysis results, using the proposed
and 4-AAP methods, are listed in Table 2.
Fig. 4. Absorption spectra of the solid product in water: . blue solid product; and B. green solid product.
For sample no. 4, the R.S.D. n s 5. was 3.1%
proposed method. and 1.7% 4-AAP method.,
respectively. The F values of the statistical analy-
sis of the two methods showed no significant
difference Ps 0.05..
3.10. Reaction mechanism
3.10.1. Synthesis of the product
Water 50 ml. was added into a 250-ml beaker
followed by addition of 5 ml of 6 mol ly1 NaOH,
to which 1.9 g of phenol and 2 g of sodium
nitroprusside were added and made to dissolve. A
solution of 1 g of hydroxylamine hydrochloride
dissolved in 10 ml of water was added dropwise
into the reaction system in the beaker. The mix-
ture was stirred continuously until the solution
became completely blue.
The blue solution was transferred into an evap-
orator and heated in a 60C water bath until a
 C. Kang et al. r Microchemical Journal 64 (2000) 161171
liquid film separated out on the surface. It was
left at room temperature until a large amount of
solid separated out, which was filtered out on a
Buchner funnel with the aid of suction.
The solid was redissolved in a minimum amount
of warm water. The filtrate was cooled gradually
to room temperature and left until a large amount
of a blue crystal separated out again. The crystals
were filtered by suction on a Buchner funnel and
dried in a desiccator.
3.10.2. Characterization of the product
The absorption spectra of the crystal of the
product in water is shown in Fig. 4 which is
similar to Fig. 1. The maximum absorbance wave-
length is also at 700 nm.
Figs. 5 and 6 are IR and Raman spectra of the
crystal of the product, respectively. In the IR
spectrum, the peak at 2057 cmy1 is assigned to
Fig. 5. The IR spectrum of the solid product KBr pellet..
167
the vibration of the cyano group CN. and it splits
into two peaks at 2102 and 2072 cmy1 in the
Raman spectrum. The peak of the nitroso group
NO. of feedstock Na 2 wFeCN.5 NOx 2H 2 O is at
1944 cmy1 w7,8x and it does not appear in Fig. 5.
So, the product does not contain an NO group.
The peak at 1628 cmy1 in the IR spectrum can
be assigned to H 2 O and it disappears in the
Raman spectrum. The solid product was synthe-
sized many times and the intensity of the peak
varied with the preservation time. The longer the
time, the weaker the intensity. In addition, the
1
H-NMR of hydrogen atoms on the benzene ring
cannot be observed while determining the 1 H-
NMR spectrum of the product. Therefore, it can
be deduced that phenol is not present in the
product.
The hysteresis loop of the product Fig. 7.
showed the weak ferromagnetism of the product.
The magnetic susceptibility calculated was 7.3=
168 C. Kang et al. r Microchemical Journal 64 (2000) 161171

Fig. 6. The Raman spectrum of the solid product power 500 mW, scanning 100 times..

10y5 cgs. The experiment of the Mossbauer

spec- also showed that the solid product has no diamag-
trum determination at room temperature 298 K. netism.

Fig. 7. The hysteresis loop of the solid product.

 C. Kang et al. r Microchemical Journal 64 (2000) 161171 169

Fig. 8. The Mossbauer spectrum of the solid product wroom temperature 298 K.x.

The ratio of mole fraction of FerNarCrN of
the product was 1:4.9:5.3:4.8, which was obtained
by elemental analysis. Hence, the theoretical ra-
tio should be 1:5:5:5. The Mossbauer spectrum of
the product at room temperature 298 K. Fig. 8.
only displayed a sharp peak and there was no
quadrupole splitting. Consequently, the ferronu-
clear site should be situated in the center of a
regular tetrahedron or an octahedron structure.
As the tetrahedron is impossible for this test, the
structure is deduced as follows:
tion, the color of the solution changed from
light-green to dark-green to green-blue to blue
with the increase in the concentration of phenol.
The blue solid product can be obtained from the
synthesis method above. If the crystal separated
out in an icesalt bath during the recrystalliza-
tion, a green crystal of the product could be
obtained. The green form had the same absorp-
tion spectrum as the blue one in aqueous solution
and the max was also at 700 nm Fig. 4.. Their IR
spectra were completely identical. So, the solid
product should have structural isomers.

3.10.3. Reaction mechanism

On the basis of the characterization of the solid
product, the mechanism of the color reaction is
deduced as follows:

Its molecular formula should be Na 10 wFe 2-

CN.10 x. Compared with Na 2 wFeCN.5 NOx 2H 2 O,
the ferro valence degraded to 0 and the absorp-
tion frequency of the cyano group CN. in the IR
spectrum also shifted significantly, from 2150 to
2057 cmy1 .
The results of elemental analysis also indicate
that the solid product contains some crystalliza-
tion water, which can also be proved by the IR
spectrum 1628 cmy1 ..
In the color reaction process in aqueous solu-
170 C. Kang et al. r Microchemical Journal 64 (2000) 161171
In the reaction, the negative oxygen ion Oy.
on the benzene ring repels the electron violently,
which makes the benzene ring very active, be-
coming a very good electron donor. As an elec-
trophile, the nitroso group in the form of qNO
attacks the benzene ring and an electrophilic
reaction occurs. It is well known that phenol is
very active and it is one of a few compounds
which can be attacked by the weak electrophilic
reagent qNO. When phenol is converted to
phenolate, the electrophilic reaction could take
place more easily. Compared with o-cresol and
m-cresol, p-cresol has a much weaker sensitivity.
This may be caused by the hindrance of the
p-substituted structure. According to pure organic
chemistry theory, the sites that the nitroso group
q NO. attacks on the benzene ring could be at
the para- or ortho-position. In the actural color
reaction, the nitroso group q NO. may be in the
larger anionic complex and the way attacking
para-position is easier. So, p-cresol is insensitive
to the color reaction. Moreover, the electrophilic
or electrophobic property of the substitutes on
the benzene ring also affects the sensitivity of the
color reaction and phenolic compounds with dif-
ferent substitutes giving different sensitivities see
Table 1.. Of the phenolic compounds tested, ni-
trophenols couldnt give the color reaction.
Maybe, this is because nitro NO 2 . is an
electron-withdrawing group and it makes the ben-
zene ring inactive, the electrophilic reaction being
inhibited.
3.11. Comparison between the modified and the
original methods
The analytical method established in this arti-
cle is similar to that reported by Nagaraj et al. w4x
mainly in amounts of the reagents used, order of
addition of the reagents, and pH range of the
reaction. But, there are significant differences in
the following aspects between the two methods.
3.11.1. Different sensiti ity
The molar absorptivity of the method reported
by Nagaraj et al. w4x is 1.68= 10 4 mol ly1 cmy1
and that of the proposed method is 1.23= 10 4
mol ly1 cmy1 .
3.11.2. Different determination range
The linear dynamic range of the determination
of the method reported by Nagaraj et al. w4x is
0.16.5 mg ly1 and that of the proposed method
is 0.055.0 mg ly1 .
3.11.3. Different effect of foreign ions
For the method reported by Nagaraj et al. w4x, it
is said that a large amount of anions does not
disturb the determination and many cations dis-
turb the determinations slightly. For the proposed
method, anions also had an effect on the determi-
nation of phenol and the errors caused by the
anions and cations are much higher than those of
the method reported by Nagaraj et al. w4x.
3.11.4. Different responses of some phenolic
compounds
For the method reported by Nagaraj et al. w4x, it
is said that o-cresol and m-cresol cannot react.
For the proposed method, o-cresol and m-cresol
not only can react, but also have a higher sensitiv-
ity. The molar absorptivities are 1.62= 10 4 mol
ly1 cmy1 and 1.35= 10 4 mol ly1 cmy1 , respec-
tively.
3.11.5. Different physical property of the solid
product
For the method reported by Nagaraj et al. w4x, it
is said that the product is diamagnetic and the
magnetic susceptibility is y1.63= 10y6 cgs. For
the proposed method, the product is weakly ferro-
magnetic and the magnetic susceptibility is 7.3=
10y5 cgs.
Also, the reaction mechanism is investigated in
this thesis, which is not given in the method
reported by Nagaraj et al. w4x.
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