Microchemical Journal 64 Ž2000.


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

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 10.6᎐11.8. The linear dynamic range is 0.05᎐5.0 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% Ž 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%
Ž n s 10.. The relative error of standard sample Ž 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,
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. 䊚 2000 Elsevier Science B.V. All rights reserved.
Keywords: Phenol; Spectrophotometry; Water

1. Introduction
Phenolic compounds have been extensively used
as pesticides, herbicides and fungicides and are
present in the wastewater from the production of


Corresponding author.
E-mail address: duyg@mail.jli.edu.cn ŽC. Kang.

plastics, organic chemicals, steel and petroleum
industries. As protoplasm toxins, they have a
strong influence on the taste and odor of water
and fish. The growing utilization and pollution of
phenolic compounds makes the development of
sensitive and specific analytical methods necessary.
The conventional method for the determination of phenols is based on the colored derivative

0026-265Xr00r$ - see front matter 䊚 2000 Elsevier Science B.V. All rights reserved.
PII: S 0 0 2 6 - 2 6 5 X Ž 9 9 . 0 0 0 2 2 - 3

. Experimental 2.3. 1.0.0 ml of 1.. 1. This procedure is not suitable for the determination of lower concentrations of phenol.5 ml. 2. The lowest detectable concentration of the method is 0. was prepared by adding 500 ml of NaH 2 PO4 solution Ž0. The crystal of the product of the colored reaction was synthesized. China. put into a series of 50-ml color tubes Žwith matching corks. a Bruker RFS 100 FT Raman Spectrophotometer ŽGermany. 0. a Bruker IFS 66V FTIR Spectrophotometer ŽGermany. sodium nitroprusside and hydroxylamine hydrochloride in a buffer medium. Instrumentation The following equipment was used: a Model 721 spectrophotometer with 2-cm glass cells ŽShanghai.1. One reported procedure w4x. and 17.25. 2. The present paper describes a new spectrophotometric determination of phenol on the basis of improvements on the method established by Nagaraj et al. is based on the blue-colored reaction between phenol. The sodium nitroprusside solution Ž1. Other disadvantages are its sensitivity to pH variation and the consequent difficulty of finding suitable buffers for the reaction.0.0 mg ly1 .162 C. and a Model 7300 vibrating sample magnetometer ŽUSA.0= 10y2 mol ly1 .. So. and diluted to the mark with distilled water. and certain p-substituted phenols show negligible response or do not react at all with 4-AAP.00 g of phenol in 1000 ml of distilled water.0= 10y2 mol ly1 sodium nitroprusside solution. This method is comparatively rapid and sensitive. grade reagents and distilled water without phenol.1. The hydroxylamine hydrochloride solution Ž4. According to the characterization of the product.R. the procedure is adding the various reagents and water sample to a 25-ml volumetric flask first and diluting to the mark with water. The stock standard solution Ž1000 mg ly1 . Severe deficiencies of this method are as follows: the enol᎐keto system disturbs phenol analysis. was prepared by dissolving 0. 2.05᎐5.. r Microchemical Journal 64 (2000) 161᎐171 formed by its coupling with 4-aminoantipyrine Ž4-AAP.0 = 10y2 mol ly1 . was prepared by dissolving 0.. China. w4x. Standard solution The appropriate volumes of the working standard solution Ž0. 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. was prepared by dissolving 1.1 mg ly1 . to which 1. the water sample was diluted..5.2.6950 g of hydroxylamine hydrochloride in 250 ml of distilled water. in the initial report. were. Procedure 2. the lowest detectable concentration was lowered to 0. the reaction products of some phenolic compounds can not be extracted by chloroform. The series of the solutions contained phenol in the range of 0. In the proposed method. 2.0.. 7. if not considering the volume of the reagents.0= 10y2 .. w1x. Therefore. various reagents were directly added into a definite volume of a water sample and there is no dilution of the water sample. a Model P-E 403 atom absorption spectrophotometer ŽUSA. So. its composition and structure have been deduced and the reaction mechanism has also been investigated.7450 g of sodium nitroprusside  Na 2 wFeŽCN. The buffer solution ŽpH 12. mg ly1 . Kang et al. worthy of more attention. the mechanism of the colored reaction is not well established. to a 1000-ml volumetric flask and diluting it to the mark with distilled water.5 NOx ⭈ 2H 2 O4 in 250 ml of distilled water.0 ml of 4. Reagents All the solutions were prepared from analytical reagent ŽA. many other spectrophotometric methods w2᎐6x have been developed in recent years.1 mol ly1 . a pHS-2C digital acidimeter ŽHangzhou. The solution should be stable for at least 30 days and kept in a refrigerator.5 ml of NaOH solution Ž6 mol ly1 . a Model MS-500 OXFORD Mossbauer ¨ spectroscope ŽEngland. a Model 1106 element analyzer ŽItaly. However. Also.. respectively. 5.

a pre-distilled step w1x was adopted before analysis. 163 maximum absorbance is at 700 nm when there is phenol in the reaction system. and 3. 3. 3. r Microchemical Journal 64 (2000) 161᎐171 mol ly1 hydroxylamine hydrochloride solution. and the blank in which there is no phenol displays a very weak absorbance at 700 nm. Results and discussion Fig. Effects of the amounts of the reagents 2.0 ml of sodium nitroprusside. and ŽB. and analyzed as described above. A 50-ml sample of the distillate was transferred into a 50-ml color tube Žwith matching cork. blank. 3. Sample analysis For water samples. Absorption spectra of the reaction system: Ž⽧.0 ml of buffer solution can produce the maximum absorbance.1. . Reaction time and stability The absorption spectra of the reaction system are shown in Fig. So. 3.0 ml of buffer solution were added. respectively. the optimum determination wavelength was selected at 700 nm. 1. Choice of the determination wa¨ elength 3.. It is worth noting that the addition of excess hydroxylamine hydrochloride could make the sensitivity drop. The absorbance at 700 nm in 2-cm cells was measured against a distilled water blank.3.2. 1.0 ml hydroxylamine hydrochloride and 3. phenol. they were allowed to stand for 15 min.2. hydroxylamine hydrochloride and buffer solution.3. It can be seen that the The color reaction goes very fast and it can occur almost completely within 5 min Žsee Fig. Fig. 2 shows the effects of the amounts of sodium nitroprusside.C. Evidently. After mixing well. 1. 1. Kang et al.

The concentrations of phenol are: 3. Effects of the amounts of the reagents: ŽB. 1. buffer.50 mg ly1 ŽB. and Ž'.C. r Microchemical Journal 64 (2000) 161᎐171 164 Fig.50 mg ly1 Ž⽧. At the range of 0᎐60⬚C. sodium nitroprusside. 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... 3. Of the reagents used in this test. but the absorbance dropped rapidly at a temperature higher than 60⬚C.00 mg ly1 Ž'. only the working standard solution of phenol must be prepared on the same day. 3. The maximum absorbance can be obtained within 15 min and it can remain stable without any obvious change within 5 h. the solutions of hydroxyl- Fig. temperature hardly showed any effect on the analysis. respectively. Effect of the reaction time. hydroxylamine hydrochloride. . 2..4. and 0. Kang et al. Ž⽧.

It is clear from Table 1 that the color intensities of o-cresol and m-cresol can match that of phenol and they are also common phenolic compounds in aquatic environments. which was obtained by the parallel determinations of two samples each day within 5 consecutive days. NHq .5. The proposed method Žmg ly1 . the linear regression equation is A s 0. but they almost have no absorbance at the range of 500᎐800 nm.7 3.4-Dichlorophenol 4-Aminophenol Green᎐blue Green᎐blue Green᎐blue Green᎐yellow Light-green Green᎐blue Green᎐yellow Green᎐blue Green᎐blue Green᎐blue 700 700 700 700 700 680 700 720 700 700 1.38= 103 amine hydrochloride and buffer can be kept for a long time and the solution of sodium nitroprusside can also be kept for a few months Žin the dark. Ba2q. Responses of other phenolic compounds Under the above optimum conditions.62= 104 1.05᎐5..93= 102 1.4-dinitrophenol all exhibit a light-yellow color.5 0.C.6.46= 103 93.35= 104 41. Fe 3q. R. Kang et al. r Microchemical Journal 64 (2000) 161᎐171 165 Table 1 The reaction features of various phenolic compounds Compounds Color ␭ma x Žnm. 3-nitrophenol. Calibration cur¨ e and precision 3.320 0. Nitrophenols including 2-nitrophenol. but its ␭ max is in the ultraviolet region.2669C y 0.282 0. ␧max Žmol 1y1 cmy1 .242 0.9996. r s 0. at room temperature. Some compounds not included in the table.323 0. Effects of the ions For phenol solutions Ž3. Cu2q. Naq. HSO 3 . Under the experimental conditions. many phenolic compounds can give the color reaction. Al 3 . 100 mg ly1 of Kq. HCOy 4 .5-mg ly1 standard solution of phenol.245 1.244 0.2 . Cly. As the Table 2 Determination of phenol in gas wastewater Sample 4-AAP method Žmg ly1 . if the relative error of 5% is permitted. the linear regression equa- 3. pre-distilled step is adopted. Under the experimental conditions.94 1. Thus.23= 104 1. CH 3 COOy did not disturb the analysis. EDTA also interfered with the analysis.67= 102 5. 1 2 3 4 5 0. it cannot be used as a masking agent.. 4nitrophenol and 2.9 1. ␭ max and molar absorptivity Žsee Table 1.275 0.01516 Ž n s 7. 3q y Fe 2q. the effects of the above-mentioned ions could be eliminated.6-dihydroxyl benzene can also exhibit a brown᎐red color.5 y2..01= 102 1. 3.0 mg ly1 .1 2.240 0.4-Dimethylphenol 1-Naphthol 4-Chlorophenol 4-Chloro-3-methyl phenol 2.0 mg ly1 . PO43y. Relative error Ž%. but they display different colors.7.D.%s 1. Phenol o-Cresol m-Cresol p-Cresol 2. SO42y.208 0.3. 2y S did disturb the analysis seriously. and 100 mg ly1 of Mg 2q. For a 0. such as 1.201 0. C s 0.S.

166 C. 69509202. respectively.1. Ž n s 5. 3.9. respectively. was added into a 250-ml beaker followed by addition of 5 ml of 6 mol ly1 NaOH.10. green solid product. using the proposed and 4-AAP methods. A solution of 1 g of hydroxylamine hydrochloride dissolved in 10 ml of water was added dropwise into the reaction system in the beaker. and ŽB.S. the R. For sample no. and 1. the R.9 g of phenol and 2 g of sodium nitroprusside were added and made to dissolve. 3.. was 3.D.0 mg ly1 phenol.E.0%. to which 1.7% Ž4-AAP method.8. Absorption spectra of the solid product in water: Ž⽧.S. blue solid product.05.9993.6% and average recovery was 98. and A s 0. The F values of the statistical analysis of the two methods showed no significant difference Ž Ps 0.394" 0.006441 Ž n s 5.10.01814 Ž n s 5.s 2. 4.8% Ž n s 10. The mixture was stirred continuously until the solution became completely blue. Precision of sample analysis For the 0. Reaction mechanism 3. r Microchemical Journal 64 (2000) 161᎐171 tions of o-cresol and m-cresol are: A s 0.9997. Comparison between the proposed and 4-AAP methods Five samples were collected from a gas waste water. The blue solution was transferred into an evaporator and heated in a 60⬚C water bath until a Fig.s 2. the R. r s 0. 4...2934C y 0.. r s 0. The analysis results.D. 3.1% Žproposed method. Synthesis of the product Water Ž50 ml.3425C q 0. are listed in Table 2. . China.. Kang et al.003-mg ly1 standard sample of phenol Žno. For South Lake water samples which were spiked with 2. the Environmental Monitoring Central Station of Jilin Province.

So. 5.8x and it does not appear in Fig. The filtrate was cooled gradually to room temperature and left until a large amount of a blue crystal separated out again. showed the weak ferromagnetism of the product.3= Fig. ¨ The solid was redissolved in a minimum amount of warm water. the weaker the intensity. the product does not contain an NO group. The hysteresis loop of the product ŽFig..5 NOx ⭈ 2H 2 O is at 1944 cmy1 w7. 3. The longer the time. r Microchemical Journal 64 (2000) 161᎐171 liquid film separated out on the surface. It was left at room temperature until a large amount of solid separated out. respectively.10. it can be deduced that phenol is not present in the product. Kang et al. of feedstock Na 2 wFeŽCN. The crystals were filtered by suction on a Buchner funnel and ¨ dried in a desiccator. The maximum absorbance wavelength is also at 700 nm. The solid product was synthesized many times and the intensity of the peak varied with the preservation time. 5. Figs. 4 which is similar to Fig. Characterization of the product The absorption spectra of the crystal of the product in water is shown in Fig. 1.C. In the IR spectrum. 5 and 6 are IR and Raman spectra of the crystal of the product. . which was filtered out on a Buchner funnel with the aid of suction. Therefore. 7. the peak at 2057 cmy1 is assigned to 167 the vibration of the cyano group ŽCN. The IR spectrum of the solid product ŽKBr pellet. The peak of the nitroso group ŽNO. The peak at 1628 cmy1 in the IR spectrum can be assigned to H 2 O and it disappears in the Raman spectrum. The magnetic susceptibility calculated was 7. In addition. and it splits into two peaks at 2102 and 2072 cmy1 in the Raman spectrum. the 1 H-NMR of hydrogen atoms on the benzene ring cannot be observed while determining the 1 HNMR spectrum of the product.2.

also showed that the solid product has no diamagnetism. The hysteresis loop of the solid product.. scanning 100 times. 10y5 cgs. The experiment of the Mossbauer spec¨ trum determination at room temperature Ž298 K. r Microchemical Journal 64 (2000) 161᎐171 Fig.168 C. The Raman spectrum of the solid product Žpower 500 mW. Kang et al. Fig. 7. . 6.

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.3:4. the theoretical ratio should be 1:5:5:5.. So. The results of elemental analysis also indicate that the solid product contains some crystallization water. 3. 8. which can also be proved by the IR spectrum Ž1628 cmy1 . which was obtained by elemental analysis. In the color reaction process in aqueous solu- . If the crystal separated out in an ice᎐salt bath during the recrystallization. The green form had the same absorption spectrum as the blue one in aqueous solution and the ␭ max was also at 700 nm ŽFig. 8. ŽFig. a green crystal of the product could be obtained. only displayed a sharp peak and there was no quadrupole splitting.8. The Mossbauer spectrum of the solid product wroom temperature Ž298 K. ¨ The ratio of mole fraction of FerNarCrN of the product was 1:4. r Microchemical Journal 64 (2000) 161᎐171 169 Fig.x. Compared with Na 2 wFeŽCN. Reaction mechanism On the basis of the characterization of the solid product. The blue solid product can be obtained from the synthesis method above. Consequently.10.10 x. the mechanism of the color reaction is deduced as follows: Its molecular formula should be Na 10 wFe 2ŽCN. Hence. the solid product should have structural isomers. from 2150 to 2057 cmy1 . 4. the structure is deduced as follows: tion. Their IR spectra were completely identical. The Mossbauer spectrum of ¨ the product at room temperature Ž298 K. Kang et al.C. the ferronuclear site should be situated in the center of a regular tetrahedron or an octahedron structure. the ferro valence degraded to 0 and the absorption frequency of the cyano group ŽCN. in the IR spectrum also shifted significantly.9:5..5 NOx ⭈ 2H 2 O.3. As the tetrahedron is impossible for this test.

3. This may be caused by the hindrance of the p-substituted structure. the electrophilic or electrophobic property of the substitutes on the benzene ring also affects the sensitivity of the color reaction and phenolic compounds with different substitutes giving different sensitivities Žsee Table 1. it is said that the product is diamagnetic and the magnetic susceptibility is y1. Lin. So. w4x mainly in amounts of the reagents used.or ortho-position. References w1x Environmental Protection Agency of China. Zhang. For the proposed method.3. respectively. it is said that o-cresol and m-cresol cannot react. As an electrophile. Kang et al. Different responses of some phenolic compounds For the method reported by Nagaraj et al. 3. When phenol is converted to phenolate. But. but also have a higher sensitivity. r Microchemical Journal 64 (2000) 161᎐171 In the reaction.J. it is said that a large amount of anions does not disturb the determination and many cations disturb the determinations slightly. the nitroso group Žq NO. nitrophenols couldn’t give the color reaction.05᎐5. on the benzene ring repels the electron violently.3= 10y5 cgs. W. Different determination range The linear dynamic range of the determination of the method reported by Nagaraj et al. 1986. the negative oxygen ion ŽOy. w4x. the electrophilic reaction could take place more easily.11. Different effect of foreign ions For the method reported by Nagaraj et al. this is because nitro ŽNO 2 . For the proposed method.2. Compared with o-cresol and m-cresol. R. becoming a very good electron donor. X. the electrophilic reaction being inhibited. w4x is 1.68= 10 4 mol ly1 cmy1 and that of the proposed method is 1.4. Yan.11. o-cresol and m-cresol not only can react. 3.0 mg ly1 . Comparison between the modified and the original methods The analytical method established in this article is similar to that reported by Nagaraj et al. anions also had an effect on the determination of phenol and the errors caused by the anions and cations are much higher than those of the method reported by Nagaraj et al.5. p-cresol has a much weaker sensitivity. The Analytical Methods of Environmental Monitoring. 158᎐167. w4x is 0.. the sites that the nitroso group Žq NO. Chem. the product is weakly ferromagnetic and the magnetic susceptibility is 7. According to pure organic chemistry theory. Moreover. Fan. 3. 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. Maybe. pp. Also. the nitroso group in the form of qNO attacks the benzene ring and an electrophilic reaction occurs. w4x. p-cresol is insensitive to the color reaction.Y.23= 10 4 mol ly1 cmy1 . In the actural color reaction. The molar absorptivities are 1. For the proposed method. the reaction mechanism is investigated in this thesis. which makes the benzene ring very active.62= 10 4 mol ly1 cmy1 and 1. w4x.35= 10 4 mol ly1 cmy1 . Ž1994. 3.1.63= 10y6 cgs. . Different physical property of the solid product For the method reported by Nagaraj et al. J. which is not given in the method reported by Nagaraj et al. w4x. Univ.170 C.11.11. attacks on the benzene ring could be at the para. is an electron-withdrawing group and it makes the benzene ring inactive. 3. Environmental Science Press of China. w4x. Of the phenolic compounds tested. may be in the larger anionic complex and the way attacking para-position is easier.1᎐6.11. and pH range of the reaction.11. 1305᎐1308. w2x H.5 mg ly1 and that of the proposed method is 0. Different sensiti¨ ity The molar absorptivity of the method reported by Nagaraj et al. order of addition of the reagents. Beijing. 15 Ž9. Chin. there are significant differences in the following aspects between the two methods.

Ž1992. 26 Ž5. 1376᎐1380. Chem.K. Anal. Inorg. Wei. L. Chin. Y. Chem. Ž1993. Chem.K. Brown. Lindner. r Microchemical Journal 64 (2000) 161᎐171 w3x J. Zhang. Bates. V.N. w7x R. Khanna. P. 37 Ž4. J. B. 9 Ž1970.C.B. 8 Ž1969.N. Chem. C.M.B.H. Khanna. Li. Achar.K. Ramachandran. w4x P. Ž1998. Gupta. R. w8x J. Anal. Bhandari.P. G. Chem. 693᎐696.P. Kang et al. 641᎐643. Indian J. 489᎐494. Jones. 32A Ž7. 171 w6x K. X. Nagaraj. Ž1992. Koch. . 2195᎐2201. 586᎐589. Inorg. Ackermann.W. J. Talanta 39 Ž6. w5x S.