Journal of the Chinese Chemical Society, 2005, 52, 575-579

575

Spectrophotometric Determination of Copper in Natural Waters and

Pharmaceutical Samples with Chloro(phenyl) glyoxime
Orhan Turkoglu and Mustafa Soylak*
Erciyes University, Faculty of Art and Science, Department of Chemistry, 38039 Kayseri, Turkey

A simple method for the determination of trace amounts of copper by spectrophotometry is described
based on the formation of the copper-chloro-(phenyl) glyoxime complex. The molar absorptivities of the
complexes at pH 4.0 at 290.5 nm were 0.8 104 l/mol cm. Optimal conditions such as reagent amounts and pH
for the copper determination were reported. The effects of the foreign ions were also investigated. The method
was successfully applied for determinations of copper in some natural waters and pharmaceutical samples.
Keywords: Spectrophotometry; Determination; Copper (II); Chloro(phenyl) glyoxime; Complex;

Pharmaceutical samples.

INTRODUCTION
Copper is one of the important and essential nutrients
for human health as well as the growth of animals and
plants.1-7 Copper is required for normal metabolic processes.
Copper combines with certain proteins to produce enzymes
that act as catalysts to help a number of body functions. Copper helps provide energy required by biochemical reactions.
Although copper is an essential micronutrient and is required
by the body in very small amounts, excess copper in the human body can cause stomach and intestinal distress such as
nausea, vomiting, diarrhea, and stomach cramps. The lowest
level at which these adverse effects occur has not been well
defined. People with Wilsons disease, a rare genetic disorder, are more sensitive to the effects of copper. Copper is also
a commonly occurring element in natural waters. Most copper contamination in drinking water happens in the water delivery system, as a result of corrosion of the copper pipes or
fittings.1-9
Uv-vis spectrophotometry is still an important instrumental method for the determination of heavy metal ions in
their inorganic or organic complexes.8-15 Various spectrophotometric methods have been proposed for the determination
of copper contents of the various samples including natural
waters and pharmaceutical samples.16-30 Chloro(phenyl) glyoxime is selected as a ligand for copper(II) ions for the present work. The chloro(phenyl) glyoxime and the electrical
properties of its Co(II), Ni(II) and Cu(II) chelates in solid

phase have been investigated by our working group.31

In the present work, a sensitive and simple method for
the determination of trace amounts of copper(II) by Uv-vis
spectrophotometry is described based on the formation of the
copper(II)-chloro(phenyl) glyoxime complex. The influences of some analytical parameters including pH of the
aqueous and reagent amounts, etc. on the complex formation
were investigated.

EXPERIMENTAL
Apparatus
A Hitachi Model 150-20 UV-VIS double beam spectrophotometer with a 10 mm optical path cell was used for spectrophotometric measurements. The pH was monitored on a
Delta 320 Mettler pH meter with a glass and calomel electrode pair.
Reagents and Solutions
All chemicals obtained from commercial sources were
of super pure grade unless otherwise stated. Freshly prepared
doubled distilled water, from a quartz still, was used in all experiments. The copper stock solution, 1000 mg/l was prepared from copper(II) nitrate (Sigma Chem. Co., St. Louis) in
10-3 M HNO3. Portions were diluted daily for obtaining reference and working solutions. Borate buffer solutions: 0.1 M
HCl solution was added dropwise to 0.05 M sodium tetra bo-

* Corresponding author. Fax: +90-352-4374933; E-mail: soylak@erciyes.edu.tr

576

J. Chin. Chem. Soc., Vol. 52, No. 3, 2005

rate solution (Merck-Darmstadt) to the required pH.

Preparation of Ligand
The ligand, chloro(phenyl) glyoxime, was prepared according to the procedure reported previously.31 5.0 g phenylglyoxime was re-ground in an agate mortar, and then it was
formed as a suspended mixture by the addition of a sufficient
amount of chloroform. Dried chlorine gas was passed through
the suspended mixture while being exposed to a beam of UV
radiation for 15 to 20 min. At the end of the processes, a white
product was obtained. After completion of the chlorination,
air was passed through the product for half an hour to remove
excess chlorine.
Chloro(phenyl) glyoxime solution (0.05% (w/v)) was
prepared daily by dissolving in water/ethanol (25/75 v/v).
The ligand was used without further purification. The structure of the ligand and its copper complex was identified in our
previous work.31 The proposed structure of the ligand and its
copper complex are given in Fig. 1.
General Procedure
Ten micrograms of copper were transferred into a 25mL calibrated flask and 1000 ml of 0.05% chloro(phenyl)
glyoxime solution was added. The pH of the solution was adjusted with borate buffer to pH 4.0. Then the whole solution
was mixed and diluted with distilled water to the mark. After
10 min, the absorbance of this solution was measured at 290.5
nm by spectrophotometer against a reagent blank.
Application of Real Samples
The method was applied to the determination of copper
contents of some vitamin samples produced in Turkey. For
this purpose, firstly the ashing procedure given by Dalman et
al. 16 was applied to the vitamin samples. Five tablets were
powdered and the required amount of the powder was ignited
in a muffle furnace at 400 C for 2 h. The ash was dissolved in
5 mL of concentrated HCl, and then the final solution was fil-

Fig. 1. Ligand (A), its copper(II) complex (B).

Turkoglu and Soylak

tered and diluted to 100 mL with distilled water. In this solution, the copper concentration was determined by the procedure given above.
Twenty mL of a drinking water sample was transferred
to a 25 mL of volumetric flask, then the General Procedure
given above was applied. The copper content was determined
spectrophotochemically at 290.5 nm.

RESULTS AND DISCUSSION

Absorption spectra
The absorption maxima was obtained for copperchloro(phenyl) glyoxime complex in water at 290.5 nm (Fig.
2). A reagent blank was used thereafter as a reference because
it showed absorption at this wavelength. The complex gives a
maximum absorption at 290.5 nm with a molar absorptivity
coefficient of 0.8 104 l/mol cm.
Effect of pH
Due to the pH of the aqueous solution being an important parameter for chelate formation, the influences of pH of
the aqueous solution on the formation of copper-chloro(phenyl) glyoxime complex were investigated spectrophotometrically. The effect of the pH on the formation of the copper-chloro(phenyl) glyoxime complex was examined at
290.5 nm using various buffer solutions of different pH values. The results are given in Fig. 3. The complex begins to
form at approximately pH 2.0, with maximum absorbance being reached at pH values between pH 4.0 to pH 6.0. In the
light of these findings, all subsequent studies were carried out
at pH 4.0 using borate buffer.
Influences of amount of chloro(phenyl) glyoxime
The influences of chloro(phenyl) glyoxime amounts on
the formation of copper-chloro(phenyl) glyoxime complex
was investigated at pH 4 containing 10 mg copper(II). It was
found that maximum absorbance values were obtained for
0.05% chloro(phenyl) glyoxime solution in the range of
800-1500 ml. In all further studies, 1000 ml of 0.05% chloro(phenyl) glyoxime solution was used.
Effect of time on the complex formation
The influence of time on the formation of the copperchloro(phenyl) glyoxime complex was also studied at the optimal working conditions. The development of the copperchloro(phenyl) glyoxime complex is complete in 10 minutes,

Spectrophotometric Determination of Copper

and the color is stable for at least 24 hours.

Calibration graphs and detection limit
A calibration curve was constructed for copper in the
range of 0.1-10.0 mg/l of copper at 290.5 nm according to the
general procedure described above. The calibration graph
can be represented by a linear regression equation:
y = 0.1047x + 0.0086 (r2 = 0.999)
(y = absorbance, x = concentration).

J. Chin. Chem. Soc., Vol. 52, No. 3, 2005

Analysis of samples
In order to highlight the utility of the proposed method,
it was used for the spectrophotometric determination of copper contents of natural water and pharmaceutical samples.
The results are given in Table 2 for pharmaceutical samples.
The copper concentration was determined by the present
method taking suitable aliquots of the above sample solution,
and the results are compared with the certified values.
The presented procedure was also applied for the determination of copper contents of two drinking water samples
from Turkey. As is seen from Table 3, good agreement was

The detection limits based on three times the standard

deviations of the blank were 10 mg/l (N = 21). The relative
standard deviations for the copper determinations were 2%
(N = 10) at 0.5 mg/l Cu(II).
Effect of diverse ions
We have also evaluated the effect of foreign ions (both
cations and anions) on the complexation of copper by measuring the absorbance of solution containing 10 mg of copper(II) in 25 mL of distilled water. Tolerable limit was defined as the highest amount of foreign ions that produced an
error not exceeding 5% in the determination of copper. The
results are summarized in Table 1. The formation of the
Cu(II)-chloro(phenyl) glyoxime complex was not influenced
by alkaline, earth alkaline ions, some transition metal ions, or
other anions. The concentrations of Co 2+ , Ni 2+ , and Cr 3+ in
our real samples were found to be within tolerable limits.
Therefore the procedure could be applied to these samples.

577

Fig. 3. Effect of pH on the formation of copper-chloro(phenyl) glyoxime complex (10 mg copper(II),

1.0 mL of 0.05% chloro(phenyl) glyoxime).

Fig. 2. Absorption spectrum of copper-chloro(phenyl) glyoxime complex (A: complex, B: ligand).

578

J. Chin. Chem. Soc., Vol. 52, No. 3, 2005

Turkoglu and Soylak

Table 1. Tolerable limits of the foreign ions on the determination

of copper as chloro(phenyl) glyoxime complex (N = 3)
Diverse Ions
+

Na
K+
Ca2+
Mg2+
SO42Acetate
ClFe3+
Pb2+
Mn2+
Cr3+
Co2+
Ni2+

Added As

Tolerable limit [Ion]/[Cu(II)]

NaCl
KCl
CaCl2
MgCl2
(NH4)2SO4
NH4CH3COO
NH4Cl
FeCl3
Pb(NO3)2
MnSO4
Cr(NO3)3
Cor(NO3)3
NiSO4

25000
1000
1000
1000
1000
1000
20000
5
5
5
2
5
5

ACKNOWLEDGEMENT
The authors are grateful for the financial support of the
Unit of the Scientific Research Projects of Erciyes University
(Kayseri-Turkey). The authors also would like to thank the
Scientific and Technical Research Council of Turkey
(TUBITAK) for financial help (TBAG-1810 and TBAG
2163).

Table 2. Determination of copper as chloro(phenyl) glyoxime

complex by spectrophotometry in pharmaceutical
samples
Sample

Certified Value (mg/g)

Our Value (mg/g)

1.0
1.0
0.07
0.2

0.97 0.09
1.06 0.09
0.069 0.006
0.19 0.02

Supradyn
Vitadiyn
Vi-mineral
Eunova

Table 3. Copper levels of drinking water samples from Turkey

(Volume: 20 mL, N: 4)
Aytac Akyudum
Added
(mg)
0
10
20
30

procedures for copper by various glyoximes given in the literature:18,20 high molar absorptivity value, low detection limit,
and easy application to the real samples. The method is easy
to perform for the determination of copper contents of natural
waters and pharmaceutical samples.

Turkuaz

Found
(mg)

Recovery,
%

Found
(mg)

Recovery,
%

N.D.
09.9 0.4
20.3 1.2
29.4 1.3

99
1020
98

N.D.
10.0 0.6
21.0 1.4
30.3 2.0

100
105
101

N.D.: not detected

obtained between the added and found analyte content using

the recommended procedure. The recoveries values calculated for the standard additions were always higher than 95%.

CONCLUSION
The proposed method is sensitive, accurate, and tolerant to many foreign substances, and all the reagents used are
stable under the conditions. The presented procedure has
some advantages according to the other spectrophotometric

Received August 31, 2004.

REFERENCES
1. Kumar, A.; Hussain, M. F.; Satake, M.; Puri, B. K. J. Chin.
Chem. Soc. 1984, 31, 55.
2. http://www.e-b-i.net/ebi/contaminants/copper.html (accessed August 2004).
3. Lin, J. L.; Chang, L. F. J. Chin. Chem. Soc. 1984, 31, 297.
4. http://www.ly-copper.com/e-tyhj.htm (accessed August
2004).
5. Lin, J. L.; Satake, M. J. Chin. Chem. Soc. 1985, 32, 105.
6. http://www.ianr.unl.edu/pubs/water/g1360.htm (accessed
August 2004).
7. Chao, M. S.; Chung, C. S. J. Chin. Chem. Soc. 1989, 36, 301.
8. Malik, A. K.; Rao, A. L. J. J. Chin. Chem. Soc. 1992, 39,
235.
9. Rauf, M. A.; Akhter, Z.; Kanwal, S. J. Trace Microprobe
Tech. 2003, 21, 577.
10. Cheng, H. S.; Pai, S. C.; Yang, M. H.; Chang, T. Y.; Ho, W.
H.; Jen, J. F.; Hsieh, C. F.; Jian, S. J.; Hsi, T. S.; Cheng, K. L.;
Lin, T. H.; Ho, S. M.; Tsai, C. M. J. Chin. Chem. Soc. 1996,
43, 237.
11. Turkoglu, O.; Soylak, M.; Colak, H. Asian J. Chem. 2002,
14, 363.
12. Ma, D. L.; Huang, Y. R.; Xia, D. S.; Wang, Y. L. J. Chin.
Chem. Soc. 1999, 46, 933.
13. Rauf, M. A.; Ikram, M.; Rauf, N. J. Chem. Soc. Pak. 2001,
23, 136.
14. Abd-Elzaher, M. M. J. Chin. Chem. Soc. 2001, 48, 153.
15. Sharma, A.; Arora, A.; Mathur, K.; Mathur, R. P. Asian J.

Spectrophotometric Determination of Copper

Chem. 2003, 15, 1091.

16. Gholivand, M. B.; Sobhani, S.; Khirdoosh, F. J. Chin. Chem.
Soc. 2002, 49, 355.
17. Molina, F.; Fernandez, D. G.; Bosque-sendra, J. M.;
Espinosa, P. J. Phar. Biomedal. Anal. 1988, 6, 1019.
18. Dalman, O.; Tufekci, M.; Nohut, S.; Guner, S.; Karabocek,
S. J. Phar. Biomedal. Anal. 2002, 27, 183.
19. Farajzadeh, M. A.; Vardast, M. R. J. Chin. Chem. Soc. 2003,
50, 245.
20. Karabocek, S.; Nohut, S.; Dalman, O.; Guner, S. Anal.
Chim. Acta 2000, 408, 163.
21. Wei, G. T.; Chen, J. C.; Yang, Z. S. J. Chin. Chem. Soc. 2003,
50, 1123.
22. Liu, J. N.; Zhang, B.; Wu, B. W.; Yu, X. Q. J. Chin. Chem.
Soc. 2004, 51, 79.
23. Yang, Y. L.; Miao, M. M.; Lin, Q.; Yang, G. Y. J. Chin.

J. Chin. Chem. Soc., Vol. 52, No. 3, 2005

579

Chem. Soc. 2004, 51, 19.

24. Soylak, M.; Erdogan, N. D.; Elci, L. J. Chin. Chem. Soc.
2004, 51, 703.
25. Sirry, S. M.; Taha, G. M. J. Chin. Chem. Soc. 2002, 49, 39.
26. Chang, C. H.; Liu, C. Y. J. Chin. Chem. Soc. 1997, 44, 231.
27. Elansary, A. L.; Issa, Y. M.; Hanna, Z. F. J. Chin. Chem. Soc.
1986, 33, 309.
28. Sinha, P. P.; Kamil, F.; Sindhwani, S. K.; Singh, R. P. J. Chin.
Chem. Soc. 1982, 29, 169.
29. Singh, S. K.; Kamini; Sindhwani, S. K. J. Chin. Chem. Soc.
1982, 29, 131.
30. Anipindi, A. R.; Jain, P.; Singh, R. P. J. Chin. Chem. Soc.
1981, 28, 165.
31. Turkoglu, O.; Soylak, M.; Belenli, I. Coll. Czech. Chem.
Com. 2003, 68, 1233.