Saudi Journal of Biological Sciences (2012) 19, 247250

King Saud University

Saudi Journal of Biological Sciences

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ORIGINAL ARTICLE

Spectrophotometric estimation of cobalt with ninhydrin

Karamat Mahmood a, Feroza Hamid Wattoo b,*,
Muhammad Hamid Sarwar Wattoo b,c, Muhammad Imran a,c,
Muhammad Javaid Asad b, Syed Ahmed Tirmizi c, Abdul Wadood a

a
Department of Chemistry, The Islamia University of Bahawalpur, Bahawalpur 63100, Pakistan
b
Institute of Biochemistry & Biotechnology, Pir Mehr Ali Shah Arid Agriculture University, Rawalpindi 46300, Pakistan
c
Department of Chemistry, Quaid-i-Azam University, Islamabad 45320, Pakistan

Received 12 February 2011; revised 4 January 2012; accepted 7 January 2012

Available online 14 January 2012

KEYWORDS Abstract A violet coloured complex was developed when cobalt metal reacts with ninhydrin at pH
Spectrophotometric 8.2, using sodium acetate buffer solution. Absorbance of the complex was measured at 395 nm. Var-
estimation; ious factors, such as volume of the ligand used, solution pH, stability of the complex with time and
Cobalt; interference of other metals, which effect the complex formation have been studied in detail. Present
Ninhydrin developed method can be used for the spectrophotometric estimation of cobalt with ninhydrin com-
plex. The method is simple, selective and cheap for the determination of cobalt in very less time.
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1. 1.Introduction
Coordinate complex is formed when ligand surrounds the metal
atom through coordinate covalent or ionic bonds. Transition
metals behave as lewis acids due to a partially lled d-orbital
and therefore are capable of accepting electron pair/s. On the
other hand ligands are capable of donating electron pair/s
and referred to as lewis bases. Stability of a particular metal
* Corresponding author. Tel.: +92 3335348510.
E-mail address: drfhwattoo@uaar.edu.pk (F.H. Wattoo).
1319-562X 2012 King Saud University. Production and hosting by
Elsevier B.V. All rights reserved.
Peer review under responsibility of King Saud University.
doi:10.1016/j.sjbs.2012.01.001
Production and hosting by Elsevier
complex depends upon the nature of the metal and ligand being
used (Kettle, 1979). Ligands having high pH usually form more
stable complexes. A higher charge on smaller sized metals and
smaller ligands with larger electron density also imparts stabil-
ity to the metal complexes (Holme and Peck, 1998). These
molecular interactions between the electron donors and accep-
tors are mostly associated with the formation of coloured
charge-transfer complexes, which absorb in the visible region.
The classical photometric methods based on molecular interac-
tions are simple and suitable due to rapid formation of these col-
oured complexes (Foster, 1969).
In the recent literature, different spectrophotometric meth-
ods (Iqbal et al., 2006, 2007) have been developed with the aid
of different complexing reagents (Zhao et al., 1999; Khedr
et al., 2005; Bingol and Atalay, 2006). Both selective (Paleolo-
gos et al., 2002; Guzar and Jin, 2008) and simultaneous (Ozturk
et al., 2000; Sozgen and Tutem, 2004; Niazi and Yazdanipour,
2008; Arain et al., 2009) spectrophotometric estimation of co-
balt was carried out using different complexing ligands. To
the best of the authors knowledge, the use of ninhydrin
248
O saturated sodium acetate solution (10% in water). On 10 ml
OH
OH
O
Figure 1 Ninhydrin (A colourless compound).
(2,2-dihydroxy-1,3-indanedione) as a complexing reagent
(Singh, 1965) for the spectrophotometric estimation of cobalt
has not yet been practised. In the present work, the possibility
of ninhydrin (Fig. 1) to act as a ligand for the complexation with
cobalt was studied. This work has provided some interesting
and useful data for investigating the ability of ninhydrin (biden-
tate ligand) used to act as an analytical reagent. Here, ninhydrin
was used as a chromogenic reagent for the spectrophotometric
determination of cobalt.
2. Materials and methods
2.1. Apparatus and reagents
Spectrophotometric measurements were performed on a Shi-
madzu (Japan), UV-1700, double beam spectrophotometer
using matched 10 mm quartz cells. A Horiba F.8 (Spain),
pH metre, calibrated with standard buffer solutions of pH 4
and 10, was used for pH measurements.
All the reagents such as ninhydrin (Fluka), cobalt chloride
hexahydrate (Merck), ethanol (Merck), sodium hydroxide
(Merck), sodium acetate (BDH), citric acid (BDH), boric acid
(BDH), diethyl butyric acid (BDH), and potassium dihydrogen
phosphate (Fluka) were of analytical reagent grade purity.
Solutions were prepared with milliQ grade water of
R = 18.2 X unless otherwise specied. Nitrogen was bubbled
into each solution to release the interfering oxygen. Glass ves-
sels were cleaned by soaking in acidied solutions of KMnO4
or K2Cr2O7, followed by washing with concentrated HNO3,
and were rinsed several times with high purity de-ionized
water.
Cobalt solution: 5 ppm working cobalt solution was pre-
pared by diluting 0.5 ml of freshly prepared 1000 ppm stock
cobalt solution with milliQ grade water to 100 ml in a volumet-
ric ask.
Cobaltninhydrin complex: 10 ml of 5 ppm cobalt solution,
containing less then 0.1 g of cobalt, was added into a 50 ml
measuring ask followed by the addition of 10 ml of the
O HO
O-
2 + Co2+
pH = 8.2
OH
O
Figure 2 Proposed reaction of ninhydrin with cobalt ion in an alkaline solution.
K. Mahmood et al.
addition of ninhydrin solution (1% in dry ethanol) gives violet
coloured cobaltninhydrin complex and the nal volume was
made up to 50 ml with milliQ grade water. Absorbance was
noted at different wavelengths ranging from 360440 nm using
water as blank to nd the kmax.
Universal buffer solution: A mixture of 6.00 g of citric acid,
3.893 g of KH2PO4, and 1.769 g of boric acid and 5.66 g of
pure diethyl butyric acid was dissolved in water and diluted
to one litre. Various volumes of 0.2 M NaOH solution were
added in this universal buffer solution to maintain different
pH values.
3. Results and discussion
In the present work ninhydrin is used as chromogenic reagent that
forms a violet coloured complex with cobalt in slightly basic med-
ium (pH 8.2). The absorption maximum (kmax = 395 nm) of the
complex was found experimentally, whereas the ninhydrin reagent
did not show any absorbance. In order to obtain high sensitivity, a
wavelength of 395 nm was chosen for the spectrophotometric
measurement of the cobalt complex against a reagent blank. The
proposed reaction of ninhydrin with cobalt ion in alkaline solution
is given in Fig. 2.
Different molar excesses of ninhydrin were added to xed
cobalt ion concentration and absorbance was measured
according to the standard Procedure. Effect of ligand concen-
tration on cobaltninhydrin complex is given in Fig. 3. The re-
sults indicate that absorbance increases with the increasing
concentrations of ninhydrin solution. After 10 ml addition of
ninhydrin (1%), the absorption remained constant indicating
ninhydrin was sufcient enough to form a complex with
5 ppm cobalt solution. Greater excesses of ninhydrin up to
22 ml addition do not show a change in the absorption.
Stability of the cobaltninhydrin complex with respect to
time is given in Fig. 4. The cobaltninhydrin complex was
formed immediately on mixing the reactants giving violet col-
our. This instantaneous reaction reveals that the complex re-
mains stable for a period of 30 min as the absorbance
remained almost constant for this time interval and then de-
creased rapidly showing decomposition of the cobaltninhy-
drin complex.
pH is always considered as quite important and critical for
complex formation and its stability. It is always inevitable to
nd the optimum pH at which the complex is most stable (Sa-
bel et al., 2010). Effects of pH and buffer solution on cobalt
ninhydrin complex are given in Fig. 5. The results show that
pH 8.2 is most suitable for the complexation of cobalt with
ninhydrin. It was also seen that in acidic media, the complex
O
O -
O
Co2+
O-
O
O OH
Spectrophotometric estimation cobalt 249

2
1.8
1.6

Absorbance (nm)
1.4
1.2
1
0.8
0.6
0.4
0.2
0
0 3 6 9 12 15 18 21 24
Ligand Volume (ml)

Figure 3 Effect of ligand concentration on cobaltninhydrin complex.

1.72

1.7
Absorbance (nm)

1.68

1.66

1.64

1.62

1.6

1.58
0 5 10 15 20 25 30 35 40 45
Time (min)

Figure 4 Stability of the cobaltninhydrin complex with respect to time.

2.5
Absorbance (nm)

1.5

0.5
3 4 5 6 7 8 9 10 11 12
Solution pH

Figure 5 Effect of pH on cobaltninhydrin complex.

Table 1 Interference study of different metal cations.

Sr. # Absorbance without metal interference Eect of dierent metals Absorbance
1 2.044 Ni 2.031
2 2.044 Ba 2.024
3 2.044 Cu 2.026
4 2.044 Cd 2.046
5 2.044 Zn 2.041
6 2.044 Na 2.040
250
was less stale. The universal buffer solution used for pH main-
tenance did not interfere in actual analysis and provided the
desired pH for complex formation.
The effect of six different cations in the determination of
cobalt has been studied and results are tabulated in Table 1.
These data describe changes in the absorbance of cobaltnin-
hydrin complex in the presence of various interferants added
in equal quantity. The results indicate that the cations like cad-
mium, zinc, and sodium do not undergo too many signicant
changes in absorbance. However barium caused the highest
change in absorbance followed by calcium and nickel,
respectively.
4. Conclusion
The present method is very simple, selective and cheaper for
the spectrophotometric determination of cobalt in very less
time. Ninhydrin is a very common and easily available reagent.
The method does not demand ultra conditions to be main-
tained. Very low concentrations of cobalt can be determined
with good accuracy at an optimum pH of 8.2. However the
complex is stable for 30 minutes only and decomposition starts
afterwards.
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