New Spectrophotometric Reagent Specific for the

Determination of Copper
Rostam H. Maghssoudi and Ahmad B. Fawzi
Department of Pharmaceutical Chemistry, College of Pharmacy, Tehran University, Tehran, Iran

Many colorimetric methods have been described for the

determination of trace amounts of copper, and those based
on bathocuproine ( I ) (2,9-dimethyl-4,7-diphenyl-1,10-
phenanthroline), neocuproine ( 2 ) (2,9-dimethyl-l,lO-phen-
anthroline), and 2,2'-diquinoline ( 3 ) are considered to be
specific for copper(1).
The use of either diethyl dithiocarbamate or dithizone in
"1
0.8

the spectrophotometric micro- and macrodetermination of

copper did not prove to be entirely satisfactory. The former
is not sufficiently specific because it is not burdened by the
presence of commonly present ions, such as iron, nickel,
and cobalt. The later is not satisfactorily selective in its ac-
tion a t a wide range of p H values, and the color of the re-
agent itself interferes with the measurements. For this rea-
son Irving et al. ( 4 ) gave a new but more complicated pro-
cedure for the use of dithizone for the determination of
copper.
Most of the other colorimetric methods (5-7) that have
been proposed for the determination of copper are of slight
importance from the practical point of view.
This paper introduces 6-phenyl-2,3-dihydro-as-triazine-
3-thione(PDTT) as a new colorimetric reagent specific for
the determination of copper(111, and describes systematic
studies on the solvent extraction of Cu(I1)-PDTT complex.
P D T T was first prepared and reported by Iraj Lalezari
(8). Then it was used for the gravimetric determination of
thallium ( 9 ) ,and for the spectrophotometric determination
of mercury(I1) (10).
WAVELENGTH, nm
EXPERIMENTAL
Figure 1. Absorption spectra of chloroform solutions of various con-
Apparatus. The absorbances were measured by a Carl-Zeiss
spectrophotometer, PMQ I1 type, with 1-cm quartz cells. centrations of Cu(II)-PDTT complex obtained following the recom-
A Pye-Unicam model SP 90A atomic absorption spectrophotom- mended analytical procedure
eter was used for atomic absorbance measurements.
The p H of the solutions were measured by using a Beckman H3
type pH meter.
All measurements were carried out a t room temperature (20 f 2 RESULTS AND DISCUSSION
"C). Absorption Curve. Figure 1 shows the absorption spec-
Reagent and Chemicals. A stock solution of copper nitrate, tra of a series of chloroform solutions containing different
containing 0.7608 mg Cu/ml solution, was prepared by dissolving
the pure metal in 5 ml concentrated nitric acid, followed by heat- amounts of the complex. The maximum absorption of the
ing until almost dry to dispel the gases, and dilution to volume. chloroform solution of Cu(I1)-PDTT complex was a t 500
This solution was standardized by the atomic absorption method nm.
vs. a standard sample of the National Bureau of Standards (Unit- Determination of the Composition of the Chelate.
ed States Department of Commerce, Washington 25, D.C.). The The stoichiometry of the compound involved in the forma-
solutions of lower concentration were prepared by volumetric dilu-
tion of the complex was ascertained by the following meth-
tion of the stock solution with distilled water.
A solution of 0.001M P D T T was prepared by adding a slight ex- ods.
cess of concentrated sodium hydroxide solution to the weighed T h e Mole Ratio Method. The method of Yoe and Jones
amount of P D T T , and diluting with distilled water to the right ( 1 1 ) was applied to aqueous solutions containing standard
volume. This solution is stable up to 5 hours. copper solutions a t a fixed concentration of 1 X lO-*M and
T o study the effect of other ions on the Cu(11)-PDTT complex, different amounts of standard PDTT solutions. Absorban-
solutions containing 10 mg/ml of the desired ion were prepared. ces were measured a t 500 nm after 10 minutes. Figure 2
All other reagents used were of Analar grade chemicals.
General Procedure. Pipet a sample of sufficient size to contain shows that the rapid increase of the absorbance occurs as
25-100 kg of copper(I1) into a 50-ml separatory funnel. Add 1 ml of the concentration of P D T T increases until a molar ratio
4N tartaric acid, 3 ml of freshly prepared 0.001M P D T T , followed [Cu(II)]:[PDTT] = 1:2. For higher concentrations of
by 5 ml of chloroform, shake vigorously, and then add 1 ml of 4N PDTT, the absorbance remains constant, suggesting the
sodium hydroxide solution. Continue shaking to extract all of the formation of 1:2 complex.
red color formed in the aqueous layer. Remove the chloroform T h e Continuous Variation Method (12, 1 3 ) . The total
layer in a 10-ml volumetric flask. Then continue the extraction
with 3 and 2 ml of chloroform, respectively. Adjust the volume of +
concentration of the mixtures [Cu(II) PDTT] was kept
the flask, and finally measure the absorbance of the solution vs. a constant, first a t 5 X 10-4M and then a t 3 X lO-*M. The
similarly prepared reagent blank at 500 nm. procedure was performed a t pH 14, with the initial use of

1694 ANALYTICAL CHEMISTRY, VOL. 47, NO. 9, AUGUST 1975

 0.6 1 I 0.6 1
W
u
z
U
m
E
0
ln
m
U

2.0 4.0 6.0 0.2 0.4 0.6 0.8 1.0

I
~PDTTI/tCu**I [Cu2+1/( tCu"I+[PDTTI)

Figure 2. Variation of the absorbance at 500 nrn. [Cu2+] = 1.0 X Flgure 3. Variation of the absorbance at 500 nrn. Concentration of
i o - 4 ~
Cu2+ and PDTT: ( 0 )5.0 X 10-4M; (0)3.0 X IOe4M

Table I. Extraction of Copper(I1)-PDTT Complex Table 11. Effect of Foreign Ionsa

with Chloroform as a Function of pH Ila\r,mun.
Copper(I1)- PDTT Ions a d d e d Sources aniomt, u 9
compleu e m - c t e d D,
PH in CHCl3, distribution ratio Hg?' H@1, 1000
1.o 18.85 0.46 Hg' HgNO?. 3HzO 1000
2 .o 35.23 1.09 Ag' -%NO, 600
4 .O 100 .oo m Fe2+ FeSO, 20000
8 .O 100 .oo cc C02' COC1, 6H?O 100
14.0 100 .oo co a Cu(I1) = 61.0pg; pH 14; PDTT = 0.001.V

tartaric acid. The absorbance measurements were made at
500 nm. Typical observations are shown in Figure 3 where
it is seen that the maxima occurs at 0.33 mole fraction of
copper indicating that the composition of the complex is
1:2.
Beer's Law, Range, a n d Precision. Three standard se-
ries of 7 samples, ranging in concentration from 2.1 to 12.3
bg/ml of cooper(II), were treated by the recommended pro-
cedure. Conformity to Beer's law was obtained in each case.
Optimum concentration range for measurement a t 500 nm
and 1.00-cm optical path is about 2.5 to 10.5 bglml of cop-
per(I1). The absorptivity a t 500 nm is 4.976 X lo3 liter
mole-' cm-l. The relative standard deviation of the calcu-
lated absorptivities of the 2 1 samples in the three standard
series is 2.7%. The relative standard deviation of the calcu-
lated absorptivities of the 15 samples in the optimum con-
centration range is 1.3%.
Effect of pH. T o prevent the precipitation of most cat-
ions, including copper, present in the form of hydroxides in
the alkaline medium, tartaric acid was added to the solu-
tion prior to the addition of PDTT. Addition of tartaric
acid forms copper tartarate salts, which eventually complex
with the PDTT. 'Following the above procedure, the pH
could be raised to as high as 14 with complete extraction of
the complex. Table I shows the extraction of Cu(11)-PDTT
complex in chloroform at pH 1 to 14. The results indicate
that the extraction is complete only between pH 4 to 14.
Stability of t h e Colored Complex. The chloroform so-
lution of Cu(I1)-PDTT complex is completely stable as a
function of time. The absorbance of the complex solution
in chloroform was measured a t elapsed intervals of 0, %, %,
1, 24,48, 72, and 120 hours, and was stable. Solutions of the
colored complex in chloroform have been kept in stored
tubes in the diffuse light of the laboratory for one year.
These solutions showed no change in the intensity when
compared with freshly prepared solutions.
Sensitivity of t h e Reaction. As little as 5 wg/ml of cop-
per could be detected visually as "red color" before the
chloroform extraction. However, the visual detection could
be improved to as low as 0.5 pg/ml, if the complex was ex-
tracted into 0.2 ml of chloroform.
Effect of Foreign Ions. In determining the effect of for-
eign ions, a known volume of standard copper(I1) solutions
containing various concentrations of the ion in question
was taken. Copper(I1) was determined as previously de-
scribed.
Thallium, nickel, magnesium, manganese, aluminum,
barium, strontium, calcium, platinum, molybdenum, anti-
mony, bismuth, lithium, potassium, sodium, tin, chromium,
iron(III), cadmium, lead, zinc, gold, fluoride, chloride, and
phosphate, were tested for their effect on the Cu(I1)-
P D T T complex, and were found to be completely without
interferences a t any concentration. Among these ions, zinc,
aluminum and antimony form white precipitates in alka-
line medium which could be dissolved by raising the pH to
14 with the addition of sodium hydroxide solution. Magne-
sium, manganese, chromium, calcium, molybdenum, bis-
muth, and barium form precipitates in alkaline medium
which could be prevented by raising the p H to 9 after the
addition of tartaric acid.

ANALYTICAL CHEMISTRY, VOL. 47, NO. 9, AUGUST 1975 1695

 Only iron(II), cobalt, silver, mercury(1) and (11) show se-
rious interferences in high concentrations. However, P D T T
dissolved in chloroform could be used to prevent the inter-
ference of iron(I1) and cobalt. Addition of silver, mercu-
ry(1) and (11) ions to the Cu(I1)-PDTT complex decompos-
es the complex giving the corresponding complexes. T o
prevent this interference by these ions, dithizone was
added to form stable complexes with silver, mercury(1) and
(11) in acidic solution, which could be extracted with chlo-
roform with no effect on the copper ion content. Then, the
copper ion content was determined by the described proce-
dure. However, the above recommendations could be omit-
ted if the concentration of the foreign ions is within the
limit shown in Table 11.
ACKNOWLEDGMENT
We thank Ali Shafiee for his valuable advice.
Application of Cryogenic Infrared Spectrometry to the
Identification of Petroleum
Patricia F. Lynch, Sheng-yuh Tang, and Chris W. Brown
Department of Chemistry, University of Rhode lsland, Kingston, Rl 0288 7
During the past two years, we have explored the applica-
bility of infrared spectrometry in identifying crude and re-
fined petroleum products (1-5), The sources of several ac-
tual oil spills involving both light ( 5 ) and heavy (2) fuels
have been correctly identified, even after the oils had been
on water for several weeks. In some cases, there were sever-
al suspect sources involved, and it was necessary to deter-
mine the correct source from as many as 14 oils.
Others (6-15) have used infrared spectra measured a t
room temperature to identify the source of petroleum but,
to our knowledge, no one has reported on the low-tempera-
ture spectra of petroleum. Herein, we discuss the feasibility
of using infrared fingerprints obtained a t 80 and 20 K to
identify the source of petroleum.
EXPERIMENTAL
Infrared spectra were measured on a Perkin-Elmer Model 521
infrared spectrometer. A typical low-temperature infrared cell
shown in Figure 1 was used for the measurements a t 80 K. CsI sub-
strate and outer windows were used on the cell. T o place a sample
of oil in the cell, one outer window was removed, the cell was held
in a horizontal position, and oil deposited on the CsI substrate (al-
ternatively, the oil can be deposited on the substrate through a
ground glass joint on the side of the cell). After depositing the
sample, cold gaseous NZwas passed into the inner chamber to cool
the substrate and sample; the outer chamber was flushed with dry
N p gas to eliminate condensation of H20.After the sample solidi-
fied, the outer window was replaced, the outer chamber evacuated
to -lo-” Torr, the cell turned upright, and the inner chamber
filled with liquid N2.Time required to prepare the sample cell and
measure the sample spectrum a t 80 K is approximately 45 min-
utes. Preparation of the sample and measurement of the spectrum
a t room temperature requires 25 to 30 minutes.
Spectra were also measured a t 20 K using a Cryotip cell (Air
Products Inc.). The sample deposition procedure was similar to
that used for the 80 K measurements, but more time consuming.
RESULTS AND DISCUSSION
Infrared spectra in the 650-1200 cm-l fingerprint region
of ( a ) a No. 2 fuel oil, ( b ) a crude oil, and ( c ) a No. 6 fuel oil
measured a t room temperature and a t 80 K are shown in
1696 ANALYTICAL CHEMISTRY, VOL. 47, NO. 9, AUGUST 1975
LITERATURE CITED
(1) G. F. Smith and D. H. Wilkins. AnalChem., 25, 510 (1953).
(2) G. F. Smith and W. H. McCurdy, Anal. Chem., 24, 371 (1952).
(3) J. G. Breckenridge. R. W. Lewis, and L. Quick, Can. J. Res., Sect. 6,17,
258 (1939).
(4) H. Irving, G. Andrew, and E. J. Risdon, Nature (London), 161, 805
(1948).
(5) J. R. Johnston and W. J. Holland, Mikro Chim. Acta, 1, 126 (1972).
(6) M. C. Patel, J. R. Shah, and R. P. Patel, J. Prakt, Chem., 314, 181
(1972).
(7) R. N. Virmani, 8. S. Gray, and R. P. Singh, lndian J. Chem., 10, 225
(1972).
(8) I. Lalezari and H. Golgolab, J. Heterocycl. Chem., 7, 689 (1970).
(9) M. Edrissi. A. Massoumi, and I. Lalezari, Talanta, 19, 814 (1972).
(10) R . H. Maghssoudi and F. A. Shamsa, Anal. Chem.. 47, 550 (1975).
(11) J. H. Yoe and A. L. Jones, lnd. Eng. Chem., Anal. Ed., 16, 111 (1944).
(12) P. Job, Ann. Chim., 9, 113 (1928).
(13) P. Job, Ann. Chim., 16, 97 (1936).
RECEIVEDfor review October 30, 1974. Accepted February
4, 1975.
Figure 2. The major differences between the low tempera-
ture spectra which are typical of all oils studied thus far
are: i) the 720 and 725 cm-I bands appear as two distinct
absorptions, increasing in intensity compared to the bands
measured a t room temperature; ii) the 740 cm-l band de-
creases in intensity at 80 K; and iii) the 890 cm-’ band in-
creases significantly in intensity compared to the band in
room temperature spectra.
The bands a t 720 and 725 cm-I are due to the in-phase
CH:! rocking mode of long-chain n-paraffins. The increase
in intensities and sharpness of these bands is most proba-
bly due to alignment of the n-paraffins into a polymeric
crystalline structure such as in polyethylene. Instead of
having a distribution of frequencies we observe only one
band, which is split into two components by intermolecular
interactions ( 1 6 ) .
There are other changes in the low temperature spectra,
but these are characteristic of each oil and cannot be gener-
alized. The distinct differences in the fingerprint a t low
temperature as compared to room temperature give an oil
sample, in effect, two “fingerprints”. The reproducibility of
the technique was tested by depositing the same oil sample
in the low temperature cell several times and measuring its
spectrum after each deposition.
Previously, we have shown that the room temperature
spectrum of,an unknown oil can be matched to that of the
correct source by ratioing the absorptivities of the bands of
oils from known sources with those of the unknown ( I ) .
Identification is made to the known having the most ratios
closest to the average ratio (currently, we are using a log-
scale). In the present study, we have used the same type of
analysis on the low temperature spectra, i.e., we have
matched unknown oils with the correct source oils by ra-
tioing absorptivities. The comparisons were made using
bands a t the same frequencies as those for the room tem-
perature spectra.
During October 1974, two major oil spills occurred and
the low temperature techniques were tested in a “real spill”