A Catalytic Method for Determining Traces of Selenium

Philip W. West and T. V. Ramakrishna

Coates ChemicaI Laboratories, Institute for the Environmental Sciences, Louisiana State University, Baton Rouge, La.
70803
A sensitive method for t he det ermi nat i on of t race
amounts of selenium based on i t s catalytic ef f ect i n t he
reduction of met hyl ene bl ue by sodium sulfide is de-
scribed. The met hod is very si mpl e and rapi d and
works satisfactorily when color comparisons ar e made
visually, i n t he range of 0.1 to 1.0 pg of selenium. Cop-
per is t he only ion tested t hat i nt erf ered seriously
when present i n excess of 10 pg: however, higher
concentration of copper can be cleanly separated
using an exchange procedure.
THE MOST COMMONLY USED METHOD for the determination of
selenium is based on the measurement of piazselenol spectro-
photometrically or fluorimetrically when Se(1V) reacts with
aromatic 0-diamines such as 3,3 -diaminobenzidine or 2,3-
diaminonaphthalene. Although these methods offer excellent
sensitivity and specificity, the procedures are lengthy and often
require close control of pH. Furthermore, the reagent solu-
tions used are generally unstable and must be prepared daily.
Based on the consideration of simplicity, sensitivity, and
specificity, catalytic methods offer an excellent alternative
choice for the determination of trace concentration of many
elements ( I , 2) . Kawashima and Tanaka (3) have recently
proposed a very sensitive procedure for selenium based on the
catalytic reduction of 1,4,6,1l-tetraazanaphthacene; but the
method is subject to interference from several ions including
tellurium. Feigl and West (4) developed a highly sensitive
spot test for selenium based on the catalytic effect of elemental
selenium on the reduction of methylene blue by sodium sulfide.
Goto, Hirayama, and Ikeda (5) applied the findings of Feigl
and West to develop a quantitative procedure for the determi-
nation of selenium. Their method, however, is neither sensi-
tive, because at least 2 pg of selenium should be present to
make satisfactory measurements, nor selective, because almost
all cations that react with sulfide interfere.
The present paper describes an investigation on the catalytic
determination of selenium. The mechanism of the reaction
has been discussed in detail by Feigl and West (4). Any
deleterious effect caused by polysulfide is readily overcome by
the addition of sodium sulfite to form thiosulfates which, like
sodium sulfite, have no interfering actions on the reduction of
methylene blue. I n the method developed it was noticed that
the addition of formaldehyde suppresses the reducing power of
sodium sulfide and, therefore, stabilizes the blank consider-
ably. Using EDTA as a general masking agent, the deter-
mination is made quite specific for selenium and by taking ad-
vantage of the inducing effect of Fe(III), as little as 0.1 pg of
selenium can be measured.
No instruments are required and the method is rapid,
accurate, and simple.
(1) K. B. Yatsimirskii, Kinetic Methods of Analysis, Pergamon
(2) P. W. West, ANAL. CHEM., 23, 176 (1951).
(3) T. Kawashima and M. Tanaka, Anal. Chim. Acra, 40, 137
(4) F. Feigl and P. W. West, ANAL. CHEM., 19,351 (1947).
(5) H. Goto, T. Hirayama, and S. Ikeda, J. Chem. SOC. Japan,
Press, Oxford, 1966.
(1968).
Pure Chem. Sect., 13,652, (1952).
0.25 -
0.20
-
-
- 0.15
-
.-
E
v
-
I- O I O -
0 0.2 0.4 0. 6 0.8 1.0
pg o f Se l e n i u m
Figure 1. Calibration curve for selenium
EXPERIMENTAL
Materials. Stoppered test tubes of 30-ml capacity, a stop
watch that registers minutes and seconds, and a suitable pH
meter are necessary.
Stock selenium solution, containing 0.5 mg of selenium
per milliliter, is prepared by dissolving 50 mg of pure selenium
metal in a few drops (minimum required) of concentrated
nitric acid, boiling gently to expel brown fumes, and making
up to 100 ml with distilled water. Standard working solu-
tions are prepared by appropriate dilution of the stock solu-
tion.
Alkaline sodium sulfide solution (ca. 0.1M) is prepared by
dissolving 2.40 grams of sodium sulfide, an equal weight of
sodium sulfite, and 4 grams of sodium hydroxide in 100 ml
of distilled water. This solution is stable for 2 days.
Conditioner solution consists of 25 grams of EDTA (di-
sodium salt), 0.4 of a gram of ferric chloride, and 50 ml of tri-
ethanolamine dissolved in distilled water and diluted to 1 liter.
Methylene blue solution is 0.05% in distilled water.
Formaldehyde solution is reagent grade (assay 36-382).
Recommended Procedure. Transfer 10 ml of sample or an
aliquot containing 0.1 to 1.0 pg of selenium into a test tube
and dilute to 10 ml with distilled water. Add 5 ml of condi-
tioner solution, 5 ml of formaldehyde solution, and 1 ml of
alkaline sulfide solution. Mix the contents of the tube well
after each addition. Add 2 drops of methylene blue solution
and shake briefly to ensure complete mixing and start the stop
watch to measure the time required for complete decoloriza-
tion of the methylene blue. The amount of selenium present
is obtained from a calibration graph constructed by plotting
T- (min-1) us. micrograms for amounts varying from 0.1 to
1.0 pg of selenium and treated as above (Figure 1).
966 ANALYTICAL CHEMISTRY
0.20
0.15
0.1 t
0.4 0.8 1.2 1.6 2. 0
mi I N NoOH
0.15 1
0 0.1 0.2 0.3 0.4
M g o f i r o n
Figure 2. Effect of pH on the reac- Figure 3. Effect of iron con-
0. 25
0. 20
i- 0.1 5
C
.-
E
-
-
i- 0.10
tion rate ceitration
0.0 f
RESULTS AND DISCUSSION
Effect of pH. The optimum pH for the reduction to take
place was established by adding different aliquots of 1N
sodium hydroxide to samples containing 1 pg of selenium
which were then appropriately diluted with distilled water be-
fore the addition of methylene blue for the recommended pro-
cedure. The final volume remained constant in each in-
stance. The pH of each of the solutions was measured after
the reaction was complete. Figure 2 shows the effect of pH
on the reaction rate from which it is evident that the rate re-
mains constant between pH 10.6 and 11.1. Subsequent ex-
periments were conducted at a pH of approximately 10.8 ob-
tained when 1 ml of 4 sodium hydroxide solution was added
to the solution. Under these conditions the blank was stable
for at least 25 minutes which is more than sufficient for the
measurement of as little as 0.05 pg of selenium.
Effect of Iron Concentration. An unexpected enhancement
in the rate of the selenium-catalyzed reaction of methylene
blue with sodium sulfide was noticed when iron was added
to the solution. It is reported that iron produces a black
precipitate in the presence of EDTA and excess sulfide (6) and
a cherry-red soluble complex, as was observed in the present
investigation, in the presence of small amounts of sulfide (7).
The color system was unstable and disappeared on standing.
However, the reappearance of the red color on shaking sug-
gested that oxygen is involved in the formation of the red com-
plex. I n the method described it is probable that upon addi-
tion of iron to the solution containing EDTA and sulfide, the
faintly colored cherry-red complex is formed with the resul-
tant removal of dissolved oxygen. Such action would create
an ideal condition to initiate the reduction of methylene blue
by seleno sulfide.
I n order to establish the optimum concentration of iron es-
sential to accelerate the rate of reduction of methylene blue,
experiments were performed as in the recommended procedure
but with varying concentrations of iron. The rate of reaction
increased with increasing amounts of iron up to 250 pg (Figure
3) and then remained constant through increased amounts of
iron up to 500 pg. It was decided to carry out the reaction in
the presence of about 300 pg of iron. It may be mentioned
here that the formation of the red complex of iron-EDTA-
sulfide was unaffected even in the presence of triethanolamine
which forms a colorless complex with iron(II1) in alkaline solu-
tions. The addition of triethanolamine was found to be bene-
ficial in stabilizing the blank, particularly in the presence of
(6) R. Pribil, Collection Czech. Chem. Comm., 16/17, 542 (1951).
(7) T. S. West, Analyst, 17,630 (1962).
J
20 2 5 30 35
Te mp e r a t u r e ' C
Figure 4. Effect of temperature
A: 1 pg Se; B: 0.5 pg Se; and C: 0.25 pg Se
large concentrations of iron, perhaps by maintaining the
higher oxidation state of iron.
Because the re-
duction of methylene blue is caused by the sulfide ion, signif-
icant change in the rate of the reaction, as anticipated, was
noticed as the concentration of sodium sulfide was varied.
Although the reaction proceeds faster at higher concentration
of sulfide solution, the blanks also became increasingly
unstable and the analytical value of the catalytic effect of
selenium was found to be of little significance. Based on
the consideration of the stability of the blank as well as the
speed of the reduction mechanism, 1 ml of a 0.1M solution was
found to be ideal for control of the reaction. Because it is
difficult to prepare accurately 0.1M solutions of sodium sulfide
without involving considerable expenditures of time, the cali-
bration curve should be checked each time before subjecting
the samples to analysis of the selenium content. However, no
significant deviation in the day to day reproducibility was
noticed when solutions containing constant weight of sodium
sulfide were used for analysis.
I n the very early stage of the investigation it was realized
that the reduction of methylene blue by sodium sulfide should
bearrested before the analytical value of the catalytic effect of
selenium could be examined satisfactorily. Attempts were
then made to achieve stabilization of the blank, without
jeopardizing the catalytic effect of selenium, by adding various
water-miscible organic solvents to the colored system. The
solvents examined included acetone, ethanol, methanol, 1,4-
dioxane, diethylene glycol, 2-propanol, methyl ethyl ketone,
and formaldehyde. Only formaldehyde was found to be
promising as the other solvents either completely prevented the
catalytic effect of selenium or were found to be ineffective.
The optimum concentration of formaldehyde required was
next established by treating 1 pg of selenium with 2,4,5,6, and
8 ml of formaldehyde solution as in the recommended proce-
dure. Solutions were appropriately diluted with water so
that the final volume remained the same in each instance.
Although higher concentrations of formaldehyde effectively
stabilized the blank, it was found that the rate of reduction of
methylene blue in the presence of selenium was also signifi-
cantly reduced. Lower concentration of formaldehyde, on
the other hand, decreases the stability of the blank and there-
Effect of Other Reagent Concentrations.
VOL. 40, NO. 6, MA Y 1968 * 967
Group
I
I1
111
IV
V
VI
VI1
VI11
Table I. Interference Studies
Li+, Na+, K+, Cu2+, Ag+.
Bet+, Mgz+, Ca2+, Sr2+, Ba2+, Zn2+, Cd2+, Hg2+.
B ~O~- , Al3+, Ce4+.
HCQ-, co32-, Zr4+, Sn2+, Pb2+.
NH~+, NO^-, Pods+, V03- , HAs012-, Sbb+, Bia+.
SO,>-, S042-, Cr3+, Te032-, M0042-, wo4'.
F-, C1-, Br-, I-, Mn2+.
Fe3+, Co2+, Ni2+.
Table 11. Precision of the Proposed Method
No. of Selenium, p g
detns. Taken Found Std dev
8 1 .o 1.09 0.25
8 0. 5 0.49 0.02
8 0 . 2 0.20 0.01
fore will affect the result when traces of selenium are involved.
Taking into account the stability of the blank as well as speed
of the reduction of methylene blue in the presence of selenium,
5 ml of formaldehyde was found to be satisfactory.
It was also interesting to note the apparent lack of effect on
the reaction when methylene blue concentrations were varied
and the other parameters were held constant. Using the
recommended procedure with 0.5 pg of selenium, the respec-
tive reaction times for the blank as well as the test reaction re-
mained constant over the range of 0.2 to 1.0 ml of 0.05% of
added methylene blue. Obviously, in the presence of excess
sulfide, the rate of reduction of the methylene blue is a direct
function of methylene blue concentration so that the time re-
quired for conversion to the leuco form remains constant.
Two or 3 drops of 0.05 % solution added by means of a drop-
per gave sufficient color for measurement of reaction time.
Effect of Temperature. Figure 4 illustrates the effect of
temperature on the catalytic reduction of methylene blue.
Although the rate of reduction increases with rise in tempera-
ture, a considerable decrease in the stability of the blank was
also observed. It was, therefore, decided to perform the
experiment at room temperature.
Because of the reactivity of sulfide
ion toward many cations, it was decided to investigate the
catalytic effect of selenium in a medium containing EDTA.
Table I lists the ions that were examined as potential inter-
ferents by the recommended procedure. The concentration
of selenium in these experiments was 1 pg and that of interfer-
ent 50 pg. With the exception of copper, silver, bismuth, and
antimony, none of the ions listed interfered in the determina-
tion. Antimony was found to inhibit the reaction slightly as
Effect of Diverse Ions.
in its presence the recovery was reduced by 25 %. However,
when the concentration of antimony was reduced to 25 pg, no
interference was noticed. Copper, silver, and bismuth, on the
other hand, interfered by giving higher recovery of selenium.
When the concentrations of silver and bismuth were reduced
to 25 pg each, and that of copper to 10 pg, no interferences
were encountered. Higher concentration of copper was
readily separated, however, from traces of selenium by passing
the solution through a IO-cmlong column prepared by filling a
25-ml buret with Dowex 50W-X2, 50 to 100 mesh, cation ex-
change resin at a flow rate of ml per minute. The eluate
and washings should be evaporated, if necessary, so that the
sample volume does not exceed 10 ml before subjecting to
analysis.
In order to study this effect a
series of solutions containing 0.5 pg of selenium were treated
with all the reagents except methylene blue as described in the
recommended procedure. The solutions were allowed to
stand different periods of time before the addition of methyl-
ene blue. Blanks were also run simultaneously. No change
in the rate of reaction was noticed for solutions that stood up
to 8 minutes after which the reduction of methylene blue be-
came rapid both in blanks and samples. It was decided to
add methylene blue as soon as other reagents have been added.
The recovery of known amounts of selenium
are shown in Table 11. The determinations were made over a
period of several days. The results indicate that the re-
covery is satisfactory in spite of the fact that the color com-
parisons were made visually.
Conclusions. The proposed procedure is almost com-
pletely free from interference from several anions and cations
and should find wide application where traces of selenium
are to be determined. Its outstanding features are that it is
simple, highly sensitive, very rapid, and no special skill is
necessary to operate the method as color comparisons can be
made visually as precisely as in indicator titrations. A
spectrophotometer was not used because there was some
difficulty in obtaining an accurate absorbance measurement
because of nonuniformity in the decolorization of the colored
system particularly when selenium concentrations are low.
I n these circumstances the accuracy of visual comparison was
unaffected because the solution can be tilted gently to com-
plete the reduction. Another practical advantage is that in a
matter of minutes several samples containing as low as 0.01
ppm of selenium can be determined simultaneously and
should, therefore, be very convenient for routine analyses.
RECEIVED for review December 27, 1967. Accepted February
28, 1968. This investigation was supported in part by Public
Health Service Research Grants No. AP 00117 and AP
00128 from the National Center for Air Pollution Control,
Bureau of State Services.
Effect of Standing Time.
Precision.
968 ANALYTICAL CHEMISTRY