Journal of Chromatographic Science, Vol.

32, November 1994

Separation of Metal Dithiocarbamate Complexes by

High-Performance Liquid Chromatography

S.F. Wang and C M . Wai*

Chemistry Department, University of Idaho, Moscow, Idaho 83843

Abstract
Analysis of metal-diethyldithiocarbamate (DDC) complexes by
high-performance liquid chromatography (HPLC) is affected by the
stability of the complexes and their interactions with the metal
components of the instrument. Using a reversed-phase stainless
steel column, good separation of Co(III)—, Cr(lll)-, Cu(ll)- Hg(ll)-,
Ni(ll)-, Pd(II)-, Pt(II)-, Se(IV)-, and Te(IV)-DDC complexes can be
achieved using ACN-H O (70:30) as a mobile phase. Ligand
2
exchange with Ni from the stainless steel column prohibits
quantitative analysis of some metal-DDC complexes with lower
stability constants relative to that of Ni. The exchange reaction can
be eliminated with a metal-free PEEK column. Separation of Bi(lll)-,
Cd(II)-, Fe(ll)-, In(III)-, Pb(II)-, Tl(l)-, and Zn(ll)-DDC complexes,
which is not feasible using a stainless steel column, can be achieved
using a PEEK column and MeOH-H O-CHCI (75:20:5) as the
2 3
mobile phase. Pyrrolidinedithiocarbamate (PDC) offers no
advantage over DDC in HPLC analysis of metal complexes.
Introduction
ments by this method (4,5). For example, Shih and Carr (6)
2+ 2 + 2 +
pointed out that the limits of detection for P b - Z n - C d -
3+
and B i - D D C complexes by reversed-phase HPLC were poor
when a stainless steel column was used because of the ligand
exchange reaction between these metal ions and nickel from
the steel surface (6). From a thermodynamic point of view, the
exchange reaction depends on the stability of a metal-DDC
complex relative to that of Ni(DDC) . The relationship between
2
the stability constants of different metal-DDC complexes and
their HPLC separation using a stainless steel column has not
been quantitatively established. The kinetics of the exchange
reaction may also play a role in determining the HPLC be­
havior of metal-DDC complexes. The limits of HPLC analysis
of DDC complexes and the interactions of the metal chelates
with the chromatographic components are not well under­
stood. This paper presents the results of our HPLC studies of 23
metal- and nonmetal-DDC complexes using a reversed-phase
stainless steel column and a nonmetal column. The factors
controlling the chromatographic behavior of these DDC com­
plexes are investigated and summarized.

Solvent extraction with a chelating agent followed by chro­

Experimental
matographic separation is one method of analyzing trace
metals in aqueous solutions. The extraction allows precon-
Reagents and solvents
centration of metal ions, elimination of matrix interferences,
All solvents used in this study were of HPLC grade purchased
and, in some cases, identification of metal species. The limita­
from EM Industries (Gibbstown, NJ). The solvents were de­
tion of this method for trace metal analysis depends on the
gassed with a sonifier (Bransonic Model 12, Branson Cleaning
chelation ability of the ligand, the stability of the metal
Equipment; Shelton, CT) in a water bath before HPLC use. All
chelates, and their interactions with chromatographic com­
reagents used were of analytical grade. Sodium-DDC and am­
ponents. Diethyldithiocarbamate (DDC) is widely used for pre-
monium pyrrolidinedithiocarbamate (APDC) were obtained from
concentration of trace elements (1). DDC reacts with a large
J.T. Baker (Phillipsburg, NJ) and Sigma Chemical (St. Louis,
number of metals and nonmetals to form complexes that are
MO), respectively. Deionized water was prepared by passing dis­
insoluble in water but easily extractable by organic solvents.
tilled water through an ultrapure water purification cartridge.
There are a number of papers in the literature that deal with
Different metal salts and standard metal ion solutions were ob­
high-performance liquid chromatographic (HPLC) analysis of
tained from J.T. Baker and EM Industries, respectively.
metal-DDC complexes (2-5). It has been reported that ligand
exchange reactions with the metal components of the column Preparation of dithiocarbamate complexes
may create severe interferences for the analysis of certain ele- Each metal-DDC complex was prepared by adding an excess
* Author to whom correspondence should be addressed. amount of DDC to a solution containing a known amount of a

506 Reproduction (photocopying) of editorial content of this journal is prohibited without publisher's permission.
Journal of Chromatographic Science, Vol. 32, November 1994

metal ion of interest. An acetate buffer was used to control the parameters were usually set as follows: mobile phase flow rate,
pH of the solution (usually between 3.5 and 5.5), according to 1.6 mL/min; detector sensitivity, 0.016 AUFS; and recorder
the extraction curve of the metal-DDC complex reported in the chart speed, 0.5 cm/min, unless otherwise stated.
literature (1). The resulting DDC complex was extracted into
chloroform in a separatory funnel. After phase separation, the
organic phase was washed with deionized water several times. Results and Discussion
Ethanol was then added to the organic phase, and the mixture
was heated with a water bath at 60°C to evaporate the chloro­
Table I summarizes the results of the 23 DDC complexes in­
form. The precipitate formed in ethanol was filtered and re-
vestigated by this study. These results were obtained using the
crystallized to obtain the pure metal-DDC complex.
stainless steel column and a 70:30 ACN-H O mixture as the 2

mobile phase. The results were unchanged when acetonitrile

Instrumentation
was replaced by methanol in the mobile phase. According to
The HPLC used in this work consisted of a Kratos Spec-
Table I, the DDC complexes can be classified into three groups.
troflow 400 solvent delivery system (Ramsey, NJ), a UV detector
The first group of DDC complexes, including As(III), Bi(III),
(Anspec AN-203) with a fixed wavelength of 254 nm, a Perkin-
Cd(II), Ga(III), In(III), Fe(II), Pb(II), Mn(II), Mo(VI), Sb(III),
Elmer recorder (Norwalk, CT) adjusted for 10-mv input, and a
T1(I), and Zn(II), was found to exhibit similar multiple peaks
50-µL loop for sample injection. At 254 nm, all the DDC com­
identical to those observed from the injection of NaDDC, as il­
plexes studied showed strong absorption. For example, the
lustrated in Figure 1. The observed peaks (b and c) in Figure 1
molar absorptivities of As(DDC) , Cu(DDC) , Ga(DDC) , 3 2 3
were identified by chromatography and atomic absorption
Ni(DDC) , Sb(DDC) , and Zn(DDC) in methanol at room tem­
2 3 2
spectroscopy to be Ni(DDC) and Fe(DDC) , respectively. Peak 2 2
perature were determined by a UV-vis spectrophotometer to be
4 4 4 4
(a) corresponds to the NaDDC ligand and its oxidation product,
4 4
1.8 ×10 ,1.6 ×10 ,3.4 ×10 ,2.2 ×10 ,7.1 ×10 , and 3.2 ×10
tetraethylthiuram disulfide (disulfiram). This oxidation product
L/mol/cm, respectively, at this wavelength. An Alltech Econosil
was previously reported by Hutchins and co-workers (3) in a
C 10-µm stainless steel column (Deerfield, IL) with dimen­
18
study of HPLC separation of metal-DDC complexes. According
sions of 250 χ 4.6-mm i.d. was used as one of the analytical
to our experiments, the retention time of peak (a) matches that
columns. The guard column was packed with silica-based C 1 8
of the pure disulfiram compound obtained from Aldrich Chem­
material in 5-µm adsorbosphere (Alltech). An Alltech Econosil
ical (Milwaukee, WI). In addition, two minor peaks that eluted
C 10-pm metal-free MF-PLUS column of the same dimensions
18
before the ligand peak were frequently observed. They corre­
as the stainless steel column but made of polyetheretherke-
spond to the decomposition products of DDC: (CH CH ) NH 3 2 2
tone (PEEK) was also used as the analytical column. In this
and CS . When relatively concentrated samples of this group of
2
case, metal-free (PTFE) column frits and a PEEK guard column
DDC complexes (e.g., Bi[DDC] and Pb[DDC] ) were injected 3 2
from Alltech were used. In addition, PEEK connecting tubing
into the column, broad and poorly defined peaks were ob­
and ferrules were used between the injector and the column and
served. In essence, no characteristic peak suitable for HPLC
between the column and the detector. The mobile phases used
analysis of this group of DDC complexes was found under our
were usually acetonitrile-water (ACN-H 0) (70:30) and 2
experimental conditions.
methanol-water (MeOH-H O) (75:25). The instrument
2
The second group of DDC complexes, which
included Co(III), Cr(III), Cu(II), Hg(II), Pd(II),
Table 1. Chromatographic Behavior of M - D D C in Reversed-Phase HPLC Pt(II), Se(IV), and Te(IV), showed characteristic
Using a Stainless Steel Column* peaks that were well-defined and separable from
each other, as shown in Figure 2. The Ni(DDC) 2
HPLC HPLC
Element (l/n)Log β elution* Element (1/n)Logp elution and Fe(DDC) peaks were not observed in the
† 2

chromatograms obtained from this group of DDC

Pd(ll) 32.5 S MoO (VI) 9.30 Μ 2
complexes. Ni(II) also belongs to this group.
Pt(II) S Cd(ll) 9.05 Μ When Ni(DDC) was injected into the column,
Au(lll)
-
27.2 L Sb(lll) 8.77 Μ only its own peak was observed. The peak height
2

Hg(ll) 20.92 S As(lll) 8.06 Μ of this group of DDC complexes was a linear func­
Ag(l) 18.72 L Cr(lll) 7.88 S tion of concentration, as expected from stable
Te(IV) 15.1 S Zn(ll) 7.42 Μ compounds. The third group includes Au(DDC) 3

Bi(lll) 12.5 Μ Co(lll) 7.4 S

and Ag(DDC), which have extremely small solu­
Cu(ll) 12.07 S Ga(lll) 7.37 Μ
bilities in acetonitrile or methanol such that their
Ni(ll) 12.0 S Fe(ll) 7.22 Μ
detection was not possible under our experi­
Se(IV) 10.7 S Tl(l) 6.55 Μ
In(lll) 9.85 Μ Mn(ll) 5.37 Μ
mental conditions.
Pb(ll) 9.70 Μ
The DDC complexes listed in Table I are in
order of decreasing stability constant (β), which is
* Mobile phase: ACN-H O (70:30).
†
2 defined as follows:
Abbreviations: S, sharp distinct peak suitable for HPLC analysis; M, multiple peaks or a broad, ill-defined
peak unsuitable for HPLC analysis; L, solubility too low in MeOH or ACN to obtain a detectable peak.
Eq1

507
 Journal of Chromatographic Science, Vol. 32, November 1994

Eq 2 ence of an Ni(DDC) peak in their chromatograms was an in­

2

dication of the ligand exchange reaction shown in Equation 3.

The β values given in Table I are actually expressed in terms The DDC complexes of Co(III), Cr(III), and Se(IV) were excep­
of (1/n)log β where η is the charge carried by the cation. The tions. These DDC complexes have lower stability constants
β value of each DDC complex represents the average of all relative to Ni, but they did not appear to exchange with Ni. The
data found in the literature (1). No β value for Pt(DDC) was 2
results suggested that kinetic factors probably also play a role
found in the literature, but we estimated that it should be in the ligand exchange reaction with Ni. The kinetic inertness
close to that of Pd(DDC) . The ligand exchange reaction be­
2
of the Co(III)- and Se(IV)-DDC complexes in exchange with
2 +
tween N i and a metal ion (M) with a charge η can be de­ other metal ions in solvent extraction experiments has been re­
scribed by the following equation: ported in the literature (7). Kinetically inert Co(III) and Cr(III)
complexes with phenanthroline are also known to exhibit
Eq 3 better chromatographic behavior (2,8).
When a metal-free PEEK column was used as the analytical
The equilibrium constant {K) of the exchange reaction is column, exchange reactions with Ni and Fe were eliminated.
given by the following equations: This was evident from the absence of the Ni(DDC) and 2

Fe(DDC) peaks in the chromatograms obtained from NaDDC or

2

Eq4 from a metal-DDC complex with a lower stability constant than

Ni. However, the peak shapes of metal-DDC complexes using the
Eq.5 PEEK column are generally broader than those observed from
the stainless steel column. This is probably because of the low
Thus, the ligand exchange reaction is thermodynamically de­ surface area of the frit and the inner surface irregularities of the
termined by the difference between (l/n)log β values of the two PEEK column. The difference in packing procedure may also
metal ions involved. From the results given in Table I, it is clear contribute to the difference in peak shape, although the supplier
that metal ions with (l/n)log β values higher than that of Ni (Alltech) claimed that the same packing procedure was used for
showed characteristic peaks suitable for HPLC analysis. both the stainless steel and the PEEK columns. According to our
Bi(DDC) was the only exception; it had a (l/n)log β value
3
experiments, the peak shape also depends strongly on the mobile
slightly higher than that of Ni(DDC) but failed to show a dis­
2
phase composition. If a methanol-water mixture was used as the
tinct peak of its own. Most of the DDC complexes with lower mobile phase, broad peaks were observed, especially for the
(l/n)log β values relative to that of Ni usually either exhibited complexes with low stability constants such as As(DDC) , 3

broad peaks or multiple peaks, as shown in Figure 1. The pres- Cd(DDC) 2, and Fe(DDC) . Using 2acetonitrile and water as a mo­
bile phase did not improve the situation appre­
ciably. However, addition of chloroform to the mo­
bile phase (methanol-water) was found to improve
the peak shape and separation of the metal-DDC
complexes. Figure 3 shows the separation of
Cd(DDC) , Pb(DDC) , Zn(DDC) , Fe(DDC) ,
2 2 2 2

In(DDC) , Tl(DDC), and Bi(DDC) using the PEEK

3 3

column with a mobile phase composition of

methanol-water-chloroform (75:20:5). These re­
sults indicate that chloroform is a good stabilizer of
the mobile phase for the separation of metal-DDC
complexes by HPLC. The detection limits (based on
a signal-to-noise ratio of 3) of Cd(II), Pb(II), Zn(II),
Fe(II), In(III), T1(I), and Bi(III), according to the
conditions specified for the PEEK column, are 11,
210,25,20,18,19, and 25 ng, respectively, using a
50-µL injection loop. Our experiments also showed
that the amount of chloroform in the mobile phase
cannot be much greater than 5% by volume. When
a mobile phase ratio of 70:20:10
MeOH-H O-CHCl was used, gas bubbles were
2 3

16 found in the elution process. This phenomenon

was not observed using a mobile phase ratio of
70:20:5 MeOH-H O-CHCl . It should be noted that
2 3

Figure 1. Chromatograms of NaDDC and some less stable metal-DDC complexes obtained some metal-DDC complexes, including those of
from the stainless steel and PEEK columns. Peaks: a, NaDDC ligand and its oxidation product,
As(III), Ga(III), Mo(VI), and Sb(III), still failed to
tetraethylthiuram disulfide (disulfiram); b, Ni(DDC) ; and c, Fe (DDC) . Conditions: flow rate,
2 2
show sharp characteristic peaks using the PEEK
1.6 mL/min; mobile phase: ACN-H O (70:30).
2

column and MeOH-H O-CHCl as a mobile phase.

2 3

508
Journal of Chromatographic Science, Vol. 32, November 1994
Although the exchange reactions can be eliminated using a
metal free column, interactions of metal-DDC complexes with
the stationary phase and instability of the complexes in mobile
phase also appear to limit the application of this method for
some trace elements, including As(III), Ga(III), Mo(VI), and
Sb(III). Spectroscopic measurements indicated that As(DDC) in
methanol tends to decompose at room temperature. The de­
composition products (CS and [CH CH ] NH with peak ab­
2 3 2 2
sorption at 200 nm) were found to increase significantly within
several hours of storage in methanol. On the other hand, stable
complexes, such as Cu(DDC) in methanol, did not show an ob­
2 phase composition of methanol-water-chloroform (75:20:5).
servable change in UV spectrum during the same period of time.
HPLC separation of metal ions complexed with pyrroli-
dine-thiocarbamate (PDC) was also studied. The stability con­
stants of metal-PDC complexes are slightly lower than those
of the corresponding DDC complexes, but follow the same
order (9). PDC is widely used in solvent extraction of trace
metals because of its stability relative to DDC in acidic media.
According to our experiments, the PDC complexes of the fol­
lowing seven elements showed characteristic peaks using the
stainless steel column: Co(III), Cr(III), Cu(II), Hg(II), Ni(II),
Pt(II), and Se(IV). The PDC complexes of Pd(II) and Te(IV)
showed extremely small solubilities in MeOH or ACN and
hence were not detectable by HPLC under our experimental
conditions. In comparison, the DDC complexes of the nine el-
Figure 2. Separation of metal-DDC complexes with a stainless steel
column with a flow rate of 1.6 mL/min and a mobile phase of ACN-H O 2
(70:30). Peaks: a, NaDDC; b, Pt(DDC) ; c, Ni(DDC) ; d, Pd(DDC) ; e,
2 2
Co(DDC) ;f, Cr(DDC) ; g, Se(DDC) ; h, Cu(DDC) ; i, Hg(DDC) .
3 3 4 2 2
ements described above were all detectable using the stainless
steel column, as shown in Figure 2. The As(III), Bi(III), Cd(II),
Fe(II), In(III), Pb(II), Sb(III), and Zn(II) complexes of PDC
that were eluted from the stainless steel column showed mul­
tiple peaks that correspond to the retention times of N i - and
3 Fe-PDC complexes and are similar to those described previ­
ously for the metal-DDC complexes. The ligand exchange
mechanism is apparently responsible for this observation.
Using the PEEK column, Bi(III)-, In(III)-, and Pb(II)-PDC
complexes show distinctive and sharp peaks using a mobile
The PDC complexes of As(III), Cd(II), Fe(II), Sb(III), and
Zn(II), which were eluted from the PEEK column, exhibited
broad peaks that were not suitable for HPLC analysis. These
metal-PDC complexes probably have strong interactions with
the stationary phase, which leads to the broadening of peaks
during elution from the column. The nature of the interac­
tions is not well understood. However, the interactions of
metal chelates with residual silanol groups of stationary
phases have been discussed by Robards and co-workers (2).
The PDC complexes are probably more reactive than the cor­
responding DDC complexes in such interactions. At least that
seems to be the case for Cd(II), Fe(II), and Zn(II). Their PDC
complexes show broad peaks, but their DDC complexes give
distinctive peaks when eluted from the PEEK column, as i l ­
lustrated in Figure 3. Based on the results obtained from
this study, we tend to conclude that PDC offers no advantage
over DDC in HPLC analysis of their metal complexes.
Conclusion
A number of factors can affect HPLC analysis of metal-DDC
complexes, including stability constants, solubility and sta-
Figure 3. Separation of less stable metal-DDC complexes using a PEEK
column with a mobile phase of MeOH-H O-CHCI (75:20:5) and a flow
2 3
2
rate of 0.7 mL/min. Peaks: a, NaDDC; b, Cd(DDC) ; c, Pb(DDC) ; d,
2 2
Zn(DDC) ; e Fe(DDC) ; f, ln(DDC) ; g, TI(DDC); h, Bi(DDC) .
2 ; 2 3 3
509

bility in the mobile phase, and interactions with the HPLC
components. By using a reversed-phase stainless steel column,
a good separation of Co(III)-, Cr(III)-, Cu(II)-, Hg(II)-, Ni(II)-
Pd(II)-, Pt(II)-, Se(IVh and Te(IV)-DDC complexes, can be
achieved with ACN-H O (70:30) as the mobile phase. Ligand
2 2. Κ. Robards, P. Starr, and E. Patsalides. Metal determination and
exchange with Ni from the stainless steel column prohibits
quantitative analysis of many metal-DDC complexes including
As(III), Bi(III), Cd(II), Ga(III), In(III), Fe(II), Pb(II), Mn(II),
Mo(VI), Sb(III), T1(I), and Zn(II). The exchange reaction can be
eliminated with a metal free column such as the PEEK column
described in this study. Separation of Bi(III)-, Cd(II)-, In(III)-,
Fe(II)-, Pb(II)-, TI(I)-, and Zn(II)-DDC complexes can be
achieved using the PEEK column and a mobile phase compo­
sition of M e O H - H O - C H C l (75:20:5). A few metal-DDC
2 3
chelates, including As(III), Ga(HI), Mo(VI), and Sb(III), still
show broad peaks not suitable for quantitative analysis using
the PEEK column. Our experiments also indicate that PDC
does not show any advantages over DDC in HPLC analysis of
metal complexes.
Acknowledgment
This work was supported by the University of Idaho Center
for Hazardous Waste Remediation Research.
510
Journal of Chromatographic Science, Vol. 32, November 1994
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Manuscript received May 27,1993;
revision received January 4,1994.