Talanta 48 (1999) 1 – 7

Studies on 2-(2-thiazolylazo)-5-diethylaminophenol as a
precolumn derivatizing reagent in the separation of platinum
group metals by high performance liquid chromatography
Hong Wang, Hua-Shan Zhang *, Jie-Ke Cheng
Department of Chemistry, Wuhan Uni6ersity, Wuhan 430072, People’s Republic of China

Received 23 June 1997; received in revised form 29 October 1997; accepted 30 October 1997

Abstract

Five platinum group metals, Pt(II), Ir(IV), Ru(III), Rh(III) and Os(IV) have been separated by high performance
liquid chromatography (HPLC) using 2-(2-thiazolylazo)-5-diethylaminophenol (TADAP) as a precolumn derivatizing
reagent. The whole analysis was completed on a C18 column in 23 min at 574 nm, with the mobile phase of
methanol–water (69.5:30.5, v:v) containing 4 mmol l − 1 tetrabutylammonium bromide (TBA Br) and 10 mmol l − 1
pH6.0 acetate buffer. The detection limits (S/N= 3) of Pt(II), Ir(IV), Ru(III), Rh(III) and Os(IV) were 0.39, 9.74,
1.64, 0.29 and 1.29 ng ml − 1, respectively. This method was rapid, sensitive and simple. © 1999 Elsevier Science B.V.
All rights reserved.

Keywords: Reversed-phase high performance liquid chromatography (RP-HPLC); Platinum group metal; 2-(2-thia-
zolylazo)-5-diethylaminophenol (TADAP)

1. Introduction [6–8]. There are S, N chelating atoms in the

The reversed-phase HPLC (RP-HPLC) spec-
trophotometric technique has been proved to be
an effective tool for the separation and determina-
tion of trace amount platinum group metal ions.
The method usually involves complex formation
of noble metals with suitable derivatizing
reagents. Many kinds of developing reagents have
been used as complexing agents, such as bipyridyl
[1], diethyl dithiocarbamate [2,3], acetylacetone
[4], 8-hydroquinolines [5] and heterocyclic azos
* Corresponding author. e-mail: lotus@wip.whcnc.ac.cn
molecules of thiazolylazo reagents, which can re-
act with platinum metals to form colored thermo-
dynamically stable complexes according to the
Lewis acid–base concept. Therefore, their appli-
cations in HPLC are focused mainly on the sepa-
ration of platinum metals. Os, Rh and Ru were
separated as 4-(2-thiazolylazo) resorcinol (TAR)
complexes [9,10]. 1-(2-Thiazolylazo)-2-naphenol
(TAN) was used in the HPLC determination of
Pd and Rh [11]. The separation of Ru, Rh and Pd
was obtained by 2-(2-thiazolylazo)-5-diethy-
laminophenol (TADAP) [12]. The 4-(2-benzothia-
zolylazo) resocinol (BTAR) chelates of Ru, Rh

0039-9140/99/$ - see front matter © 1999 Elsevier Science B.V. All rights reserved.
PII S0039-9140(97)00360-3
2 H. Wang et al. / Talanta 48 (1999) 1–7

Table 1
Spectrophotometric properties of TADAP (R) metal chelates

Chelate Pt–R Ir–R Ru–R Rh–R Os–R Pd–R

omax (nm) 574 573 595 581 590 574

omax×10−4 (l mol−1 cm−1) 3.15 1.73 4.14 5.36 1.67 3.26
o574×10−4 (l mol−1 cm−1) 3.15 1.73 3.16 5.15 1.37 3.26

and Os were separated on a C18 column, [13]. RuCl3, RhCl3 and (NH4)2OsCl6, respectively.
Recently, a new reagent, 2-(6-methyl-2-benzothia- Pt(II) standard solution was prepared from
zolylazo)-5-diethylaminophenol (MBTAE) was K2PtCl6 (AR) standard solution by reduction with
synthesized and used as a derivatizing reagent in sodium sulphite.
HPLC separation [14,15]. After the separation of Buffer solutions were adjusted to the required
Pt, Rh, Pd and Os [14], five noble metals Ru, pH with 1 mol l − 1 acetic acid and 1 mol l − 1
Rh(Ir), Pd, Os and Pt were determined within sodium acetate solution using the acidimer. Tetra-
about 40 min [15], which is the most thus far. In butylammonium bromide was purchased from the
order to find a rapid, simple and conventional Medical Company of Beijing. A 0.05% solution of
method for separating noble metals as much as TADAP (synthesized in our lab) in absolute
possible, a common reagent, TADAP, was used in ethanol was used and other reagents were used as
this study and its chelates of five platinum group purchased. Doubly distilled water was used for all
metals, Pt, Ir, Ru, Rh and Os were eluted in 23 solution preparation.
min with the mobile phase of methanol–water
(69.5:30.5, v:v) containing 4 mmol l − 1 tetrabuty-
lammonium bromide (TBA Br) and 10 mmol l − 1
pH 6.0 acetate buffer at 574 nm on a C18 column.
The detection limits (S/N =3) were 0.39, 9.74,
1.64, 0.29 and 1.29 ng ml − 1, respectively. The
proposed method gave a rapid, sensitive and sim-
ple method for the simultaneous determination of
multi-platinum group metals by HPLC.

2. Experimental

2.1. Apparatus

An LC-10 HPLC apparatus (Japan Analytical)

with an ODS column (5 mm, 150× 4.6 mm i.d.;
Beijing Analytical Instrument Factory) and an
S-3702 UV-VIS detector (AIC, Japan) was used.

2.2. Reagents
Stock solutions of Ir(IV), Ru(III), Rh(III) and
Os(IV) in 1 mol l − 1 hydrochloric acid were pre-
pared from analytical-reagent grade (NH4)2IrCl6,
Fig. 1. Effect of methanol concentration on k% mobile phase:
methanol – water in different ratios, 4 mmol l − 1 TBA Br and
10 mmol l − 1 pH 6.0 acetate buffer; column, C18 (5 mm,
150 × 4.6 mm i.d.); detection wavelength, 574 nm; flow rate,
1.0 ml min − 1; injection, 20 ml; detector sensitivity, 0.02 AUFS.
 H. Wang et al. / Talanta 48 (1999) 1–7
Fig. 2. Effect of TBABr concentration on k% mobile phase:
methanol – water= 60.5:30.5 (v:v), 10 mmol l − 1 pH 6.0 acetate
buffer, TBA Br with different concentrations; other conditions
as shown in Fig. 1.
2.3. Procedure
Acetate buffer (5.0 ml of 1 mol l − 1, pH 5.0)
and TADAP solution (5 ml, 0.05%) were trans-
Fig. 3. Effect of pH on k% mobile phase: methanol:water=
60.5:30.5 (v:v), 4 mmol l − 1 TBA Br, 10 mmol l − 1 acetate
buffer with different pHs; other conditions as shown in Fig. 1.
3
Fig. 4. Separation of Pt – , Ir – , Ru – , Rh – and Os – TADAP
chelates by HPLC mobile phase: methanol:water =60.5:30.5
(v:v), 4 mmol l − 1 TBA Br, 10 mmol l − 1 pH 6.0 acetate
buffer; other conditions as shown in Fig. 1.
ferred into an aqueous solution containing 3, 30,
10, 5 and 28.5 mg of Pt, Ir, Ru, Rh and Os,
respectively. The mixture was heated in a water
bath at 80–85°C for 1 h, after which 10 ml of 95%
alcohol was added immediately. After having
cooled to room temperature, the solution was
diluted to 25.00 ml with water. A 20 ml aliquot of
this solution was injected into the chromato-
graphic column for analysis.
For HPLC analysis, the ODS column was pre-
equilibrated with the mobile phase for 30 min.
The complexes were eluted at a flow-rate of 1.0
4 H. Wang et al. / Talanta 48 (1999) 1–7

Table 2
Regression equation, calibration range and detection limit of metal ions

Metal chelate Regression equationa r Calibtation range (ng ml−1) Detection limit (ng ml−1; S/N=3)

Pt–R y= 4.198×103+1.943 0.9933 8–800 0.37

5
× 10 x
Ir–R y= −1.180×102+1.618 0.9989 80–2000 9.74
×104x
Ru–R y= 1.771×103+8.595 0.9940 12–800 1.64
× 104x
Rh–R y= −5.187×103+6.282 0.9983 4–800 0.29
5
×10 x
Os–R y= −4.510×102+9.050 0.9995 57–1140 1.29
4
×10 x

a
x, Concentration of the metal ion (mg ml−1); y, peak area of the corresponding chelate (arbitrary unit).

ml min − 1 and detected at 574 nm. The detector
sensitivity was set at 0.02 absorbance unit for
full scale deflection (AUFS). The peak areas
were measured for quantitative calculations.
3. Results and discussion
3.1. Spectrophotometric properties
In a weak acidic solution, platinum group
metal ions can form stable and colored chelates
in the present of TADAP (the absorption data
are given in Table 1). The maximum absorption
wavelengths of these chelates were in the range
573–595 nm, with 574 nm being used in this
experiment.
3.2. Chromatographic separation
In the literature cited above, methanol–water
or acetonitrile – water comprised the basic com-
position of the mobile phase. The methanol–wa-
ter system was chosen by test (the effect of the
methanol content on the capacity factors (k%) is
shown in Fig. 1). The k% value of each chelate
fell rapidly with increased methanol content and
the change patterns of the retention times of
these chelates were remarkably different. This
even caused the alteration of the elution order of
the chelates. Therefore, it was much more
difficult to select a suitable methanol content for
a good separation. When the methanol content
was below 68.0%, the TADAP peak affected that
of Os–TADAP. If the methanol content above
70.0%, the peaks of Rh–TADAP and Os–
TADAP interfered with each other. In the
methanol content range of 68.5–69.0%, Rh and
Ru overlapped. The separation could only be
obtained in a narrow range of methanol content.
The ratio of methanol to water was set at
69.5:30.5 (v:v) for further studies.
The retention times of these chelates were long
and the peak shapes poor, except for Pt–
TADAP in the mobile phase of methanol–water
(69.5:30.5, v:v) containing acetate buffer only.
With the addition of the ion-interaction reagent,
TBA Br, the elution rates of most of these
chelates were accelerated, while that of Pt–
TADAP was almost unchangeable (Fig. 2). It
could be concluded, based on the ion-interaction
model proposed by Bidlingmeyer [16,17], that the
chelates of Ir–, Ru–, Rh– and Os–TADAP
were positively-charged. The effect of TBA Br
could also be interpreted by the ion-interaction
model (4 mmol l − 1 of TBA Br was available in
this experiment).
Table 3
Tolerance limits of foreign ions (mg)

Metal ion Amount of foreign ion (mg)

Cr(III) Cr(VI) Mn(II) Pd(II) Hg(II) Pb(II) Fe(III) Ni(II) Cu(II) Zn(II) Co(II) V(V)

Pt 160 160 160 160 160 160 20 20 40 160 20 0.2

Ir 160 160 160 160 160 160 5 5 40 160 0.2 20
Ru 160 160 160 160 160 160 20 20 40 160 0.2 40
Rh 160 160 160 160 160 160 20 20 40 160 20 40
Os 160 160 160 160 160 160 20 20 40 160 20 40
H. Wang et al. / Talanta 48 (1999) 1–7
5
6 H. Wang et al. / Talanta 48 (1999) 1–7

Table 4
Comparison of thiazolylazo reagents used in the determination of platinum group metals by HPLC

Derivatizing Noble metal de- Mobile phase Detection limit (ng Reference
reagent termined ml−1; S/N=3)

TAR Os, Rh, Ru Acetonitrile–water, TBA Br, buffer — [9]

Os, Rh, Ru Methanol–water, TBA Br, acetate buffer 5–17 [10]
TAN Pd, Rh Actonitrile–water, acetate buffer 50, 19 nga [11]
TADAP Pd, Rh, Ru Methanol–water, buffer, cetyltrimethylammonium 1000, 200, 300 [12]
bromide
BTAR Ru, Rh, Os Methanol–water, TBA Br, buffer 3.7–108 [13]
MBTAE Pt, Rh, Pd, Os Methanol–n-butanol–water, TBA Br, ethylenedi- 1.5, 0.54, 0.6, 1.0 [14]
amine diacetic acid diethyl acetate, acetate buffer
Ru, Rh(Ir), Pd, Methanol–water, malic acid, acetate buffer 1.77, 0.057 (0.47), [15]
Os, Pt 0.17, 0.89, 0.55b
TADAP Pt, Ir, Ru, Rh, Methanol–water, TBA Br, acetate buffer 0.37, 9.74, 1.64, 0.29, This paper
Os 1.29

a
The determination limit.
b
S/N= 2.

The acidity of the mobile phase was controlled
by acetate buffer solution. The acidity and con-
centration of the buffer had a complicated influ-
ence in the chromatographic system, which would
affect not only the solubilities and dissociations of
the chelates, but also the effect of TBA + . In
order to separate these chelates well, the effect of
acetate buffer was investigated. With pHB 5.5,
the Os–TADAP peak could not be detected (Fig.
3). Within the range 5 – 20 mmol l − 1, the buffer
concentration had no obvious effect on the reten-
tion times of the chelates (in this experiment, 10
mmol l − 1 of pH 6.0 acetate buffer was used).
3.3. Calibration
Under optimum conditions, separation of the
TADAP chelates of Pt, Ir, Ru, Rh and Os was
achieved with the mobile phase of methanol–wa-
ter (69.5:30.5, v:v) containing 4 mmol l − 1 TBA Br
and 10 mmol l − 1 pH 6.0 acetate buffer (Fig. 4). It
was interesting to note that the elution sequence
was related to the valencies of the metal ions,
except in the case of Ir. Ions having the same
valences showed an elution sequence consistent
with their positions in the periodic table. The
whole analysis was fulfilled in 23 min, with detec-
tion limits (S/N =3) of 0.39, 9.74, 1.64, 0.29 and
1.29 ng ml − 1, respectively. The regression equa-
tions, calibration ranges and detection limits of
these metal ions are listed in Table 2.
3.4. Interference
In order to examine the selectivity of the pro-
posed method, the interference of foriegn ions was
investigated. Under the precolumn derivatization
conditions, Ni, Co, Fe, V, Cu and Pd reacted with
TADAP to form colored chelates. Cu could not
be eluted, V interfered with the determination of
Pt and Co interfered with Ir and Ru. The peaks of
Fe and Ni could be separated with the elements
determined. Eight folds of Pd were allowed. When
5.0 ml of 0.05% TADAP was used, with the
amounts of Pt, Ir, Ru, Rh and Os being 3, 20, 10,
5 and 14 mg in 25 ml respectively, the allowance of
the foreign ions was obtained by the experiment
(see Table 3). This was expressed as the maximum
amount to be determined within an error of 9
5%.
4. Conclusion
A comparison of the proposed method with
others using thiazolylazo reagents in the determi-
nation of the platinum group metals by HPLC is
given in Table 4. This method appears to be more
 H. Wang et al. / Talanta 48 (1999) 1–7
sensitive, convenient and rapid, providing a sim-
ple method for the determination of a multi-noble
metal by HPLC.
Acknowledgements
This research was supported by the National
Natural Science Foundation of China.
References
[1] S.J. Valenty, P.E. Behnken, Anal. Chem. 50 (1978) 834.
[2] B.J. Mueller, R.J. Lovett, Anal. Chem. 59 (1987) 1405.
[3] B.J. Mueller, R.J. Lovett, Anal. Chem. 57 (1985) 2693.
[4] R.C. Gurira, P.W. Carr, J. Chromatogr. Sci. 20 (1982)
461.
[5] B. Wenclawiak, F. Bickmann, Bunseki Kagaku 33 (1984)
E67.
7
[6] H.S. Zhang, W.Y. Mou, J.K. Cheng, Talanta 41 (1994)
1459.
[7] N. Uehara, Y. Annoh, T. Shimizu, Y. Shijo, Anal. Sci. 5
(1989) 111.
[8] Y.S. Nikitin, N.B. Morozova, S.N. Lanin, T.A. Bal’shova,
V.M. Ivanov, E.M. Basova, Talanta 34 (1987) 223.
[9] E.M. Basova, T.A. Bol’shova, E.N. Shapovalova, V.M.
Ivanov, Zh. Anal. Khim. 45 (1990) 1947.
[10] Q. Liu, H. Zhang, J. Cheng, Talanta 38 (1991) 669.
[11] N.A. Baketova, E.M. Basova, V.M. Ivanov, T.A. Bol’-
shova, Zh. Anal. Khim. 45 (1990) 2178.
[12] E.N. Shapovalova, I.V. Mishenina, E.M. Bosova, T.A.
Bol’shova, O.A. Shpigun, Zh. Anal. Khim. 46 (1991) 1503.
[13] H.S. Zhang, Z.D. Shi, J.K. Cheng, J. Wuhan Univ. (Nat.
Sci. Ed.) 80 (1993) 18.
[14] Q. Liu, J. Liu, Y. Tong, J. Cheng, Anal. Chim. Acta 269
(1992) 223.
[15] Q. Liu, Y. Wang, J. Liu, J. Cheng, Anal. Sci. 9 (1993) 523.
[16] B.A. Bidlingmeyer, S.N. Deming, W.P. Price Jr., B.
Sachok, M. Petrusek, J. Chromatogr. 186 (1979) 419.
[17] B.A. Bidlingmeyer, J. Chromatogr. Sci. 18 (1980) 525.