Journal of the Chinese Chemical Society, 2005, 52, 863-871 863

Complexation Equilibria of Zn(II), Pb(II) and Cd(II) with Reduced

Glutathione (GSH) and Biologically Important Zwitterionic Buffers

Zeinab Mohamed Anwar

Department of Chemistry, Faculty of Science Suez Canal University, Ismailia, Egypt

The interaction of zinc(II), lead(II), and cadmium(II) with Glutathione (d-L-glutamyl-Lcysteinylglycine)

as primary ligand and zwitterionic buffers (N-[2-Hydroxyethyl]piperazine-N¢-[2-ethanesulfonic acid])
(HEPES) and (N-Hydroxyethyl]piperazine-N¢-[2-hydroxy-propanesulfonic acid]) (HEPPSO) as secondary
ligands were studied by potentiometric-pH titration in 1:1:1 ratio at 25.0 °C and I = 0.1 mol.dm-3 (KNO3). The
formation constants of different normal and protonated binary and ternary complex species were calculated.
Formation constants for the monohydroxy, and dihydroxy complexes for the binary systems M(II) +
HEPES and M(II) + HEPPSO have been evaluated. The distribution curves for the various complex species as
a function of pH were constructed.

Keywords: Glutathione; Zwitterionic buffers; Binary complexes; Ternary complexes; Hydroxy com-
plexes; Zn(II), Pb(II), and Cd(II) complexes.

INTRODUCTION or amide coordination.

Sulphur donor atoms are one of the most important
binding sites of metalloproteins; therefore, widespread inves-
tigations have been carried out to reveal the coordination
chemistry of amino acids and peptides containing sulphur do-
nors. One of the biologically important ligands of this type is
the tripeptide glutathione.
Glutathione (d-L-glutamyl-Lcysteinylglycine) is one
of the most important naturally occurring tripeptides. It has
been proved that glutathione reduces copper(II) to give
copper(I) complexes and disulphide. 1 A number of models
were proposed for the relation between hexokinase enzyme,
Glutathione reduced (GSH), and heavy metal toxicity in
vivo.2,3
Glutathione, which is the most common biological thiol,
is directly involved in the cellular metabolism of genotoxic
and carcinogenic Cr(VI) compounds.4 Chromium (VI/V/IV)
intermediates, formed during the reduction of Cr(VI) to
Cr(III) by GSH, are thought to act as DNA-damaging species
in Cr(VI)-induced genotoxicity.5 A. Levina et al.6 have per-
formed a detailed study of the intermediates formed in Cr(VI)
reactions with GSH using a combination of electrospray mass
spectrometry and UV-visible, EPR, and x-ray absorption
spectroscopies.
Interactions of nickel(II), cobalt(II), and zinc(II) with
glutathione start at the amino acid locus in slightly acidic or
medium pH-range.7-9 In the case of nickel(II) this is followed
by the formation of square-planar complexes due to sulphur
Williams et al.8 concluded that deprotonation of the am-
ide in glutathione occurs at around pH ~ 8, while the forma-
tion of sulphur-coordinated polynuclear species was sug-
gested.
Nickel(II) and palladium(II) – promoted amide binding
was presumed to occur above pH ~ 12 and 9, respectively.9
The possible roles of glutathione in Zn(II) physiology
have been studied.10 Zn(II) complexes of glutathione and dif-
ferent analogues have been studied by potentiometry and
1D-NMR. The formation of Zn(II) (GSH) (His) ternary com-
plex is of particular interest due to its stability. The results
suggest that GSH may be a regulatory factor for some, but not
all, zinc enzymes and structural proteins.
Attention has been drawn to the use of zwitterionic
amino acids and zwitterionic-N-substituted amino sulfonic
acids as buffers of biological interest.11,12 Those compounds
are all ampholytes (with zwitterionic structures) and are use-
ful buffers compatible with most media of physiological in-
terest. Two of the potentially useful zwitterionic buffers for
use in biochemistry are N-2-hydroxyethyl piperazine-N-eth-
anesulfonic acid (HEPES) and N-2-hydroxethyl piperazine-
N¢-2-hydroxypropanesulfonic acid (HEPPSO).
The interaction of La(III), Ce(III), Pr(III), and Eu(III)
with adenosine 5¢-monophosphate, guanosine 5¢-monophos-
phate, cytidine 5¢-monophosphate and HEPES in addition to
MES, ACES and MOPSO was studied potentiometrically at
25.0 ± 0.1 °C. Ionic strength I = 0.1 mol.dm-3 KNO3. The for-
mation constants of various normal 1:1:1 and protonated
864 J. Chin. Chem. Soc., Vol. 52, No. 5, 2005
mixed ligand complex species were calculated.13
Azab et al.14-16 have studied the interaction of different
types of zwitterionic buffers and many biologically important
molecules including nucleobases, nucleosides and nucleo-
tides, leading to the investigation of the ability of formation
of various ternary complexes in solutions as these systems
may serve as models for metalloenzyme reactions in biologi-
cal systems.
The aim of the present investigation is to obtain quanti-
tative information on the complexes of three biologically im-
portant metal ions Zn(II), Pb(II), and Cd(II) with glutathione
as binary metal complexes, and in the presence of two of the
biologically important zwitterionic buffers HEPES and
HEPPSO as ternary complexes. GSH is thought to be in-
volved in the transport of amino acids across cell membranes
via the d-glutamyl cycle17 and also seems to play a role in de-
termining the behavior of some metal ions in the body. Thus
the systems M(II) + GSH + Z [M(II) = Zn(II)] may be consid-
ered as model systems mimicking interesting biological in-
teractions. Also, the Pb(II) + GSH + Z or Cd(II) + GSH + Z
may be considered as models which may account for the toxi-
cology of Pb(II) and Cd(II) ions. Potentiometric methods are
applied to compare the stability of the formed complexes in
the presence of buffers. The compounds used and their termi-
nology are shown in Chart 1.
Chart 1
Anwar
EXPERIMENTAL
Materials
All reagents were analytical grade. Reduced gluta-
thione (GSH), HEPES, and HEPPSO were purchased from
Sigma Chemical Co. and were used without purification.
All solutions were made up under a N2 atmosphere in
H2O which was decarbonated and doubly distilled. The stan-
dard solutions of KOH (0.200 mol.dm-3) were kept for no lon-
ger than three weeks; stock solutions of KNO 3 (1.0 mol.
dm -3 ), Zn(NO 3 ) 2 .6H 2 O (0.1 mol.dm -3 ), Pb(NO 3 ) 2 (0.1 mol.
dm-3) and Cd(NO3)2.3H2O (0.1 mol.dm-3) were kept under N2.
Solutions of the ligands (0.1 mol.dm-3) were made up prior to
use. The exact concentrations of the metal salt, nitric acid and
KOH solutions were determined titrimetrically (EDTA, KOH,
potassium hydrogen phthalate).
Measurements
All KOH titrations were done under N2 in the thermo-
stated vessels at 25.0 ± 0.1 °C. The fully automatic pH titra-
tion unit consisted of a combined glass electrode, and a digi-
tal pH-meter (Fisher Accumet 825 MP). The exact calibration
was done daily by first using commercial buffer solutions
(Merck) of pH’s 4.0, 7.0, 9.18 and then titrating 50 mL of 1 ´
10 -3 mol.dm -3 tris[tris(hydroxymethyl)methylamine)] and
HNO 3 (2 ´ 10 -3 mol.dm -3 ) with KOH (0.200 mol.dm -3 ).
Values of the apparent activity coefficient a (0.957), pka of
tris (8.167), were taken from literature18 and used for final re-
adjustment of the experimental pH readings.
A constant ionic strength was obtained with 0.1 mol.
-3
dm KNO3, and the total volume was kept constant at 10 mL.
The solutions titrated can be presented according to the
following scheme: HNO3 (4 ´ 10-3 mol.dm-3) + glutathione (1
´ 10-3 mol.dm-3) (a); HNO3 (4 ´ 10-3 mol.dm-3) + gluathione
(1 ´ 10-3 mol.dm-3) + M(II) (1 ´ 10-3 mol.dm-3) (b); HNO3 (4 ´
10-3 mol.dm-3) + zwitterionic buffer (1 ´ 10-3 mol.dm-3) (c);
HNO 3 (4 ´ 10 -3 mol.dm -3 ) + zwitterionic buffer (1 ´ 10 -3
mol.dm -3 ) + M(II) (1 ´ 10 -3 mol.dm -3 ) (d); HNO 3 (4 ´ 10 -3
mol.dm -3 ) + gluathione (1 ´ 10 -3 mol.dm -3 ) + zwitterionic
buffer (1 ´ 10-3 mol.dm-3) + M(II) (1 ´ 10-3 mol.dm-3) (e). At
least four titrations were performed for each system.
RESULTS AND DISCUSSION
The chemical and biological activity of proteins and
peptides would be expected to vary with the degree of ioniza-
tion. These molecules may exist in a great number of distinct
Complexes of Zn(II), Pb(II), and Cd(II) with GSH, HEPES and HEPPSO
ionization states. The acidic groups are uncharged in strongly
acidic solutions and negatively charged in sufficiently alka-
line solutions. The basic groups are positively charged (pro-
tonated) in a strongly acidic solution and are uncharged in a
sufficiently alkaline solution.
Such molecules have macroscopic constants which of-
ten describe a composite of the processes which are actually
occurring in solution. On the other hand, microconstants are
the equilibrium constants for equilibria involving individual
species in solution.
At the molecular level, the acid-base chemistry of GSH
is described by eight microscopic constants. 20-21 Reduced
glutathione (GSH) has eight donor sites of hetero atoms. In its
interactions with metal ions, GSH presents eight potential
binding sites.
The concentration of free hydrogen ion, C H + , at each
point of the titration is related to the measured emf, E°, of the
cell by the Nernst equation.
E = E° + Q log CH+ (1)
where E° is a constant which includes the standard potential
of the glass electrode and Q is the slope of the glass electrode
response. The value of E° for the electrode was determined
from a Gran plot derived from titration of tris buffer and nitric
acid with a standard KOH solution under the same tempera-
ture and medium conditions as for the test solution titration.
The results so obtained were analyzed by the nonlinear least-
squares computer program ESAB2M19 to refine E° and the
autoprotolysis constant of water, Kw.
During these calculations Kw was refined until the best
value for Q was obtained. The results obtained indicated the
reversible Nernstian response of the glass electrode used.
It was found that the major macroscopic ionization con-
stant for this molecule is [HA]5- and found to be 9.82 ± 0.01.
The formation constants for the different binary and
ternary complexes were refined with the SUPERQUAD com-
puter program.22 The stability constant for the binary com-
plexes of metal ions with the reduced glutathione in the form
Mp (A)q (H)r (OH)s and those for the complexes of metal ions
with zwitterionic buffers (HEPES or HEPPSO) in the form of
Mp (Z)n (H)r (OH)s were calculated according to the follow-
ing equilibria (charges are omitted for clarity).
pM + qA + rH + sOH ƒ Mp (A)q (H)r (OH)s (2)
or pM + nZ + rH + sOH ƒ Mp (A)n (H)r (OH)s (3)
where p, q, n, r and s are moles of M, A, Z, H and OH in the
J. Chin. Chem. Soc., Vol. 52, No. 5, 2005 865
formed complexes Mp (A)q (H)r (OH)s or Mp (Z)n (H)r (OH)s.
While for ternary complexes of the type Mp (A)q (H)r
the equilibrium could be defined as:
pM + qA + nZ + rH ƒ Mp (A)q (Z)n (H)r (4)
The data points collected in the pH range 3-10 were
used for the calculations. The constants were refined by mini-
mizing, U, defined by:
U = Si Wi (Eobs - Ecalc)2 (5)
where Eobs and Ecalc refer to the measured potential and that
calculated from.
E = E° + Q log CH+
The weighting factor Wi is defined as the reciprocal of
the estimated variance of the measurement.
Wi = 1/[sE2 + (dE/dV)2 sv2] (6)
where sE and sv are the estimated variances of the potential
and volume readings respectively. The quality of the fit was
judged by the values of the sample standard deviation S and
the goodness of fit X2 (Pearson’s test). At sE = 0.1 mV (0.001
pH error) and sv = 0.005 mL, the values of S in different sets
of titrations were between 1.0 and 1.8, and X2 was between
12.0 and 13.0.
The scatter of residuals (Eobs - Ecalc) versus pH was rea-
sonably random, without any significant systematic trends,
thus indicating a good fit of experimental data.
The major species formed of Zn(II), Cd(II), and Pb(II)
complexes with reduced glutathione could be described ac-
cording to the following equilibria,
[HA]5- + M2+ ƒ [MA]3- + H+
[HA]5- + M2+ ƒ [MHA]2+.
The concentration of the dimeric complex [M 2 A] - is
very low in equimolar solutions.
Formation constants for Zn(II), Cd(II) or Pb(II) com-
plexes of GSH are listed in Table 1. The values were calcu-
lated and refined using the SUPERQUAD computer pro-
gram.22 The data are in good agreement with that found in the
literature.23,24 In all cases the major species [MA]3- is present
predominantly at neutral pH. Inspection of the values of for-
mation constants for binary complexes for Zn(II), Cd(II), and
866 J. Chin. Chem. Soc., Vol. 52, No. 5, 2005 Anwar

Table 1. Formation constants for normal and protonated binary plexes. The chemical shift data in 13C NMR previous study of
M(II) + glutathione complexes at 25.0 ± 0.1 °C and I = Pb(II)-GSH system indicates no detectable binding to the
0.1 mol.dm-3 KNO3
amino group with the acid-base properties of the amino group
Metal ion 2-
in Pb(HL) identical to the acid-base properties of the amino
M(II)
Log K[M(II)(HA)]2- Log K[M(II)(HA)]
[M(II)(A)]3- group of GSH.
M(II)
Tables 2 and 3 include the formation constants for the
Zn(II) 4.83 ± 0.02 7.72 ± 0.02 different complex species formed by Zn(II), Pb(II), and Cd(II)
Pb(II) 6.66 ± 0.03 8.48 ± 0.03
ions with the zwitterionic buffers HEPES and HEPPSO ac-
Cd(II) 5.70 ± 0.03 10.65 ± 0.030
cording to the following equilibria:
± Refers to three times standard deviation (3s).

M(II) +HZ+ + nH2O ƒ [M(II) (HZ) (H2O)2]3+ (7)

Pb(II) with glutathione as collected in Table 1 reveal that
Cd(II) forms more stable complexes with GSH either in the
[CdA]3- or the protonated [CdHA]2- form than does Zn(II).
The extra stability of Cd(II) complexes may be attributed to
the fact that Cd(II) binds more strongly to the sulfhydryl
group than does Zn(II), and that is consistent with the results
of 13C NMR studies of the complexation of GSH with Zn(II)
and Cd(II).
As shown in Table 1, the binding of Pb(II) by GSH is
considerably stronger than is the binding of Zn(II) or Cd(II).
This may be attributed to the nature of binding of Pb(II) to
GSH in which the ion is binding only to the sulfhydryl group
and, to a lesser extent, to the glycine carboxylate group. The
extent of binding to the carboxylate group decreases at pH >
8, presumably due to the displacement by hydroxide ions
with the formation of Pb(II)-hydroxo-GSH mixed com-
¯ -H+
[M(II) (Z) (H2O)2]2+
¯ -H+
[M(II) (Z) (OH) (H2O)]+
¯ -H+
[M(II) (Z) (OH)2]
Inspection of the data reveals that generally the trend of
stability to be Pb(II) > Zn(II) > Cd(II) for HEPES buffer
ligand. While for HEPPSO ligand the stability order is Pb(II)
> Cd(II) > Zn(II). The higher stability of HEPPSO ate com-
plexes of Zn(II), Cd(II) and Pb(II) compared to those of
HEPES ate may be attributed to the basicity effect as well as
the participation of hydroxo groups of HEPPSO in the coor-
dination with metal ions, especially with Pb(II).
Figs. 1-3 represent sets of experimental titration curves

Table 2. Formation constants for the normal, protonated and hydroxo binary M(II) + HEPES
(Z) complexes at 25.0 ± 0.1 °C and I = 0.1 mol.dm-3 KNO3

Metal ion M(II) +

Log K[M(II)(HZ)]+ Log K[M(II)(HZ)
[M(II)(Z)] Log K[M(II)(Z)]
[M(II)(Z)(OH)]
Log K[M(II)(Z)(OH)]
[M(II)(Z)(OH)2 ]
M(II)

Zn(II) 6.73 ± 0.02 3.27 ± 0.03 3.47 ± 0.02 7.05 ± 0.02

Pb(II) 7.90 ± 0.03 3.14 ± 0.03 3.65 ± 0.03 7.88 ± 0.03
Cd(II) 4.16 ± 0.01 3.25 ± 0.02 3.29 ± 0.02 5.73 ± 0.02
± Refers to three times standard deviation (3s).

Table 3. Formation constants for the normal, protonated and hydroxo binary M(II) +
HEPPSO (Z) ligand complexes at 25.0 ± 0.1 °C and I = 0.1 mol.dm-3 KNO3

Metal ion M(II) +

Log K[M(II)(HZ)]+ Log K[M(II)(HZ)
[M(II)(Z)] Log K[M(II)(Z)]
[M(II)(Z)(OH)]
Log K[M(II)(Z)(OH)]
[M(II)(Z)(OH)2 ]
M(II)

Zn(II) 3.77 ± 0.01 3.25 ± 0.02 3.32 ± 0.02 6.36 ± 0.02

Pb(II) 7.98 ± 0.02 3.18 ± 0.03 3.92 ± 0.03 7.14 ± 0.03
Cd(II) 4.53 ± 0.02 3.28 ± 0.02 3.41 ± 0.03 6.71 ± 0.03
± Refers to three times standard deviation (3s).
Complexes of Zn(II), Pb(II), and Cd(II) with GSH, HEPES and HEPPSO
Fig. 1. Potentiometric pH-titration curves for Zn(II) +
Glutathione (GSH) + Z system, I = 0.1 mol.dm-3
KNO 3 and at 25.0 ± 0.1 o C. 4 ´ 10 -3 mol.dm -3
HNO3 + 1 ´ 10-3 mol.dm-3 Glutathione ( + ), 4 ´
10 -3 mol.dm -3 HNO 3 + 1 ´ 10 -3 mol.dm -3
Glutathione + 1 ´ 10-3 mol.dm-3 Zn(II) (n ), 4 ´
10-3 mol.dm-3 HNO3 + 1 ´ 10-3 mol.dm-3 Z (=),
4 ´ 10-3 mol.dm-3 HNO3 + 1 ´ 10-3 mol.dm-3 Z +
1 ´ 10-3 mol.dm-3 Zn(II) (¶), 4 ´ 10-3 mol.dm-3
HNO 3 + 1 ´ 10 -3 mol.dm -3 Glutathione + 1 ´
10-3 mol.dm-3 Z + 1 ´ 10-3 mol.dm-3 Zn(II) (à).
(a) Z = HEPES, (b) Z = HEPPSO.
J. Chin. Chem. Soc., Vol. 52, No. 5, 2005 867
Fig. 2. Potentiometric pH-titration curves for Pb(II) +
Glutathione (GSH) + Z system, I = 0.1 mol.dm-3
KNO 3 and at 25.0 ± 0.1 o C. 4 ´ 10 -3 mol.dm -3
HNO3 + 1 ´ 10-3 mol.dm-3 Glutathione ( + ), 4 ´
10 -3 mol.dm -3 HNO 3 + 1 ´ 10 -3 mol.dm -3
Glutathione + 1 ´ 10-3 mol.dm-3 Pb(II) (n ), 4 ´
10-3 mol.dm-3 HNO3 + 1 ´ 10-3 mol.dm-3 Z (=),
4 ´ 10-3 mol.dm-3 HNO3 + 1 ´ 10-3 mol.dm-3 Z +
1 ´ 10-3 mol.dm-3 Pb(II) (¶), 4 ´ 10-3 mol.dm-3
HNO 3 + 1 ´ 10 -3 mol.dm -3 Glutathione + 1 ´
10-3 mol.dm-3 Z + 1 ´ 10-3 mol.dm-3 Pb(II) (à).
(a) Z = HEPES, (b) Z = HEPPSO.
868 J. Chin. Chem. Soc., Vol. 52, No. 5, 2005
Fig. 3. Potentiometric pH-titration curves for Cd(II) +
Glutathione (GSH) + Z system, I = 0.1 mol.dm-3
KNO 3 and at 25.0 ± 0.1 o C. 4 ´ 10 -3 mol.dm -3
HNO3 + 1 ´ 10-3 mol.dm-3 Glutathione ( + ), 4 ´
10 -3 mol.dm -3 HNO 3 + 1 ´ 10 -3 mol.dm -3
Glutathione + 1 ´ 10-3 mol.dm-3 Cd(II) (n ), 4 ´
10-3 mol.dm-3 HNO3 + 1 ´ 10-3 mol.dm-3 Z (=),
4 ´ 10-3 mol.dm-3 HNO3 + 1 ´ 10-3 mol.dm-3 Z +
1 ´ 10-3 mol.dm-3 Cd(II) (¶), 4 ´ 10-3 mol.dm-3
HNO 3 + 1 ´ 10 -3 mol.dm -3 Glutathione + 1 ´
10-3 mol.dm-3 Z + 1 ´ 10-3 mol.dm-3 Cd(II) (à).
(a) Z = HEPES, (b) Z = HEPPSO.
Anwar
Table 4. Formation constants for mixed ligand complexes M(II)
+ glutathione (A) + zwitterionic buffer (Z) at 25.0 ± 0.1
°C and I = 0.1 mol.dm-3 KNO3
Z = HEPES Z = HEPPSO
Metal ion
M(II) Log K M(II)(A)
M(II)(A)(HZ) Log K M(II)(A)
M(II)(A)(Z)
Zn(II) 5.56 ± 0.02 6.59 ± 0.02
Pb(II) 4.94 ± 0.03 7.18 ± 0.03
Cd(II) 3.57 ± 0.01 6.02 ± 0.01
± Refers to three times standard deviation (3s).
obtained according to the sequence described in the experi-
mental section for M(II) + GSH + HEPES and M(II) + GSH +
HEPPSO systems, where M(II) = Zn(II), Pb(II) and Cd(II).
The complexation equilibria for these ternary com-
plexes can be postulated as follows:
M(II) + [HA]5- + [HZ]+ ƒ [M(II) (A)5- (HZ)+] (8)
¯
[M(II) (A) (HZ)]2-
where [HA]5- represents the coordinated anion of GSH and
[HZ]+ the protonated zwitterionic buffer ligand.
The metal ion will coordinate firstly with the peptide
molecule forming a stable complex, then the secondary ligand
coordinates with one of the zwitterionic molecules forming
the ternary complex species. The formation constant of the
ternary complexes formed in solution according to the previ-
ous stepwise manner follows the order Zn(II) > Pb(II) >
Cd(II).
In the case of HEPPSO as the secondary ligand mole-
cule, the equilibria can be presented as:
M(II) + [HA]5- + [HZ]+ ƒ [M(II) (A) (Z)]3- (9)
where the secondary ligand HEPPSO combines with the
formed binary complex as a deprotonated one forming stable
complexes with the stability order: Pb(II) > Zn(II) > Cd(II)
which indicate the ability of HEPPSO ligand to isolate Pb(II)
GSH binary complex which may induce toxicity in the body.
The previous trend of stability for the ternary com-
plexes may be explained that upon the interaction of HEPES
ate ligand to the binary metal ion-GSH complex, the second-
ary ligand basic center of coordination is through the nitrogen
atom of the piperazine ring, hence with nitrogen donors Pb(II)
binds more weakly than Cd(II) and Zn(II).26
Complexes of Zn(II), Pb(II), and Cd(II) with GSH, HEPES and HEPPSO
On the other hand for HEPPSO ate ligand, the sites of
coordination are the nitrogen of piperazine ring and oxygen
of OH group side chain. Hence Pb(II) moves up the ruler to
the strength or more than Cd(II) and Zn(II).26-28
The most unexpected result for the ternary systems of
the type M(II) + GSH + Z is certainly the high stability of the
ternary Zn(II) + GSH + HEPES complex compared to Pb(II)
+ GSH + HEPES and Cd(II) + GSH + HEPES. This means
that the equilibrium [4] favours the products:
Zn(GSH) + Zn(HEPES) ƒ
Zn(GSH) (HEPES) + Zn (10)
Fig. 4. Distribution curves for Zn(II) + zwitterionic
ionic buffer (Z). (a) Zn(II) + HEPES, (b) Zn(II)
+ HEPPSO.
J. Chin. Chem. Soc., Vol. 52, No. 5, 2005 869
This behaviour may be attributed to the ability of Zn(II) to
easily switch from coordination number 6 to 4 or 5.29,30 This
behaviour may explain if the primary ligand GSH binds to an
octahedral Zn(II) with a very high probable structure. Further
binding of the HEPES ate anion should reduce the coordina-
tion to 4 or 5 with two water molecules released upon the co-
ordination of the secondary ligand. Such a process is entro-
pically favoured which could not be expected with Pb(II) and
Cd(II) ions.
The SUPERQUAD computer program which employs
the same algorithm as BEST, was used to calculate the spe-
cies distribution curves shown in Fig. 4-6.
Fig. 5. Distribution curves for Pb(II) + zwitterionic
ionic buffer (Z). (a) Pb(II) + HEPES, (b) Pb(II)
+ HEPPSO.
870 J. Chin. Chem. Soc., Vol. 52, No. 5, 2005
Fig. 4a shows all species present in the 1:1 Zn(II)-
HEPES system. The major species becomes the monopro-
tonated form, which reaches its maximum concentration at
pH » 5, while the normal complex of the type Zn(II)-Z is
probably formed in the pH range from 6-8; exactly the maxi-
mum formation is obtained at the pka value of HEPES (7.60).
On the other hand, for Zn(II)-HEPPSO (Fig. 4b) the major
species also is found to be formed maximally in the pH range
of 6-8. On releasing the proton of HEPPSO (pka = 7.8) the
neutral complex species is predicted.
The protonated species [PbHZ]+ where Z = HEPES or
HEPPSO are also the predominant species in the pH range
Fig. 6. Distribution curves for Cd(II) + zwitterionic
ionic buffer (Z). (a) Cd(II) + HEPES, (b) Cd(II)
+ HEPPSO.
Anwar
2-6, while in the range of 6-8, the neutral species are formed,
as shown in Fig. 5.
The pH range of 5.5-7.0 is the preferred one for the for-
mation of [CdHZ]+, while near pH = 8.0 the neutral species
[CdZ] are most probable, as depicted in Fig. 6.
Received April 12, 2004.
REFERENCES
1. Goodgame, D. M. L.; Joy, M. A. J. Inorg. Biochem. 1986,
26, 219.
2. Louie, Y. A; Meade, J. T. Chem. Rev. 1999, 2711.
3. Kiezel, A.; Bal, W. Acta Biochim. Polon, 1999, 46, 567.
4. Ning, J.; Grant, H. M. Toxicol. In Vitro 2000, 14, 329.
5. Connett, H. P.; Wetterhahn, E. K. J. Am. Chem. Soc. 1985,
107, 4282.
6. Levina, A.; Zhang, L.; Lay, A. P. 35th International Confer-
ence on Coordination Chemistry, Heidelberg, 2002, 313.
7. Martin, B. R.; Edsall, T. J. J. Am. Chem. Soc. 1959, 81, 4044.
8. Kozlowska, F. G.; May, P. R.; Williams, R. D. Inorg. Chim.
Acta 1980, 46, L51.
9. Sovago, I.; Martin, B. R. J. Inorg. Nucl. Chem. 1981, 43,
425.
10. Krezel, A.; Wojcik, J.; Maciegczyk, M.; Bal, W. 35th Interna-
tional Conference on Coordination Chemistry, Heidelberg,
2002, 30.
11. Good, E. N.; Winget, D. G.; Winter, W.; Connoly, N. T.;
Izawa, S.; Singh, M. M. R. Biochemistry 1966, 5, 4677.
12. Ferguson, J. W.; Braunschweiger, I. K.; Braunschweiger, R.
W.; Smith, R. J.; McCormic, J. J.; Wasmann, C. C.; Jarvis, P.
N.; Bell, H. D.; Good, E. N. Anal. Biochem. 1980, 104, 300.
13. Anwar, M. Z.; Azab, A. H. J. Chem. Eng. Data 2001, 46,
613.
14. Anwar, M. Z.; Azab, A. H. J. Chem. Eng. Data 2001, 46(1),
34.
15. Azab, A. H.; Deghaidy, S. F.; Orabi, S. A.; Farid, Y. N. J.
Chem. Eng. Data 2000, 45, 709.
16. Azab, A. H.; Orabi, S. A.; Abdel-Salam, T. E. J. Chem Eng.
Data 2001, 46, 346.
17. Arias, M. I.; Jakoly, B. W. eds. Glutathione: Metabolism and
Function, Raven Press: New York, 1976, p. 35.
18. Sundaralingam, M. A. J. Am. Chem. Soc. 1973, 95, 2333.
19. Destefane, C.; Princi, P.; Rigano, C.; Sammartano, S. Ann.
Chim. (Rome), 1987, 77, 643.
20. Rabenstein, L. D. J. Am. Chem. Soc. 1973, 95, 2797.
21. Martin, B. R.; Edsall, T. J. Bull. Soc. Chim. Biol. 1958, 40,
Complexes of Zn(II), Pb(II), and Cd(II) with GSH, HEPES and HEPPSO J. Chin. Chem. Soc., Vol. 52, No. 5, 2005 871

1763. 83-86.
22. Gans, P.; Sabatini, A.; Vacca, A. J. Chem. Soc., Dalton 27. Martin, B. R. Met. Ions Biol. Syst. 1986, 20, 21-65.
Trans. 1958, 1195. 28. Martin, B. R. In Encyclopedia of Molecular Biology and
23. Perrin, D. D.; Watt, E. A. Biochim. Biophys. Acta 1971, 230, Molecular Medicine; Meyers, A. R., Ed.; VCH: Weinheim,
96. 1966; vol. 1, pp 125-134.
24. Corrie, M. A.; Walker, D. M.; Williams, R. D. J. Chem. Soc. 29. Martin, B. R. In Metal ions in biological systems; Sigel, H.,
Dalton Trans. 1976, 1012. Ed.; Marcel Dekker: New York, 1986; vol. 20, pp 21-65.
25. Fuhr, J. B.; Rabenstein, L. D. J. Am. Chem. Soc. 1973, 95, 30. Fausto da Silva, R. J. J.; Williams, P. J. R. The biological
6944. chemistry of the elements; Clarendon: Oxford, U.K., 1991;
26. Martin, B. R. In Molecular Biology and Biotechnology; pp 307-311.
Meyers, A. R., Ed.; VCH Publishers: New York, 1995; pp