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Interaction of Serotonin and Melatonin With Sodium, Potassium, Calcium, Lithium and Aluminium
Interaction of Serotonin and Melatonin With Sodium, Potassium, Calcium, Lithium and Aluminium
2001; 31:102–108
1993; Limson and Nyokong, 1997; Limson et al., ing solutions of the ligands with an excess of the
1998]. metal ions and freeze drying the solution. Nuclear
Magnetic Resonance (1HNMR) data in de-uter-
Materials and methods ated dimethylsulphoxide (DMSO [D6]) was
recorded with the Bruker EMX 400 NMR
All reagents were of analytical grade, and distilled spectrometer.
deionised water was used for the experiments.
Solutions of Na + , K + , Ca2 + , Li + and Al3 + were
Results
prepared from the corresponding metal chloride
salts. Serotonin (as a creatine sulphate complex; The reduction peak for serotonin in the absence of
Sigma-Aldrich, St Louis, MO, USA) and mela- metal ions was observed in this study at −1.76 V
tonin (Sigma-Aldrich) solutions were prepared versus AgAgCl. For melatonin, the AdCSV peak
daily; serotonin was dissolved in distilled water, in the absence of metal ions was observed at
while melatonin was dissolved in absolute ethanol
−2.18 V versus AgAgCl. The shapes of the peaks
and subsequently diluted with distilled water. The
shown in Figs. 2 and 3 for metal– serotonin and
final ethanol concentration in the solution was
metal –melatonin complexes, respectively, are sim-
0.2% (v/v). A solution of 0.05 mol dm − 3 tetra-
butylamoniumbromide (TBABr; Sigma-Aldrich) ilar to those of the metal alone, thus suggesting
was used as an electrolyte for the electrochemical that reduction of the metal –serotonin or metal –
experiments (Fig. 1). melatonin complex occurs at the metal. Reduction
Stripping voltammograms were obtained with peaks, as a result of serotonin or melatonin alone,
the BioAnalytical Services (BAS, Lafayette, IN, are not evident in Figs. 2 and 3 at concentrations
USA) CV-50 W voltammetric analyser. A Con- of serotonin and melatonin employed.
trolled Growth Mercury Electrode (CGME; BAS The enhancement in the reduction currents,
Model MF-9058) was employed as the working upon addition of serotonin to the metal containing
electrode. A silver/silver chloride (3 mol dm − 3 solution, is an indication of the in situ formation
KCl) and a platinum wire were employed as the and accumulation of the metal –serotonin or
reference and auxiliary electrodes, respectively. So- metal– melatonin complex onto the Hg electrode.
lutions containing the metal ion and the ligand The shift in the reduction peaks to more negative
(serotonin or melatonin) were de-aerated for a potentials upon addition of serotonin or mela-
minimum period of 5 min, after which a flow of tonin to solutions containing the metal ion,
nitrogen gas was maintained over the solution suggests that the metal – serotonin or metal –
throughout the measurement. The metal –ligand melatonin complexes formed are more difficult to
complex is expected to have formed at this stage. reduce than the free metal. The extent of the shift
A deposition potential ( − 1,200 mV for Al3 + and in reduction peak potential of the metal on addi-
Li + , and −1,400 mV for Na + , Ca2 + , K + ) was tion of serotonin or melatonin is a good measure
applied for a deposition time of 120 s. The adsorp- of the stability of the metal –serotonin or metal –
tion of the metal –ligand complex onto the elec- melatonin complex formed in situ. Table 1 lists the
trode occurs at this stage. The deposition potential
shifts in peak potentials, DEp, for the reduction of
values given above were obtained from optimisa-
the metals on addition of serotonin or melatonin.
tion of the deposition potential for each metal–lig-
and complex at fixed deposition time and at
constant concentrations of the metals and ligands.
The deposition time was similarly optimised using
the optimal deposition potentials and constant
concentrations of the metal and the ligand. The
voltammograms were recorded in a negative direc-
tion from the accumulation potential to at least
0.2 V beyond the reduction peak of the metal
complex. A scan rate of 25 mV s − 1 was applied
during the stripping step. AdCSV experiments
were also performed for the metal ion in the
absence of the ligand or for the ligand in the
absence of the metal ion.
Solid complexes formed between the metal ion
and serotonin or melatonin were prepared by mix- Fig. 1. Molecular structure of serotonin and melatonin.
103
Lack et al.
Fig. 2. Adsorptive cathodic stripping voltammograms obtained for 6.7×10 − 8 mol dm − 3 serotonin in the presence of (A) Ca2 + ,
(B) K + , (C) Na + , (D) Li + and (E) Al3 + . (a) Voltammograms for metals alone and (b) voltammograms in the presence of
serotonin. Metal concentration=3.3×10 − 9 mol dm − 3. Scan rate=25 mV s − 1. Electrolyte = 0.05 mol dm − 3 TBABr.
104
Interactions of serotonin and melatonin
Fig. 3. Adsorptive cathodic stripping voltammograms obtained for 6.7×10 − 8 mol dm − 3 melatonin in the presence of (A) Ca2 + ,
(B) K + , (C) Na + , (D) Li + and (E) Al3 + . (a) Voltammograms for metals alone and (b) voltammograms in the presence of
melatonin. Metal concentration=3.3×10 − 9 mol dm − 3. Scan rate=25 mV s − 1. Electrolyte =0.05 mol dm − 3 TBABr.
105
Lack et al.
Serotonin −1760
Sodium −2246 −41 1.97 (0.995)
Potassium −2254 −77 1.04 (0.985)
Calcium −2266 +27 0.88 (0.991)
Lithium −2348 −123 0.59 (0.967)
Aluminium −2360 −71 0.81 (0.955)
Melatonin −2181
Sodium −2290 −85 1.39 (0.996)
Potassium −2318 −141 1.29 (0.991)
Calcium −2309 −16 1.02 (0.986)
Lithium −2323 −97 0.39 (0.982)
Aluminium −2333 −44 0.65 (0.987)
[serotonin]=[melatonin] =6.7×10−8 mol dm−3. Ep = peak potentials of the metal–ligand complexes. Ep is the difference
between the peak potentials of the metal ions and those of the metal–ligand complexes. a Metal concentration=3.3×
10−9 mol dm−3. b Regression coefficients shown in brackets.
For serotonin, the high negative shift observed The current increased with increases in concentra-
(DEp = −123 mV) on addition of serotonin to tion of melatonin and serotonin for concentrations
Li + solutions reflects the stability of the resulting ranging from 1 ×10 − 9 to 1 ×10 − 8 mol dm − 3.
lithium–serotonin complex when compared to the The 1HNMR spectra of melatonin and sero-
other metal –serotonin complexes. The high nega- tonin in the presence and absence of the metal ions
tive DEp value also shows that the resulting com- suggests that an interaction occurs between the
plex is more difficult to reduce when compared to ligands and the metal ions (Tables 2 and 3). For
the reduction of the free metal and the other both serotonin and melatonin, the data presented
metal–serotonin complexes. For melatonin, a high here do not give clear evidence of the actual coor-
negative Ep value was observed for the potassium – dination site of the metals.
serotonin complex showing that a more stable
complex is formed between potassium and mela-
tonin when compared to the other metal –mela-
tonin complexes.
Based on the magnitudes of the shifts in reduc-
tion potentials of the metals on addition of sero-
tonin, the relative stability of the in situ complexes
formed between the metals and serotonin de-
creases with the metal as follows: Li + \ K + \
Al3 + \Na + \Ca2 + . For the complexes between
melatonin and the metals, the stability of the com-
plexes increases with the metal as follows: K + \
Li + \Na + \Al3 + \ Ca2 + . Thus, both Li + and
K + form more stable complexes with serotonin
and melatonin when compared to the other metal
complexes under consideration. Both melatonin
and serotonin do not favour coordination to Ca2+.
Peak currents for the metal–serotonin or
metal– melatonin complexes increased with in-
creases in the concentration of the metal as shown
in Fig. 4 for serotonin and melatonin complexes,
respectively. The plots are linear with regression
coefficients listed in Table 1. The slopes of the
plots shown in Fig. 4 are listed in Table 1. Fig. 4. (A) Variation of currents with metal ion concentra-
tion for 6.7× 10 − 8 mol dm − 3 serotonin and (B) for 6.7×
AdCSV currents increased with increases in the 10 − 8 mol dm − 3 melatonin. Ca2 + ( ), K + (
), Na + (),
concentrations of serotonin and melatonin as Li + () and Al3 + ( ). Scan rate= 25 mV s − 1. Electrolyte =
shown in Fig. 5 at constant concentration of K + . 0.05 mol dm − 3 TBABr.
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Interactions of serotonin and melatonin
1
Table 3. HNMR peaks observed for NH2 and OH resonances
for serotonin in the presence of the metal ions
d (ppm)*
Metal OH NH2
107
Lack et al.
108