Synthesis, aggregation and spectroscopic studies of novel water soluble metal free, zinc, copper and magnesium phthalocyanines and investigation of their anti-bacterial properties

Article abstract of DOI:10.1016/j.saa.2014.05.075

In this study, novel phthalonitrile derivative (3) was synthesized by the reaction between 4-nitrophthalonitrile (2) and a triazole derivative (1) containing pyridine moiety. Crystal structure of compound (3) was characterized by X-ray diffraction. New metal free and metallo-phthalocyanine complexes (Zn, Cu, and Mg) were synthesized using the phthalonitrile derivative (3). Cationic derivatives of these phthalocyanines (5, 7, 9, and 11) were prepared from the non-ionic phthalocyanines (4, 6, 8, and 10). All proposed structures were supported by instrumental methods. The aggregation behaviors of the phthalocyanines (4-11) were investigated in different solvents such as dimethylsulfoxide (DMSO), N,N-dimethylformamide (DMF), chloroform and water. Water soluble cationic Pcs (5, 7, 9, and 11) aggregated in water and sodium dodecyl sulfate was used to prevent the aggregation. The second derivatives of the UV-Vis spectra of aggregated Pcs were used for analyzing the Q and B bands of aggregated species. Thermal behaviors of the phthalocyanines were also studied. In addition, anti-bacterial properties of the phthalocyanines were investigated. We used four gram negative and two gram positive bacteria to determine antibacterial activity of these compounds. Compound 7 has the best activity against the all bacteria with 125 μg/mL of MIC value. Compounds 4, 6, and 10 have the similar effect on the bacteria with 250 μg/mL of MIC value.

Full text of DOI:10.1016/j.saa.2014.05.075

Accepted Manuscript
Synthesis, aggregation and spectroscopic studies of novel water soluble metal
free, zinc, copper and magnesium phthalocyanines and investigation of their
anti-bacterial properties
Rıza Bayrak, Hakkı Türker Akçay, Fatih Şaban Beriş, Ertan Şahin, Hacer
Bayrak, Ümit Demirbaş
PII:
S1386-1425(14)00866-X
http://dx.doi.org/10.1016/j.saa.2014.05.075
SAA 12237
DOI:
Reference:
Spectrochimica Acta Part A: Molecular and Biomo-
lecular Spectroscopy
To appear in:
Received Date:
Revised Date:
Accepted Date:
31 March 2014
15 May 2014
28 May 2014
Please cite this article as: R. Bayrak, H.T. Akçay, F. Beriş, E. Şahin, H. Bayrak, Ü. Demirbaş, Synthesis, aggregation
and spectroscopic studies of novel water soluble metal free, zinc, copper and magnesium phthalocyanines and
investigation of their anti-bacterial properties, Spectrochimica Acta Part A: Molecular and Biomolecular
Spectroscopy (2014), doi: http://dx.doi.org/10.1016/j.saa.2014.05.075
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Synthesis, aggregation and spectroscopic studies of novel
water soluble metal free, zinc, copper and magnesium
phthalocyanines and investigation of their anti-bacterial
properties
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Rıza Bayraka*, Hakkı Türker Akçayb, Fatih Şaban Berişc, Ertan Şahind, Hacer Bayrake, Ümit
Demirbaşe
aDepartment of Chemistry, Faculty of Arts and Sciences, Sinop University, 57000 Sinop,
Turkey
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bDepartment of Chemistry, Faculty of Art and Sciences, Recep Tayyip Erdoğan University,
53100 Rize, Turkey
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cDepartment of Biology, Faculty of Sciences, Recep Tayyip Erdogan University, 53100
Rize, Turkey
dDepartment of Chemistry, Faculty of Science, Atatürk University, 2524 Erzurum, Turkey
eDepartment of Chemistry, Faculty of Sciences, Karadeniz Technical University, 61100
Trabzon, Turkey
* Author for correspondence:
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Dr. Rıza BAYRAK, Department of Chemistry, Sinop University, 57000 Sinop, Turkey
Fax: 00 90 368 271 55 24
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E-mail: bayrakriza@ktu.edu.tr
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Abstract
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In this study, novel phthalonitrile derivative (3) was synthesized by the reaction between 4-
nitrophthalonitrile (2) and a triazole derivative (1) containing pyridine moiety. Crystal structure of
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compound (3) was characterized by X-ray diffraction. New metal free and metallo-phthalocyanine
complexes (Zn, Cu, and Mg) were synthesized using the phthalonitrile derivative (3). Cationic
derivatives of these phthalocyanines (5, 7, 9, and 11) were prepared from the non-ionic
phthalocyanines (4, 6, 8, and 10). All proposed structures were supported by instrumental methods.
The aggregation behaviors of the phthalocyanines (4-11) were investigated in different solvents such
as dimethylsulfoxide (DMSO), N,N-dimethylformamide (DMF), chloroform and water. Water soluble
cationic Pcs (5, 7, 9, and 11) aggregated in water and sodium dodecyl sulfate was used to prevent the
aggregation. The second derivatives of the UV-Vis spectra of aggregated Pcs were used for analyzing
the Q and B bands of aggregated species. Thermal behaviors of the phthalocyanines were also studied.
In addition, anti-bacterial properties of the phthalocyanines were investigated. We used four gram
negative and two gram positive bacteria to determine antibacterial activity of these compounds.
Compound 7 has the best activity against the all bacteria with 125 µg/mL of MIC value. Compounds
4, 6, and 10 have the similar effect on the bacteria with 250 µg/mL of MIC value.
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Keywords: Cationic phthalocyanine; X-ray diffraction; aggregation; sodium dodecylsulfate; thermal
stability; anti-bacterial activity.

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1. Introduction
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Phthalocyanines (Pcs) have many potential applications in many areas such as
electrochemistry [1-6], chemical sensors [7, 8], electrochromic devices, electrocatalysts [9],
liquid crystals [10], antimicrobial agents and photodynamic therapy [11-14]. Insolubility is
one of the most common problems in applications of Pcs. While non-substituted Pcs are
insoluble in common organic solvents, nonionic peripherally and non-peripherally Pcs mainly
are soluble in DMSO, DMF, pyridine, tetrahydrofuran (THF) – slightly soluble methanol,
ethanol, chloroform – and are not soluble in water, exceptionally polyethyleneglycol,
carbonhydrate, polyhydroxylated substituted Pcs [14]. Cationic (pyridinium) and anionic
(carboxylate, sulfonate) forms of peripherally and non-peripherally Pcs usually are soluble in
water. Synthesis of water soluble Pcs is important in terms of many technological
applications. Another problem limiting the use of Pcs in tech-applications is aggregation
phenomena resulting from co-planar interaction between the Pc rings. As a consequence of
this interaction, monomeric Pcs change to their dimeric or higher order forms. Aggregation
causing broadening and decreasing of the Q band absorption, depends on the concentration of
solution, type of the solvent, nature of the substituent, the type of the metal ion in central
cavity and temperature. [14-19].
In this work, novel peripherally substituted metal free, zinc, copper and magnesium Pc
complexes bearing triazole moiety (4, 6, 8, 10) and their cationic complexes (5, 7, 9, 11) have
been synthesized and investigated their spectroscopic properties, aggregation behaviors and
thermal stabilities. The aim of this work is not only determination of aggregation behaviors of
the novel Pc derivatives (4-11) and effect of sodium dodecyl sulfate (SDS) –anionic
surfactant- on the aggregation behavior of them in water, but also are how to effect on
bacterial growth or to determine their anti-bacterial activity.
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2. Experimental
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All reactions were carried out under dry and oxygen free nitrogen atmosphere using
schlenk system. DMF was dried and purified as described by Perrin and Armarego [20], 4-
Phenyl-5-pyridin-4-yl-4H-1,2,4-triazole-3-thiol (1) [21] and 4-nitrophthalonitrile (2) [22]
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were prepared as described in the literature. H-NMR/ C-NMR spectra were recorded on a
Varian XL-200 NMR spectrometer in CDCl3, DMSO d6, and chemical shifts were reported
(δ) relative to Me4Si as internal standard. IR spectra were recorded on a Perkin-Elmer
Spectrum One FT-IR spectrometer with ATR technique. The MS spectra were measured with
a Thermo Quantum Access Mass spectrometer with H-ESI probe. Methanol and chloroform
were used as solvents in mass analysis and all mass analysis were conducted in positive ion
mode. Elemental analysis was performed on a Costech ECS 4010 instrument, UV-Vis spectra
were recorded by Perkin Elmer Lambda 25 spectrometer, using 1 cm path length cuvettes at
room temperature. Thermal gravimetric analysis was performed with SII-TGA 6400
instrument in N2 atmosphere and gas flow rate was set up to 20 ml/min. Temperature rate was
set up to 10 ºC /min. Melting points were measured by an electrothermal apparatus.
2.1. Synthesis of the compounds (3-11)
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2.1.1. 4-Phenyl-5-pyridin-4-yl-4H-1,2,4-triazole-3-mercapto phthalonitrile (3)
4-Phenyl-5-pyridin-4-yl-4H-1,2,4-triazole-3-thiol (1) (0.81g, 3.2 mmol) and 4-
nitrophthalonitrile (2) (0.55 g, 3.2 mmol) were dissolved in dry N,N-dimethylformamide
(DMF) (10 ml) under N2 atmosphere. After stirring for 10 min at 55 ºC, dry finely powdered
potassium carbonate (0.48 g, 3.5 mmol) was added portion wise within 2 hours with efficient
stirring. The reaction mixture was stirred under N2 atmosphere at 55 ºC for 5 days. Then the
reaction mixture was poured into 250 ml ice-water and precipitate was filtered off, washed
with water, diethylether and dried in vacuum. Residue was crystallized from

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acetone/petroleum ether solvent system. Pale yellow solid was obtained. Yield 0,86 g
(%70,6), IR (ATR tablet) υmax / cm-1: 3098, 3070, 3037, 2228, 1582, 1546, 1493, 1460, 1428,
1364, 1328, 1315, 1288, 1257, 1215, 1126, 1071, 1007, 991, 916, 864, 838, 773, 739, 722,
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689. H-NMR (DMSO-d6), (δ: ppm): 8.63-8.58 (d, 2H/Ar–H), 8.04-7.99 (t, 3H/Ar–H), 7.74-
7.72 (d, 2H/Ar-H), 7.56-7.49 (m, 3H/Ar-H), 7.37-7.34 (d, 2H/Ar-H). 13C-NMR (DMSO-d6),
(δ: ppm): 154.31, 150.63, 147.46, 141.73, 135.51, 134.76, 134.21, 133.89, 132.60, 132.30,
131.05, 130.36, 128.23, 122.23, 116.13, 116.02, 115.74. MS (ESI), (m/z): [M+H]+
Calculated: 381.1 Found: 381.3.
2.1.2. General procedure for the synthesis of nonionic metal-free and
metallophthalocyanines (4, 6, 8, 10).
Compound (3) (0.38 g, 1 mmol), dry N,N-dimethylaminoethanol (DMAE) (4 ml), 1,8-
diazabicyclo [4.5.0]undec-7-ene (DBU) (3 drops) and for metallophthalocyanines (MPcs)
equivalent amounts of corresponding anhydrous metal salts (Zn(OAc)2, CuCl2, and MgCl2)
were added in a schlenk tube. The mixtures were heated at reflux temperature for 24 h under
N2 atmosphere. Then the mixtures were refluxed in methanol (30 ml) to precipitate the
products which were then filtered off. The green solid products were washed with hot ethanol
acetone and diethyl ether and dried in vacuo. The raw products were purified by silica gel
column chromatography.
2.1.2.1. Metal free Pc (4)
Elution solvent system: Chloroform: methanol (100:2) as eluent. Yield: 113 mg (30%).
Calc. for C84H50N24S4: C, 66.21; N, 22.24; S, 8.42 Found: C, 66.01; N, 23.95; S, 7.88 IR
(ATR) υmax / cm-1: 3288, 3057, 1601, 1497, 1429, 1327, 1007, 827, 746, 695, 1H-NMR
(DMSO-d6), (δ: ppm): 8.42-8.32 (bm, 8H/Ar–H), 8.10-8.02 (bm, 16H/Ar–H), 7.84-7.68 (bs,

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4H/Ar–H), 7.38-7.28 (bm, 16H/Ar-H), 7.10-7.24 (bs 4H/Ar-H). MS (ESI), (m/z): [M+H] +.
Calculated: 1511.7 (100%); Found: 1512.8. UV-vis (CHCl3) λmax/nm: [(10-5 ε, dm3 mol-1 cm-1
)] : 706 (5.09), 670 (5.05), 655 (4.71), 628 (4.57), 345 (4.92).
2.1.2.2. Zn(II)Pc (6)
Elution solvent system: chloroform: methanol (100:3) as eluent. Yield: 150 mg
(%37.8). IR (ATR νmax/cm−1): 3061, 1602, 1495, 1431, 1384, 1307, 1098, 908, 828, 749,
695. Calc. for C84H48N24S4Zn: C, 63.57; N, 21.18; S 8.08 Found: C, 63.48; N 21.49; S, 8.88
1H-NMR (DMSO-d6), (δ: ppm):8.50-8.38 (bm, 8H/Ar–H), 8.12-8.02 (bm, 16H/Ar–H), 7.86-
7.7 (bs, 4H/Ar–H), 7.42-7.30 (bm, 16H/Ar-H), 7.12-7.22 (bs 4H/Ar-H) MS (ESI), (m/z)
[M+H]+ calculated : 1585.3 (100%) ; found 1585.6. UV-vis (DMSO) λmax/nm: [(10-5 ε, dm3
mol-1 cm-1 )] : 687 (4.98), 620 (4.72), 362 (4.98).
2.1.2.3. Cu(II)Pc (8)
Elution solvent system: chloroform:methanol (100:5) as eluent. Yield: 123 mg (31%).
IR (ATR) νmax/cm−1:3056, 1602, 1496, 1430, 1396,1310, ,1257, 1143, 1100, 919, 745, 691
Calc. for C84H48N24S4Cu: C 63.64; N 21.21; S, 8.09 Found: C 64.49; N, 21.12; S 8.42 MS
(ESI), (m/z) [M+H]+ calculated : 1584.3 100% ; found 1584.4. UV-Vis (DMSO) λmax/nm:
[(10-5 ε, dm3 mol-1 cm-1)]: 683 (4.97), 648 (4.69), 362 (4.70).
2.1.2.4. Mg(II)Pc (10)
Elution solvent system: chloroform:methanol (100:2) as eluent. Yield: 114 mg (%28.5).
IR (ATR) νmax/cm−1: 3043, 1604, 1496, 1431, 1329, 1134, 1092, 1009, 970, 856, 828, 771,
748, 695 Calc. for C84H48N24S4Mg: C 65.26, N 21.74, S 8.30 Found: C 66.02, N 21.55, S
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8.50. H-NMR (DMSO-d6), (δ: ppm):8.36-8.22 (bm, 8H/Ar–H), 8.01-7.88 (bm, 20H/Ar–H),

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7.38-7.29 (bm, 16H/Ar-H), 7.04-7.24 (bs 4H/Ar-H) MS (HESI+), m/z [M+H]+ calculated :
1545.32 100% ; found 1545.66. UV-vis (DMSO) λmax/nm: [(10-5 ε, dm3 mol-1 cm-1 )] : 687
(4.99), 620 (4.25), 364 (4.55).
2.1.3.General Procedure for the synthesis of quarternized metal-free and
metallophthalocyanines (5, 7, 9, 11)
The nonionic metal free and metallophthalocyanines were dissolved in chloroform (10
ml) and excess amount of methyl iodide was added to these solutions. The reaction mixtures
were stirred at ambient temperature for two days. The green precipitates were filtered off,
washed with ethanol, acetone, diethyl ether and chloroform, and dried.
2.1.3.1. Quaternized metal free Pc (5)
IR (ATR) νmax/cm−1: 3292, 3041, 2925, 1642, 1605, 1495, 1451, 1367, 1309, 1008, 832,
746, 697 Calc. for C88H62I4N24S4: C, 50.54; N, 16.07; S,16.17; Found: C, 49.93; N, 16.73; S,
16.72, UV-Vis (DMSO) λmax/nm: [(10-5 ε, dm3 mol-1 cm-1)]: 702 (4.64), 666 (4.73), 643
(4.61), 346 (4.86)] Yield: %85.
2.1.3.2. Quaternized Zn(II) Pc (7)
IR (ATR) νmax/cm−1: 3048, 2923, 1641, 1488, 1418, 1383, 1332, 1308, 1139, 1097, 907,
744, 697, Calc. for C88H60I4N24S4Zn: C, 49.05; N, 15.60; S, 5.95 Found: C, 48.58; N 15.08; S,
5.91. UV-vis (DMSO) λmax/nm: [(10-5 ε, dm3 mol-1 cm-1 )] : 685 (4.82), 619 (4.52), 360 (4.83)
Yield: %91.

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2.1.3.3. Quaternized Cu(II) Pc (9)
IR (ATR) νmax/cm−1: 3033, 2925, 1639, 1602, 1493, 1449, 1365, 1340, 1141, 1098,
1043, 979, 836, 770, 745, 695 Calc. for C88H60I4N24S4Cu: C 49.04; N 15.61; S, 5.96 Found:
C 48.88; N,15.21; S 6,88 UV-vis (DMSO) λmax/nm: [(10-5 ε, dm3 mol-1 cm-1 )] : 684 (4.98),
616 (4.57), 352 (4.86) Yield: %88.
2.1.3.4. Quaternized Mg(II) Pc (11)
IR (ATR) νmax/cm−1: 3049, 2927, 1641, 1603, 1481, 1418, 1380, 1307, 1140, 1042,
1042, 908, 831, 770, 747, 696, Calc. for C88H60I4N24S4Mg: C 50.00, N 15.90, S 6.07 Found:
C 49.72, N 15.48, S 6.54. UV-vis (DMSO) λmax/nm: [(10-5 ε, dm3 mol-1 cm-1 )] : 687 (5.27),
618 (4.50), 364 (4.84) Yield: %88.
2.2. Crystal Structure of the Compound (3)
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For the crystal structure determination, the single crystal of the compound 3 was used
for data collection on a four-circle Rigaku R-AXIS RAPID-S diffractometer (equipped with a
two-dimensional area IP detector). The graphite-monochromatized Mo Kα radiation
(λ=0.71073 Å) and oscillation scans technique with ∆w= 5° for each image were used for data
collection. The lattice parameters were determined by the least-squares methods on the basis
of all reflections with F2>2σ(F2). Integration of the intensities, correction for Lorentz and
polarization effects and cell refinement was performed using CrystalClear (Rigaku/MSC Inc.,
2005) software [23]. The structures were solved by direct methods using SHELXS-97 [24]
and refined by a full-matrix least-squares procedure using the program SHELXL-97 [24]. H
atoms were positioned geometrically and refined using a riding model. The final difference
Fourier maps showed no peaks of chemical significance. Crystal data for compound 3:
C21H12N6S, crystal system, space group: monoclinic, P21; (no:4); unit cell dimensions:
a

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=10.8828(3), b = 7.0379(7), c = 12.3509(4) Å, β = 104.370(5); volume: 916.38(6) Å3; Z = 2;
calculated density: 1.38 g/cm3; absorption coefficient: 0.196 mm-1; F(000): 392; θ-range for
data collection 2.2-26.4°; refinement method: full-matrix least-square on F2; data/parameters:
3625/253; goodness-of-fit on F2: 1.023; final R indices [I > 2σ(I)]: R1 = 0.062, wR2 = 0.135; R
indices (all data): R1=0.098, wR2=0.152; largest diff. peak and hole: 0.298 and -0.254 e Å3;
Crystallographic data were deposited in CSD under CCDC registration number 941101.
These data can be obtained free of charge from The Cambridge Crystallographic Data Center
via www.ccdc.cam.ac.uk/data_request/cif.
2.3. Antibacterial activity
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The qualitative screening of the susceptibility spectra of different microbial strains to
the complexes was performed by the quantitative assay of minimal inhibitory concentration
(MIC, µg/mL) value was based on liquid medium serial micro-dilutions [25, 26]. The MIC
assays were performed in LB medium at pH 7.2. The stock solutions of compounds were
prepared with dimethyl sulfoxide (DMSO). Dilution series of chemical compounds to be
tested were prepared from 5000 to 39 µg/mL concentrations in 100 µL medium. The
microplates (Grainer, Germany) were incubated at 37.0
1 °C for 18–24 h.
Dimethysulphoxide, LB medium with or without antibiotic, ampicillin, were used as solvent
control, positive, and negative controls, respectively. The MIC was taken to be the last well in
the dilution series that did not exhibit growth as determined on the basis of turbidity.
The determination of minimum inhibitory concentration [26] was done with two of
Gram-positive bacterial strains i.e. Bacillus subtilis ATCC 66333 and Staphylococcus aureus
ATCC 25923, and seven Gram-negative bacterial strains i.e. Klebsiella pneumonie ATCC
13883, Pseudomonas aeruginosa ATCC 27853, Proteus vulgaris ATCC 13315, and
Enterococcus faecalis ATCC 29212 which were inoculated into Luria broth medium

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containing 1% tryptone, 0.5% yeast extract, 0.5% sodium chloride. The pH of the medium
was adjusted to 7.2 and incubated at 37 ºC for 18-24 h. The optical density of the bacteria
from mid-log phase of growth was measured at 600 nm and diluted in fresh medium so as to
get an optical density of 0.004 (corresponding to 5 x 105 colony forming units/mL). At the
end of incubation, the effect of the compounds on the growth of microorganisms was
determined. The MIC was defined as the lowest concentration of the compounds allowing no
visible growth.
3. Results and Discussion
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3.1. Synthesis and spectroscopic characterization of the compound (3)
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Synthesis of the phthalonitrile derivatives from 4-nitrophthalonitrile are reactions based
on nucleophilic aromatic substitution reaction with many examples in literature [27]. These
reactions are performed in polar aprotic solvents (DMSO or DMF) in the presence of
potassium carbonate as basic catalyst. Reaction temperature is kept at 55 ºC for five days. In
our study, the same reaction conditions were used for the synthesis of compound 3. The
synthesis of the compound 3 was shown at figure 1. Crude product was crystallized in
acetone/ethanol solvent system and yield of the reaction was moderate. After several
crystallization steps, single crystal suitable for X-ray analysis was obtained. In addition to the
analysis of the single crystal, spectral characterization was performed by IR, mass analysis.
All spectral data of the compound 3 were satisfactory. In IR spectral data, new vibration
obtained at 2228 cm-1 showed that compound 3 has the nitrile group. Mass spectral analysis of
compound 3 was supported the theoretical mass spectral data by [M+H]+ ion peak at 439.29.

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3.2. Description of the crystal structure of the compound (3)
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The ORTEP plot of the molecule and stacked geometry with the unit cell along the a-
axis of the structure 3 are shown in Fig. 2 and 3 respectively. The compound crystallizes in
the monoclinic space group P21 (no:4), with 2 molecules in the unit cell. It consists of 4-
Phenyl-5-pyridin-4-yl-4H-1,2,4-triazole-3-thiol and phthalonitrile units. The bond lengths of
cyano, C≡N in the structure are in range of 1.144-1.148 Å. Heterocyclic and phenyl rings are
not co-planar. The triazole ring is planar, maximum deviation from mean plane for N2 is
0.008 Å. Its plane forms dihedral angles of 62.4 (2), 83.7(2) and 31.1 (2)° with the mean
planes of the phenyl, phthalonitrile and pyridine rings, respectively. Triazole ring is in the
thiol tautomer form. Double bond between N3=C7=1.319 Å and N4=C14=1.312 Å and single
bond between C13-S1=1.743 Å is corresponding properties for its thiol form. These values
are comparable to similar molecules which contain the thione substituted 1,2,4-triazole group
[28]. The crystal structure is further stabilized by the π···π stacking interactions between the
symmetry related (-x,-1/2+y,2-z and 1-x,1/2+y,2-z) pyridin rings; the distance between
centroids of pyridin rings are the same [3.782(4) and 3.783(4) Å ]. In addition, C-H···π
[C18···Cgi(phenyl) =3.552(6) Å, C18-H..Cgi =132° symmetry code: (i) x,1+y,z ] interaction is
also observed.
3.3. Synthesis and spectroscopic properties of novel phthalocyanines (4-9)
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We used the most preferred method for the synthesis of phthalocyanines that includes
the cyclotetramerization of the 4-substituted phthalonitrile derivative in DMAE and DBU at
reflux temperature. This reaction results in the formation of four isomers but any separation
technique wasn’t carried out. [29]. ZnPc (6) and CuPc (8) and MgPc (10) metal complexes
were synthesized the same method using Zn(CH3COO)2 and CuCl2 and MgCl2 salts. Cationic
derivatives of these new phthalocyanines (5, 7, 9, 11) were synthesized using methyl iodide in

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chloroform. All phthalocyanine derivatives were characterized by using elemental analysis,
NMR, IR, UV-Vis and mass spectroscopic techniques and proposed structures were
supported the spectroscopic data.
In the IR spectra of the phthalocyanines (4-11), the proof of the cyclotetramerizations
-1
≡N
were the absence of the -C
stretching vibration observed at 2228 cm of the compound 3.
In addition, aliphatic C-H stretching vibrations in the IR spectra of the compounds (5, 7, 9,
11) showed that proposed cationic structures were synthesized. The main differences between
metal-free (4, 5) and metallophthalocyanines (6-11) are inner core N-H stretching vibrations
observed at 3288, 3292 cm-1 in the IR spectra, respectively. The rest of the spectra are similar
to that of compound 3.
The 1H-NMR spectra of CuPc (8) and its cationic derivative (9) couldn’t be determined
1
because of paramagnetic nature of Cu(II) cation [30]. H-NMR spectra of the other
phthalocyanine derivatives were obtained in DMSO-d6 at room temperature. The inner core
N-H protons couldn’t be observed because of strong aggregation between phthalocyanine ring
systems [31].
3.4. UV-Vis absorption spectra and aggregation studies of the compounds (4-11)
289
290
291
292
293
294
295
296
The UV-Vis spectrophotometry is a powerful technique for determination of the
structural properties and aggregation characteristics of the phthalocyanines. The absorption
spectra of the phthalocyanines give information about the symmetry of HOMO and LUMO
orbitals and aggregation type of molecule. Metallo phthalocyanines have two important bands
(Q and B) in their absorption spectra. The Q bands appear at the range of 600-750 nm and
consist of one sharp and intense band and two low-intense shoulders. The B bands appear at
the range of 250-400 nm as a broad band. In absorption spectra of metallo-phthalocyanines,
Q bands form the electronic transition between the HOMO and LUMO orbitals and B bands

13
297
298
299
300
301
302
303
304
305
306
307
308
309
310
311
312
313
314
315
316
317
318
319
320
321
form the electronic transition between deeper level of HOMO to LUMO orbitals. Symmetry
of the metallophthalocyanines is D4h and in case of metal free phthalocyanines, LUMO
orbital splits to two energy level called LUMO and LUMO+1 due to the absence of the
central metal. This splitting result in the formation of two sharp and intense bands (Qx, Qy) in
absorption spectrum and symmetry reduces to D2h.
All of the non-ionic Pc compounds (4, 6, 8, 10) were soluble in common organic
solvents such as chloroform, dichloromethane, DMSO, DMF, while the cationic Pcs (5, 7, 9,
11) were soluble in water, DMSO and DMF. The Q bands of the metal free phthalocyanine
(4) were observed at 707 nm (Qx) - 672 (Qy) nm and 656 nm with shoulder at 626 nm in
chloroform. The B band described the transition of deeper π levels to LUMO were observed
at 353 nm. This situation shows that the LUMO orbitals corresponding transition have D2h
symmetry (figure 4) [32].
UV-Vis absorption properties of non-ionic ZnPc (6) and MgPc (10) were studied in
DMSO, DMF, CHCl3, CH2Cl2 and THF. The compounds (6, 10) didn’t aggregate in these
solvents at 1 x 10-5 mol/dm3. The Q and Q(vib) bands were obtained at 687 nm and 620 nm,
respectively. The B band described the transition of deeper π levels to LUMO were obtained
at 362 and 364 nm (figure5). On the other hand, CuPc (8) aggregated in these solvents, so
determination of the Q and B bands was made by using second derivative of UV-Vis
spectrum in DMSO at 1 x 10-5 mol/dm3 concentration (figure 5). Second derivative of the UV-
Vis spectrum gives much enlightening data for the analysis of spectrum [32]. In the second
derivative of the UV-Vis spectrum of the CuPc (8), Q band and shoulder was obtained at 683,
643 and 615 nm, B band was obtained at 353 nm (figure 6).
While cationic metal free phthalocyanine (5) aggregated in DMSO, DMF, MeOH and
water, cationic Zn-, Cu-, Mg- Pcs (7, 9, 11) didn’t aggregate in DMSO and DMF at 1 x 10-5
mol/dm3 (figure 7). These phthalocyanine complexes (7, 9, 11) gave Q bands and the

14
322
323
324
325
326
327
328
329
330
331
332
333
334
335
336
337
338
339
340
shoulders at 685, 684, 687 nm and 619, 616, 618 nm respectively and the B bands of these
phthalocyanines (7, 9, 11) were obtained at 360, 352 and 363 nm, respectively. However, they
showed aggregation behavior at same concentration in water (figure 8a, 8b, 9a). Typically,
cationic phthalocyanines tend to form aggregates in water [33-36]. Surface-active materials
like Triton X, SDS (sodium dodecylsulfate), CTAB (cetyltrimethylammonium bromide) can
be used to prevent the aggregation in water. In this study, SDS was used as surface-active
material to prevent the aggregation and gave positive results for the cationic Zn and MgPc (7,
11) (figure 8a, 8b), but did not for the cationic metal-free and CuPcs (5, 9) (figure 9a and
10a).
Second derivative of the UV-Vis spectra of the cationic metal free and Cu Pcs (5, 9)
were used for analysis of Q and Q(Vib) bands, too. As can be seen in the UV-Vis spectra of the
cationic CuPc (9) in water (figure 9a), although SDS couldn’t prevent the aggregation, the
second derivative of the UV-Vis spectrum shows the Q band at 683 nm and Q(vib) bands at
642 and 621 nm (figure 9b). Similarly, for the cationic metal free phthalocyanine Pc (5),
aggregation couldn’t be prevented, although the addition of SDS. However the Q band are
seen at 708, 673 and Q(vib) bands are shown at 641, 617 nm, in the second derivative of the
UV-Vis spectrum (figure 10b). All spectral data are listed in table 1.
3.5. Thermal properties of the phthalocyanines (4-11)
341
342
343
344
345
346
Thermal behaviour of the synthesized phthalocyanine compounds were studied by
TGA/DT in N2 atmosphere and gas flow rate was set up to 20 ml/min. Temperature rate was
set up to 10 ºC /min. TG/DTA curves of cationic metal free Pc was shown in figure 11.
Generally, phthalocyanines are known as compounds which have high thermal stability
[37,38]. Synthesized phthalocyanines showed good thermal stability as expected. While the
cationic phthalocyanines (5, 7, 9, 11) decomposed at two step, non-ionic phthalocyanines (4,

15
347
348
349
350
351
352
6, 8 ,10) decomposed at one step. In the first step of the decomposition of cationic
phthalocyanine, it can be supposed that loss of mass belonged to methyl iodide. It could not
be made prediction about other decomposition steps. Thermal gravimetric data were shown in
table 2.
3.6. Antibacterial properties of the compounds (4-11)
353
354
355
356
357
358
359
360
361
362
363
364
365
366
367
368
369
370
371
All the synthesized compounds were tested against three Gram-positive and negative
bacterial strains according to the literature protocol [25, 26]. The results were compared with
standard drug ampicillin as depicted in table 3. The investigation of antibacterial activity
(Table 3) revealed that all the newly synthesized compounds showed good to moderate
minimum inhibitory concentration at 1.95-4000 µg/mL in DMSO and double distilled water.
First of all, we used precursor compound (3) that the other substances to be
synthesized were differentiated from it, against all bacteria were used. According to
antibacterial activity, compound 3 has the same activity with 1 mg/mL against to all bacteria
except for K. pneumonie ATCC 13883. Compound 3 has MIC value of 500 µg/mL against K.
pneumonie ATCC 13883. We showed that the lowest concentration to effect on all bacteria
belongs to compound 7. Thus, compound 7 was the better activity with 125 µg/mL.
Compounds 3, 6, and 10 have the same activity against the all bacteria with MIC value of 250
µg/mL. Compounds 5 and 9 have the same activity with MIC value of 250 µg/mL against
gram-negative bacteria. But these compounds have different MIC values against the gram-
positive bacteria with MIC value of 500 µg/mL to B. subtilis and 250 µg/mL S. aureus of
compound 5, although compound 9 has the reverse activity on the bacteria, as MIC value of
250 µg/mL to B. subtilis and 500 µg/mL S. aureus. Compound 11 has two MIC value of 250
µg/mL against Pseudomonas aeruginosa ATCC 27853, Proteus vulgaris ATCC 13315,
Bacillus subtilis ATCC 66333 and Staphylococcus aureus ATCC 25923, and 500 µg/mL

16
372
373
374
375
376
377
378
379
against to others. Compound 8 has two MIC value with different effects on bacteria like as
compound 11. It has 250 µg/mL against Enterococcus faecalis ATCC 29212, Proteus
vulgaris ATCC 13315, and Bacillus subtilis ATCC 66333, and 500 µg/mL against to others.
In all results of antimicrobial activity of compounds, compound 7 has the best activity to all
bacteria we used.
According to antibacterial activity of compound 3 and its derivatised substances, all
substances were more effective than compound 3. Only compound 1 and 2 were the same
activity against K. pneumonie ATCC 13883 as compound 3. Expecially the lowest
concentration of compound 6 can influence than the others.
380
381
382
Conclusion
383
384
385
386
387
388
389
390
391
392
393
394
395
In this study, novel phthalonitrile derivative (3) was synthesized and its single crystal
suitable for X-ray analysis was obtained. Novel phthalocyanine derivatives (4-11) were
synthesized by using the compound (3). The cationic derivatives of the phthalocyanines were
synthesized by the reaction of non-ionic phthalocyanines (4, 6, 8, 10) and methyl iodide in
chloroform. All spectral analysis supported the proposed structures. Aggregation studies of
the phthalocyanines were performed in different solvents. Thermal gravimetric analysis and
anti-bacterial properties of the phthalocyanines were performed.
Although metal free phthalocyanine (4) aggregated in DMSO, didn’t aggregate in
chloroform. CuPc (8) aggregated in all solvents, so second derivative of the UV-Vis spectra
of the CuPc (8) was calculated for analyzing the absorbtion bands. Q and B bands were
obtained clearly at 683, 643, 615 nm and 353 nm respectively. Zn and Mg Pcs were not
aggregate in DMSO. All water soluble phthalocyanines aggregated in water. SDS was used to
prevent the aggregation. The aggregation could be prevented for cationic Zn and MgPcs (7,

17
396
397
398
399
400
401
402
11), but it couldn’t be prevented for cationic metal free and CuPcs (5, 8). The analysis of
absorbtion bands were performed by their second derivatives of the UV-Vis spectra.
The thermal gravimetric analysis of the phthalocyanines was performed by TG/DTA
system. In cationic phthalocyanines (5, 7, 9, 11), it was proposed that methyl iodide moieties
were separated from the phthalocyanine ring system at below the expected main
decomposition temperature. We see that all compounds have moderately effect on bacteria
strains used in our study. In all results of antimicrobial activity of compounds, compound 7
has the best activity to all gram negative and positive bacteria we used.
403
404

18
405
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409
410
411
412
413
414
415
416
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429
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Acknowledgement
Authors thank to Sinop University, Recep Tayyip Erdoğan University and Karadeniz
Technical University.

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Table 1. The UV-Vis spectral data of the phthalocyanines in different solvents
Compounds
Free Pc
Q bands (nm) B bands (nm)
Solvents
CHCl3
4
5
6
707 - 672
708 - 673
687
353
348
362
360
352
353
Cat. Free Pc
ZnPc
Water/SDS*
DMSO
685
DMSO
7
8
Cat. ZnPc
CuPc
Water/SDS
DMSO*
684
683
DMSO
684
352
9
Cat. CuPc
MgPc
683
687
687
687
348
364
363
356
Water/SDS*
DMSO
10
11
DMSO
Cat. MgPc
Water/SDS
(* the spectral data were obtained the second derivatives of the corresponding UV-Vis spectra)
486
487
488
489
490
491

22
492
493
494
495
496
Table 2. Thermal gravimetric parameters for synthesized phthalocyanines (4-11)
Decomp. Range
Temp ºC
Main Decomp.
Temp ºC
369ºC
Weight Loss %
Weight Loss
Compounds
(measured)
% (calculated)
4
Free Pc
Cationic
Free Pc
ZnPc
275-522ºC
170-260ºC
259-538ºC
292-613ºC
173-265ºC
313-449ºC
265-484ºC
171-220ºC
327-454ºC
275-470ºC
160-297ºC
297-442ºC
38
28
32
36
25
28
31
24
26
34
30
24
---
27.1
---
209ºC
5
6
371ºC
351ºC
---
Cationic
ZnPc
212ºC
26.3
---
7
372ºC
8
CuPc
392ºC
---
Cationic
CuPc
205ºC
26.3
---
9
373ºC
10
11
MgPc
382ºC
---
Cationic
MgPc
230ºC
26.8
---
366ºC
497
498
499
500
501

23
502
503
504
505
Table 3. In-Vitro Antibacterial Activity Data of Compounds (4-11)
Microorganisims and Minimal Inhibition Concentration
(MIC) Value
Stock
Number
Compound
Solution
(µg/mL)
Gram-Positive
Bacteria
Gram-Negative Bacteria
Ef
Kp
Pa
Pv
Bs
Sa
Free Pc
4
10000
250
250
250
250
250
250
6.56 E-4
Free cationic Pc
5
6
7
8
9
10000
10000
10000
10000
10000
250
250
125
250
250
250
250
125
500
250
250
250
125
500
250
250
250
125
250
250
500
250
125
250
250
250
250
125
500
500
4.76. E-4
Zn Pc
6.3 E-4
Zn cationic Pc
4.64. E-4
Cu Pc
6,3. E-4
Cu cationic Pc
4,64. E-4
Mg Pc
10
10000
10000
250
250
250
250
250
250
6,46. E-4
Mg cationic Pc
11
DMSO
PC + amp.
PC
500
NE
-
500
NE
-
250
NE
-
250
NE
-
250
NE
-
250
NE
-
100
+
+
+
+
+
+
Ef: Enterococcus faecalis ATCC 29212, Kp: Klebsiella pneumonie ATCC 13883, Pa: Pseudomonas aeruginosa ATCC
27853, Pv: Proteus vulgaris ATCC 13315, Bs: Bacillus subtilis ATCC 66333, Sa: Staphylococcus aereus ATCC 25923.
PC: positive control as broth medium without chemical component, amp: ampicilline, NE: not effected for growing.
506
507
508

24
509
510
511
512
513
514
515
516
517
518
519
520
521
522
523
524
525
526
527
528
529
530
531
532
FIGURE CAPTURES
Figure 1. Synthetic route of novel compounds.
Figure 2. The molecular structure of 3 and the atom numbering scheme. Ellipsoids for non-
hydrogen atoms are drawn at the 50% probability level.
Figure 3. Stacked geometry with the unit cell along the a-axis of the structure 3
Figure 4. Absroption spectra of metal free Pc (4) in CHCl3 and DMSO at 1x10-5 mol/dm3
Figure 5.Absorbtion spectra of Zn and MgPcs (6, 10) in DMSO at 1x10-5 mol/dm3
Figure 6. (a) Absrobtion spectra CuPc (8) in different solvent at 1x 10-5 mol/dm3
(b) Second derivative of the UV-Vis spectrum of the CuPc (8) in DMSO
Figure 7. UV-Vis spectra of the Zn, Cu, Mg cationic phthalocyanines (7, 9, 11) in DMSO at
1 x 10-5mol dm-3
Figure 8. (a) UV-Vis spectra of the cationic ZnPc(7) in water (1x10-5mol/dm3) at different
SDS concentration,
(b) UV-Vis spectra of the cationic MgPc(11) in water (1x10-5mol/dm3) at
different SDS concentrations.
Figure 9. (a) UV-Vis spectra of the cationic CuPc (9) in water (1x10-5mol/dm3) and effect of
SDS
(b)Second derivative of the UV-Vis spectrum of the cationic CuPc (9) in water
(SDS concentration: 1.67x10-3 mol/dm3).
Figure 10. (a) UV-Vis spectra of the cationic metal free phthalocyanine (5) in water (1x10-
5mol/dm3) at different SDS concentrations, (b) Second derivative of the UV-Vis
spectrum of the cat.metal free phthalocyanine (5) in water (SDS concentration=
2.04x10-3 mol/dm3).

25
533
534
Figure 1. Synthetic route of novel compounds.

26
535
536
537
538
539
540
541
542
543
544
545
Figure 2. The molecular structure of 3 and the atom numbering scheme. Ellipsoids for non
hydrogen atoms are drawn at the 50% probability level.
546
547
548
549
550
551
552

27
553
554
555
556
557
558
559
Figure 3. Stacked geometry with the unit cell along the a-axis of the structure 3
560
561
562
563
564
565
566

28
567
568
569
570
571
572
573
574
Figure 4. Absroption spectra of metal free Pc (4) in CHCl3 and DMSO at 1x10-5 mol/dm3
575
576
577
578
579
580
581

29
582
583
584
Figure 5. Absorbtion spectra of Zn and MgPcs (6,10) in DMSO at 1x10-5 mol/dm3