Dyes and Pigments 102 (2014) 35e45

Contents lists available at ScienceDirect

Dyes and Pigments

journal homepage: www.elsevier.com/locate/dyepig

A pH sensitive and selective ratiometric PAMAM wavelength-shifting

bichromophoric system based on PET, FRET and ICT
Nikolai I. Georgiev a, Abdullah M. Asiri b, c, Abdullah H. Qusti b, Khalid A. Alamry b,
Vladimir B. Bojinov a, b, *
a
Department of Organic Synthesis, University of Chemical Technology and Metallurgy, 8 Kliment Ohridsky Str., 1756 Sofia, Bulgaria
b
Chemistry Department, Faculty of Sciences, King Abdulaziz University, P.O. Box 80203, Jeddah 21589, Saudi Arabia
c
Center of Excellence for Advanced Materials Research (CEAMR), King Abdulaziz University, P.O. Box 80203, Jeddah 21589, Saudi Arabia

a r t i c l e i n f o a b s t r a c t

Article history: A fluorescent poly(amidoamine)-based wavelength-shifting bichromophore was synthesized using

Received 30 August 2013 divergent strategy and characterized photophysically. Novel compound was configured as a light-har-
Received in revised form vesting antenna where the systems surface is labelled with yellow-green emitting 4-(N-piperazinyl)-1,8-
30 September 2013
naphthalimide “donor” units capable of absorbing light and efficiently transferring the energy to a focal
Accepted 3 October 2013
Available online 25 October 2013
Rhodamine 6G “acceptor”. The energy transfer was calculated to 91%. Moreover, the 1,8-naphthalimide
periphery of the system was designed on the “fluorophoreespacerereceptor” format able to act as a
molecular fluorescence photoinduced electron transfer based probe. Due to the both effects, photoin-
Keywords:
1,8-Naphthalimide
duced electron transfer in the periphery and pH dependent spirolactam to ring-opening amide equi-
Rhodamine 6G librium of rhodamine core, novel bichromophoric system acts as a selective ratiometric pH fluorescence
PAMAM dendrimer probe. Thus, the distinguishing features of light-harvesting (fluorescence resonance energy transfer)
Selective fluorescence pH probes systems were successfully combined with the properties of classical photoinduced electron transfer and
Light-harvesting FRET-based system ring-opening sensing systems.
Photoinduced electron transfer (PET) Ó 2013 Elsevier Ltd. All rights reserved.

1. Introduction two different dye molecules in which excitation is transferred from

a donor molecule to an acceptor molecule without emission of a
In the past few decades, a great interest has been focused on a photon [16,17]. Fluorescent properties of the FRET-based multi-
supramolecular fluorescent systems in which fluorescence fluorophoric systems are very promising because of the long
switches between “on” and “off” states as these can be modulated, communication wavelengths exhibited by these molecules. Long-
or tuned, by employing external sources such as ions, molecules, wavelength excitation reduces problems of autofluorescence and
light, etc. [1,2]. scattering during fluorescent sensing within many biological and
The widespread use of fluorescent sensors was prompted by the industrial matrices [18]. Also it would be possible to fabricate a
increasing need of fast and reliable sensing of chemical species in ratiometric probe based on the FRET mechanism, in which the ratio
many areas of human activity [3e9]. Different design strategies are of the fluorescent intensities at two different wavelengths provides
being employed in the development of molecular fluorescent sen- a built-in correction for environmental effects, and stability under
sors, for instance PET (photoinduced electron transfer) based sensors illumination [19]. This method allows precise and quantitative
[10,11], CT (charge transfer) sensor [12], ET (energy transfer) sensors analysis and imaging even in complicated systems [20].
[13], ring-opening sensors [14,15]. PET using the “fluorophoree Molecular systems capable of light-harvesting and efficiently
spacerereceptor” format is the most commonly exploited approach transferring absorbed radiation unidirectionally over a nanometer
for the design of molecular fluorescent sensors and switches [1]. distance are of great interest [21]. The most attractive artificial
Fluorescence resonance energy transfer (FRET) is a distance- light-harvesting systems are the dendritic assemblies because of
dependent interaction between the electronic excited states of their unique structures, reminiscent of the architecture of natural
light-harvesting complexes [21e27]. The globular shape of den-
dritic architectures provides a large surface area that can be deco-
* Corresponding author. Department of Organic Synthesis, University of Chemical
rated with chromophores, resulting a large absorption cross section
Technology and Metallurgy, 8 Kliment Ohridsky Str., 1756 Sofia, Bulgaria. Tel.: þ359
2 8163206. and efficient capture of photons. Furthermore, the use of the flex-
E-mail address: vlbojin@uctm.edu (V.B. Bojinov). ible aliphatic PAMAM bone as a scaffold for light-harvesting

0143-7208/$ e see front matter Ó 2013 Elsevier Ltd. All rights reserved.
http://dx.doi.org/10.1016/j.dyepig.2013.10.007
36 N.I. Georgiev et al. / Dyes and Pigments 102 (2014) 35e45

antennae could give new systems with high efficiency of energy recorded on a Scinco FS-2 fluorescence spectrophotometer. The
transfer [28]. excitation source was a 150 W Xenon lamp. Excitation and emission
Particularly useful for PAMAM antennae are fluorophores such slits width were 5 nm. Fluorescence measurement was carried out
as Rhodamine 6G and 1,8-naphthalimide [29e35]. Because of their in right angle sample geometry. A 1  1 cm quartz cuvette was used
excellent fluorescence properties and good photostability, 1,8- for the spectroscopic analysis. Relative fluorescence quantum yields
naphthalimide dyes were used extensively in a number of areas, (FF) were measured using Rhodamine 6G (FF ¼ 0.95 in ethanol
including chemosensing materials [36e58]. On the basis of the [71]) or Coumarin 6 (FF ¼ 0.78 in ethanol [72]) as standards. All
spirolactam (non-fluorescent) to ring-open amide (fluorescent) experiments were performed at room temperature. The spectral
equilibrium of rhodamine, series of rhodamine-based dyes with data were collected using FluoroMaster Plus 1.3 and further pro-
excellent “offeon” switching of fluorescence upon encountering cessed by OrginPro 6.1 software. A pH meter Metrohm 704 coupled
the correct target have been synthesized [20,59e69]. with combined pH electrode was used for pH measurements. The
In this paper, we report on the design, synthesis and photo- commercial standard buffers for pH 2, 7 and 10 (Aldrich) were used
physical properties of a novel ratiometric fluorescence “offeon” for calibration. TLC was performed on silica gel, Fluka F60 254,
light-harvesting antenna based on a core and peripherally func- 20  20, 0.2 mm. The melting points were determined by means of
tionalized PAMAM dendron (Scheme 1). The decorated with a Kofler melting point microscope.
yellow-green emitting 1,8-naphthalimide units dendron periphery The absorption and fluorescence properties were studied as a
is capable of absorbing light and efficiently transferring the energy function of pH by multiple additions of NaOH and HCl aqueous
to a focal Rhodamine 6G. Also, the peripheral 1,8-naphthalimides solutions to 400 mL 1  105 M solution of examined compounds in
were designed on a “fluorophoreespacerereceptor” format thus water/DMF (4:1, v/v). The addition was limited to 1 mL so that
providing PET based sensing properties of the novel light-har- dilution remains insignificant. The solution pH, absorption and
vesting antenna. fluorescence spectra were recorded at each addition. The effect of
the metal cations was examined by addition of 0.2 mL of ammonia/
2. Experimental ammonium chloride buffer (pH 10) solution to 2 mL water/DMF
(4:1, v/v) solution of the corresponding fluorophore (1  105 M)
2.1. Materials and 20 mL of the metal stock solution (1  102 M).

Commercially available Rhodamine 6G 1, ethylenediamine, N- 2.3. Synthesis of Rhodamine 6G core (2)

methylpiperazine and methyl acrylate (Aldrich, Merck) were used
without purification. The starting 4-nitro-1,8-naphthalic anhydride To a solution of Rhodamine 6G 1 (2.3 g, 4.6 mM) in 90 mL of
5 was prepared according to the reported procedure [70]. All sol- absolute ethyl alcohol, 1.8 mL of ethylenediamine (28 mM) was
vents (Fluka, Merck) were pure for analysis or of spectroscopy added dropwise at room temperature. The resulting solution was
grade. NaOH and HCl were supplied by Merck (Germany). stirred at reflux for 5 h. After cooling to room temperature the solid
Ammonia buffer solution (ammonia/ammonium chloride, pH 10, precipitated was filtered off, washed with water and dried to give
Aldrich), metal stock solutions (1  104 M) of Zn(NO3)2, Cu(NO3)2, 1.9 g 1.9 g (88%) of 2 as pale pink crystals (m.p. >250  C, Rf ¼ 0.45 in
Ni(NO3)2, Co(NO3)2, Pb(NO3)2, Fe(NO3)3, Cd(NO3)2, AgNO3 and a solvent system chloroform/ethylacetate/ethanol ¼ 1:1:1). FT-IR
Hg(NO3)2 in DMF (all Aldrich salts at p.a. grade) and working dye (KBr) cm1: 3220 (nNH and nNH2); 2942, 2848 (nCH); 1678 (nC]
solutions (1  105 M) were prepared daily. O); 1634, 1528 and 1484 (nAr]CH). 1H NMR (CDCl3-d, 250.13 MHz)
ppm: 7.93 (m, 1H, 9-Ph H-3); 7.47 (m, 2H, 9-Ph H-4 and 9-Ph H-5);
2.2. Methods 7.06 (m, 1H, 9-Ph H-6); 6.34 (s, 2H, Rhodamine H-4 and H-5); 6.22
(s, 2H, Rhodamine H-1 and H-8); 3.51 (br.s, 2H, 2  ArNH); 3.21 (m,
FT-IR spectra were recorded on a Varian Scimitar 1000 spec- 6H, 2  CH2CH3 and CH2NCO); 2.35 (t, 2H, J ¼ 6.8 Hz, CH2NH2); 1.90
trometer. The 1H NMR spectra (chemical shifts are given as d in (s, 6H, 2  ArCH3); 1.32 (m, 8H, 2  CH2CH3 and NH2). Elemental
ppm) were recorded on a Bruker DRX-250 spectrometer, operating analysis: Calculated for C28H32N4O2 (MW 456.58) C 73.66, H 7.06, N
at 250.13 MHz. Absorption spectra were recorded on a Hewlett 12.27%; Found C 74.19, H 7.08, N 12.36%.
Packard 8452A spectrophotometer. Fluorescent spectra were
2.4. Synthesis of ester-functionalized Rhodamine 6G (3)

To a suspension of 2 (1.82 g, 4 mM) in 40 mL of cooled to 0  C

N N
N O O N
methanol, a solution of methyl acrylate (3.6 mL, 40 mM) in 4 mL of
methanol was added dropwise over a period of 30 min. The reac-
N N
tion mixture was allowed to warm slowly to room temperature and
O O then stirred for 3 days. The final product was obtained after evap-
NH HN
oration of methyl acrylate and methanol under vacuum as white
O O
crystals (yield e 2.46 g (98%), m.p. 134e136  C, Rf ¼ 0.66 in a solvent
N
NH system toluene/ethanol ¼ 2:1). FT-IR (KBr) cm1: 3324 (nNH); 2942,
2896 (nCH); 1728 (nCOOMe); 1670 (nC]O); 1622, 1518 and 1450
N O (nArCH). 1H NMR (CDCl3-d, 250.13 MHz) ppm: 7.89 (m, 1H, 9-Ph H-
O 3); 7.44 (m, 2H, 9-Ph H-4 and 9-Ph H-5); 7.05 (m, 1H, 9-Ph H-6);
6.35 (s, 2H, Rhodamine H-4 and H-5); 6.20 (s, 2H, Rhodamine H-1
and H-8); 3.59 (s, 6H, 2  OCH3); 3.48 (br.s, 2H, 2  ArNH); 3.21 (q,
NH 4H, J ¼ 7.1 Hz, 2  ArNHCH2); 2.58 (t, 4H, J ¼ 7.2 Hz,
2  CH2COOCH3); 2.22 (t, 2H, J ¼ 7.2 Hz, CH2NCOAr); 2.19 (m, 6H,
7 N(CH2)3); 1.89 (s, 6H, 2  ArCH3); 1.32 (t, 6H, J ¼ 7.1 Hz, 2  CH2CH3).
Scheme 1. PAMAM light-harvesting dendron 7, core and peripherally functionalized Elemental analysis: Calculated for C36H44N4O6 (MW 628.76) C
with Rhodamine 6G and 4-(N-methylpiperazinyl)-1,8-naphthalimides. 68.77, H 7.05, N 8.91%; Found C 68.96, H 6.99, N 8.96%.
 N.I. Georgiev et al. / Dyes and Pigments 102 (2014) 35e45 37

2.5. Synthesis of amino-functional Rhodamine 6G dendron (4) The choice of 1,8-naphthalimide for the fluorescence modification
of PAMAM periphery was prompted by its chemical stability and
To a solution of ethylenediamine (12.5 mL, 18 mM) in 10 mL of high fluorescence efficiency [73]. On the basis of the spirolactam
cooled to 0  C methanol, a suspension of ester-functionalized (non-fluorescent) to ring-open amide (fluorescent) equilibrium of
rhodamine 3 (1.88 g, 3 mM) in 15 mL of methanol was added rhodamine, rhodamine-based dyes are excellent “offeon” fluores-
over a period of 30 min. The resulting mixture was stirred for 7 days cence sensing molecules [59e68]. A requirement for efficient en-
at room temperature. Then the solvent and the ethylenediamine ergy transfer is that there should be a spectral overlap between the
excess were distilled under vacuum. Final traces of excess ethyl- emission of the peripheral donor dyes and the absorbance of the
enediamine were removed azeotropically using a 9:1 toluene/ focal acceptor chromophore. It is well-known that rhodamines are
methanol (v/v) solution. The final compound was obtained as white orange-red emitting fluorophores with maximal absorption in the
crystals (yield - 1.98 g (96%), m.p. 88e90  C, Rf ¼ 0.27 in a solvent visible region at about lA ¼ 520e530 nm, where the emission of 4-
system n-propanol/ammonium hydroxide ¼ 1:1). FT-IR (KBr) cm1: alkylamino-1,8-naphthalimides is appeared [30,31]. Consistent
3314 and 3204 (nNH and nNH2); 2922, 2878 (nCH); 1644 (nC]O); with this requirement, 4-alkylamino-1,8-naphthalimides and
1636 and 1490 (nAr]CH). 1H NMR (CDCl3-d, 250.13 MHz) ppm: 7.88 rhodamine dyes are suitable fluorescence donor-acceptor pair for
(m, 1H, 9-Ph H-3); 7.47 (m, 2H, 9-Ph H-4 and 9-Ph H-5); 7.34 (t, 2H, construction of wavelength-shifting bichromophoric systems
J ¼ 5.7, 2  NHCO); 7.06 (m, 1H, 9-Ph H-6); 6.36 (s, 2H, Rhodamine [19,61,66].
H-4 and H-5); 6.16 (s, 2H, Rhodamine H-1 and H-8); 3.64 (m, 4H, In order to impart “offeon” properties to the novel light-har-
2  CONHCH2CH2NH2); 3.55 (br.s, 2H, 2  ArNH); 3.21 (m, 8H, vesting dendron, yellow-green emitting 4-(N-methylpiperazinyl)-
2  NH2 and 2  ArNHCH2); 3.11 (m, 2H, CH2NCOAr); 2.73 (m, 4H, 1,8-naphthalimides were used as peripheral donors. The 4-(N-
2  CH2NH2); 2.55 (t, 4H, J ¼ 6.1 Hz, 2  CH2CH2CONH); 2.15 (m, 6H, methylpiperazinyl)-1,8-naphthalimides are well known “offeon”
N(CH2)3); 1.90 (s, 6H, 2  ArCH3); 0.93 (t, 6H, J ¼ 7.2 Hz, fluorescent probes based on “fluorophoreespacerereceptor”
2  CH2CH3). Elemental analysis: Calculated for C38H52N8O4 (MW format, which fluorescence is quenched and can be switched “on”
684.87) C 66.64, H 7.65, N 16.36%; Found C 66.96, H 7.49, N 16.58%. by coupling with the analyte (protons or different metal ions)
[73,74]. In this particular case, it could be predicted that a PET
2.6. Synthesis of light-harvesting dendron (7) process (an electron transfer from the receptor to the excited state
of the peripheral fluorophore) would quench fluorescence of the
To a solution of 0.97 g (4 mM) of 4-nitro-1,8-naphthalic anhy- peripheral 1,8-naphthalimide unit. Because of the quenched fluo-
dride 5 in 40 mL of boiling methanol, a solution of 1.37 (2 mM) of rescence of the periphery the energy transfer in light-harvesting
Rhodamine 6G dendron 4 in 25 mL of methanol was added drop- dendron 7 is disabling and this would represent the “off-state” of
wise under stirring over a period of 2 h. The resulting solution was the system. Protonation of the N-methylpiperazine receptor would
refluxed for 6 h. After cooling the brown precipitate was filtered off, increase its oxidation potential, and as such, thermodynamically
treated with 50 mL of 5% aqueous sodium hydroxide to give after disallow the electron transfer to the peripheral 1,8-naphthalimides.
filtration and drying an intermediate 8. Then to the solution of the Consequently the energy transfer in light-harvesting dendron 7
obtained intermediate 8 (0.68 g, 0.6 mM) in 12 mL of DMF, N- would be “switched on” (Scheme 2).
methylpiperazine (0.48 g, d ¼ 0.91, 4.8 mM) was added portion wise Due to the “offeon” sensing properties of yellow-green emitting
at room temperature. After 48 h the resulting solution was poured periphery and pH sensitive character of Rhodamine 6G acceptor
into water. The crude product that precipitate was filtered off, dye, we expect the fluorescence signal of the novel system to be
washed with fresh water and dried. Silica gel column chromatog- function of pH. At pH >6 the Rhodamine 6G derivatives are in
raphy (methanol/chloroform ¼ 1:2, v/v) afforded 0.60 g (81%) yel- colourless spirolactam closed form and the energy transfer from
low solid of light-harvesting dendron 7 (81%, m.p. 188e190  C, the peripheral 1,8-naphthalimes to the rhodamine core in the light-
Rf ¼ 0.54 in a solvent system n-propanol/ammonium harvesting antenna is not feasible (Scheme 2). It could be expected
hydroxide ¼ 1:1). FT-IR (KBr): 3384 (nNH); 2966, 2936 (nCH); 1692 that under alkaline conditions due to the spirolactam closed form of
(nasNeC]O); 1654 (nsNeC]O). 1H NMR (CDCl3-d, 250.13 MHz) Rhodamine 6G and PET effect in the periphery the emission of
ppm: 8.30 (d, 2H, J ¼ 7.5 Hz, 2  naphthalimide H-5); 8.22 (d, 2H, light-harvesting dendron 7 will be quenched. In acid media
J ¼ 8.6 Hz, 2  naphthalimide H-2); 8.09 (d, 2H, J ¼ 8.4 Hz, (pH ¼ 2e6) the Rhodamine 6G spirolactam ring is opened and the
2  naphthalimide H-7); 7.84 (m, 1H, 9-Ph H-6); 7.56 (m, 2H, PET process in periphery is disallowed. Thus the energy of the
2  NHCO); 7.45 (m, 4H, 2  naphthalimide H-6, 9-Ph H-4 and 9-Ph peripheral 1,8-naphthalimides in antenna 7 will be transferred to
H-5); 7.01 (m, 3H, 2  naphthalimide H-3 and 9-Ph H-3); 6.34 (s, the focal Rhodamine and the systems will emit orange-red fluo-
2H, Rhodamine H-4 and H-5); 6.14 (s, 2H, Rhodamine H-1 and H-8); rescence signal.
4.17 (t, 4H, J ¼ 6.4 Hz, 2  (CO)2NCH2); 3.56 (m, 4H, 2  CONHCH2); The novel light-harvesting antenna was prepared in three basic
3.20 (m, 16H, 2  Rhodamine ArNHCH2, 2  Rhodamine ArNHCH2, steps (Scheme 3): synthesis of amino-functional Rhodamine 6G
CH2NCOAr and 4  piperazine ArNCH2); 2.70 (m, 8H, 4  piperazine core, PAMAM dendronization of the amino-functional core to an
MeNCH2); 2.48 (m, 10H, 2  CH2CH2CONH and 2  piperazine CH3); amino-terminated Rhodamine 6G PAMAM dendron and peripheral
2.16 (m, 6H, N(CH2)3); 1.84 (s, 6H, 2  Rhodamine ArCH3); 1.28 (t, decoration of the latter with yellow-green emitting 4-(N-methyl-
6H, J ¼ 7.0 Hz, 2  Rhodamine CH2CH3). Calculated for C72H80N12O8 piperazinyl)-1,8-naphthalimide units to the desired antenna.
(MW 1241.48) C 69.66, H 6.50, N 13.54%; Found C 70.01, H 6.67, N In brief, the synthesis of Rhodamine 6G core 2 was performed by
13.78%. reaction of Rhodamine 6G 1 with ethylenediamine under reflux in
absolute ethyl alcohol. Then the rhodamine core 2 was subse-
3. Results and discussion quently converted into the PAMAM dendron 4 via divergent strat-
egy involves initial Michael addition of 2 with methyl acrylate
3.1. Design and synthesis of light-harvesting antenna 7 followed by exhaustive amidation of the resulting ester 3 with a
large excess of ethylenediamine. The target light-harvesting an-
The desired PAMAM dendron 7 was configured as a FRET-based tenna 7 was synthesized in two steps as shown in Scheme 3. First,
light-harvesting antenna, core and peripherally decorated with a the intermediate dendron 6 with 4-nitro-1,8-naphthalimide pe-
rhodamine acceptor and 1,8-naphthalimide donors, respectively. riphery was obtained by reaction of 4-nitro-1,8-naphthalic
38 N.I. Georgiev et al. / Dyes and Pigments 102 (2014) 35e45

FRET N FRET NH
X - X e-

X
N e N

O O
HN O NH N HN O NH N
O O O O
NH NH
N H (pH 6) N
N FLUORESCENCE N FLUORESCENCE
O O
NH OH NH
O O O O
N N
O O
FLUORESCENCE FLUORESCENCE
X X
FRET N e- FRET N e-

X
7 7
N NH

FRET NH

e-

X
N

O H (pH 2-6)
HN O NH N
O O OH
NH
HN
N FLUORESCENCE
O
NH
O O
N
O
FLUORESCENCE

FRET N e-
X

7
NH

Scheme 2. Photophysical behaviour of antenna 7 as a function of pH after excitation within the spectral region of maximal absorption of the peripheral fluorophores.

anhydride 5 and PAMAM dendron 4 under reflux in methanol so- nitro-1,8-naphthalimide units in the intermediate dendron 6 were
lution. Finally, the yellow-green emitting periphery of the system completely converted in yellow-green emitting donor periphery.
was obtained after substitution of the nitro groups in the inter- Moreover the 1H NMR spectra contain all requisite peaks for
mediate 6 with N-methylpiperazinyl moieties in DMF at room rhodamine and 1,8-naphthalimide moieties as well as peaks in the
temperature. range of 2.3e3.5 ppm, attributed to the protons in the peripheral N-
The synthesized compounds were characterized (melting point, methylpiperazine.
TLC retention value Rf) and identified by conventional techniques e
elemental analysis data, UVeVis, fluorescence, FT-IR and 1H NMR 3.2. Photophysical characterization of antenna 7
spectroscopy. For instance, in the 1H NMR (CDCl3-d, 250.13 MHz)
spectrum of antenna 7 a resonance at 4.17 ppm (Fig. 1, A) was Photophysical properties of the synthesized compounds 2e4
observed. This is characteristic for the methylene protons coupled and 6e7 were determined in water/DMF (4:1, v/v) solution. Under
to N-position of 1,8-naphthalimide moiety, which proves the these conditions the rhodamine moiety adopts a closed, non-
presence of 1,8-naphthalimide units in the dendron periphery. Also fluorescent spirolactam form. At ca. pH 2 the spirolactam ring of
the 1H NMR spectrum of antenna 7 contains two resonances at rhodamine is opened, which results in a new absorption (rhoda-
6.34 ppm and 6.14 ppm that are typical for the core Rhodamine 6G mine) band between 450 and 575 nm with maximum at 530e
protons at position C-1, C-4, C-5 and C-8 (Fig. 1, C). The resonances 532 nm. The listed in Table 1 absorption data for compounds 2e4
at 4.17 ppm, 6.14 ppm and 6.34 ppm are in ratio 2:1:1 suggesting and 6 are common for Rhodamine 6G derivatives [76,77]. The
that in the novel dendron rhodamine core is surrounded by two presented data show that the different alkylamino substituents in
1,8-naphthalimide units. Furthermore, a resonance at 7.01 ppm the 9-phenyl amide of Rhodamine 6G (compounds 2e6) have a
(Fig. 1, B) appears which is characteristic for the proton in position small effect on the energy and the shape of the dyes’ absorption
C-3 of the yellow-green emitting 1,8-naphthalimide, substituted in bands.
position C-4 with an electron-donating piperazine group. In alkaline solution antenna 7 do not emit light due to the PET
This resonance is rather different from the corresponding value for quenching process in peripheral 1,8-naphthalimides and non-
a non-substituted 4-nitro-1,8-naphthalimide moieties (8.35e fluorescent spirolactam form of the focal rhodamine (Scheme 2).
8.70 ppm) [17,75], which is a solid evidence that the peripheral 4- This was the reason to investigate the photophysical properties of
 N.I. Georgiev et al. / Dyes and Pigments 102 (2014) 35e45 39

HN O NH Cl HN O NH HN O NH
O
H2N O
OMe
NH2 OMe
O N N
EtOH, Reflux NH2 MeOH, 20 C o N
O O O
OMe
O
1 NO2 2 3

O
NO2
O O
HN O NH N HN O NH NH2
O O O O
NH NH NH2
N 5 N H2N
N N
O MeOH, Reflux O MeOH, 20 oC
NH NH
O O O
4
N NH2
6 O
N

N
NO2

O
HN O NH N
O O
HN N NH
N
N
DMF, 20oC O
NH
O O
N
7 O

Scheme 3. Synthesis of PAMAM light-harvesting antenna 7.

antenna 7 at pH 6.5 where the PET is not feasible and also at pH 2 It is well known, that the spectral overlap is the main factor that
where the rhodamine is in ring-opened form. At pH 6.5 light- predetermines the efficiency of FRET. Peripheral 1,8-naphthalimide
harvesting dendron 7 showed only one longest-wavelength ab- fluorophores in antenna 7 showed a broad emission band in the
sorption band in a range of about 340e520 nm, which is attributed visible region (450e650 nm) that practically covers completely a
to an internal charge transfer process in the 1,8-naphthalimide rhodamine absorption band (Fig. 2).
chromophores. After acidification to ca. pH 2 where the rhoda- The efficiency of the energy transfer ET in antenna 7 was
mine spirolactam form is opened, as expected the absorption calculated to be 91% by applying Eq. (1) [16,78], where FDA and FD
spectrum of light-harvesting system 7 showed two bands (Table 1) are the normalized to the optical density fluorescence intensities of
corresponding to the absorption location of the peripheral 1,8- the donor in the presence of acceptor (1,8-naphthalimide part in
naphthalimide donor chromophores (lA ¼ 394 nm) and the focal the examined antenna at pH 2) and of the free donor without
rhodamine acceptor unit (lA ¼ 534 nm). acceptor (1,8-naphthalimide part at pH 6.5 e FRET process to the
At pH 2 and excitation at 510 nm, compounds 2e4, 6 and 7 show acceptor is not faceable), respectively. The calculated energy
typical for Rhodamine 6G fluorescence spectra with maxima at transfer clearly shows that the novel light-harvesting antenna is
about 560 nm [51,52], suggesting that the substituents at 9-phenyl able to act as an efficient wavelength-shifting chromophore.
amide do not affect the energy of the dyes’ fluorescence maximum.
The fluorescence spectra of light-harvesting antenna 7 in water/ ET ¼ 1  ðFDA =FD Þ (1)
DMF (4:1, v/v) solution at pH 6.5, recorded after excitation within
the spectral region of maximal absorption of the donor fluo- The Stokes’ shift (nA  nF) is an important parameter for the
rophores (lex ¼ 400 nm), showed emission band at 522 nm, cor- fluorescent compounds that indicates the differences in the prop-
responding to the emission band of the donor 1,8-naphthalimide erties and structure of the fluorophores between the ground state
fragments in the donor-acceptor systems. In contrast, when the S0 and the first excited state S1. The calculated Stokes’ shift (nA  nF)
fluorescence spectra was recorded at ca. pH 2, the observed emis- values [57] for compounds 2e4 and 6 as well as for the individual
sion was shifted to 561 nm (Table 1), which can be attributed to the fluorophores in antenna 7 are typical for the rhodamines and 1,8-
energy transfer from the donor 1,8-naphthalimide to the ring- naphthalimides [76,77]. The value of 6220 cm1 for the periph-
opened (fluorescence) form of the rhodamine acceptor. eral 1,8-naphthalimides and 876e947 cm1 for the focal
40 N.I. Georgiev et al. / Dyes and Pigments 102 (2014) 35e45

Fig. 1. 1H NMR spectrum (CDCl3-d, 250.13 MHz) of antenna 7.

Table 1 As can be seen from the data in Table 1, the quantum yield of
Photophysical characteristics of compounds 2e4 and 6 (lex ¼ 510 nm) and 7 fluorescence of the rhodamine core is decreasing with the increase
(lex ¼ 400 nm) in water/DMF (4:1, v/v).
of the molecular weight of the compounds under study. A similar
Compound lA (nm) 3 (l mol1 cm1) lF (nm) nA  nF (cm1) FF effect was reported before for the core functional PAMAM dendrons
2 530a 56 308a 558a 947a 0.92a using 1,8-naphthalimide and perylene-3,4,9,10-tetracarboxylic
3 532a 46 738a 558a 876a 0.72a diimide units [29,79]. This is probably due to the more flexible
4 532a 42 988a 560a 940a 0.55a PAMAM scaffold in the larger molecules which induces energy
6 532a 42 764a 560a 940a 0.52a loses thus reducing the quantum yield of fluorescence.
7 394a,b 17 564a,b 522a 6 220a 0.28a
534b 48 735b 561b 900b 0.43b
a
3.3. Influence of pH on the photophysical properties of antenna 7
Photophysical data recorded at pH 6.5.
b
Photophysical data recorded at pH 2.0 to avoid a rhodamine spirolactam closed
(non-fluorescent) form. The light-harvesting system under study was designed as a
molecular fluorescence sensor for determination of pH changes
rhodamine, do not indicate remarkable changes in the fluorophore over a wider pH scale. This was the reason to investigate their
excited state due to their incorporation in the dendritic systems. photophysical behaviour in water/DMF (4:1, v/v) solution at
The quantum yields of fluorescence were calculated using different pH values. In order to receive a more complete compar-
Rhodamine 6G (FF ¼ 0.95 in ethanol [71]) or Coumarin 6 (FF ¼ 0.78 ative picture for the influence of the dendron bone to the focal
in ethanol [72]) as standards according to Eq. (2), where Aref, Sref, nref rhodamine, compounds 2e4 were involved in the present study.
and Asample, Ssample, nsample represent the absorbance at the exited Fig. 3 presents the changes of absorption spectra of 3 at different pH
wavelength, the integrated emission band area and the solvent values as a typical example for the influence of pH on the absorp-
refractive index of the standard and the sample, respectively. tion spectra of the examined compounds 2e4. As can be seen, the
! ! ! decrease of pH results in increase of the absorbance at about
Ssample Aref n2sample 532 nm due to the ring opening reaction of rhodamine core.
FF ¼ Fref (2) Taking the part of the graphs located between pH 2 and 6, the
Sref Asample n2ref
pKa values of 2e4 have been calculated by Eq. (3) [18]. The calcu-
lated pKa value for amino-functional rhodamine 2 was 4.0 and 3.9
for the branching compounds 3 and 4. Probably the decrease of pKa
values is a result of the core protective role of the PAMAM scaffold.

log½ðAmax  AÞ=ðA  Amin Þ ¼ pH  pKa (3)

The absorption spectrum of light-harvesting antenna 7 does not
show significant pH-dependent changes in a pH window 8e10 as
the 1,8-naphthalimide fluorophores do not affect their ICT excited
states. In contrast, upon acidification from pH 8 to ca. pH 2 the
absorption maximum of peripheral 1,8-naphthalimides is blue
shifted of about 20 nm (Fig. 4). One of the reasons for this effect is
that the protonation of the amine receptors exerts some weak
charge repulsion on the 4-amino moiety of the fluorophores. On the
other hand in very acidic conditions the pushepull character of the
ICT state is partially reduced due to the protonation of the aromatic
4-amino moiety.
When the pH decreases from 6 to ca. 2 a novel band corre-
sponding to the absorption of the focal rhodamine due to the ring
opening reaction is appeared. From the absorption changes at
Fig. 2. Normalized absorption spectrum of dendron 4 at pH 2 and emission spectrum 534 nm the titration curve of antenna 7 was obtained (Inset in
of antenna 7 at pH 6.5 (lex ¼ 400 nm) in water/DMF (4:1, v/v). Fig. 4). Analysis of these changes according to Eq. (3) gives two pKa
 N.I. Georgiev et al. / Dyes and Pigments 102 (2014) 35e45 41

Family of fluorescence spectra of antenna 7 (lex ¼ 400 nm) at pH

window 6.5e10 in water/DMF (4:1, v/v) are presented in Fig. 6. As
could be expected in alkaline media the novel compound showed
very weak fluorescence intensity in range of 450e650 nm with
maximum at 522 nm (1,8-naphthalimide emission). However, upon
acidification an emission signal was gradually increased. The in-
crease in fluorescence intensity on transition from alkaline to
slightly acidic pH indicates that the peripheral fluorescence of the
examined antenna is being quenched by PET process from the
tertiary amine receptor to the 1,8-naphthalimide fluorophore. Upon
recognition of the analyte (protons) the oxidation potential of the
receptor is increased thermodynamically thus disallowing PET
process and as such, the fluorescence of the periphery is recovered.
Furthermore, as demonstrated experimentally by Liu and de
Silva, only the receptor that is directly attached to the 1,8-
naphthalimide 4-amino moiety (the “lower” moiety) is capable of
quenching the fluorophore excited state [80]. The different
behaviour of the receptors in C-4 and N-position of the peripheral
Fig. 3. Absorption changes of 3 as a function of pH at concentration 1  105 mol L1 in 1,8-naphthalimides can be easily rationalized according to Scheme
water/DMF (4:1, v/v). Inset: Titration plots of compounds 2e4 in a pH range of ca. 10e2. 4. The 4-aminonaphthalimide fluorophore is a “pushepull” p
electron system with the C-4 amine group as a donor and the 1,8-
values (3.8 and 2.5). The pKa of 3.8 is a typical for the rhodamine naphthalimide as an acceptor. This leads to strong internal charge
core, while pKa of 2.5 suggests reversible aggregation processes at transfer (ICT) in the lowest excited singlet state and considerable
alkaline pH values. dipole character (positive pole at the 4-amino terminus). A large
The fluorescence spectra of compounds 2e4 and 7 were also dipole moment in an excited state gives rise to a strong photo-
recorded in water/DMF (4:1, v/v) solution at different pH values. In generated electric field. Such a molecular electric field can,
alkaline solution rhodamines 2e4 are in spirolactam closed form depending on its sign and magnitude, inhibit or accelerate a tran-
and do not emit light. However, upon acidification an emission siting electron in the 1,8-naphthalimide compounds. Thus the
signal in range between 500 and 700 nm was gradually increased fluorescence quenching PET process is observed only if the electron
(Fig. 5). leaving the unprotonated amine donor can enter the space of the 4-
Analysis of the fluorescence changes at 560 nm according to Eq. aminonaphthalimide fluorophore across C-4 position with its
(4) [6] gives the pKa values 4.0 for amino-functional rhodamine 2 attractive electric field (Scheme 4). The corresponding PET path
and 3.9 for the compounds 3 and 4. These pKa values are the same from the unprotonated amino receptor in N-position is just as
as these calculated according to the absorption changes and should feasible thermodynamically but requires the electron to enter the
be attributed to the spirolactam opening reaction. fluorophore across the imide moiety with its repulsive electric field
and is not observed.
log½ðIFmax  IF Þ=ðIF  IFmin Þ ¼ pH  pKa (4) The enhancement of the fluorescence emission FE was used as a
qualitative parameter for the evaluation of the sensing potential of
As can be seen (Figs. 3 and 5), the increase of rhodamine the synthesized antenna 7. The FE ¼ I/Io was determined as ratio
absorbance and fluorescence intensity occurs at pH values less than between the maximum of fluorescence intensity I at pH ca. 6.5 and
5, which is only due to the usual for the rhodamine derivatives ICT the minimum fluorescence intensity Io at about pH 10. Upon acid-
ring opening reaction. Obviously, the presence of a PET process ification the fluorescence intensity of the yellow-green emitting
would reflect in emission changes at pH values more than 5, but periphery had enhanced by over an order of magnitude (FE ¼ 11).
actually such changes are absent. These changes are of such magnitude that they can be considered as

Fig. 4. Absorption changes of antenna 7 as a function of pH at concentration

1  105 mol L1 in water/DMF (4:1, v/v). Inset: Titration plot of antenna 7 in a pH Fig. 5. Fluorescence changes (lex ¼ 510 nm) of rhodamines 2e4 as a function of pH (ca.
range of ca. 10e2. 7e2) at concentration 1  105 mol L1 in water/DMF (4:1, v/v).
42 N.I. Georgiev et al. / Dyes and Pigments 102 (2014) 35e45

Fig. 6. Fluorescence changes (lex ¼ 400 nm) of antenna 7 as a function of pH (ca. 10e
6.5) at concentration 1  105 mol L1 in water/DMF (4:1, v/v). Inset: Titration plot of
antenna 7 in a pH range of ca. 6.5e10.

representing two different “states”, where the fluorescence emis-

sion is “switched off” in alkaline solution and “switched on” in
neutral to slightly acidic media.
As can be seen from the Inset of Fig. 6, peripheral 1,8-
naphthalimide fluorophores switch between “off” and “on” states
in pH interval of ca. 8.5e6.5. The calculated using Eq. (4) pKa value
of 7.15 for antenna 7 is in agreement with the data for 4-
piperazinyl-1,8-naphthalimide derivatives [7,34,48,50].
Further acidification from pH 6.5 to ca. pH 2 opens the rhoda-
mine spirolactam form, which allows the energy transfer from
periphery to the acceptor moiety (lex ¼ 400 nm). This results in
fluorescence quenching of the peripheral fluorescence and
remarkable fluorescence intensity enhancement (FE ¼ 32) in the
rhodamine emission region at 560 nm (Fig. 7).
Analysis of the fluorescence changes of antenna 7 at 520 nm and Fig. 7. Fluorescence changes of antenna 7 as a function of pH (6e2) at concentration
560 nm after excitation at 400 nm (Inset of Fig. 7A) according to Eq. 1  105 mol L1 in water/DMF (4:1, v/v) excited at lex ¼ 400 nm (A) and lex ¼ 510 nm
(B). Insets: Titration plot of antenna 7 in a pH range of ca. 6e2.
(4) gives pKa values of 3.8 and 2.5. The same results were obtained
after direct core excitation at 510 nm (Inset of Fig. 7B) suggesting
that only the spirolactam reaction of the rhodamine core is probe for pH determination. We expect that the method will serve
responsible for the pH sensing properties of the novel FRET system. as a practical tool for analysis of environmental samples and bio-
Thus, the changes in the fluorescence intensity of antenna 7 in the logical studies.
presence of protons are due to the PET process in the periphery of
the system and of the rhodamine core ICT ring opening reaction.
These results suggest that the novel wavelength-shifting 3.4. Influence of metal cations on the fluorescence intensity of the
bichromophoric system is able to act as a ratiometric fluorescent antenna 7

With regard to the potential application of antenna 7 as a

Repulsion Attractive fluorescent probe for recognition of different cations, its signalling
Field Field
properties in the presence of transition metal ions were studied in
e X
e DMF and in water/DMF (4:1, v/v) solution. DMF has been chosen
δ− δ+ since it is able as a polar solvent to stabilize the dye charge sepa-
N O rated state thus favouring the fluorescence switching by PET pro-
O cess. Also DMF guarantees a good solubility of the dye ligands, used
HN N N
O N metal salts and the respective complexes. Experiments have been
performed in the presence of different metal cations: Cd2þ, Co2þ,
O Cu2þ, Fe3þ, Ni2þ, Pb2þ, Zn2þ, Hg2þ and Agþ.
In DMF solution of antenna 7, considerable photophysical
X

changes were observed only upon addition of Cu2þ and Pb2þ, while
hν1 hν2 (fluorescence) the other metal ions produced a negligible effect (Fig. 8). The
presence of Cu2þ and Pb2þ induces a hypsochromic shift of the 1,8-
fluorophore sp. receptor
naphthalimide absorption maximum of DlA ¼ 10 nm for Cu2þ and
Scheme 4. Feasible photoinduced electron transfer in the 1,8-naphthalimide periph- 12 nm for Pb2þ (Fig. 8A). This means that the both nitrogen atoms of
ery of antenna 7. the N-methylpiperazine substituent at C-4 of the peripheral 1,8-
 N.I. Georgiev et al. / Dyes and Pigments 102 (2014) 35e45 43

The examined metal ions do not induced any changes in the

photophysical properties of the antenna core even in a higher
cation concentration (1  102 M), suggesting that only protons are
able to open the spirolactam cycle. These results showed that
during the experiments the metal salts do not generate protons
from partial hydrolysis of the salts and as such do not provoke
unusual fluorescence enhancement of the rhodamine core.
It is well known that in aqueous solutions, especially in the
presence of an anionic buffer, coordination of the metal ions with
the receptor fragments competes with a number of other anions
and nucleophiles. That is why it was of interest to investigate the
emission behaviour of antenna 7 in the presence of metal ions in
aqueous medium in regard to its potential for practical application.
To avoid the effect of proton generation due to the partial hydro-
lysis of metal salts the experiments were conducted in buffered
solution. The influence of various metal ions on the fluorescence
behaviour of antenna 7 in water/DMF (4:1, v/v) solution in the
presence of ammonia/ammonium chloride buffer (pH 10) is pre-
sented in Fig. 9. As can be seen, the addition of metal ion solution
(even up to 1  102 mol L1) to the antenna solution
(1  105 mol L1) does not caused any noticeable changes, sug-
gesting that in water/DMF (4:1, v/v) the metal cations are not able
to coupling with the receptors of the peripheral PET based
naphthalimides.
The results obtained show excellent selectivity of antenna 7
toward protons over representative metal ions in aqueous medium.
The high selectivity and ratiometric pH sensitivity of the novel
antenna may be beneficially for monitoring pH variations in com-
plex samples.

4. Conclusions

A PAMAM light-harvesting dendron, core and peripherally

functionalized with Rhodamine 6G and 1,8-naphthalimide fluo-
rophores, respectively, was synthesized based on a divergent
approach. Novel compound was designed as a wavelength-shifting
Fig. 8. Absorption (A) and fluorescence (B) changes of antenna 7 (1  105 mol L1) in chromophore capable of absorbing light by its periphery and effi-
the presence of metal cations (1  104 mol L1) in DMF. ciently transferring the energy to a single acceptor dye in the focal
point of the system. The calculated energy transfer of 91% indicates
naphthalimides are subjected to coordination with metal cations that the selected fluorophores are suitable donor-acceptor pair for
[66]. The addition of Cu2þ and Pb2þ also results in enhancement constructing light-harvesting molecules. Furthermore, the yellow-
only of the peripheral fluorescence intensity (1,8-naphthalimides) green emitting periphery of the system was designed on the “flu-
of antenna 7. The enhancement of the fluorescence emission was orophoreespacerereceptor” format, making the antenna capable
FE ¼ 16.0 for Cu2þ and FE ¼ 16.7 for Pb2þ (Fig. 8B). to act as a molecular fluorescence PET-based pH sensor. After
excitation within the spectral region of maximal absorption of
peripheral 1,8-naphthalimides the synthesized antenna enhanced
its peripheral fluorescence intensity by over an order of magnitude
(FE ¼ 11) in a pH range of ca. 6.5e10, while upon acidification to pH
2 the fluorescence intensity of the core was enhanced more than 30
times with the decrease of the peripheral emission. Novel antenna
also showed excellent selectivity toward protons over representa-
tive metal ions in aqueous medium. Thus the distinguishing fea-
tures of the synthesized light-harvesting dendron were
successfully combined with the properties of classical PET and ring-
opening sensor systems. These results suggest that the novel an-
tenna is able to act as a highly selective and pH sensitive ratiometric
fluorescence probe, indicating that it could be used for analysis of
environmental samples and biological studies.

Acknowledgements

This work was supported by the National Science Foundation of

2 1
Fig. 9. Effect of metal ions (1  10 mol L ) on the fluorescence intensity of antenna
Bulgaria (project DDVU-02/97). Authors also acknowledge the
7 (1  105 mol L1) in water/DMF (4:1, v/v) in the presence ammonia/ammonium Science Foundation at the University of Chemical Technology and
chloride buffer (pH 10). Metallurgy (Sofia, Bulgaria).
44 N.I. Georgiev et al. / Dyes and Pigments 102 (2014) 35e45

References [31] Georgiev N, Bojinov V. Design, synthesis and photostability of novel 1,8-
naphthalimide PAMAM light-harvesting dendrons. J Fluoresc 2011;21:51e63.
[32] Grabchev I, Staneva D, Bojinov V, Betcheva R, Gregoriou V. Spectral investi-
[1] de Silva AP, Fox D, Huxley A, Moody T. Combining luminescence, coordi-
gation of coordination of cuprum cations and protons at PAMAM dendrimer
nation and electron transfer for signalling purposes. Coord Chem Rev
peripherally modified with 1,8-naphthalimide units. Spectrochim Acta Part A
2000;205:41e57.
2008;70:532e6.
[2] Balzani V. Photochemical molecular devices. Photochem Photobiol Sci 2003;2:
[33] Bojinov V, Georgiev N, Nikolov P. Synthesis and photophysical properties of
459e76.
fluorescence sensing ester- and amidoamine-functionalized 1,8-
[3] Badugu R, Lakowicz J, Geddes C. Cyanide-sensitive fluorescent probes. Dye
naphthalimides. J Photochem Photobiol A Chem 2008;193:129e38.
Pigment 2005;64:49e55.
[34] Georgiev N, Bojinov V. The design and synthesis of a novel 1,8-naphthalimide
[4] Jun M, Roy B, Ahn K. “Turn-on”’ fluorescent sensing with “reactive” probes.
PAMAM light-harvesting dendron with fluorescence “off-on” switching core.
Chem Commun 2011;47:7583e601.
Dye Pigment 2010;84:249e56.
[5] Xu Z, Pan J, Spring D, Cui J, Yoon J. Ratiometric fluorescent and colorimetric
[35] Georgiev N, Bojinov V, Marinova N. Novel PAMAM light-harvesting antennae
sensors for Cu2þ based on 4,5-disubstituted-1,8-naphthalimide and sensing
based on 1,8-naphthalimide: synthesis, energy transfer, photophysical and pH
cyanide via Cu2þdisplacement approach. Tetrahedron 2010;66:1678e83.
sensing properties. Sens Actuators B Chem 2010;150:655e66.
[6] Tal S, Salman H, Abraham Y, Botoshansky M, Eichen Y. Sensitive and selective
[36] Konstantinova T, Miladinova P. Synthesis and properties of some fluorescent
photoinduced-electron-transfer-based sensing of alkylating agents. Chem Eur
1,8-naphthalimide derivatives and their copolymers with methyl methacry-
J 2006;12:4858e64.
late. J Appl Polym Sci 2009;111:1991e8.
[7] Marinova N, Bojinov V, Georgiev N. Design, synthesis and pH sensing prop-
[37] Jin W, Jiang J, Wang X, Zhu X, Wang G, Song Y, et al. Continuous intra-arterial
erties of novel 1,8-naphtalimide-based bichromophoric system. J Photochem
blood pH monitoring in rabbits with acid-base disorders. Respir Physiol
Photobiol A Chem 2011;222:132e40.
Neurobiol 2011;177:183e8.
[8] Dai H, Xu H. A water-soluble 1,8-naphthalimide-based “turn on” fluorescent
[38] Bojinov V, Simeonov D, Georgiev N. A novel blue fluorescent 4-(1,2,2,6,6-
chemosensor for selective and sensitive recognition of mercury ion in water.
pentamethylpiperidin-4-yloxy)-1,8-naphthalimide pH chemosensor based
Bioorg Med Chem Lett 2011;21:5141e4.
on photoinduced electron transfer. Dye Pigment 2008;76:41e6.
[9] Kim H, Guo Z, Zhu W, Yoon J, Tian H. Recent progress on polymer-based
[39] Xie J, Chen Y, Yang W, Xu D, Zhang K. Water soluble 1,8-naphthalimide
fluorescent and colorimetric chemosensors. Chem Soc Rev 2011;40:79e93.
fluorescent pH probes and their application to bioimagings. J Photochem
[10] de Silva AP, Uchiyama S. Molecular logic gates and luminescent sensors based
Photobiol A Chem 2011;223:111e8.
on photoinduced electron transfer. Top Curr Chem 2011;300:1e28.
[40] Ott I, Xu Y, Qian X. Fluorescence properties and antiproliferative effects of
[11] Parkesh R, Lee T, Gunnlaugsson T. Fluorescence imaging of bone cracks
mono-, bis-, and tris thiophenylnaphthalimides: results of a comparative pilot
(microdamage) using visibly emitting 1,8-naphthalimide-based PET sensors.
study. J Photochem Photobiol B Biol 2011;105:75e80.
Tetrahedron Lett 2009;50:4114e6.
[41] de Souza M, Correa R, Filho V, Grabchev I, Bojinov V. 4-Nitro-1,8-
[12] Kaur P, Kaur S, Singh K. A fluoride selective dipyrromethane-TCNQ colori-
naphthalimides exhibit antinociceptive properties. Pharmazie 2002;56:430e1.
metric sensor based on charge-transfer. Talanta 2011;84:947e51.
[42] Bojinov V, Konstantinova T. Fluorescent 4-(2,2,6,6-tetramethylpiperidin-4-
[13] Fabbrizzi L, Licchelli M, Pallavicini P, Perotti A, Taglietti A, Sacchi D. Fluores-
ylamino)-1,8-naphthalimide pH chemosensor based on photoinduced elec-
cent sensors for transition metals based on electron-transfer and energy-
tron transfer. Sens Actuators B Chem 2007;123:869e76.
transfer mechanisms. Chem Eur J 1996;2:75e82.
[43] Zhu W, Song L, Yang Y, Tian H. Novel bisthienylethene containing ferrocenyl-
[14] Sasaki H, Hanaoka K, Urano Y, Terai T, Nagano T. Design and synthesis of a
substituted naphthalimide: a photo- and redox multi-addressable molecular
novel fluorescence probe for Zn2þ based on the spirolactam ring-opening
switch. Chem Eur J 2012;18:13388e94.
process of rhodamine derivatives. Bioorg Med Chem 2011;19:1072e8.
[44] Bojinov V, Panova I, Chovelon J-M. Novel blue emitting tetra- and pentam-
[15] Chen X, Pradhan T, Wang F, Kim J, Yoon J. Fluorescent chemosensors based on
ethylpiperidin-4-yloxy-1,8-naphthalimides as photoinduced electron transfer
spiroring-opening of xanthenes and related derivatives. Chem Rev 2012;112:
based sensors for transition metal ions and protons. Sens Actuators B Chem
1910e56.
2008;135:172e80.
[16] Li W-S, Teng M-J, Jia X-R, Wang B-B, Yeh J-M, Wei Y. Synthesis and energy-
[45] Wang H, Yang L, Zhang W, Zhou Y, Zhao B, Li X. A colorimetric probe for
transfer properties of poly(amidoamine) dendrons modified with naphthyl
copper(II) ion based on 4-amino-1,8-naphthalimide. Inorg Chim Acta
and dansyl groups. Tetrahedron Lett 2008;49:1988e92.
2012;381:111e6.
[17] Georgiev N, Bojinov V, Venkova A. Design, synthesis and pH sensing prop-
[46] Bojinov V, Panova I, Simeonov D, Georgiev N. Synthesis and sensor activity of
erties of novel PAMAM light-harvesting dendrons based on rhodamine 6G
photostable blue emitting 1,8-naphthalimides containing s-triazine UV
and 1,8-naphthalimide. J Fluoresc 2013;23:459e71.
absorber and HALS fragments. J Photochem Photobiol A Chem 2010;210:89e99.
[18] Daffy L, de Silva AP, Gunaratne H, Huber C, Lynch P, Werner T, et al. Arene-
[47] Wang Y, Zhang X, Han B, Peng J, Hou S, Huang Y. The synthesis and photo-
dicarboximide building blocks for fluorescent photoinduced electron transfer
luminescence characteristics of novel blue light-emitting naphthalimide de-
pH sensors applicable with different media and communication wavelengths.
rivatives. Dye Pigment 2010;86:190e6.
Chem Eur J 1998;4:1810e5.
[48] Bojinov V, Georgiev N, Marinova N. Design and synthesis of highly photo-
[19] Liu Y, Lv X, Zhao Y, Chen M, Liu J, Wang P, et al. A naphthalimide-rhodamine
stable fluorescence sensing 1,8-naphthalimide-based dyes containing s-
ratiometric fluorescent probe for Hg2þ based on fluorescence resonance en-
triazine UV absorber and HALS units. Sens Actuators B Chem 2010;148:6e16.
ergy transfer. Dye Pigment 2012;92:909e15.
[49] Bojinov V, Panova I. Novel 4-(2,2,6,6-tetramethylpiperidin-4-ylamino)-1,8-
[20] Zhu S, Lin W, Yuan L. Development of a ratiometric fluorescent pH probe for
naphthalimide based yellow-green emitting fluorescence sensors for transi-
cell imaging based on a coumarin-quinoline platform. Dye Pigment 2013;99:
tion metal ions and protons. Dye Pigment 2009;80:61e6.
465e71.
[50] Bojinov V, Georgiev N, Bosch P. Design and synthesis of highly photostable
[21] Nantalaksakul A, Reddy D, Bardeen C, Thayumanavan S. Light harvesting
yellow-green emitting 1,8-naphthalimides as fluorescent sensors for metal
dendrimers. Photosyn Res 2006;87:133e50.
cations and protons. J Fluoresc 2009;19:127e39.
[22] Dichtel WR, Hecht S, Frèchet J. Functionally layered dendrimers: a new
[51] Georgiev N, Yaneva I, Surleva A, Asiri AM, Bojinov V. Synthesis, sensor activity
building block and its application to the synthesis of multichromophoric light-
and logic behavior of a highly water-soluble naphthalimide derivative. Sens
harvesting systems. Org Lett 2005;7:4451e4.
Actuators B Chem 2013;184:54e63.
[23] Mètivier R, Kulzer F, Weil T, Müllen K, Basch T. Energy transfer rates and
[52] Georgiev N, Bojinov V. Design, synthesis and sensor activity of a highly
pathways of single donor chromophores in a multichromophoric dendrimer
photostable blue emitting 1,8-naphthalimide. J Lumin 2012;132:2235e41.
built around a central acceptor core. J Am Chem Soc 2004;126:14364e5.
[53] Ferreira R, Remon P, Pischel U. Multivalued logic with a tristable fluorescent
[24] Ceroni P, Bergamini G, Marchioni F, Balzani V. Luminescence as a tool to
switch. J Phys Chem C 2009;113:5805e11.
investigate dendrimer properties. Prog Polym Sci 2005;30:453e73.
[54] Georgiev N, Bojinov V, Nikolov P. Design, synthesis and photophysical prop-
[25] Flomenboma O, Amir R, Shabat D, Klaftera J. Some new aspects of dendrimer
erties of two novel 1,8-naphthalimide fluorescent pH sensors based on PET
applications. J Lumin 2005;111:315e25.
and ICT. Dye Pigment 2011;88:350e7.
[26] Thomas K, Thompson A, Sivakumar A, Bardeen C, Thayumanavan S. Energy
[55] Georgiev N, Lyulev M, Bojinov V. Sensor activity and logic behavior of PET
and electron transfer in bifunctional non-conjugated dendrimers. J Am Chem
based dihydroimidazonaphthalimide diester. Spectrochim Acta Part A
Soc 2005;127:373e83.
2012;97:512e20.
[27] Li S, Zhu W, Xu Z, Pan J, Tian H. Antenna-functionalized dendritic b-diketo-
[56] Bojinov V, Georgiev N. Molecular sensors and molecular logic gates [Review]
nates and europium complexes: synthetic approaches to generation growth.
J Univ Chem Technol Metal (Sofia) 2011;46:3e26.
Tetrahedron 2006;62:5035e48.
[57] Marinova N, Georgiev N, Bojinov V. Facile synthesis, sensor activity and logic
[28] Bar-Haim A, Klafter J. Dendrimers as light harvesting antennae. J Lumin
behaviour of 4-aryloxy substituted 1,8-naphthalimide. J Photochem Photobiol
1998;76e77:197e200.
A Chem 2013;254:54e61.
[29] Bojinov V, Georgiev N, Nikolov P. Design and synthesis of core and periph-
[58] Sun Y, Wei S, Yin C, Liu La, Hu Ci, Zhao Y, et al. Synthesis and spectroscopic
erally functionalized with 1,8-naphthalimide units fluorescent PAMAM den-
characterisation of 4-butoxyethoxy-N-octadecyl-1,8-naphthalimide as a new
dron as light harvesting antenna. J Photochem Photobiol A Chem 2008;197:
fluorescent probe for the determination of proteins. Bioorg Med Chem Lett
281e9.
2011;21:3798e804.
[30] Georgiev N, Bojinov V, Nikolov P. Design and synthesis of a novel pH sensitive
[59] Dong M, Ma T-H, Zhang A-J, Dong Y-M, Wang Y-W, Peng Y. A series of highly
core and peripherally 1,8-naphthalimide-labeled PAMAM dendron as light
sensitive and selective fluorescent and colorimetric “off-on” chemosensors for
harvesting antenna. Dye Pigment 2009;81:18e26.
Cu (II) based on rhodamine derivatives. Dye Pigment 2010;87:164e72.
 N.I. Georgiev et al. / Dyes and Pigments 102 (2014) 35e45 45

[60] Mab Q-J, Zhang X-B, Zhao X-H, Jin Z, Mao G-Ji, Shen G-L, et al. A highly se- [71] Kubin R, Fletcher A. Fluorescence quantum yields of some rhodamine dyes.
lective fluorescent probe for Hg2þ based on a rhodamine-coumarin conjugate. J Lumin 1982;27:455e62.
Anal Chim Acta 2010;663:85e90. [72] Reynolds G, Drexhage K. New coumarin dyes with rigidized structure for
[61] Ahamed B, Ghosh P. An integrated system of pyrene and Rhodamine-6G for flashlamp-pumped dye lasers. Opt Commun 1975;13:222e5.
selective colorimetric and fluorometric sensing of mercury(II). Inorg Chim [73] Niu C, Zeng G, Chen L, Shena G, Yu R. Proton “off-on” behaviour of methyl-
Acta 2011;372:100e7. piperazinyl derivative of naphthalimide: a pH sensor based on fluorescence
[62] Lei Y, Su Y, Huo J. Photophysical property of rhodamine-cored poly(amido- enhancement. Analyst 2004;129:20e4.
amine) dendrimers: simultaneous effect of spirolactam ring-opening and PET [74] Gan J, Chen K, Chang C, Tian H. Luminescent properties and photo-induced
process on sensing trivalent chromium ion. J Lumin 2011;131:2521e7. electron transfer of naphthalimides with piperazine substituent. Dye
[63] Lv H-S, Liu J, Zhao J, Zhao B-X, Miao J-Y. Highly selective and sensitive pH- Pigment 2003;57:21e8.
responsive fluorescent probe in living hela and HUVEC cells. Sens Actuators [75] Bojinov V, Panova I. Synthesis and absorption properties of new yellow-
B Chem 2013;177:956e63. green emitting benzo[de]isoquinoline-1,3-diones containing hindered
[64] Zhu H, Fan J, Xu Q, Li H, Wang J, Gao P, et al. Imaging of lysosomal pH changes amine and 2-hydroxyphenylbenzotriazole fragments. Dye Pigment 2007;74:
with a fluorescent sensor containing a novel lysosome-locating group. Chem 551e60.
Commun 2012;48:11766e8. [76] Bakkialakshmi S, Menaka T. A study of the interaction between rhodamine 6G
[65] Li H, Guan H, Duan X, Hu J, Wang G, Wang Q. An acid catalyzed reversible and hydroxy propyl b-cyclodextrin by steady state fluorescence. Spectrochim
ring-opening/ring-closure reaction involving a cyano-rhodamine spirolactam. Acta Part A 2011;81:8e13.
Org Biomol Chem 2013;11:1805e9. [77] Zakerhamidi M, Moghadam M, Ghanadzadeh A, Hosseini S. Anisotropic and
[66] Bojinov V, Venkova A, Georgiev N. Synthesis and energy-transfer properties of isotropic solvent effects on the dipole moment and photophysical properties
fluorescence sensing bichromophoric system based on Rhodamine 6G and of rhodamine dyes. J Lumin 2012;132:931e7.
1,8-naphthalimide. Sens Actuators B Chem 2009;143:42e9. [78] Fortier J, Even-Hernandez P, Baros F, Poulain S, Martinet N, Donner M, et al.
[67] Lv H-S, Huang S-Y, Zhao B-X, Miao J-Y. A new rhodamine B-based lysosomal The synthesis, photophysical properties and energy transfer of a coumarin-
pH fluorescent indicator. Anal Chim Acta 2013;788:177e82. based bichromophoric compound. Dye Pigment 2009;80:115e20.
[68] Lei Y, Zhang C, Lei H, Huo J. Visible light photocatalytic activity of aromatic [79] Georgiev N, Sakr A, Bojinov V. Design and synthesis of novel fluorescence
polyamide dendrimer/TiO2 composites functionalized with spirolactam-based sensing perylene diimides based on photoinduced electron transfer. Dye
molecular switch. J Colloid Interface Sci 2013;406:178e85. Pigment 2011;91:332e9.
[69] Lei Y, Su Y, Huo J. A novel fluorescent sensor for Cr3þ based on rhodamine-cored [80] Liu J, de Silva AP. Path-selective photoinduced electron transfer (PET) in a
poly(amidoamine) dendrimer. Spectrochim Acta Part A 2011;81:149e54. membrane-associated system studied by pH-dependent fluorescence. Inorg
[70] Grabchev I, Moneva I, Bojinov V, Guittonneau S. Synthesis and properties of Chim Acta 2012;381:243e6.
fluorescent 1,8-naphthalimide dyes for application in liquid crystal displays.
J Mater Chem 2000;10:1291e6.