Eu3+- and Tb3+-Based Coordination Complexes of Poly(N-Isopropyl,N-methylacrylamide-stat-N,N-dimethylacrylamide) Copolymer: Synthesis, Characterization and Property

This contribution reports the syntheses, structural analyses and properties of europium (Eu3+)- and terbium (Tb3+)-based coordination complexes of poly(N-isopropyl,N-methylacrylamide-stat-N,N-dimethylacrylamide) (poly(iPMAm-stat-DMAm)) copolymer, named as poly-Eu(III) and poly-Tb(III), respectively. In greater detail, poly(iPMAm85-stat-DMAm15) is first prepared by random copolymerization of N-isopropyl,N-methylacrylamide (iPMAm) and N,N-dimethylacrylamide (DMAm) via group transfer polymerization (GTP). Next, poly(iPMAm85-stat-DMAm15) is used as the polymer matrix for chelating with Eu3+ and Tb3+ cations at its side amide groups, to produce poly-Eu(III) and poly-Tb(III). Their structural characterizations by FT-IR spectroscopy and XPS confirm the formation of polymeric complexes. The study on their fluorescence emission characteristics and luminescence lifetime demonstrates that Poly-Eu(III) shows four strong emission peaks at 578, 593, 622, and 651 nm, which are responsible for the electron transitions from the excited 5D0 state to the multiplet 7FJ (J = 0, 1, 2, 3) states, respectively, and poly-Tb(III) also displays four emission peaks at 489, 545, 588, and 654 nm, mainly due to the electron transitions of 5D4 → 7Fi (i = 6, 5, 4, 3). The luminescence lifetimes of poly-Eu(III) (τpoly-Eu(III)) and poly-Tb(III) (τpoly-Tb(III)) are determined to be 4.57 and 7.50 ms, respectively. In addition, in aqueous solutions, poly-Eu(III) and poly-Tb(III) are found to exhibit thermoresponsivity, with their cloud temperatures (Tcs) locating around 36.4 and 36.8 °C, respectively. Finally, the cytotoxicity study on the human colon carcinoma cells LoVo and DLD1 suggests that the luminescent Eu3+ and Tb3+ in the chelated state with poly(iPMAm-stat-DMAm) show much better biocompatibility and lower toxicity than their inorganic salts.


Introduction
Luminescent rare earth complexes have become the research hotspot of new rare earth functional materials, because of their unique luminescence properties, such as extremely sharp emission bands [1,2], long excited-state lifetimes [3][4][5], and potential high internal quantum efficiency [6][7][8]. When rare earth metals are complexed with appropriate organic ligands, their improved solubility and dispersion would make their luminescence properties more prominent than they are in states of inorganic salts [9,10]. Furthermore, rare earth metals complexed with a polymeric matrix, namely, the so-called polymer-rare earth complexes, possess the combined properties of luminescent rare earth metals and the polymer >99%), methanol (MeOH, >98%), toluene (99%), triethylamine (>99.0%), 3,5-dibromoaniline, triisopropylsilylacetylene, trimethylsilylacetylene and sodium hydride (60 wt.% in mineral oil) were purchased from TCI Chemicals. Dichloromethane (CH 2 Cl 2 , >99.5%; extra dry, with molecular sieves, water < 50 ppm) and tetrahydrofuran (THF, >99.5%; extra dry, with molecular sieves, water < 50 ppm) were purchased from Energy Chemical Co., Ltd., Shanghai, China. Toluene and THF were distilled over Na/benzophenone under an argon atmosphere and degassed by three freeze-pump-thaw cycles prior to use. All other chemicals were purchased from available suppliers and used without further purification.
Polymerization was carried out in an MIKROUNA stainless-steel glove box equipped with a gas purification system under a dry argon atmosphere (H 2 O, O 2 < 0.01 ppm). The moisture and oxygen contents in the glove box were monitored by an MB-MO-SE 1 and an MB-OX-SE 1 sensor, respectively. The 1 H and 13 C NMR spectroscopy was recorded using a Bruker AVANCE III HD 400, Billerica, MA USA. Size exclusion chromatography (SEC) in DMF was performed at 40 • C at a flow rate of 0.35 mL min −1 using a TosohHLC-8320 GPC System equipped with two TSK gel Super Multipore HZ-M columns (4.6 mm I.D. × 15 cm × 2), an EcoSEC GPC System with an RI detector (+, 0.5 s), a UV-8320 detector (254 nm, +0.5 s), a GPC workstation EcoSEC-WS, and an auto sampler, to which 10 µL of sample was injected in a concentration of 0.2 wt.%. The number-average molecular weight (M n,SEC ) and polydispersity index (
Polymerization was carried out in an MIKROUNA stainless-steel glove box equipped with a gas purification system under a dry argon atmosphere (H2O, O2 < 0.01 ppm). The moisture and oxygen contents in the glove box were monitored by an MB-MO-SE 1 and an MB-OX-SE 1 sensor, respectively. The 1 H and 13 C NMR spectroscopy was recorded using a Bruker AVANCE III HD 400, Billerica, MA USA. Size exclusion chromatography (SEC) in DMF was performed at 40 °C at a flow rate of 0.35 mL min −1 using a TosohHLC-8320 GPC System equipped with two TSK gel Super Multipore HZ-M columns (4.6 mm I.D. × 15 cm × 2), an EcoSEC GPC System with an RI detector (+, 0.5 s), a UV-8320 detector (254 nm, +0.5 s), a GPC workstation EcoSEC-WS, and an auto sampler, to which 10 µL of sample was injected in a concentration of 0.2 wt.%. The number-average molecular weight (Mn,SEC) and polydispersity index (Ɖ) of the polymers were calculated based on PMMA standards. FT-IR spectra were recorded on a Perkin Elmer Frontier, and samples fashioned into KBr pellet disks were placed into a holder for transmission IR spectral analysis. The fluorescence spectra were recorded using a fluorescence spectrophotometer (RF-5301PC, Shimadzu, Kyoto, Japan). Quantum yields were determined by comparison of the total light emitted from the solutions to the total light emitted from a known standard [Ru(bipy)3]Cl2 [33]. XPS spectra (Al Kα) were recorded with a Thermo Fisher Scientific K-Alpha instrument, Waltham, MA USA. The cloud point measurements were performed on the ultraviolet-visible (UV-vis) spectra by passing through a 10 mm path-length cell using a Jasco V-770 spectrophotometer equipped with a temperature controller (Jasco CTU-100, Tokyo, Japan). The hydrodynamic radii (Rhs) of the obtained polymer in deionized water were analyzed using a dynamic light scattering (DLS) detector (Wyatt Technology, Dyna Pro Nanostar ® , Santa Barbara, CA, USA). Cell viability was evaluated by MTT, and the optical density (OD) was measured at 490 nm with a microplate reader (Bio-Rad, Hercules, CA, USA). Cell viability was determined as a percentage of the negative control (untreated cells).

Synthesis of Statistical Copolymer Poly(iPMAm85-Stat-PDMAm15)
iPMAm (246 µL, 1.7 mmol), DMAm (33 µL, 0.3 mmol), SKA Et (40 µL, 20.0 µmol; 0.50 mol L −1 CH2Cl2), and CH2Cl2 (1.60 mL) were added to a test tube, followed by the addition of the B(C6F5)3 stock solution in CH2Cl2 (80 µL, 4.0 µmol; 0.05 mol L −1 ) for 2 h in a glove box. The polymerization was quenched by adding a small amount of a 2-PrOH/pyridine mixture to the polymerization solution. Aliquots were removed from the reaction mixture to determine the conversion of iPMAm and DMAm using 1 H NMR measurements. The polymer product was purified by dialysis against methanol using a cellophane tube, after which the product was lyophilized to give poly(iPMAm85-stat-PDMAm15) as a white solid.
) of the polymers were calculated based on PMMA standards. FT-IR spectra were recorded on a Perkin Elmer Frontier, and samples fashioned into KBr pellet disks were placed into a holder for transmission IR spectral analysis. The fluorescence spectra were recorded using a fluorescence spectrophotometer (RF-5301PC, Shimadzu, Kyoto, Japan). Quantum yields were determined by comparison of the total light emitted from the solutions to the total light emitted from a known standard [Ru(bipy) 3 ]Cl 2 [33]. XPS spectra (Al Kα) were recorded with a Thermo Fisher Scientific K-Alpha instrument, Waltham, MA USA. The cloud point measurements were performed on the ultravioletvisible (UV-vis) spectra by passing through a 10 mm path-length cell using a Jasco V-770 spectrophotometer equipped with a temperature controller (Jasco CTU-100, Tokyo, Japan). The hydrodynamic radii (R h s) of the obtained polymer in deionized water were analyzed using a dynamic light scattering (DLS) detector (Wyatt Technology, Dyna Pro Nanostar ® , Santa Barbara, CA, USA). Cell viability was evaluated by MTT, and the optical density (OD) was measured at 490 nm with a microplate reader (Bio-Rad, Hercules, CA, USA). Cell viability was determined as a percentage of the negative control (untreated cells).
Polymerization was carried out in an MIKROUNA stainless-steel glove box equipped with a gas purification system under a dry argon atmosphere (H2O, O2 < 0.01 ppm). The moisture and oxygen contents in the glove box were monitored by an MB-MO-SE 1 and an MB-OX-SE 1 sensor, respectively. The 1 H and 13 C NMR spectroscopy was recorded using a Bruker AVANCE III HD 400, Billerica, MA USA. Size exclusion chromatography (SEC) in DMF was performed at 40 °C at a flow rate of 0.35 mL min −1 using a TosohHLC-8320 GPC System equipped with two TSK gel Super Multipore HZ-M columns (4.6 mm I.D. × 15 cm × 2), an EcoSEC GPC System with an RI detector (+, 0.5 s), a UV-8320 detector (254 nm, +0.5 s), a GPC workstation EcoSEC-WS, and an auto sampler, to which 10 µL of sample was injected in a concentration of 0.2 wt.%. The number-average molecular weight (Mn,SEC) and polydispersity index (Ɖ) of the polymers were calculated based on PMMA standards. FT-IR spectra were recorded on a Perkin Elmer Frontier, and samples fashioned into KBr pellet disks were placed into a holder for transmission IR spectral analysis. The fluorescence spectra were recorded using a fluorescence spectrophotometer (RF-5301PC, Shimadzu, Kyoto, Japan). Quantum yields were determined by comparison of the total light emitted from the solutions to the total light emitted from a known standard [Ru(bipy)3]Cl2 [33]. XPS spectra (Al Kα) were recorded with a Thermo Fisher Scientific K-Alpha instrument, Waltham, MA USA. The cloud point measurements were performed on the ultraviolet-visible (UV-vis) spectra by passing through a 10 mm path-length cell using a Jasco V-770 spectrophotometer equipped with a temperature controller (Jasco CTU-100, Tokyo, Japan). The hydrodynamic radii (Rhs) of the obtained polymer in deionized water were analyzed using a dynamic light scattering (DLS) detector (Wyatt Technology, Dyna Pro Nanostar ® , Santa Barbara, CA, USA). Cell viability was evaluated by MTT, and the optical density (OD) was measured at 490 nm with a microplate reader (Bio-Rad, Hercules, CA, USA). Cell viability was determined as ), 1.17.

Synthesis of Poly-Eu(III) and Poly-Tb(III) Complexes
A typical method for the synthesis of poly-Eu(III) and poly-Tb(III) complexes is described as follows: EuCl 3 (100.0 mg, 3.87 mmol), poly(iPMAm 85 -stat-PDMAm 15 ) (1.0 g, 0.07 mmol), and ethanol (15 mL) were added into a flask, then stirred at 40 • C for 24 h. The product was purified by dialysis against methanol using a cellophane tube (Spectra/Por 6 Membrane; MWCO: 1000), after which the product was lyophilized to produce the poly-Eu(III) complex as a white solid.

Determination of the Cloud Point (T c )
An aqueous solution of the polymer (1 wt.%) was sonicated for several minutes, and the resulting clear solution was then transferred to a 10 mm quartz cell. The transmittance of the aqueous solution at 500 nm was recorded by a UV-vis spectrophotometer equipped with a temperature controller. The solution was heated at a heating rate of 1.0 • C min −1 .

Determination of Hydrodynamic Radius (R h )
A sonicated aqueous solution of the polymer (1 wt.%) was filtrated into glass cells using a 0.45 µm PTFE filter. The relaxation time (τ) distribution and particle size distribution were obtained by the CONTIN analysis of the autocorrelation function. The apparent diffusion coefficients D were calculated using the following equation: where Γ is the relaxation frequency (Γ = τ −1 ) and q is the wavevector defined by the following equation: where λ is the wavelength of the laser beam (532 nm), θ is the scattering angle, and n is the refractive index of the media. Consequently, the hydrodynamic radius (R h ) was calculated from the Stokes-Einstein relation as follows: where k B is the Boltzmann constant, T is the temperature, and η is the viscosity of the medium.

Cell Viability Study
Typically, cell viability was investigated using the human colon carcinoma cells LoVo and DLD1 in culture. After incubation for 24 h in 96-well plates (8 × 10 4 cells mL −1 per well) using Dulbecco's modified Eagles medium (DMEM) in an incubator (36 • C, 5% CO 2 ), the culture medium was mixed with 200 µL of DMEM containing a sample of poly(iPMAm 85stat-PDMAm 15 ), poly-Eu(III), poly-Tb(III), EuCl 3 and TbCl 3 under concentrations from 0.1 to 1000.0 µg mL −1 . The mixture was further incubated for 48 h [34]. Each sample was tested in five replicates per plate, then 20 µL of MTT solution was added to the mixture in each well, which was incubated for an additional 4 h. Next, 200 µL of DMSO was added and the mixtures were shaken at room temperature. Five replicate wells were used for the control and test concentrations for each microplate. The cell viability (%) was calculated by the following equation: where A sample was the absorbance of the cells incubated in DMEM and mixture, and A control was the absorbance of the cells incubated in DMEM [35].

Synthesis of Poly-Eu(III) and Poly-Tb(III)
For the preparation of poly-Eu(III) and poly-Tb(III), we first synthesized poly(iPMAm85-stat-DMAm15) as the polymer matrix, according to the previously reported B(C6F5)3-catalyzed GTP method, using 1-methoxy-2-methyl- The monomers, iPMAm and DMAm, were completely consumed after 2 h, and the structure of the obtained polymer was verified using 1 H NMR, as shown in Figure 1. The methyl and methoxy protons, as the α-terminal group of the polymer, were observed at 1.16-1.20 (signal a) and 3.67 ppm (signal b), respectively. The signals due to the methyl protons of the N-isopropyl group in PiPMAm (signal f) and the N-methyl group in PDMAm (signal h) were observed at 0.98-1.20 and 2.81-3.19 ppm, respectively. The methenyl protons of the N-isopropyl group in PiPMAm (signal g) were observed at 4.69-4.92 ppm. The monomer composition in the copolymer could be regarded as the same as the monomer feed ratio of 85/15, due to the quantitative monomer conversion confirmed by the 1 H NMR spectrum, i.e., the obtained copolymer was poly(iPMAm85-stat-DMAm15). The monomers, iPMAm and DMAm, were completely consumed after 2 h, and the structure of the obtained polymer was verified using 1 H NMR, as shown in Figure 1. The methyl and methoxy protons, as the α-terminal group of the polymer, were observed at 1.16-1.20 (signal a) and 3.67 ppm (signal b), respectively. The signals due to the methyl protons of the N-isopropyl group in PiPMAm (signal f) and the N-methyl group in PDMAm (signal h) were observed at 0.98-1.20 and 2.81-3.19 ppm, respectively. The methenyl protons of the N-isopropyl group in PiPMAm (signal g) were observed at 4.69-4.92 ppm. The monomer composition in the copolymer could be regarded as the same as the monomer feed ratio of 85/15, due to the quantitative monomer conversion confirmed by the 1 H NMR spectrum, i.e., the obtained copolymer was poly(iPMAm 85 -stat-DMAm 15 ).

Synthesis of Poly-Eu(III) and Poly-Tb(III)
For the preparation of poly-Eu(III) and poly-Tb(III), we first synthesized poly(iPMAm85-stat-DMAm15) as the polymer matrix, according to the previously reported B(C6F5)3-catalyzed GTP method, using 1-methoxy-2-methyl-  The monomers, iPMAm and DMAm, were completely consumed after 2 h, and the structure of the obtained polymer was verified using 1 H NMR, as shown in Figure 1. The methyl and methoxy protons, as the α-terminal group of the polymer, were observed at 1.16-1.20 (signal a) and 3.67 ppm (signal b), respectively. The signals due to the methyl protons of the N-isopropyl group in PiPMAm (signal f) and the N-methyl group in PDMAm (signal h) were observed at 0.98-1.20 and 2.81-3.19 ppm, respectively. The methenyl protons of the N-isopropyl group in PiPMAm (signal g) were observed at 4.69-4.92 ppm. The monomer composition in the copolymer could be regarded as the same as the monomer feed ratio of 85/15, due to the quantitative monomer conversion confirmed by the 1 H NMR spectrum, i.e., the obtained copolymer was poly(iPMAm85-stat-DMAm15). The SEC trace of poly(iPMAm85-stat-DMAm15) exhibited a unimodal distribution, as shown in Figure 2. The SEC-determined number-average molar mass (Mn,SEC) and FT-IR spectra were recorded on a Perkin Elmer Frontier, and les fashioned into KBr pellet disks were placed into a holder for transmission IR ral analysis. The fluorescence spectra were recorded using a fluorescence rophotometer (RF-5301PC, Shimadzu, Kyoto, Japan). Quantum yields were mined by comparison of the total light emitted from the solutions to the total light ed from a known standard [Ru(bipy)3]Cl2 [33]. XPS spectra (Al Kα) were recorded a Thermo Fisher Scientific K-Alpha instrument, Waltham, MA USA. The cloud point urements were performed on the ultraviolet-visible (UV-vis) spectra by passing gh a 10 mm path-length cell using a Jasco V-770 spectrophotometer equipped with perature controller (Jasco CTU-100, Tokyo, Japan). The hydrodynamic radii (Rhs) of btained polymer in deionized water were analyzed using a dynamic light scattering ) detector (Wyatt Technology, Dyna Pro Nanostar ® , Santa Barbara, CA, USA). Cell lity was evaluated by MTT, and the optical density (OD) was measured at 490 nm a microplate reader (Bio-Rad, Hercules, CA, USA). Cell viability was determined as centage of the negative control (untreated cells). Poly(iPMAm85-Stat-PDMAm15) ) were 13.0 kg mol −1 and 1.17, respectively. All the results confirm the desired synthesis of this polymer matrix.  Poly-Eu(III) and poly-Tb(III) were prepared by coordination reactions between poly(iPMAm85-stat-DMAm15) and the rare earth chlorides EuCl3 and TbCl3, respectively. The structural analyses of poly-Eu(III) and poly-Tb(III) were first commenced by FT-IR spectroscopy. Figure 3 shows the FT-IR spectra of poly(iPMAm85-stat-PDMAm15), poly- Poly-Eu(III) and poly-Tb(III) were prepared by coordination reactions between poly(iPMAm 85 -stat-DMAm 15 ) and the rare earth chlorides EuCl 3 and TbCl 3 , respectively. The structural analyses of poly-Eu(III) and poly-Tb(III) were first commenced by FT-IR spectroscopy. Figure 3 shows the FT-IR spectra of poly(iPMAm 85 -stat-PDMAm 15 ), poly-Eu(III), and poly-Tb(III). For poly(iPMAm 85 -stat-PDMAm 15 ), the characteristic adsorptions due to the stretching and bending vibrations of the acylamino group (ν N-H and δ N-H ) and stretching vibration of carbonyl (ν C=O ) appear at 3301, 1550, and 1674 cm −1 , respectively. In comparison, the same bond vibrations in poly-Eu(III) are observed at 3293, 1538, and 1657 cm −1 , and those for poly-Tb(III) appear at 3295, 1540, and 1658 cm −1 . The redshift of these characteristic absorptions after the coordination reactions resulted from the formation of coordination bonds between Eu 3+ /Tb 3+ and the carbonyl groups in poly(iPMAm 85 -stat-PDMAm 15 ). Namely, part of the lone pair electrons of the O atom transfers to the outer orbitals of the rare earth ion, which weakens the σ covalent bond and force constant of the C=O. In addition, the inductive effect along the NH-C=O → Eu 3+ /Tb 3+ also lowers the electron density of the N atom, leading to the redshift phenomenon [36]. Poly-Eu(III) and poly-Tb(III) were prepared by coordination reactions poly(iPMAm85-stat-DMAm15) and the rare earth chlorides EuCl3 and TbCl3, resp The structural analyses of poly-Eu(III) and poly-Tb(III) were first commenced b spectroscopy. Figure 3 shows the FT-IR spectra of poly(iPMAm85-stat-PDMAm1 Eu(III), and poly-Tb(III). For poly(iPMAm85-stat-PDMAm15), the chara adsorptions due to the stretching and bending vibrations of the acylamino group ( δN-H) and stretching vibration of carbonyl (νC=O) appear at 3301, 1550, and 16 respectively. In comparison, the same bond vibrations in poly-Eu(III) are observed 1538, and 1657 cm −1 , and those for poly-Tb(III) appear at 3295, 1540, and 1658 c redshift of these characteristic absorptions after the coordination reactions result the formation of coordination bonds between Eu 3+ /Tb 3+ and the carbonyl gr poly(iPMAm85-stat-PDMAm15). Namely, part of the lone pair electrons of the transfers to the outer orbitals of the rare earth ion, which weakens the σ covale and force constant of the C=O. In addition, the inductive effect along the NH Eu 3+ /Tb 3+ also lowers the electron density of the N atom, leading to the phenomenon [36]. The structural characterizations of poly-Eu(III) and poly-Tb(III) were implemented by X-ray photoelectron spectroscopy (XPS). Figure 4a shows profiles of poly(iPMAm85-stat-PDMAm15), poly-Eu(III), and poly-Tb(III), an The structural characterizations of poly-Eu(III) and poly-Tb(III) were further implemented by X-ray photoelectron spectroscopy (XPS). Figure 4a shows the XPS profiles of poly(iPMAm 85 -stat-PDMAm 15 ), poly-Eu(III), and poly-Tb(III), and their average binding energies of O 1s, N 1s, Eu 4d, and Tb 4d are listed in Table 1. Poly(iPMAm 85 -stat-DMAm 15 ) shows peaks at 283.71, 530.51, and 399.12 eV, corresponding to the C 1s, N 1s, and O 1s, respectively, while, in addition to these peaks, poly-Eu(III) and poly-Tb(III) display additional Eu 4d and Tb 4d peaks at 139.68 and 145.12 eV, respectively. The average binding energies of O 1s and N 1s in poly-Eu(III) are 531.25 and 399.58 eV, respectively, showing a +0.74 and +0.46 eV shift, in comparison with those in poly(iPMAm 85 -stat-PDMAm 15 ). Meanwhile, the average binding energy of Eu 4d in poly-Eu(III) (139.68 eV) is 5.44 eV smaller than that of EuCl 3 (145.12 eV). The change in average binding energy before and after the coordination reaction between poly(iPMAm 85 -stat-PDMAm 15 ) and TbCl 3 shows the similar tendency, i.e., the average binding energies of O 1s and N 1s in poly-Tb(III) are +0.70 and +0.32 eV greater than those in poly(iPMAm 85 -stat-PDMAm 15 ), respectively, while the average binding energy of Tb 4d in poly-Tb (148.97 eV) is 2.86 eV smaller than that of TbCl 3 (151.83 eV). In summary, the increase in the average binding energies of O 1s and N 1s of poly-Eu(III) and poly-Tb(III) is caused by the decrease in electron density for the O 1s and N 1s atoms. On the contrary, the decrease in the average binding energy of Eu 4d and Tb 4d in the polymer-rare earth complexes is due to the enhanced electron density around the rare earth metal cations after the lone pair electrons of O and N atoms of the acylamino group are coordinated to the outer orbital of Eu 3+ or Tb 3+ . These results strongly indicate the formation of the target poly-Eu(III) and poly-Tb(III) complexes [37].

ynthesis of Statistical Copolymer
poly(iPMAm85-stat-PDMAm15). Meanwhile, the average binding energy of Eu 4d in po Eu(III) (139.68 eV) is 5.44 eV smaller than that of EuCl3 (145.12 eV). The change in avera binding energy before and after the coordination reaction between poly(iPMAm85-st PDMAm15) and TbCl3 shows the similar tendency, i.e., the average binding energies of 1s and N 1s in poly-Tb(III) are +0.70 and +0.32 eV greater than those in poly(iPMAm85-st PDMAm15), respectively, while the average binding energy of Tb 4d in poly-Tb (148 eV) is 2.86 eV smaller than that of TbCl3 (151.83 eV). In summary, the increase in t average binding energies of O 1s and N 1s of poly-Eu(III) and poly-Tb(III) is caused the decrease in electron density for the O 1s and N 1s atoms. On the contrary, the decrea in the average binding energy of Eu 4d and Tb 4d in the polymer-rare earth complexes due to the enhanced electron density around the rare earth metal cations after the lo pair electrons of O and N atoms of the acylamino group are coordinated to the ou orbital of Eu 3+ or Tb 3+ . These results strongly indicate the formation of the target po Eu(III) and poly-Tb(III) complexes [37].

Luminescence Properties of Poly-Eu(III) and Poly-Tb(III)
The luminescence properties of the poly-Eu(III) and poly-Tb(III) complexes a shown in Figure 5. The excitation spectrum of poly-Eu(III) exhibits wide-range absorpti

Luminescence Properties of Poly-Eu(III) and Poly-Tb(III)
The luminescence properties of the poly-Eu(III) and poly-Tb(III) complexes are shown in Figure 5. The excitation spectrum of poly-Eu(III) exhibits wide-range absorption peaks from 350 to 472 nm, and the peak at 355 nm shows the maximum excitation intensity, while only a negligible excitation peak is observed for EuCl 3 in all the excitation fluorescence spectra (Figure 5a). This phenomenon can be attributed to the π-π* transition by exciting the carbonyl and amide groups of the complexes. In addition, the conjugated structure increases the electron delocalization and the absorption of ultraviolet light [33], which makes the intensity of the excitation peak increase sharply. Figure 5b indicates that EuCl 3 exhibits very weak emission peaks, but poly-Eu(III) gives four strong emission peaks at 578, 593, 622, and 651 nm, which are responsible for the transition from an excited 5 D 0 state to multiplet 7 F J , (J = 0, 1, 2, 3) states, respectively. Moreover, the 4f orbital is shielded by the outer shell of the 5s and 5p orbitals, and the f -f absorption bands are very narrow, which makes the intensity of the emission peak at 622 nm ( 5 D 0 → 7 F 2 ) 13.2 times that of EuCl 3 . appears at 350 nm. When compared with poly-Tb(III), TbCl3 shows very weak absorption, mainly due to the extremely low solubility of TbCl3 in water. Figure 5d is the emission spectra of poly-Tb(III) and TbCl3. Similarly to poly-Eu(III), poly-Tb(III) has four emission peaks at 489, 545, 588, and 654 nm, due to the transitions of 5 D4 → 7 Fi (i = 6,5,4,3). The intensity at the maximum emission (545 nm) of poly-Tb(III), due to the 5 D4 → 7 F5 transition, is 13.4 times that of TbCl3, which originated from the transitions between the 4f states in poly-Tb(III). The fluorescent lifetimes of the poly-Eu(III) and poly-Tb(III) complexes are calculated according to the luminescence decay curves in Figure 6. Poly-Eu(III) and poly-Tb(III) have τpoly-Eu(III) = 4.57 ms and τpoly-Tb(III) = 7.50 ms, respectively. The photoemission efficiency of The excitation spectra of the poly-Tb(III) complex and TbCl 3 are shown in Figure 5c. Poly-Tb(III) displays wide-range absorption from 280 to 390 nm, and the maximum peak appears at 350 nm. When compared with poly-Tb(III), TbCl 3 shows very weak absorption, mainly due to the extremely low solubility of TbCl 3 in water. Figure 5d is the emission spectra of poly-Tb(III) and TbCl 3 . Similarly to poly-Eu(III), poly-Tb(III) has four emission peaks at 489, 545, 588, and 654 nm, due to the transitions of 5 D 4 → 7 F i (i = 6,5,4,3). The intensity at the maximum emission (545 nm) of poly-Tb(III), due to the 5 D 4 → 7 F 5 transition, is 13.4 times that of TbCl 3 , which originated from the transitions between the 4f states in poly-Tb(III).
The fluorescent lifetimes of the poly-Eu(III) and poly-Tb(III) complexes are calculated according to the luminescence decay curves in Figure 6. Poly-Eu(III) and poly-Tb(III) have τ poly-Eu(III) = 4.57 ms and τ poly-Tb(III) = 7.50 ms, respectively. The photoemission efficiency of the two rare earth complexes is increased, because the much higher coordination ability of the poly(iPMAm 85 -stat-DMAm 15 ) matrix than the chloride anion further stabilizes the Eu 3+ or Tb 3+ . The more stable coordination in the rare earth complexes can largely enhance the absorption coefficient. Therefore, the initial strong absorption of ultraviolet energy excites the amide ligand to the excited singlet (S1) state, and the transition from the S1 state to the triplet (T) state, and then the energy transfers non-radiatively from the lowest triplet state of the ligand to the resonance state of Eu 3+ or Tb 3+ [38,39]. The energy undergoes multiphoton relaxation and subsequent emission in the visible light region.
of the poly(iPMAm85-stat-DMAm15) matrix than the chloride anion further stabilizes the Eu 3+ or Tb 3+ . The more stable coordination in the rare earth complexes can largely enhance the absorption coefficient. Therefore, the initial strong absorption of ultraviolet energy excites the amide ligand to the excited singlet (S1) state, and the transition from the S1 state to the triplet (T) state, and then the energy transfers non-radiatively from the lowest triplet state of the ligand to the resonance state of Eu 3+ or Tb 3+ [38,39]. The energy undergoes multiphoton relaxation and subsequent emission in the visible light region.

Thermoresponsive Properties
The thermal phase transition behavior of poly(iPMAm85-stat-DMAm15), poly-Eu(III), and poly-Tb(III) is studied by a UV-vis spectrophotometer and dynamic light scattering (DLS) measurements. The temperature at light transmittance of 1 wt.% polymer aqueous solution = 50% on the transmittance-temperature curve is used as the cloud temperature (Tc). DLS is used to monitor the polymer assembly state, by determining the hydrodynamic radius (Rh) of a polymer in the same aqueous solution. Figure 7 shows the transmittance dependence of poly(iPMAm85-stat-DMAm15), poly-Eu(III), and poly-Tb(III) on the temperature. Each transmittance-temperature curve shows a sharp transmittance decrease at 32-40 °C, indicating that the polymer undergoes a quick thermal phase transition. The Tcs of poly(iPMAm85-stat-DMAm15), poly-Eu(III), and poly-Tb(III) are 36.1, 36.4, and 36.8 °C, respectively ( Table 2). It seems that the complexes have a slightly higher Tc than poly(iPMAm85-stat-DMAm15), but the difference in Tc is so small that we cannot exclude the possibility that it may be caused by experimental error. In general, the incorporation of highly hydrophilic metal cations into thermoresponsive polymers would increase the Tc value.

Thermoresponsive Properties
The thermal phase transition behavior of poly(iPMAm 85 -stat-DMAm 15 ), poly-Eu(III), and poly-Tb(III) is studied by a UV-vis spectrophotometer and dynamic light scattering (DLS) measurements. The temperature at light transmittance of 1 wt.% polymer aqueous solution = 50% on the transmittance-temperature curve is used as the cloud temperature (T c ). DLS is used to monitor the polymer assembly state, by determining the hydrodynamic radius (R h ) of a polymer in the same aqueous solution. Figure 7 shows the transmittance dependence of poly(iPMAm 85 -stat-DMAm 15 ), poly-Eu(III), and poly-Tb(III) on the temperature. Each transmittance-temperature curve shows a sharp transmittance decrease at 32-40 • C, indicating that the polymer undergoes a quick thermal phase transition. The T c s of poly(iPMAm 85 -stat-DMAm 15 ), poly-Eu(III), and poly-Tb(III) are 36.1, 36.4, and 36.8 • C, respectively ( Table 2). It seems that the complexes have a slightly higher T c than poly(iPMAm 85 -stat-DMAm 15 ), but the difference in T c is so small that we cannot exclude the possibility that it may be caused by experimental error. In general, the incorporation of highly hydrophilic metal cations into thermoresponsive polymers would increase the T c value. the polymer chain undergoes a coil-to-globule transition from the hydrated unimolecular state at 28 °C to the dehydrated aggregated state at 42 °C. It should be noted that the Rh size of poly(iPMAm85-stat-DMAm15) at 42 °C was almost the same in our previous work [32], while poly-Eu(III) and poly-Tb(III) had a 2.4 times larger Rh than poly(iPMAm85-stat-DMAm15). This phenomenon could also be caused by the polyelectrolyte nature of poly-Eu(III) and poly-Tb(III) [40,41].

Assessment of Cell Viability
Since the Tc values of poly-Eu(III) and poly-Tb(III) are very close to the human body temperature, they are of great potential to be used as drug delivery systems (DDSs). Precisely for this, the assessment of cell viability is further carried out to evaluate their cytotoxicity for DDS materials. LoVo (CCL-229 TM ) is a cell line isolated in 1971 from the large intestine of a 56-year-old male, Caucasian, grade IV Dukes C colorectal cancer patient. LoVo cells can be used for cancer, toxicology, and immuno-oncology research, and high-throughput screening. DLD1 (CCL-221 TM ) is a colorectal adenocarcinoma cell line isolated from the large intestine of a colon adenocarcinoma patient. It can be used for cancer research. The cytotoxicity towards the human colon carcinoma cells LoVo and DLD1 is studied by comparing the biocompatibility of poly(iPMAm85-stat-DMAm15), poly-Eu(III), and poly-Tb(III), as well as EuCl3 and TbCl3. The percentage of cell viability is determined by comparing them with the control group, in which the cells are not The R h values of poly(iPMAm 85 -stat-DMAm 15 ), poly-Eu(III), and poly-Tb(III) in Table 2 are 3.1, 5.2, and 6.9 nm, respectively, at 28 • C lower than T c . In this case, all of them should be in a unimolecular hydrated state. It is interesting that poly-Eu(III) and poly-Tb(III) have much greater R h values than the parent poly(iPMAm 85 -stat-DMAm 15 ). It is rational to assume that poly-Eu(III) and poly-Tb(III) incorporated with rare earth metal cations, in a sense, are polyelectrolytes, which have stretched chain structures and, thus, hydrated dimensions, i.e., greater R h values. When the temperature is enhanced to 42 • C higher than T c , the R h values are 1758.4, 4216.3, and 3952.5 nm, respectively. Undoubtedly, the polymer chain undergoes a coil-to-globule transition from the hydrated unimolecular state at 28 • C to the dehydrated aggregated state at 42 • C. It should be noted that the R h size of poly(iPMAm 85 -stat-DMAm 15 ) at 42 • C was almost the same in our previous work [32], while poly-Eu(III) and poly-Tb(III) had a 2.4 times larger R h than poly(iPMAm 85stat-DMAm 15 ). This phenomenon could also be caused by the polyelectrolyte nature of poly-Eu(III) and poly-Tb(III) [40,41].

Assessment of Cell Viability
Since the T c values of poly-Eu(III) and poly-Tb(III) are very close to the human body temperature, they are of great potential to be used as drug delivery systems (DDSs). Precisely for this, the assessment of cell viability is further carried out to evaluate their cytotoxicity for DDS materials. LoVo (CCL-229 TM ) is a cell line isolated in 1971 from the large intestine of a 56-year-old male, Caucasian, grade IV Dukes C colorectal cancer patient. LoVo cells can be used for cancer, toxicology, and immuno-oncology research, and high-throughput screening. DLD1 (CCL-221 TM ) is a colorectal adenocarcinoma cell line isolated from the large intestine of a colon adenocarcinoma patient. It can be used for cancer research. The cytotoxicity towards the human colon carcinoma cells LoVo and DLD1 is studied by comparing the biocompatibility of poly(iPMAm 85 -stat-DMAm 15 ), poly-Eu(III), and poly-Tb(III), as well as EuCl 3 and TbCl 3 . The percentage of cell viability is determined by comparing them with the control group, in which the cells are not exposed to polymer samples. When exposed to EuCl 3 and TbCl 3 , the cell viability of LoVo and DLD1 drops sharply, as shown in Figure 8. Furthermore, the lethality resulting from EuCl 3 and TbCl 3 increases when increasing the salt concentration from 0.1 to 1000.0 µg mL −1 , indicating a dose-dependent cytotoxic effect. In contrast to the rare earth chlorides, poly(iPMAm 85stat-DMAm 15 ), poly-Eu(III), and poly-Tb(III) turn out to be non-toxic in both LoVo and DLD1 cells, with nearly 100% vitality in a wide concentration range (0.1 to 1000.0 µg mL −1 ), and no obvious difference in cytotoxicity is observed between the parent polymer matrix and its complexes. These results suggest that the complexation of a polymer matrix, poly(iPMAm 85 -stat-DMAm 15 ), and Eu 3+ or Tb 3+ greatly reduces the toxicity of inorganic rare earth chlorides. These results confirm that the coordination of poly(iPMAm 85 -stat-DMAm 15 ) with Eu 3+ and Tb 3+ to form complexes can enhance the biocompatibility and eliminate the cell. These new types of complexes are potential luminescent probing materials for targeted applications in cytology and immune omics, and anti-tumor therapy. mL −1 , indicating a dose-dependent cytotoxic effect. In contrast to the rare earth chlorides, poly(iPMAm85-stat-DMAm15), poly-Eu(III), and poly-Tb(III) turn out to be non-toxic in both LoVo and DLD1 cells, with nearly 100% vitality in a wide concentration range (0.1 to 1000.0 µg mL −1 ), and no obvious difference in cytotoxicity is observed between the parent polymer matrix and its complexes. These results suggest that the complexation of a polymer matrix, poly(iPMAm85-stat-DMAm15), and Eu 3+ or Tb 3+ greatly reduces the toxicity of inorganic rare earth chlorides. These results confirm that the coordination of poly(iPMAm85-stat-DMAm15) with Eu 3+ and Tb 3+ to form complexes can enhance the biocompatibility and eliminate the cell. These new types of complexes are potential luminescent probing materials for targeted applications in cytology and immune omics, and anti-tumor therapy.

Conclusions
Poly-Eu(III) and poly-Tb(III) complexes are synthesized by a coordination reaction between poly(iPMAm85-stat-DMAm15) and their parent chloride salts. The complexes have coordination interactions between the O and N atoms of the acylamino group with Eu 3+ and Tb 3+ , which bring about the strong emission peaks at 622 and 545 nm, respectively. Poly-Eu(III) and poly-Tb(III) have Tcs near the human body temperature and slightly higher than that of the parent polymer matrix. The assessment of cell viability verifies that poly-Eu(III) and poly-Tb(III) can greatly enhance the biocompatibility and reduce the cell toxicity. These PAm rare earth complexes are expected to be used as luminescent probes in the biomedical field.