Synergistic co-delivery of doxorubicin and melittin using functionalized magnetic nanoparticles for cancer treatment: loading and in vitro release study by LC–MS/MS

Figures & data

Figure 1. XRD patterns of (a) uncoated Fe₃O₄ NPs and (b) CA-MNPs.

Figure 2. FTIR spectra of (a) uncoated Fe₃O₄ NPs, (b) citric acid and (c) CA-MNPs.

Figure 3. (a) TEM image, (b) FESEM image and (c) hydrodynamic size distribution of CA-MNPs by DLS analysis.

Figure 4. Magnetization curves of (a) uncoated Fe₃O₄ NPs and (b) CA-MNPs.

Figure 5. (a) Full scan mass spectrum of DOX and (b) product ion spectrum of the mass-selected [M + H]⁺ ion of m/z 544.1.

Figure 6. (a) Full scan mass spectrum of MEL and (b) product ion spectrum of the mass-selected [M + 5H]⁵⁺ ion of m/z 570.2.

Figure 7. Representative SRM chromatograms of DOX and MEL.

Table 1. Cytotoxicity of different treatment combinations of DOX and MEL against MCF-7 cells after 48 h incubation.

Treatment samples	IC50 (ppm)	IC50 (µM)	CI50
Free DOX	0.3726	0.6855
Free MEL	7.8023	2.7410
DOX/MEL (1:5)	0.1397/3.6602	0.2570/1.2858	0.8440
DOX/MEL (1:4)	0.1002/2.0997	0.1843/0.7376	0.5380
DOX/MEL (1:3)	0.1914/3.0085	0.3521/1.0569	0.8991
DOX/MEL (1:2)	0.2091/2.1908	0.3847/0.7696	0.8418
DOX/MEL (1:1)	0.2244/1.1750	0.4128/0.4128	0.7528

Figure 8. DOX and MEL loading efficiency (binding isotherm of DOX and MEL with CA-MNP).

Figure 9. (a) FESEM image and (b) hydrodynamic size distribution of (DOX/MEL)-loaded CA-MNPs by DLS analysis.

Figure 10. In vitro release profiles of (a) DOX and (b) MEL from (DOX/MEL)-loaded CA-MNPs at different pH values.

Table 2. Release kinetics data of DOX and MEL from CA-MNPs at different pH values.

Drug	pH	Zero-order		First order		Hixson–Crowell		Higuchi
		K	R^2	K × 10^2	R^2	K × 10^2	R^2	K	R^2
DOX	4.5	0.45	0.88	0.51	0.90	1.32	0.89	4.87	0.92
DOX	5.5	0.30	0.90	0.19	0.90	0.61	0.90	3.28	0.94
DOX	7.4	0.12	0.75	0.06	0.75	0.21	0.75	1.36	0.81
MEL	4.5	0.40	0.93	0.41	0.97	1.10	0.96	5.20	0.98
MEL	5.5	0.08	0.94	0.11	0.94	0.15	0.94	0.90	0.97

Figure 11. Viabilities of MCF-7 cells after 48 h treatment with medium containing different concentrations of CA-MNP.

Figure 12. Viabilities of MCF-7 cells after 48 h treatment with medium containing different formulations at various concentrations of DOX and MEL. Statistical significance between groups: free DOX and free MEL vs. (DOX/MEL)-loaded CA-MNPs *p<.05, free DOX and free MEL vs. free (DOX/MEL) (1:4) ^#p<.05.