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. 2021 Apr 1;11:7347. doi: 10.1038/s41598-021-86762-6

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© The Author(s) 2021

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/.

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¹H-NMR analysis to characterize PTX-imidazole carbamate formation: (a) Schematic diagram of reactions involved for PTXNR functionalization and TTZ conjugation on the surface of PTXNRs. (b) ¹H-NMR spectra of hydrolyzed carbonyldiimidazole (CDI), PTX-imidazole carbamate (PTXNR-CDI), and PTXNR. The NMR spectra were acquired at room temperature. For characterization of PTXNR and PTXNR-CDI deuterated chloroform (CDCl₃, δ 7.24) and hydrolyzed CDI deuterated oxide (D₂O, δ 4.65) were used as reference carrier solvents. (c) The fluorescence intensity of PTXNR-TTZ confirms the conjugation of TTZ on the surface of PTXNRs. The F.I. of bare PTXNRs (grey) and Alexa 594 fluorophore bound TTZ conjugated PTXNR (PTXNR-TTZ) (green) shows a significant increase in emission spectra. (d) The quantitative mean F.I. of PTXNR-TTZ increases by ~ 14,000 fold (2.54 logarithmic fold) compared to bare PTXNRs indicating TTZ is effectively conjugated on the surface of PTXNRs using a CDI linker.