Abstract

A pre-column derivatization method was developed to analyze polyamidoamine (PAMAM) dendrimers. The fluorescein isothiocyanate (FITC) derivation method improved the retention of PAMAM dendrimers on the reversed-phase column of high performance liquid chromatography (RP-HPLC). The derivation benefited the UV-Vis detection of PAMAM, although the fluorescence of FITC was quenched by PAMAM dendrimers. The primary amine-terminated PAMAM dendrimers, generation 3 (PAMAM-NH2 (G3)), were quantitatively determined by UV-Vis spectroscopy over the concentration range of 10–80 μg mL−1, with the calibration curve expressed as A = 131.6C (μg mL−1) − 295.6 (R2 = 0.9901) and the detection limit of 2.7 μg mL−1. The binding ratio of FITC to PAMAM-NH2 (G3) was determined as 8 : 1 by liquid chromatography-electrospray ionization-ion trap mass spectrometry (LC-ESI-IT-MS).

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