FITC (0.39 g, 1.00 mmol) was dissolved in 20-ml anhydrous DCM under the ice bath. Then, NH2-EO4-N3 (0.24 g, 1.1 mmol) was added. The solution was allowed to slowly rise to room temperature and reacted overnight in the dark. After reaction, the solution was concentrated, and the product was purified by column chromatography. Fls-EO4-N3 was obtained as a brown oil with a yield of 41%.

1H NMR (500 MHz, DMSO): δ (ppm) 8.30 (s, 1H, Ph), 7.75 (d, 1H, Ph), 7.17 (d, 1H, Ph), 6.67 (s, 2H, Ph), 6.57 (m, 4H, Ph), 3.70 to 3.45 (m, 14H, CSNH─(CH2CH2O)3CH2─), and 3.25 (t, 2H, ─CH2─N3). MALDI-TOF (positive ion): Calcd m/z for C29H29O8N5S: 607.17; found, 608.21 (+H+).

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