Reaction #336805

Reaction equation

fluorescein isothiocyanate

Et3N

→

fluorescein

Solvents

DMF

CH2Cl2

Conditions

Workup

1. 1
   OtherAn orange colored precipitate was immediately formed
2. 2
   Otherremained during the entire period of the reaction
3. 3
   OtherAfter completion of the reaction
4. 4
   Filtrationthe insoluble precipitate was filtered off
5. 5
   Otherand the solvent was removed under reduced pressure
6. 6
   workup.ADDITIONThe residue was diluted with CH2Cl2 (15 mL)
7. 7
   Washwashed with 1N HCl (2×15 mL) and H2O (2×15 mL)
8. 8
   DryingThe organic phase was dried over anhydrous Na2SO4
9. 9
   Otherevaporated under reduced pressure

Procedure

The starting amine (5) (0.15 g, 0.12 mmol) and fluorescein isothiocyanate (FLITC) (0.056 g, 0.14 mmol) were dissolved in a mixture of DMF and CH2Cl2 (6 mL, 1:5 ratio) at room temperature. The reaction was cooled to 0° C., and then an excess amount of Et3N (66 μL, 0.48 mmol) was added dropwise. An orange colored precipitate was immediately formed and remained during the entire period of the reaction. After completion of the reaction, the insoluble precipitate was filtered off; and the solvent was removed under reduced pressure. The residue was diluted with CH2Cl2 (15 mL), washed with 1N HCl (2×15 mL) and H2O (2×15 mL). The organic phase was dried over anhydrous Na2SO4 and evaporated under reduced pressure to afford 0.18 g (92%) of fluorescein-labeled guanidine (17) as an orange colored foam-shaped solid. 1H NMR (CD3OD): δ 8.20 (s, 1H, ArH), 7.86 (m, 1H, ArH), 7.22 (m, 2H, ArH), 6.64 (m, 3H, ArH), 6.56 (m, 2H, ArH), 3.86 (m, 2H, CH2NHCS), 3.66 (m, 12H, CH2O), 3.46 (m, 6H, CH2NHCO), 3.38 (m, 6H, CH2NH), 2.58 (m, 2H, CH2CO), 2.42 (m, 6H, CH2CO), 1.48 (s, 27H, t-Bu), 1.46 (s, 27H, t-Bu).

Source