Reaction #54905

ord-0c1c367d3ea64a0d92066fd5febe77bf

Reaction equation

CN[C@@H]1[C@@H](O)[C@@H](O[C@@H]2[C@@H](O)[C@H](O[C@H]3O[C@H]([C@@H](C)NC)CC[C@H]3N)[C@@H](N)C[C@H]2N)OC[C@]1(C)O
Gentamicin
O=C1OC2(c3ccc(O)cc3Oc3cc(O)ccc32)c2ccc(N=C=S)cc21
fluorescein isothiocyanate
→
O=C(O)c1ccccc1-c1c2ccc(=O)cc-2oc2cc(O)ccc12
fluorescein

Reactants

—
Sephadex
CN[C@@H]1[C@@H](O)[C@@H](O[C@@H]2[C@@H](O)[C@H](O[C@H]3O[C@H]([C@@H](C)NC)CC[C@H]3N)[C@@H](N)C[C@H]2N)OC[C@]1(C)O
Gentamicin
O=C1OC2(c3ccc(O)cc3Oc3cc(O)ccc32)c2ccc(N=C=S)cc21
fluorescein isothiocyanate
—
reaction mixture

Reagents

None

Solvents

O=C([O-])O.O=C([O-])[O-].[Na+].[Na+]
sodium carbonate bicarbonate

Conditions

Detailed conditions
See reaction.notes.procedure_details.

Workup

  1. 1
    Washeluted with carbonate/bicarbonate buffer as above, with a flow rate of 1.8ml/h
  2. 2
    workup.WAITUnreacted gentamicin left the column first
  3. 3
    Otherwell separated from the gentamicin peak
  4. 4
    Washwas not eluted

Procedure

Gentamicin (1mM) and fluorescein isothiocyanate (FITC) (1.25mM) were allowed to react in 0.05M sodium carbonate/bicarbonate buffer, pH 9.0, for 2 hours at room temperature. 2ml of the reaction mixture was applied to a column (1 × 97cm) of G-15 grade Sephadex and eluted with carbonate/bicarbonate buffer as above, with a flow rate of 1.8ml/h. Column fractions (1.8ml) were assayed for gentamicin content by radio-immunoassay and for fluorescein content by fluorimetry. FIG. 1 of the accompanying drawings shows the result. Unreacted gentamicin left the column first. The FTC-G product exited next, well separated from the gentamicin peak. Unreacted FITC was very strongly held on the G-15 column and was not eluted.

Source

DOI: 10.6084/m9.figshare.5104873.v1Patent: US04150949uspto-grants-1979_04