Sodium Hydroxide → 2-(6-Hydroxy-3-oxo-3H-xanthen-9-yl)benzoic acid

HRID 1440978Notes

Reaction equation

HRID 1440978 reaction equation
[OH-].[Na+].[CH:3]1[C:8](N=C=S)=[CH:7][C:6]2[C:12]([O:14][C:15]3([C:25]4[CH:26]=[CH:27][C:28]([OH:30])=[CH:29][C:24]=4[O:23][C:17]4[CH:18]=[C:19]([OH:22])[CH:20]=[CH:21][C:16]3=4)[C:5]=2[CH:4]=1)=[O:13]>O.CN(C)C=O>[CH:3]1[CH:8]=[CH:7][C:6]([C:12]([OH:14])=[O:13])=[C:5]([C:15]2[C:16]3[CH:21]=[CH:20][C:19]([OH:22])=[CH:18][C:17]=3[O:23][C:24]3[C:25]=2[CH:26]=[CH:27][C:28]([CH:29]=3)=[O:30])[CH:4]=1

Participants

5

Reactant 2

HCID 14798Sodium Hydroxide2×HCID 18730Fluorescein 5-isothiocyanate2×

Product 1

HCID 33832-(6-Hydroxy-3-oxo-3H-xanthen-9-yl)benzoic acid

Reagents, catalysts & solvents 2

HCID 962WaterSolventHCID 6228DimethylformamideSolvent

Procedure

Bovine serum albumin (100 mg) was dissolved in distilled water (2.0 ml) and the mixture was adjusted to a pH of 9 with 1N NaOH. Fluorescein isothiocyanate (FITC, 100 mg) in dimethylformamide (1.0 ml) was added by drops with stirring while maintaining the pH at 9 by addition of 1N NaOH. After all of the FITC was added, the mixture was stirred at room temperature for two hours. The solution was then dialysed in a cellulose dialyzing tube (MW 12,000-14,000) against 10 mM phosphate buffer (1.0 liter, pH 7) for two hours. The dialyzing tube was then dialysed against two changes of 0.9% aqueous sodium chloride (4.0 liters), for 19 hours each time, and then against three changes of 10% dimethylsulfoxide in 0.9% aqueous sodium chloride, for 24 hours each time. The final solution showed no sign of fluorescein in the dialysis solution. The resulting immunogen solution was then removed from the dialyzing tube and lyophilized to yield an orange solid. The immunogen was used according to standard procedures to produce a rabbit anti-fluorescein antibody capture reagent. The antibody was purified according to the procedure described in Example 1.c. above.

Workup

  1. temperaturewhile maintaining the pH at 9
  2. stirringthe mixture was stirred at room temperature for two hours
  3. customThe solution was then dialysed in a cellulose dialyzing tube (MW 12,000-14,000) against 10 mM phosphate buffer (1.0 liter, pH 7) for two hours
  4. customThe dialyzing tube was then dialysed against two changes of 0.9% aqueous sodium chloride (4.0 liters), for 19 hours each time
  5. customThe resulting immunogen solution was then removed from the dialyzing tube
  6. customto yield an orange solid

Sources & evidence

ORD record
ord-454f22e766744204a6fa80d9f505a5c9
Source dataset
uspto-grants-1996_03
Patent number
US05501985

Reaction notes

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