Preparative purification of tetrabromotetrachlorofluorescein and phloxine B by centrifugal counter-current chromatography.
A centrifugal counter-current chromatographic method for preparative purification of commercial tetrabromotetrachlorofluorescein and Phloxine B (D&C Red Nos. 27 and 28, respectively) was developed. Ethyl acetate-n-butanol-0.01 M ammonium acetate (1:1:2) was used as the two-phase solvent system. Each purification trial involved 50 mg of sample and yielded 22 mg (+/- 2 mg) of pure dye. The purity of the product was measured by high-performance liquid and thin-layer chromatography and was found to be 99.9%. The partition coefficients of these compounds were found to be highly concentration-dependent in the two-phase solvent system used. If this problem can be circumvented, then the counter-current chromatographic method can be extended for use with gram quantities of dye.
Weisz
A
A
Division of Colors and Cosmetics, Food and Drug Administration, Washington, DC 20204.
Langowski
A J
AJ
Meyers
M B
MB
Thieken
M A
MA
Ito
Y
Y
D020001:1-Butanol; D000085:Acetates; D000440:Butanols; D002498:Centrifugation; D002851:Chromatography, High Pressure Liquid; D002853:Chromatography, Liquid / Q000295:instrumentation / Q000379:methods*; D002855:Chromatography, Thin Layer; D010697:Eosine I Bluish / Q000302:isolation & purification*; D012997:Solvents
D000085:Acetates; D000440:Butanols; D012997:Solvents; C007650:ethyl acetate; D020001:1-Butanol; D010697:Eosine I Bluish; C018824:ammonium acetate