Separation/Extraction/Detection of Chloramphenicol Using Binary Small Molecule Alcohol-Salt Aqueous Two-phase System Coupled with High-performance Liquid Chromatography

A green and sensitive sample separation and purification method coupled with high-performance liquid chromatography(HPLC) was developed for the analysis of chloramphenicol(CAP). One element small molecule alcohol-salt aqueous two-phase system(ATPS) can’t effectively adjust the polarity of the system, but binary small molecule alcohol-salt ATPS can adjust the polarity and improve the extraction efficiency of antibiotics. A binary aqueous two-phase system based on 1-propanol+2-propanol and NaH 2 PO 4 system was formed and applied to the separation and purification of trace CAP in real samples. The influence factors on partition behaviors of CAP were discussed, including the types and mass of salts, the volume ratio of alcohol, the pH, temperature and the standing time. The optimal condition was found at pH=5.0, 2.5 g of NaH 2 PO 4 , 3.0 mL of 1-propanol and 2-propanol(volume ratio 1:1) and 30 °C by using response surface methodology. Under this optimal condition, the extraction efficiency of CAP reached 98.91%, and partition coefficient of CAP was 17.31.