Application Of Aqueous Two-Phase Extraction And Aqueous Two-Phase Floatation On Bioseparation And Analysis Of Natural Product

Aqueous Two-Phase Extraction (ATPE) is a new extraction method based on the different distribution coefficiency of components in two incompatible substances that are both water-soluble. Since it does not use toxic organic solvent, and has mild operation conditions with simple steps and time-saving, it has demonstrates a good prospect in biological separation, extraction and determination of active ingredients in Chinese herbal medicine.Aqueous Two-Phase Flotation (ATPF) is a brand-new technology in separation and concentration which combines the advantages of aqueous two-phase extraction and solvent sublation, and it overcomes the problems of low concentration factor and easy emulsification in aqueous two-phase extraction, and the secondary pollution brought by toxic organic solvent used in solvent sublation.In this paper, ATPE was for the first time used to separate and concentrate ergosterol in the yeast, Magnolol and Honokiol in Magnolia officinalis Rehd., and determine the concentration. Also ATPF was for the first time used to separate and concentrate lincomycin. Specific studies are described below. In the first part, ethanol used during saponification process was also used as one phase during ATPE to extract ergosterol for determination. The paper examined the effects of KOH content, the type and content of inorganic salts, ethanol content and concentration of ergostrol, and under the optimized conditions, ergosterol was extracted from yeast and determined by HPLC. The concent of ergostrol in yeast sample was 2.74 mg/g, RSD was 2.08%(n=5), recoveries were 91.6%～96.9%.In the second part, ATPE was used to analyse Magnolia officinalis Rehd. Which were determined by UV spectrophotometry afterwards. The paper examined the effects of the type and content of inorganic salts, ethanol content, and pH values, and under the optimized conditions, Magnolol and Honokiol are extracted from Magnolia officinalis Rehd. and determined by UV-spectrophometer. The total concents of Magnolol and Honokiol in samples from Yunnan and Sichuan were 47.67 mg/g and 55.44 mg/g, RSD were 2.42% and 3.36%, and the recoveries were 80.70%～117.87%.In the third part, ATPF was used to separate and concentrate lincomycin. In the ATPF system of polyethylene glycol (PEG)/inorganic salt, the effects of solution pH, PEG molecular weight, concentration of inorganic salt in aqueous solution, nitrogen flow rate and floatation time were investigated in details, and the optimal conditions for ATPF were obtained. Under optimal conditions, the efficiency of ATPF was 85%, the distribution ratio is 118 and the concentration coefficiency is 16, and those results were compared with ATPE. The preliminary kinetics of the process of lincomycin in ATPF was also studied.