[Method of separating and determining deacetylcephalosporin C in the fermentation media of Acremonium chrysogenum]

During fermentation of Acremonium chrysogenum, known cephalosporin-producing organism, there is formed, in addition, the main metabolite cephalosporin C. Its 3-desacetyl derivative, desacetylcephalosporin (DACS). The admixture of this derivative is not desirable. Such factors as the value of pH aeration level and concentration of free glucose influence formation of DACS. A method of descending chromatography with the use of Whatman paper No. 4 and the system of n-butanol-acetic acid-water (60:15:25) was elaborated for separation of the cephalosporins. The spot of DACS with Rf 0.15 was developed in UV light at 254 nm and eluated with phosphate buffer. The eluate absorption was estimated at a wave length of 260 nm and the amount of DACS was determining by the standard curve. The effect of various fermentation conditions on formation of DACS was compared. The highest levels of DACS was compared. The highest levels of DACS formation were observed in 500 ml flasks with unfavourable aeration and agitation conditions. Determination of the DACS levels during the fermentation process provides control of pH, aeration rate and sugar levels which results in minimum formation of DACS. Close chemical structures of DACS and cephalosporin C impede their separation during the process of cephalosporin C recovery.