反应详情

To 50 ml. of a medium comprising Brewer modified throgylcollate broth (Baltimore Biological Laboratories), a stock culture of Clostridium perfringens ATCC 19574 is inoculated under anaerobic conditions. The medium is incubated under anaerobic conditions, at 37° C for a period of 96 hours and the resultant culture is employed to inoculate 1 liter of the same medium. At the time of inoculation, 2 gm. of 3α, 7α, 12α-trihydroxy-5β-cholanic acid dissolved in 15 ml. of ethenol and sterilized through a Seitz pad is added. The flask is incubated anaerobically for 72 hours and the broth is then filtered on a Buchner funnel with the aid of filter cel and the filtrate is then acidified to pH 1-2 with HCl, extracted 3 times with ether and the combined ether extracts washed with water, dried over sodium sulfate, and evaporated to dryness in vacuo. The residue is then crystalized from ethyl acetate-heptane to yield crude 3α, 7α,-dihydroxy-5β-cholanic acid. If further purification is required, the resultant product may be further treated in accordance with the procedures set forth in my copending application, Ser. No. 417,170, filed Nov. 19, 1973.

1. additionis added
2. waitThe flask is incubated anaerobically for 72 hours
3. filtrationthe broth is then filtered on a Buchner funnel with the aid of filter cel
4. extractionextracted 3 times with ether
5. washthe combined ether extracts washed with water
6. dry with materialdried over sodium sulfate
7. customevaporated to dryness in vacuo
8. customThe residue is then crystalized from ethyl acetate-heptane