2.5. Estimation of aflatoxin content of basal medium by HPLC
After 21 days of incubation period, the cultures present in the medium were subjected to aflatoxin analysis by HPLC using Kobra cell.
20 g of the culture medium
and 5 g of sodium chloride
were blended
with
100 ml of 80% methanol for 1 min
. After filtration, the blended contents were filtered through a fluted filter paper into a 250 ml beaker.
The 25 ml of filtrate was made up to 50 ml with distilled water. The contents were allowed to pass through a glass fiber filter
. About 10 ml of the filtrate was allowed to absorb on the Aflatest column used for cleanup, (AflaprepR
R-Biopharm Rhone Ltd.,
Glasgow, UK, previously conditioned with 10 ml of PBS) and rinsed with 10 ml of HPLC grade water thrice.
The 1 ml methanol was added to the column and the methanol eluate was collected in a vial.
The 25 ll of meth- anol filtrate was injected into HPLC system for analysis.