Ce. Tsai et F. Kondo, SIMPLE CONTINUOUS AND SIMULTANEOUS DETERMINATION OF MULTIPLE SULFONAMIDE RESIDUES, Journal of food protection, 56(12), 1993, pp. 1067-1072
A new continuous separation method was developed for the determination
of nine different sulfonamides (sulfaguanidine, sulfamethazine, sulfa
pyridine, sulfadiazine, sulfathiazole, sulfamethizole, sulfamethoxazol
e, sulfadimethoxine, and sulfaquinoxaline). Bioassay on minimum medium
seeded with Bacillus subtilis ATCC 6633 was carried out for detection
. An extract taken from an agar block of the clear inhibition zone on
minimum medium produced by a mixture of sulfonamides was then subjecte
d to high-performance liquid chromatography. For identification, high-
performance liquid chromatography analyses were performed using two di
fferent columns and analytical conditions. Using a mu-Bondapak C18 col
umn, the sulfonamides were separated at room temperature using a mobil
e phase of methanol: 0.1 M KH2PO4 (30:70, vol/vol) at a flow rate of 1
.0 ml/min. A variable wavelength detector set at 265 nm and recorder s
et at 4 mm/min were used for the detection. The entire mixture was res
olved as eight peaks from 4.68 to 50.78 min. When an Asahipak GS-320 c
olumn was employed, nine peaks were separated with retention times ran
ging from 12.62 to 54.43 min using a mobile phase of acetonitrile: 1%
acetic acid (25:75, vol/vol) at a flow rate of 2.0 ml/min. Correlation
coefficients of standard curves for individual sulfonamides were line
ar (>0.99) with recoveries ranging from 25.2 +/- 8.6% to 64.1 +/- 8.6%
for a concentration range of 1.0-25 mug/ml.