Ce. Tsai et F. Kondo, SIMPLE CONTINUOUS AND SIMULTANEOUS DETERMINATION OF TETRACYCLINE RESIDUES, Research in Veterinary Science, 56(3), 1994, pp. 277-283
A new continuous separation method was developed for the determination
of five different tetracyclines (oxytetracycline, tetracycline, chlor
tetracycline, methacycline and doxycycline). A bioassay using minimum
medium (MM) seeded with Bacillus subtilis ATCC 6633 was carried out fo
r the detection, with simple extraction from the agar block of the cle
ar inhibition zone on the MM produced by the mixed tetracyclines, The
extract was subjected to continuous identification by high performance
liquid chromatography using a mu-Bondapack C-18 column, The tetracycl
ines were separated at ambient temperature using a mobile phase of 0.0
1 M oxalic acid: acetonitrile: N,N-dimethylformamide (74:18:8, v/v/v)
at a flow rate of 1.0 ml min(-1). A variable-wavelength detector set a
t 355 nm and recorder set at 4 mm min(-1) were used for the detection.
The entire mixture was resolved as five peaks with retention times ra
nging from 2.75 to 9.65 minutes. This continuous, simple and rapid met
hod of detection, extraction and identification may be useful for rout
ine laboratory testing of residual antimicrobial agents in food.