High pressure liquid chromatographic determination of sulfamethazine in pork tissue

A high pressure liquid chromatographic (HPLC) method has been developed for determination of sulfamethazine residues in pork liver, kidney, muscle, and fat. The sample was extracted with acetone-chloroform, concentrated in the presence of dilute HCl, and partitioned between dilute HCl and hexane. The acid solution was washed with methylene chloride and then buffered with trisodium citrate and sodium hydroxide to pH 5.8-5.9. Sulfamethazine was extracted from the aqueous mixture with methylene chloride, concentrated, dissolved in buffer, and eluted from XAD-2 resin with methanol. Sulfamethazine was reliably quantitated at 0.1 ppm by HPLC on a Zorbax ODS column with detection at 254 nm with no interference from tissues or reagents. The average recovery from the edible tissues, i.e., liver, muscle, kidney, and fat, fortified at 0.1-0.4 ppm was 85.6 +/- 3.7%.