Role of zwitterionic structures in the solid-phase extraction based method development for clean up of tetracycline and oxytetracycline from honey

Solid-phase extraction (SPE) based sample clean up of tetracycline (TC) and oxytetracycline (OTC) from honey samples was evaluated using a neutral polymeric sorbent (strata-X), a weak cation exchange polymeric sorbent (strata-X-CW) and a combination of neutral and strong cation exchange sorbents (strata-X plus Strata-Screen-C or strata-X-C). Both TC and OTC are recovered in low yields from the strong cation exchange sorbents strata-X-C and Strata-Screen-C under basic (pH >9) or acidic (pH ≥2) elution conditions. The cleanest extracts along with quantitative recoveries were obtained with strata-X-CW under either acidic or basic elution conditions, as demonstrated by analysis of eluates by both LC/UV and LC/MS–MS. On the other hand, the neutral sorbent (strata-X) was less efficient in eliminating the honey matrix constituents, since it could not retain the tetracyclines with the strong organic wash needed to remove other compounds. The differences in the elution behavior of the strong and weak cation exchange sorbents is rationalized on the basis of divergent deprotonation mechanisms of the tetracyclines, which exist in the zwitterionic structures under the SPE conditions.