Simultaneous Determination Of16Sulfonamide Residues In Animal Food Products

The antibacterial medicines were used for preventing and treatingbacterical infection by inhibiting or killing bacterical. Sulfa drugs is one ofthe most commonly used antimicrobial agents，having a good antibacterialactivity against most Gram-positive bacteria and Gram-negative bacteria.SAs are easy to accumulate in the body and cause harm to humans, such ascrystal urine, hematuria, neonatal jaundice et al. Therefore, many countrieshave developed limited provisions for SAs maximum residue in food,HPLC and HPLC-MS/MS are widely used for SAs residue detection. Toavoid using large amounts of organic solvents, which seriously pollute theenvironment, and tedious operations, this study established a simple, fast,safe and accurate sample treatment method to simultaneously purify16kinds of SAS in animal derived food, detected by HPLC andHPLC-MS/MS respectively. The full text is composed of the two parts. Part ⅠDetermination of16sulfonamide residues in animalfood products by HPLCObjective: An immune affinity chromatography purification and highperformance liquid chromatography simultaneous determination of16sulfonamide residues in animal food products has been developed.Compared with liquid-liquid extraction and conventional solid phaseextraction，immune affinity chromatography purification was better，applicable to trace determination for sulfonamides in animal food products.Methods: Samples were extracted with80%ethanol, cleaned-up withimmune affinity column. Separation was performed on an Agilent C18column (150mm×4.6mm,3.5μm) with a gradient elution using methanol(A)-1.1%acetic acid10mmol/L PBS (B) mobile phase gradient elution ata flow rate of1.0mL/min at30℃. The detection wavelength was270nmand injection volume was50μL.Results: The resolutions of16kinds sulfonamides were greater than1.5.Good linear relationship (r＞0.999, n=5) was observed within theconcentration range of0.01-0.2mg/L. Estimated limits of detection (S/N=3)varied from2to5ng/mL. Recoveries of the analytes in animal foodproducts were59.6%～111.7%with RSDs ranging between0.57and7.21%.Conclusion: An immune affinity chromatography purification and highperformance liquid chromatography simultaneous determination of16 sulfonamide residues in animal food products has been developed. Theadvantages of immune affinity column purification over existing methodsare its decreased organic solvents and specificity for sulfonamides.Part Ⅱ Determination of16sulfonamide residues in animalfood products by HPLC-MS/MSObjective: To develop an HPLC-MS/MS method for the determination of16kinds of sulfonamides residues in animal food．Methods: The analytes were separated on an Shimadzu VP-ODSC18(2.0×150mm，5μm), eluted by gradient with mobile phase ofacetonitrile and2mmol/L ammonium acetic contained0.2%formic acid ata flow rate at0.3mL/min. The column temperature was40℃and theinjection volume was10μL. MS/MS parameters as flow: ion-sprayvoltage,5500V, atomizing gas pressure,50psi; air curtain gas pressure,35psi; auxiliary gas pressure,60psi; source temperature,550℃; collisionchamber entrance voltage,10V, collision chamber outlet voltage,12V.Results: Calibration curves resulting from spiked blank samples at the1～200ng/mL (r≥0.999) level showed good linear correlation. Recoveriesranged from57.5to107.8%for all compounds at three different spikinglevels with RSDs ranging between3.54and9.08%and the limits ofdetection (S/N=3) ranged from0.1to1.6ng/mL depending on variouscompounds.Conclusions: An immune affinity chromatography purification and HPLC-MS/MS simultaneous determination of16sulfonamide residues inanimal food products has been developed. The advantages of this methodare its decreased sample preparation and analysis time, high sensitivity andhigh accuracy, which make the method more suitable for trace analysis.