Sulfamethazine analysis

Most immunochemical methods published for the determination of sulfonamides in edible animal products, serum, and urine concern sulfamethazine analysis (Table 28.3). Early methods for screening sulfamethazine in swine blood (54) necessitated extraction of the antibiotic from the sample and application of long assay protocols that rendered them impractical for routine analysis in hog slaughterhouses. Later methods developed for the detection of sulfamethazine residues in swine serum (55), urine and muscle (8), and in milk (9) addressed the extraction and assay problems of previous methods. 

Stemesjo A., Mellgren C., Bjorck L., Analysis of Sulfamethazine in Milk by an Immunosensor assay Based on Surface Plasmon Resonance, Immunoassays For Residue Analysis, ACS Symposium Series, 621 463-470 (1996). 

As occurred with the other antibiotics, commercial immunoassay formats, also available as kits for tetracyclines and penicillins such as the Parallux, the LacTek, or the Charm II, have also been placed on the market for the analysis of sulfonamides (see Table 4). Thus, the Parallux detects sulfamethazine and sulfadimethoxine in raw milk with a LOD of 10 pg L1. The Charm II detects almost all sulfonamides in honey and milk with a LOD in the range from 1 to 10 pg L, whereas LacTek is able to detect sulfamethazine. Moreover, the 5101SULlp and 5101SUDAlp tests reach LOD values for sulfamethazine and sulfadiazine of around 0.2 pg L 1 and they have been applied to the analysis of urine, milk, and plasma. These tests have proved to be efficient as a point of care for on-site applications on farms. Moreover, commercially available antibodies can be found from several sources such as Silver Lake Research, US Biological, Cortex Biochem. Inc., Accurate Chemical Scientific, Fitzgerald Industries International Inc., and Biotrend Chemikalien GmbH. 

Tatavarti AS et al. Assessment of NIR spectroscopy for nondestructive analysis of physical and chemical attributes of sulfamethazine bolus dosage forms. Pharm Sci Tech 2005. 

Analysis of the 1994 tissue residue data revealed that FSIS reported 2514 animals containing violative residues. FDA, in cooperation with participating states, conducted follow-up investigation on 1076 (45%) of the reported violations. The drugs most frequently identified as causing antibiotic residues included penicillin (21%), oxytetracycline (10%), sulfamethazine (10%), streptomycin (6%), tetracycline (5.2%), neomycin (4.1%), gentamicin (3.7%), and sulfadimeth-... 

Berge and Deye studied the effect of column surface area on the retention of polar solutes [18]. They found that there was a linear relationship between retention and the surface area. 4-Hydroxy benzoic acid was used as a model acidic compound, and sulfamethazine, sulfanilamide, sulfi-somidine, and sulfapyridine were used as the model basic compounds. The separations were carried out on a packed Nucleosil Diol column with a methanol-modified carbon dioxide as the mobile phase. The UV detector was used for the analysis. It was observed that 0.1% acetic acid for the acidic solutes and 0.1% isopropylamine for basic solutes was required in the methanol to achieve the separations. The efficiency was found to be similar for 100-, 300-, and 500-A packing materials. 

The simplest and most straightforward application of thermal analysis is concerned with studies of the relative stability of polymorphic forms. For example, DTA thermograms enabled the deduction that one commercially available form of chloroquine diphosphate was phase pure, while another consisted of a mixture of two polymorphs. DTA analysis was used to demonstrate that in spite of the fact that different crystal habits of sulfamethazine could be obtained, these in fact consisted of the same anhydrous poly-morph.f In a study aimed at profiling the dissolution behavior of the three polymorphs and five solvates of spironlactone, DTA analysis was used in conjunction with powder X-ray diffraction to establish the character of the various materials. ... 

G. Bartolucci, G. Pieraccini, F. ViUanelli, G. Moneti, A. Triolo, LC-MS-MS quantitation of sulfamethazine and its metabolites direct analysis of swine urine by triple-quadrupole and ion-trap MS, Rapid Commun. Mass Spectrom., 14 (2000) 967. 

Sample clean-up can be done rapidly using a short LC column operated under gravity flow or under vacuum, centrifuge or syringe pressure. The most popular phases include silica and bonded phases. Disposable and inexpensive columns are available and can be found under the following trade names Sample Enrichment and Purification, PreSep Extraction, Bond-Elute, and solid-phase extraction. For example, purification of sulfamethazine in milk by SPE followed by quantitative analysis by TLC has been described earher... 

Stolker et al. " described an analytical method based on TFC-LC-MS/MS for the direct analysis of 11 veterinary drugs (belonging to seven different classes) in milk. The method was applied to a series of raw milk samples, and the analysis was carried out for albendazole, difloxacin, tetracycline, oxytetracycline, phenylbutazone, salinomycin-Na, spiramycin, and sulfamethazine in milk samples with various fat contents. Even without internal standards, results proved to be linear and quantitative in the concentration range of 50-500 (xg/1, as well as repeatable (RSD<14% sulfamethazine and difloxacin <20%). The limits of detection were between 0.1 and 5.2 xg/l, far below the maximum residue limits for milk set by the EU. While matrix effects, namely, ion suppression or enhancement, were observed for all the analytes, the method proved to be useful for screening purposes because of its detection limits, linearity, and repeatability. A set of blank and fortified raw milk samples was analyzed and no false-positive or falsenegative results were obtained. 

AOAC, Sulfamethazine residues raw bovine milk-liquid chromatographic method, 992.21, in Official Methods of Analysis, 18th ed. (revised), AOAC International, Gaithersburg, MD (available at http //www.eoma.aoac.org/ gateway/readFile. asp id=992 21. pdf accessed 3/24/10). 

Table Hi Analysis of Pork Urine, Liver and Muscle Tissues for Sulfamethazine Residues by the EZ-Screen Test, Thin-Layer Chromotography (TIC) or the Sulpha-on-Site (SOS) Procedures...

Antibiotics The AOAC has listed methods for sulfamethazine residues in swine tissues with determination either by GC-MS or GC-ECD of methylated derivatives and for sulfamethazine (and for the class of sulfonamides) in milk with determination by HPLC-UV. There is an AOAC method for the class of sulfonamide antimicrobials in animal tissues using solvent extraction and liquid partitioning with determination by TLC and fluorimetric scanning. For analysis of tetracyclines, AOAC describes methods based on buffer extraction from tissue samples and SPE (Cis) cleanup, or metal chelate affinity binding from milk samples, with determination in both cases by HPLC-UV. USDA/FSIS methods include (1) a method (similar to the AOAC GC-MS method for sulfamethazine) for confirmation of sulfonamide residues in edible tissues using solvent extraction and multiple liquid partitioning with determination of the methylated derivatives by GC-MS (2) methods for determination and confirmation of chloramphenicol in muscle by solvent extraction, liquid partitioning, and determination of the trimethylsilane (TMS) derivative by GC-ECD and GC-MS, respectively and (3) a method for determination of the beta-lactam antibiotic amoxicillin by aqueous extraction, cleanup by tricarboxylic acid precipitation, and ether extraction and formation of a fluorescent derivative for determination by LC. 

McGrane, M. O Keeffe, M. Smyth, M.R. The analysis of sulphonamide drug residues in pork muscle using automated dialysis, Anal.Lett., 1999, 32, 481-495. [LOD 40 ng/g sulfadiazine sulfathiazole sulfapyridine sulfamerazine suRamethizole sulfamethazine sulfamethoxypyridazine sulfachlorpyridazine sulfisoxazole]... 

Wutz et al. (2011) developed for the first time a multianalyte immunoassay based on an automated flow-through CL microarray technique for identification and quantification of antibiotic derivatives in honey samples using regenerable antigen microarrays, an indirect competitive immunoassay format using horseradish peroxidase (HRP)-labeled antibodies and CL read-out with a CCD camera. The method allows the analysis of four analytes (enrofloxacin, sulfadiazine, sulfamethazine, and streptomycin) simultaneously in 8 min with adequate recoveries and without purification or extraction. Due to the regenerability of the microarray each chip could be individually calibrated before the analysis and allowed more than 40 assays, which reduces the costs per analysis and permits an automated work flow in routine laboratories. 

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