Sulfamethazine methods

A/ -HeterocycHc derivatives can be formed in some cases by a ring closure to give the heterocycle. Sulfadiazine, sulfamethazine, sulfamerazine, and sulfathiazole have been prepared in this fashion, but also by the usual procedure from the sulfonyl chloride and heterocycHc amine. The synthesis of sulfamethazine from sulfaguanidine is an example of the ring closure method. 

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. 

In each of these methods, undiluted serum, urine, acid-deproteinized milk, or a buffered saline extract of muscle was mixed with sulfamethazine-horseradish peroxidase and added to antibody-coated wells of a microtiter plate. A sulfamethazine-bovine serum albumin conjugate prepared by the glutaraldehyde procedure (56) was used for antibody production. Results showed that screening of serum was of value since sulfamethazine concentrations in serum directly correlated with those in swine tissues. Thus, for example, a level of 100 ppb of sulfamethazine in... 

In another method for the determination of sulfamethazine in muscle and liver tissues (59), the extraction problem was successfully addressed by applying a matrix solid-phase dispersion procedure for rapid and efficient purification of the tissue extracts. Determination was made by ELISA on the basis of antibodies raised against sulfamethazine-diazo-bovine serum albumin conjugates. 

SPECTROMETRIC METHODS Sulfamethazine Bovine CHCls/acetone 18 - Vis 545 nm 100 ppb/ 252... 

GAS CHROMATOGRAPHIC METHODS Sulfamethazine Swine CHCb/acetone... 

Simeonidou et al. (238) reported an ion-pair liquid chromatographic method for the determination of sulfadiazine, sulfamethazine, sulfadimethoxine, and sulfaquinoxaline residues in chicken muscle. According to this method, a 3 g ground tissue sample is extracted with 30 ml chloroform. Following centrifugation the supernatant is filtered and a 10 ml aliquot is extracted with 1 ml 3N hydrochloric acid and submitted to precolumn derivatization with fluorescamine. Liquid chro-... 

C. Mellgren, A. Sternesjo, P. Hammer, G. Suhren, L. Bjorck and W. He-eschen, Comparison of biosensor, microbiological, immunochemical and physical methods for detection of sulfamethazine residues in raw milk, J. Food Prot., 59(11) (1996) 1223-1226. 

Grant, G.A., S.L. Frison, and P. Spoms. 2003. A sensitive method for detection of sulfamethazine and N4-acety lsulfamethazine residues in environmental samples using solid phase immunoextraction coupled with MALDI-TOF MS. J. Agric. Food Chem. 51 5367-5375. 

HPLC on pBondapak Ph column with UV detection at 280 nm has been used for the determination of sulfacetamide, sulfabenzamide and sulfathiazole in a triple sulfa cream. The mobile phase consists of a 7 3 mixture of 0.01 M ammonium hydrogen phosphate and methanol (pH 7.2). The proposed method has been shown to give better results for sulfacetamide assay compared with those of the USP method (97). Sulfacetamide, sulfadiazine, sulfamerazine and sulfamethazine in mixtures and in pharmaceuticals have been determined by HPLC using a nonpolar column with methanol-0.02 M potassium dihydrogen phosphate in water as the mobile phase and a variable wavelength UV detector. The method is precise with relative standard deviations of 2.1, 0.6,1.9 and 1.6% respectively for the four compounds. The preservatives do not interfere with the method (98). 

Macek, J. Vacha, J. Rapid and sensitive method for determination of piroxicam in human plasma by high-performance liquid chromatography. J.Chromatogr., 1987,420, 445-449 [column temp 35 glass column LOD 150 ng/mL pharmacokinetics simultaneous antipyrine, diclofenac, hydroxychloroquine, ibuprofen, isoxicam, plaquenil, sulfamethazine]... 

Long, A.R. Hsieh, L.C. Malbrough, M.S. Short, C.R. Barker, S.A. A multiresidue method for the isolation and liquid chromatographic determination of seven sulfonamides in infant formula. J.Liq.Chromatogr., 1989, 12, 1601-1612 [extracted sulfadiazine, sulfadimethoxine, sulfamerazine, sulfamethazine, sulfathiazole, sulfisoxazole column temp 45 SPE]... 

Sulfonamides having a free j )-amino group are readily assayed by titration with nitrous acid. The sulfonamide function may also be titrated with base, such as lithium methoxide. The majority of the sulfas listed in the U.S. Pharmacopeia XXII, however, are assayed by chromatographic methods, particularly high performance liquid chromatography (49). Sulfonamides for which assays are listed in the U.S. Pharmacopeia XXII-National Formulary XVII include the following sulfacetamide, sulfabenzamide, sulfadiazine, sulfadoxine, sulfamerazine, sulfamethazine, sulfamethizole, sulfamethoxazole, sulfapyridine, sulfasalazine, sulfathiazole, sulfinpyrazone, sulfis ox azole, sulfisoxazole acetyl, sulfisoxazole diolamine, sulfoxone, triple sulfa, dapsone, and various combinations with prednisolone, pyrimethamine, and trimethoprim. 

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). 

Figure 9.2 Schematic diagram of a method for the determination of sulfamethazine.

A study was undertaken to evaluate the performance of the EZ-Screen Quik-C ard T est for sulfamethazine in pork urine, liver and muscle tissue (Table III). Initially, the EZ-Screen Quik Card was compared with the current USDA Sulfa-on-Site TLC method (14) for the screening of 569 pork urine samples at the plant level. Eighty-one urine samples were found positive by the EZ-Screen and only 36 by SOS. The corresponding 81 pork liver samples were then analyzed by both EZ-Screen and TLC. ... 

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