Liquid chromatographic determination of zoalene in medicated feeds

A rapid, reliable separation and quantitation of zoalene (3,5-dinitro-o-toluamide) from feeds is accomplished by using reverse phase liquid chromatography (LC) and ultraviolet detection. An extraction technique which is similar to the present AOAC official colorimetric method is used before chromatographic analysis. This extraction is followed by an activated alumina cleanup and LC to separate zoalene from feed matrix. The methodology was applied to a variety of spiked feed matrices, and yielded good recoveries. Liquid chromatographic results were shown to correlate with colorimetric determinations.     

Liquid chromatographic determination of oxfendazole in swine feeds

A relatively simple analytical method is presented for determination of oxfendazole (2-(methoxycarbonylamino)-5-phenylsulfinyl-benzimidazole) at levels as low as 0.012% in swine feeds, using cation exchange liquid chromatography (LC). The sample was extracted with a solvent mixture of methanol-glacial acetic acid (90 + 10) at 45 degrees C, using a gyrorotory shaker. Plant pigments and other feed excipients were removed using zinc acetate treatment and pH-controlled extraction. Oxfendazole was further separated from the remaining interferences and quantitatively determined by LC on a Partisil SCX column with acetonitrile-0.01M phosphate buffer as mobile phase. The method is stability-specific, linear, precise, and accurate at 80-120% labeled strength (relative standard deviation 0.9-1.7 with mean recovery of 98-99%). Supporting data at a level of 0.0135% oxfendazole in swine feed indicated that this method is capable of complete recovery of oxfendazole from medicated swine feeds.     

Determination of dimetridazole and ipronidazole in feeds at cross-contamination levels

A rapid method for the determination of dimetridazole and ipronidazole in feeds is described. The compounds are extracted from a borate buffer (pH 8.65) with benzene, partitioned into 1N HCl, and then partitioned back into benzene from a basic aqueous phase. The benzene extract is concentrated and injected onto a nonpolar (Apiezon L) gas chromatographic column for determination by 63Ni electron-capture detection. Recoveries from feeds of various composition, spiked at 0.2 ppm with both dimetridazole and ipronidazole, ranged from 70 to 115%; for the same feeds spiked at 1 ppm or more, the recoveries were greater than 80%. Carbadox, furazolidone, levamisole, oxytetracycline, chlortetracycline, sulfamethazine, sulfaquinoxaline, arsanilic acid, piperazine, penicillin, and commonly added vitamins and minerals do not interfere. A 2-dimensional thin layer chromatographic system is presented as a means of additional identification.     

Quantitative colorimetric method for sulfamethazine in swine feeds

Sulfamethazine (SM) in swine feeds is quantitated colorimetrically by using a relatively simple procedure. The drug is isolated from an extract of feed by trapping it on a small bed of anion-exchange resin buffered at pH 7.9 following the removal of possible interfering compounds by pre-columns. SM is then eluted, diazotized, and coupled using the Bratton-Marshall reagents. The intensity of color developed is determined spectrophotometrically at 540 nm and the concentration of SM is calculated using a molar absorptivity of 52 500. Feeds containing from 1 to 130 ppm can be analyzed with very good accuracy and precision. The method has satisfactory specificity, is inexpensive, and can be applied to a relatively large number of samples per day. A simple and safe method for preparing N1-methyl SM at the low microgram level is described together with a thin layer chromatographic system for tentatively verifying the presence of SM in the feed both as the underivatized drug and as N1-methyl SM.     

Liquid chromatographic determination of olaquindox in medicated feeds and in contents of porcine gastrointestinal tract

A liquid chromatographic method for the determination of olaquindox in both medicated feeds and porcine gastrointestinal tract is described. Samples are extracted with water and cleaned on a disposable reverse-phase column. The eluate is chromatographed on a reverse-phase column under isocratic conditions. Olaquindox is detected by UV absorption at 260 nm. The minimum amount detected by this method was 0.075 ng. The corresponding minimum detectable concentration in a 1 g sample was 0.3 mg/kg. The detector response was linear within the interval of 0-500 ng. Mean recovery of olaquindox in spiked gastrointestinal samples was 89 +/- 5% (mean +/- standard deviation, n = 43). Concentration profiles of olaquindox in the gastrointestinal tract of pigs fed medicated feed were used to evaluate the preventive potency against Treponema hyodysenteriae. The presence of some N-O reduced metabolites of olaquindox in the gastrointestinal tract was assessed.     

Liquid chromatographic determination of carbadox and desoxycarbadox in medicated feeds and in porcine gastrointestinal tract

A liquid chromatographic method for the assay of carbadox and desoxycarbadox in medicated feeds and porcine stomach and intestinal contents is described. Samples were extracted with dimethylformamide-water and cleaned up on an alumina column. The eluate was chromatographed using either gradient elution for simultaneous assay of both compounds or isocratic elution for carbadox only. Detection of carbadox by its native fluorescence yielded a sensitive and specific assay without interferences by metabolites or matrix components. The optimal UV absorption of desoxycarbadox was at 280 nm. Mean recoveries of carbadox in spiked feed and stomach contents were 104 and 97%, respectively; mean recovery of desoxycarbadox in stomach contents was 106%. The day-to-day reproducibility for carbadox in different feed samples and stomach contents samples had a coefficient of variation of 6-13%.     

Determination of virginiamycin in feeds.

Virginiamycin was extracted from the feed by ethanol-pH 2.5 phosphate buffer (1 + 1). The pH during extraction was adjusted (when necessary) to between 4 and 5. Sample dilutions and the standard dose response line were prepared to contain ethanol pH 6 phosphate buffer (2 + 8), and the test organism was Sarcina lutea. Three feeds (a poultry ration, a swine finishing ration, and a swine starter ration) showed virginiamycin recovery of 88.8--108.9% when standard solutions were added at concentrations of 4.54--90.8 g/ton. The coefficient of variation (4--20%) was larger for low potency feeds (10 g/ton) compared to the higher feeds (100 g/ton). Similarly, excellent recovery was obtained when the swine starter feed was fortified by a commercial premix. Amprolium, roxarsone, and monensin can be present at 20 times the concentration of virginiamycin with little or no interference in the antibiotic determination. Lasalocid at 10 times the concentration of virginiamycin caused a slightly positive bias (recovery, 107.4%).     

Sulfamethazine and sulfathiazole determination at residue levels in swine feeds by reverse-phase liquid chromatography with post-column derivatization

Twenty g sample, to which sulfamerazine has been added as internal standard, is extracted with 0.3N HCl + 1.5% diethylamine in 25% methanol. The sample extract is chilled (to aid clarification), centrifuged, and filtered. The sulfonamides are separated from each other and from co-extracted materials on a C-18 reverse-phase column and detected at 450 nm following post-column derivatization with dimethylaminobenzaldehyde. Two isocratic mobile phases have been tested: (1) acetonitrile-2% acetic acid (17 + 83), with an analysis time of 13 min; and (2) acetonitrile-methanol-2% acetic acid (4 + 16 + 80), with an analysis time of 20 min but an improved analysis for some samples. As many as 40 samples have been analyzed at one time unattended with the aid of an autosampler. A total of about 1500 field samples have been assayed using the method. Method sensitivity is 0.1 ppm for either analyte in a hog finishing fed. Linearity for each of the analytes is satisfactory over a range of 0.4-25 ppm in spiked feeds. Coefficients of variation range from 13% at 0.5 ppm to 2% at 13 ppm as tested over a period of time in naturally contaminated samples. The absolute recovery of sulfamerazine varies with sample matrix, but, in the presence of sulfamerazine as internal standard, recovery has been 96.7-99.7% over the range of 0.1-10 ppm. Sulfamerazine and sulfamoxole were tested for their suitability as internal standards. Sulfamerazine is a good internal standard for sulfamethazine; neither is ideal for sulfathiazole.(ABSTRACT TRUNCATED AT 250 WORDS)     

Determination of amprolium in feeds: collaborative study.

The official final action method, 42.028--42.032, for determining amprolium in feeds was modified by a change in the preparation of aluminum oxide for chromatography. A premix containing 0.5% amprolium was collaboratively studied by the modified and the official methods. Compared with the modified method, 87.7% of the drug was recovered from the premix by using the official method. The modification makes possible the assay of premixes as well as finished feeds. The official final action method has been modified to incorporate this change.     

Liquid chromatographic determination of chlortetracycline hydrochloride in ruminant and poultry/swine feeds.

A liquid chromatographic (LC) method is described for the determination of chlortetracycline hydrochloride (CTC) in poultry/swine and ruminant feeds in the 10-100 ppm range and in premix. CTC is extracted from ground feed/premix with acidified acetone, and the extract is filtered through a Millex-HV filter or disposable C18 column. The filtrate is partitioned with methylene chloride when additional cleanup is necessary. A Nova-Pak C18 column is used for LC separation with determination at 370 nm. The average recovery of CTC from premix was 95% with a standard deviation (SD) of 1.70 and a coefficient of variation (CV) of 1.79%. The overall average recovery from feeds was 77% with an SD of 3.18 and a CV of 4.10%.     

Determination of polybrominated biphenyl residues in dry animal feeds.

A new procedure is described for the determination of polybrominated biphenyls (PBBs) in dry animal feeds and developmental results are discussed. Finely ground feed is packed into a chromatographic column containing Celite and then eluted with methylene chloride. The concentrated extract is cleaned up by elution with petroleum ether through Florisil before gas-liquid chromatographic quantitation. Chromatograms thus obtained were essentially free of the interfering peaks encountered when using AOAC methods for pesticide residues in dry products. Results of feed analyses by the proposed procedure averaged 30% higher than those obtained by AOAC procedures. Recoveries of PBBs from samples fortified at levels of 0.04 to 0.4 ppm ranged from 90 to 104%, with an average of 98%.     

Determination of water-insoluble cell walls in feeds: interlaboratory study

A collaborative study was conducted to test a new rapid procedure for determination of water-insoluble cell wall (WICW) content in feeds. In the method, starch is solubilized near boiling temperature with Termamyl, a heat-stable alpha-amylase, and proteins are solubilized at 40 degrees C with sodium dodecylsulfate and Pronase. Then, the organic matter of the residue is determined by incineration. Three hours were required to treat 12 different samples, including solubilization treatments, filtrations, and rinses. Eleven unknown products including 9 common feedstuffs of various origin and 2 mixed diets for poultry were analyzed by 7 analysts in France. Coefficients of variation ranged from 2.3 to 6.1%. The results were compared to those for water-insoluble dietary fiber (WIDF), total dietary fiber, and neutral detergent fiber. Agreement was best with the water-insoluble dietary fiber procedure. For most samples, the ratios of WIDF/WICW ranged from 0.981 to 0.842. The differences between WICW and WIDF values correspond to cell wall protein which is accounted for in WICW, but not in WIDF.     

Spectrophotometric determination of pyrantel tartrate in swine feeds by method of standard additions: collaborative study

The spectrophotometric method for pyrantel tartrate in swine feeds was collaboratively studied. Twenty-seven laboratories assayed feeds containing 0.0103, 0.0965, and 0.7902% pyrantel tartrate. Repeatability (sigmao) and reproducibility (sigmax) standard deviations were: sigmao = 0.00068%, sigmax = 0.00105% (10% of grand mean) for 0.0103% pyrantel tartrate level; sigmao - 0.0065%, sigmax = 0.0090% (10% of grand mean) for 0.0965% pyrantel tartrate level; and sigmao = 0.0415%, sigmax = 0.0743% (10% of grand mean) for 0.7902% pyrantel tartrate level. The mean theoretical recovery values for feeds containing 0.0103, 0.0965, and 0.7902% were 100, 97, and 96%, respectively. The method was adopted as official first action for feeds or concentrates containing 0.0106-0.8811% pyrantel tartrate.     

Preliminary collaborative study of modified spectrophotometric determination of pyrantel tartrate in swine feeds by the method of standard additions

A modified spectrophotometric method for determining pyrantel tartrate in swine feeds was subjected to a preliminary collaborative study. Two small-scale commercial pyrantel-medicated feed samples (0.0881 and 0.0106%) were assayed in replicate by 4 collaborators. The mean results of all laboratories were 0.0862 and 0.0112%. The mean coefficients of variation were 10.57 and 6.48%, repectively. Suggestions for improving recovery include the following: complete dissolution of standard, use of analytical grade KI, careful phase separation, thorough mixing, and minimum exposure of compound to light.