Progress on the development and single-laboratory validation of a high-performance liquid chromatographic method for the determination of carbadox and pyrantel tartrate in type B and C medicated feeds.

Carbadox, an antimicrobial agent, and pyrantel tartrate, an anthelmintic, are feed additives that are often used in combination in the United States. The current AOAC methods for these analytes are spectrophotometric, using standard addition techniques. These methods are labor-intensive and prone to variability as well as matrix interferences. Published methods for both analytes that use high-performance liquid chromatography were evaluated and a test method was developed. The method uses a water prewetting step to enhance extraction of pyrantel followed by extraction with acetonitrile-ethanol (50 + 50). Sample extracts are filtered through a glass fiber filter and purified using alumina solid-phase extraction columns. Chromatography is performed on a C18 column with a gradient mobile phase of dibutylamine acetate and acetonitrile. The data show that both analytes exhibit acceptable peak shape when a C18 column that is both acid- and base-deactivated is used. Linearity has been established and initial recovery studies on medicated swine feeds are promising.

Liquid chromatographic determination of pyrantel tartrate in medicated formulations.

The validation of a novel liquid chromatographic (LC) method for the determination of pyrantel tartrate in feed is presented. The method provides a significant improvement over the efficiency and precision of AOAC Official Method 978.30. The method was shown to be accurate, precise, linear, and robust for medicated articles. Unlike the official method, the LC method was shown to be a superior stability-indicating method. After the method was validated by using laboratory blends, the effectiveness of the method was demonstrated with marketed product as well.

A collaborative study: high-pressure liquid chromatographic determination of carbadox and pyrantel tartrate in animal feeds.

Carbadox (CBX), an antibacterial agent, and pyrantel tartrate (PT), an anthelmintic, are formulated either separately or together in swine feeds. The official Association of Official Analytical Chemists (AOAC) spectrophotometric methods for both drugs are long, nonspecific, and require standard addition techniques. Results by this technique are positively biased. A simple, direct, specific, high-pressure liquid chromatographic (HPLC) method to determine either one or both drugs simultaneously with apparent accuracy and precision is developed. Drugs are released from feed matrices by water, extracted with dimethylformamide (DMF), cleaned up on alumina, and quantitated by direct comparison to standards using a Whatman Partisil 10 ODS-3 column and a mobile solvent containing 23.5 +/- 1.5% DMF in phosphate buffer (pH 2.0). Fourteen laboratories participated in a collaborative study of this method for determination of CBX and PT in animal feeds.

High pressure liquid chromatographic determination of carbadox and pyrantel tartrate in swine feed and supplements.

A rapid yet reliable procedure for the simultaneous extraction and assay of carbadox and pyrantel tartrate is described. The feed is extracted with water-acetonitrile-methanol and cleaned up on a short alumina column. The eluant is separated by high pressure liquid chromatography and the compounds are detected at different wavelengths. The drugs of interest are well resolved in all feeds studied. The procedure has also been applied to a wide range of feeds which contained either one of the drugs or both in combination. No significant interferences were observed. Spiked sample recoveries were 97% for carbadox and 101% for pyrantel tartrate. Ruggedness test coefficients of variation were 2.0% for carbadox and 2.1% for pyrantel tartrate.

High performance liquid chromatographic method fo pyrantel tartrate in swine feeds and supplements.

A new method for the determination of pyrantel tartrate in swine feed an supplements has been developed because the current official AOAC method is not applicable to feeds co-medicated with tylosin. The new method involves: (a) leaching of drug from feed with methanolic NaCl solution, (b) removal of interfering substances by ion pair liquid-liquid extraction and high performance liquid chromatography, and (c) quantitation of pyrantel tartrate by monitoring the ultraviolet absorption of the effluent stream at 313nm. The method of standard addition is used to compensate for the effect of the feed matrix on drug recovery. No interference is encountered from tylosin, carbadox, lincomycin, non-drug components of feeds and supplements, or potential degradation products of pyrantel tartrate, i.e., cis isomer of pyrantel tartrate and (E)-N-(3-methylaminopropyl)-2-thiopheneacrylamide. Results for the assay of 3 lots each of feeds and supplements containing 0.0106 and 0.106% pyrantel tartrate, respectively, were within +/-4% of label claim. Coefficients of variation ranged from 1.6 to 1.8% for feeds and from 1.9 to 3.9% for supplements.

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.