Determination of the banned growth promoter moenomycin A in feed stuffs by liquid chromatography coupled to electrospray ion trap mass spectrometry.

Flavomycin complex is an antibiotic banned in the European Union as an additive in feed stuffs. As a consequence, the monitoring programmes for official control within the Community require analysis of feeds for possible illegal use of flavomycin. A method for unambiguous identification and quantification of moenomycin A, the main pharmacologically active component of flamomycin complex, in several feeds by liquid chromatography coupled to electrospray ion trap mass spectrometry (LC/ESI-MS/MS) is herein described for the first time. The method was developed to be used as a confirmative analytical tool for the network of Italian official control laboratories; both the singly and doubly charged molecular ions were observed as precursor ions, from which four product ions were selected for both quantitative analysis and unambiguous identification of moenomycin A. The method was in-house validated for feeds in the concentration range 0.50-30.0 microg/g, according to the Regulation 882/2004/EC requirements. Mean recoveries ranging between 83.9-94.2% and relative standard deviations <23% account for method trueness and repeatability, respectively. Moreover, other analytical performance parameters, i.e. method specificity, ruggedness, the linearity of detector response, the limit of quantification (LOQ), the limit of detection (LOD), and measurement uncertainty were evaluated and reported. The ion trap LC/ESI-MS/MS method is highly selective and reliable; high drug recovery, good reproducibility and an LOQ down to 0.10 microg/g guarantee its applicability for confirmatory purposes in the official control activity in Italy.

Determination of the antimicrobial growth promoter moenomycin-A in chicken litter.

This study aimed to develop and optimize a method for the extraction and analysis of moenomycin antibiotics (a.k.a. flavomycin) in corn-based feed premix and in chicken litter. Moenomycin-A was isolated from chicken litter using pressurized liquid extraction followed by a solid-phase extraction (SPE) clean-up step. The highly lipophilic nature of moenomycin necessitated the use of the less hydrophobic sorbent, C(4)-based SPE cartridge, and a higher temperature elution solvent, methanol at 50 degrees C, in order to obtain satisfactory percent recoveries. After clean-up, the sample was analyzed by liquid chromatography-electrospray ionization-mass spectrometry (LC-ESI-MS). Various reversed-phase columns were examined, including C(18), CN, perfluorinated C(6), and a porous graphitic carbon. The set of conditions that gave the highest separation efficiency while still maintaining symmetrical peak shape was the C(18) column using the H(2)O+0.3% HCOOH and acetonitrile mobile phase.

Bioefficacy of enzyme preparations containing beta-glucanase and xylanase activities in broiler diets based on barley or wheat, in combination with flavomycin.

The objective of the study was to determine the effects of two enzyme preparations containing beta-glucanase and xylanase activities on barley- and wheat-based diets, respectively, for broilers, in combination with flavomycin. In addition, the stability of the enzyme preparations after pelleting was measured. Temperatures recorded during the pelleting process reached 75 to 80 C, and the activities recovered with respect to the amounts present in the mash feed before pelleting were 80% or higher. Two performance experiments were conducted simultaneously under the same conditions over 6 wk. In addition, intestinal viscosity and incidence of vent pasting were measured and carcasses were eviscerated to determine abdominal fat, carcass yield, and percentage weight of intestines and viscera. Twenty-four pens (12 per sex), each containing 75 chickens were used in each experiment. Wheat- or barley-based diets were supplemented with flavomycin and a xylanase or a beta-glucanase preparation, respectively, in a 2 x 2 factorial arrangement of treatments. In the wheat diets, xylanase and flavomycin improved feed efficiency, in parallel with a reduction of intestinal viscosity. Xylanase reduced the incidence of vent pasting and the percentage viscera, especially of intestines, and increased abdominal fat. In the barley diets, beta-glucanase and flavomycin improved feed conversion. beta-Glucanase also reduced intestinal viscosity and vent pasting. Both beta-glucanase and flavomycin reduced percentage intestines, but the effects were not additive. In general, the effects of the enzyme preparations and flavomycin were independent, except for percentage intestines with beta-glucanase.

[Ergotrophic action of flavophospholipol during the fattening of geese]. / Ergotropnoto deistvie na flavofosfolipola pri ugoiavane na guski.

Experiments were carried out with a total of 144 geese of the Benkovska breed divided into 6 groups of 24 birds each. The average weight was 3.5 kg, the birds being compulsory fattened with steamed maize over an average period of 27 days. Flavophospholipol (flavofarm-NIHFI) was given at rates of 0, 4, 8, 12, and 16 ppm. The method of positive control check was employed--a group which was offered zinc-bacitracin--(bacifarmin) at 30 ppm. Under the effect of flavophospholipol the weight gain was 2.2 to 21.2 per cent higher, and the weight of the liver rose by 1.4-23.1 per cent. The intake of maize per kg of gain dropped by 2.5 to 15.7 per cent, and it was 1.8 to 16.9 per cent lower per kg of liver. When flavophospholipol was given at 16 ppm, a maximum effect was produced--21.0 percent higher gain, 23.1 per cent increase of the liver weight, 15.7 per cent drop of maize intake per kg of gain, and 16.9 per cent drop of maize intake per kg of liver. With geese that were offered bacifarmin these indices were 3.3, 11.9,, 4.8, and 10.7 per cent, respectively. The amount of flavophospholipol used correlated positively with its ergotropic effect. The liver quality with geese treated with flavophospholipol and bacifarmin was higher. No residual amounts of flavophospholipol were found in the meat, liver and viscera of the treated birds.