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3 Bulletin of the Veterinary Institute in Puławy

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DNA based approaches for the detection and identification of bovine meat and bone meal in feeds

100%

Pietron W. J. , Golebiowska A. , Paprocka I. , Kwiatek K.

Bulletin of the Veterinary Institute in Puławy

2013

tom 57

nr 4

The aim of this paper was to present the results of comparative evaluation of the usefulness of PCR method for the detection and identification of bovine DNA in feeds. In the validation study, the limit of detection for PCR was determined as 0.05% for bovine meat and bone meal (MBM). Among 132 feed samples, bovine DNA was detected in eight (6.06%) samples. In the next stage of the study, sediment and flotate from the investigated samples were examined with PCR. Out of 132 sediment and flotate samples, bovine DNA was detected in eight (6.06%) and nine (6.82%) samples, respectively. On the basis of the results obtained with the use of the PCR, it is possible to state that the molecular biology methods can, at present, be used as supplementary tools for detection and identification of bovine MBM.

Amphenicols stability in medicated feed – development and validation of liquid chromatography method

88%

Pietron W. J. , Wozniak A. , Pasik K. , Cybulski W. , Krasucka D.

tom 58

nr 4

A liquid chromatography-ultraviolet detection method for the determination of florfenicol (FF) and thiamphenicol (TAP) in feeds is presented. The method comprises the extraction of analytes from the matrix with a mixture of methanol and acetonitrile, drying of the extract, and its dissolution in phosphate buffer. The analysis was performed with a gradient programme of the mobile phase composed of acetonitrile and buffer (pH = 7.3) on a Zorbax Eclipse Plus C18 (150 × 4.6 mm, 5 µm) analytical column with UV (λ = 220 nm) detection. The analytical procedure has been successfully adopted and validated for quantitative determination of florfenicol and thiamphenicol in feed samples. Sensitivity, specificity, linearity, repeatability, and intralaboratory reproducibility were included in the validation. The mean recoven of amphenicols was 93.5% within the working range of 50-4000 mg/kg. Simultaneous determination of chloramphenicol, which is banned in the feed, was also included within the same procedure of FF and TAP stability studies. Storing the medicated feed at room temperature for up to one month decreased concentration in the investigated drugs even by 45%. These findings are relevant to successful provision of therapy to animals.

Determination of five sulfonamides in medicated feedingstuffs by liquid chromatography with ultraviolet detection

75%

Pietron W. J. , Cybulski W. , Krasucka D. , Mitura A. , Kos K. , Antczak M.

Bulletin of the Veterinary Institute in Puławy

2013

tom 57

nr 4

A fast and reliable method of liquid chromatography and ultraviolet detection of sulfaguanidine, sulfadiazine, sulfamethazine, sulfamethizole, and sulfamethoxazole in feedingstuffs was described. The method involves THE procedure of preparation of spiked samples, and extraction of sulphonamides from the matrix using a mixture of methanol and acetonitrile, followed by drying the extract and dissolving it in a phosphate buffer. The analysis uses octadecyl (C18) analytical column with UV detection at λ = 260 nm and a gradient programme of mobile phase composition. The analytical procedure has been successfully adopted and validated for quantitative determination of the sulfonamides in feedingstuff samples. Validation included sensitivity, specificity, linearity, repeatability, and intra-laboratory reproducibility. The mean recovery of sulfonamides was 84%, within the working range of 200-2000 mg/kg. Direct, simple sample preparation and HPLC-UV analysis allow the method to be successfully included in the scope of routine analyses. The presented results could be an answer to a need of simple and easy method for sulfonamide determination applicable in medicated feedingstuffs analysis.