<abstract xmlns="http://www.w3.org/1999/xhtml">

<sec id="j_jvetres-2020-0044_s_005">
<h3>Introduction</h3>
<p>A high-performance liquid chromatographic–diode array detector (HPLC-DAD) method for the determination of amoxicillin in medicated feedingstuffs was developed and validated. The method was used to investigate the quality requirements of animal feedingstuffs (declared content of active substance and feed homogeneity).</p>
</sec>
<sec id="j_jvetres-2020-0044_s_006">
<h3>Material and Methods</h3>
<p>Two-gram samples were extracted by potassium phosphate buffer solution. Extracts were filtered and directly analysed by HPLC-DAD without further clean-up. Amoxicillin was separated by acetonitrile and 0.01M phosphate buffer (pH 5.0) on a Phenomenex Luna C18 column.</p>
</sec>
<sec id="j_jvetres-2020-0044_s_007">
<h3>Results</h3>
<p>This method provided average recoveries of 76.1 to 81.6% with coefficients of variation (CV, %) for repeatability and reproducibility in the ranges of 3.7–7.2% and 5.3–7.6%, respectively. The limit of detection was 51.2 mg/kg and limit of quantification was 103.0 mg/kg.</p>
</sec>
<sec id="j_jvetres-2020-0044_s_008">
<h3>Conclusion</h3>
<p>The method was successfully validated and proved to be efficient, precise, and useful for quantification of amoxicillin in medicated feedingstuffs.</p>
</sec>
</abstract>

Introduction
A high-performance liquid chromatographic–diode array detector (HPLC-DAD) method for the determination of amoxicillin in medicated feedingstuffs was developed and validated. The method was used to investigate the quality requirements of animal feedingstuffs (declared content of active substance and feed homogeneity).


Material and Methods
Two-gram samples were extracted by potassium phosphate buffer solution. Extracts were filtered and directly analysed by HPLC-DAD without further clean-up. Amoxicillin was separated by acetonitrile and 0.01M phosphate buffer (pH 5.0) on a Phenomenex Luna C18 column.


Results
This method provided average recoveries of 76.1 to 81.6% with coefficients of variation (CV, %) for repeatability and reproducibility in the ranges of 3.7–7.2% and 5.3–7.6%, respectively. The limit of detection was 51.2 mg/kg and limit of quantification was 103.0 mg/kg.


Conclusion
The method was successfully validated and proved to be efficient, precise, and useful for quantification of amoxicillin in medicated feedingstuffs.

In-house validation method for quantification of amoxicillin in medicated feedingstuffs with the use of HPLC-DAD technique

Journal of Veterinary Research

This work is licensed under the Creative Commons Attribution-NonCommercial-NoDerivatives 3.0 License.

Introduction
A high-performance liquid chromatographic–diode array detector (HPLC-DAD) method for the determination of amoxicillin in medicated feedingstuffs...

Validation parameters	Results
Solvent STD regression equations	y = 80.695x + 392.76

Coefficient of determination (R²)	0.99

Recovery (%)

150 mg/kg	76.1

300 mg/kg	78.2

500 mg/kg	81.6

Repeatability (%)

150 mg/kg	7.2

300 mg/kg	5.8

500 mg/kg	3.7

Within-laboratory reproducibility (%)

150 mg/kg	7.6

300 mg/kg	6.8

500 mg/kg	5.3

LOD (mg/kg)	51.2

LOQ (mg/kg)	103.0

U (%)	19.4

In-house validation method for quantification of amoxicillin in medicated feedingstuffs with the use of HPLC-DAD technique

Publicado en línea: 20 jun 2020

Páginas: 433 - 438

Recibido: 09 dic 2019

Aceptado: 10 jun 2020

DOI: https://doi.org/10.2478/jvetres-2020-0044

Palabras clave
high performance liquid chromatography, diode array detector, medicated feedingstuffs, amoxicillin, validation

© 2020 E. Patyra, K. Kwiatek et al. published by Sciendo

This work is licensed under the Creative Commons Attribution-NonCommercial-NoDerivatives 3.0 License.

Fig. 1

Fig. 2

Fig. 3

Validation parameters of optimised HPLC-DAD method

In-house validation method for quantification of amoxicillin in medicated feedingstuffs with the use of HPLC-DAD technique

Publicado en línea: 20 jun 2020

Páginas: 433 - 438

Recibido: 09 dic 2019

Aceptado: 10 jun 2020

DOI: https://doi.org/10.2478/jvetres-2020-0044

Palabras clavehigh performance liquid chromatography, diode array detector, medicated feedingstuffs, amoxicillin, validation

© 2020 E. Patyra, K. Kwiatek et al. published by Sciendo

This work is licensed under the Creative Commons Attribution-NonCommercial-NoDerivatives 3.0 License.

Fig. 1

Fig. 2

Fig. 3

Validation parameters of optimised HPLC-DAD method

Palabras clave
high performance liquid chromatography, diode array detector, medicated feedingstuffs, amoxicillin, validation