Sample Application

Determining Coccidiostats in Poultry Feed

Solvent Front Position Extraction (SFPE) is a novel sample preparation technique developed for LC-MS/MS analysis. Fresh Chromato created the world’s first laboratory robot utilizing SFPE. This article shows how effectively it can be used for the determination of coccidiostats in animal feed.

In this study, the technique was used for isolation and determination of 7 coccidiostats from both main groups: ionophores (Lasalocid, Salinomycin, Monensin, Narasin and Maduramycin) and chemical coccidiostats (Robenidine, Nicarbazin).

The effectiveness of the SFPE-LC-MS/MS procedure was evaluated by comparing it with the SPE-LC-MS/MS procedure. The results obtained with Fresh Chromato’s robot were very satisfactory:

  • Linearity R2: ≥ 0.971
  • Recovery: 90.1% - 111.1%
  • RSDr: 8.7% - 16.6%,
  • RSDR: 9.0% - 17.7%

These results have met requirements described in the EU Commission Regulation 2021/808 of 22 March 2021, showing great potential of the SFPE technique in sample preparation of poultry feed for the determination of coccidiostats.

Procedure 1 - New Technique

Steps performed when utilizing Solvent Front Position Extraction (SFPE):

  1. Solvent and internal standard addition
  2. Mixing
  3. Extracting solutes of interest from sample (with Fresh Chromato's robot*)
  4. Determination with LC-MS/MS

* First, the robot applies sample solutions to the chromatographic (TLC) plate and it runs chromatogram development in a manner that focuses solutes of interest into one zone of the chromatographic plate. Finally, the solutes of interest are extracted from the focused zone of the TLC plate and loaded into vials. For more information, see the video on the main page.

Procedure 2 - Traditional Technique

Steps performed when utilizing Solid-Phase Extraction (SPE):

  1. Solvent and internal standard addition
  2. Mixing
  3. Centrifugation
  4. SPE cleanup
  5. Centrifugation and filtration
  6. Determination with LC-MS/MS
Determination of Coccidiostats in Poultry Feed by LC-MS/MS

1. Average determination coefficients (R2) of calibration curves of coccidiostats in feed samples:

Coccidiostats Determination Coefficients

2. Recovery of coccidiostats in samples for each procedure:

Coccidiostats Recovery

3. Within-laboratory repeatability and reproducibility of determination of coccidiostats for each procedure:

Coccidiostats Repeatability

4. Sample preparation time per sample:

Coccidiostats Determination Time
Chromatogram

LC–MS scan mode chromatogram of poultry feed sample prepared using the SFPE sample preparation procedure. The visible peaks belong to coccidiostats only, demonstrating successful removal of sample matrix components.

Coccidiostats Determination Chromatogram
References
  • Maciej Jan Rybicki, Ilse Becue, Els Daeseleire, Anna Klimek‐Turek & Tadeusz Henryk Dzido. “Solvent Front Position Extraction and some conventional sample preparation techniques for the determination of coccidiostats in poultry feed by LC–MS/MS”. Scientific Reports. 12:3786 (2022).
  • Link to publication: https://www.nature.com/articles/s41598-022-07587-5