Validation of a Simple HPLC-Based Method for Lysine Quantification for Ruminant Nutrition
João Albuquerque1,2, Susana Casal1, Rebeca Cruz1
1LAQV, REQUIMTE, Departamento de Ciências Químicas, Faculdade de Farmácia, Universidade do Porto, Rua Jorge Viterbo Ferreira n.° 228, 4050-313 Porto, Portugal.
Molecules (Basel, Switzerland)
|July 24, 2021
Summary
A new method accurately quantifies lysine, an essential amino acid for dairy cows. This technique aids in analyzing feed supplementation effectiveness for improved ruminant nutrition and milk production.
Area of Science:
- Animal Science
- Analytical Chemistry
- Biochemistry
Background:
- Accurate quantification of specific amino acids is crucial for evaluating feed supplementation strategies in animal nutrition.
- Lysine is a key limiting amino acid in ruminant diets, essential for milk production and overall animal health.
Purpose of the Study:
- To develop and validate a robust and selective method for quantifying lysine.
- To assess the method's applicability across various matrices relevant to ruminant nutrition studies.
Main Methods:
- Reversed-phase high-performance liquid chromatography (RP-HPLC) with fluorescence detection was employed.
- The method was validated using Li2CO3 buffer, phosphate buffer solution, and rumen inoculum.
- Sample stability was assessed at different storage temperatures (room temperature, 4°C, -20°C).
Main Results:
- The developed RP-HPLC method demonstrated selectivity, accuracy (92% ± 2%), and precision across tested matrices.
- Low detection and quantification limits ( < 1.24 µM and < 4.14 µM) were achieved.
- Lysine quantification in real samples confirmed the method's utility for ruminant feeding studies.
Conclusions:
- The validated RP-HPLC method provides a reliable tool for free lysine quantification in ruminant nutrition research.
- This method can significantly benefit dairy cow nutrition, aiding in supplementation optimization and feed effectiveness analysis.
