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Due to the essentiality of lysine for fish, its availability is commonly used as a predictor of the protein nutritional quality of fish feed. The objective of this work was to establish a high-throughput analytical method for protein quality control in fish feed through the measurement of available lysine by planar chromatography. Sample was first incubated with 1-fluoro-2,4-dinitrobenzene to obtain the dinitrophenyl-lysine derivative and then hydrolyzed with hydrochloric acid in order to release the derivative from proteins. Chromatography was performed on silica gel 60 F254 HPTLC plates using n-propanol25 ammonia (7 + 3, v/v) as mobile phase. Quantitative analysis was performed by densitometry in the absorbance mode at 360 nm. Calibration showed a polynomial relationship with an R2 of 0.9991 in the range of 25 to 125.0 ng/band. Repeatability relative standard deviation (RSD) and intermediate precision (RSD) in matrix were 0.8 and 3.6, respectively. Recoveries of spiked samples at two levels ranged from 72 to 85 with RSD from 3 to 8. This method provides the salmon feed industry with a reliable, high-throughput, and low-cost means for routine quality control of available lysine.

Determination of Available Lysine by Planar Chromatography: A Useful Tool for Protein Quality Evaluation in Fish Feed