Quantitative HPLC Analysis of Riboflavin and Aromatic Amino Acids in Three Forms of Shrimp Hydrolysates

Abstract

This paper presents a high performance liquid chromatography (HPLC) method for the determination of riboflavin and free aromatic amino acids. To determine riboflavin, the method includes an acid hydrolysis (HCl 0.1 N) followed by an enzymatic digestion and a protein precipitation with trichloroacetic acid. An analytical column, Chrom SEP SS C18 5 µm (4.6 mm × 150 mm) was used with an isocratic flow using a mobile phase consisting of 5 mM ammonium acetate-methanol (72:28, vv⁻¹), at a flow rate of 1.0 mL min⁻¹ to 28°C. The method showed adequate linearity, repeatability, reproducibility, accuracy, and limits of detection. The method was used to quantify the cited analytes in three different forms of hydrolysate obtained from fermented shrimp remnants.

Keywords:

• Aromatic amino acids
• Fluorescence detection
• HPLC
• Hydrolysate
• Riboflavin
• Shrimp remnants

Notes

x, amount (µg mL⁻¹); y, peak area; r ², determination coefficient.

^a Results expressed as dry mass.

^a Results expressed as dry mass.

^b Percent recovery = (amount found−mean value)/(amount added) × 100.

^a Results expressed as dry mass.