Validation of vitamin B5 (pantothenic acid) and B6 (pyridoxine, pyridoxal, and pyridoxamine) analyses in seafood

Abstract

Vitamin B5 (pantothenic acid) and B6 (pyridoxine, pyridoxal, and pyridoxamine) contents of various seafoods, were analyzed by high-performance liquid chromatography coupled with UV and fluorescence detection, respectively. Analytes were extracted using 20 mM potassium phosphate and 10 mM ammonium formate. The pantothenic acid content of seafood ranged from 0.14 ± 0.01 mg/100 g (fusiformis) to 10.19 ± 0.46 mg/100 g (salted pollack roe), and total vitamin B6 content in seafood varied from 0.01 ± 0.00 mg/100 g (seersucker) to 1.39 ± 0.04 mg/100 g (salted pollack roe). Applied analytical method was validated using multiple method validation parameters, including detection limit (B5, 0.0003 mg/100 g; B6, 0.04–0.07 µg/100 g), quantification limit (B5, 0.0008 mg/100 g; B6, 0.10–0.12 µg/100 g), linearity (R2 = 0.9998–1.000), precision (B5, repeatability: RSD 1.5%; reproducibility: RSD 3.8%, B6, repeatability: RSD 2.6%; reproducibility: RSD 5.0%), and accuracy. Accuracy was evaluated based on the recovery percentages from certified reference materials and standard spiked samples. Vitamin B5 was recovered from SRM-1849a (102.0 ± 0.2%), SRM-2387 (99.7 ± 1.8%), and standard spiked sample (81.1 ± 2.5%–103.9 ± 6.7%). Recoveries of vitamin B6 obtained from SRM 3290 (100.0–103.4%) and standard spiked sample (84.1–114.7%) were within the acceptable range established by the AOAC guidelines.