Reversed-Phase Liquid Chromatographic Determination of Riboflavin in Feeds

Author:

Britton Nancy L1,Riter Ken L1,Smallidge Robert L1,Hillebrandt Joseph2

Affiliation:

1. Office of Indiana State Chemist, 1154 Biochemistry, Purdue University, West Lafayette, IN 47907

2. Cornell University Diagnostic Laboratory, Nutritional and Environmental Analytical Services, 777 Warren Rd, Ithaca, NY 14850

Abstract

Abstract A method for determination of riboflavin in animal feeds using liquid chromatography (LC) was developed for feed samples fortified with riboflavin at 1 mg/lb or greater (up to 10 000 mg/lb). Feed samples were extracted in 0.1N HCl with heating on a steam bath for 30 min, followed immediately by mechanical shaking for 30 min. Sample extracts were diluted to target volume with 2%acetic acid and filtered; riboflavin was determined by LC on a reversed-phase C18 column with 2% acetic acid–acetonitrile (85 + 15) mobile phase for separation and fluorescence detection with excitation at 460 nm and emission at 530 nm. The extraction was compared with that of the AOAC Official Method for riboflavin in food and feed premixes. The 2 method extractions were not significantly different from each other at the 95% confidence level. The developed method also had good linearity over 4 orders of magnitude, recovery of 95–99% from spiked feed samples, a limit of detection of riboflavin at 0.00034 μg/mL in solution, a limit of quantitation of 0.023 mg/lb in feed, and good ruggedness.

Publisher

Oxford University Press (OUP)

Subject

Pharmacology,Agronomy and Crop Science,Environmental Chemistry,Food Science,Analytical Chemistry

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