Determination of riboflavin by high-performance liquid chromatography with riboflavin-depleted urine as calibration and control matrix.
A simple method, exposure to natural-light, was developed to remove riboflavin from urine to enhance its use as the biological matrix for the preparation of calibration and control samples. Riboflavin-depleted urine containing less than 1 ng/ml of riboflavin was used to validate a high-performance liquid chromatography with fluorescence detection method for the determination of urinary riboflavin. The linearity of the assay (r2=0.999) was acceptable over the range of 10-5000 ng/ml. The intra-assay and inter-assay CVs were 3.3% and 9%, respectively. Subsequent stability studies found that urine riboflavin was stable for up to 6 months at 4 or -20 degrees C.
Chen
Meng
M
Center for Human Toxicology, Department of Pharmacology and Toxicology, University of Utah, 20 South 2030 East Room 490, Salt Lake City, UT 84112, USA. mc4@utah.edu
Andrenyak
David M
DM
Moody
David E
DE
Foltz
Rodger L
RL
D000328:Adult; D002138:Calibration / Q000592:standards; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D004355:Drug Stability; D006801:Humans; D008027:Light; D012256:Riboflavin / Q000528:radiation effects / Q000652:urine*
D012256:Riboflavin