TY - JOUR
T1 - Determination of febuxostat in human plasma by high performance liquid chromatography (HPLC) with fluorescence-detection
AU - Kamel, Bishoy
AU - Williams, Kenneth M.
AU - Graham, Garry G.
AU - Norris, Ross L.G.
AU - Stocker, Sophie L.
AU - Carland, Jane E.
AU - Pile, Kevin D.
AU - Day, Richard O.
PY - 2019
Y1 - 2019
N2 - Febuxostat prevents gout attacks by lowering serum urate. Aspects of the pharmacokinetic-pharmacodynamic relationship of febuxostat concentrations to urate in gout patients need further elucidation. In order to undertake these studies, the assay methodology for febuxostat has been enhanced and validated to meet FDA standards. An HPLC method with fluorescence-detection has been modified to increase sensitivity, reduce complexity, shorten the sample preparation process and improve the inter-day coefficient of variation of the lowest quality control sample (0.03"¯Î¼g/L). Protein in plasma samples (200"¯Î¼L) is now precipitated with acetonitrile (400"¯Î¼L) containing the internal standard (2-naphthoic acid). The supernatant is analysed at excitation and emission wavelengths of 320 and 380"¯nm, respectively as in the previous method. A Luna C18 column (Phenomenex, Australia) at 40"¯Ã‚°C with mobile phase of glacial acetic acid (0.032%) in acetonitrile:water (60:40, v:v), an injection volume of 10"¯Î¼L and a flow rate of 1.5"¯mL/min is employed. Analysis time is 8"¯min. Calibration curves in drug-free plasma range from 0.005 to 10.00"¯Î¼g/mL. Data points are fitted using linear regression with a weighting factor of 1/concentration. The inter-day accuracy and imprecision of the quality control samples (0.0075, 0.015, 3.00 and 9.80"¯Î¼g/mL) is 90-115% and"¯ ≤"¯14.5%, respectively.
AB - Febuxostat prevents gout attacks by lowering serum urate. Aspects of the pharmacokinetic-pharmacodynamic relationship of febuxostat concentrations to urate in gout patients need further elucidation. In order to undertake these studies, the assay methodology for febuxostat has been enhanced and validated to meet FDA standards. An HPLC method with fluorescence-detection has been modified to increase sensitivity, reduce complexity, shorten the sample preparation process and improve the inter-day coefficient of variation of the lowest quality control sample (0.03"¯Î¼g/L). Protein in plasma samples (200"¯Î¼L) is now precipitated with acetonitrile (400"¯Î¼L) containing the internal standard (2-naphthoic acid). The supernatant is analysed at excitation and emission wavelengths of 320 and 380"¯nm, respectively as in the previous method. A Luna C18 column (Phenomenex, Australia) at 40"¯Ã‚°C with mobile phase of glacial acetic acid (0.032%) in acetonitrile:water (60:40, v:v), an injection volume of 10"¯Î¼L and a flow rate of 1.5"¯mL/min is employed. Analysis time is 8"¯min. Calibration curves in drug-free plasma range from 0.005 to 10.00"¯Î¼g/mL. Data points are fitted using linear regression with a weighting factor of 1/concentration. The inter-day accuracy and imprecision of the quality control samples (0.0075, 0.015, 3.00 and 9.80"¯Î¼g/mL) is 90-115% and"¯ ≤"¯14.5%, respectively.
KW - blood plasma
KW - gout
KW - high performance liquid chromatography
UR - http://handle.westernsydney.edu.au:8081/1959.7/uws:52804
U2 - 10.1016/j.jchromb.2019.121764
DO - 10.1016/j.jchromb.2019.121764
M3 - Article
SN - 1570-0232
VL - 1126-1127
JO - Journal of Chromatography B
JF - Journal of Chromatography B
M1 - 121764
ER -