[Measurement of serum amiodarone by protein precipitation followed by high performance liquid chromatography with ultraviolet detection].
Our aim was to develop a quantitative method for serum amiodarone measurement using high performance liquid chromatography with ultraviolet photometric detection. We studied previous reports in the literature in order to obtain a simpler method to be used routinely in our TDM unit. Sample preparation was based on protein precipitation adding 2 parts of acetonitrile to 1 part of serum, followed by vortex-agitation for 45 s, incubation at 24 degrees C for 5 min, and centrifugation at 6000 x g for 2 min. Twenty microliters of the supernatant was directly injected into the chromatographic system. A microBondapak CN RP column (3.9 x 150 mm) at 45 degrees C, with a mobile phase consisting of KH2PO4 10 mM/methanol/acetonitrile (40:37:23 v/v/v), pH 3.5, were used. Eluting with a flow rate of 0.6 mL/mm the retention time of amiodarone was approximately 4.9 min. Detection was performed at 242 nm and the quantification was made by peak height comparison with external standards. The mass/response ratio is linear (r2 > 0.99) within a mass range of 2.96 to 18,930 ng of injected amiodarone, which exceeds the requirements for the monitoring of serum levels (0.3 to 6.0 micrograms/mL). Sample storage should be done with acetonitrile-extracted sera at -16 degrees C to avoid degradation. The method is very efficient, linear, sensitive and specific but it also simpler and cheaper than others reported in the literature.
Valencia
M
M
Unidad de Detección de Medicamentos y Química Clínica, Instituto de Medicina Experimental, Caracas, Venezuela.
Zapata
V
V
Bider
A
A
Pérez-González
M
M
Lamanna
V
V
D000638:Amiodarone / Q000097:blood*; D000889:Anti-Arrhythmia Agents / Q000097:blood*; D011232:Chemical Precipitation; D002851:Chromatography, High Pressure Liquid / Q000379:methods; D003365:Costs and Cost Analysis; D006801:Humans; D011506:Proteins / Q000737:chemistry*; D012680:Sensitivity and Specificity
D000889:Anti-Arrhythmia Agents; D011506:Proteins; D000638:Amiodarone