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Nonextractive Procedure and Precolumn Derivatization with 7-Chloro-4-nitrobenzo-2-oxa-1,3-diazole for Trace Determination of Trimetazidine in Plasma by High-Performance Liquid Chromatography with Fluorescence Detection

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A highly sensitive high-performance liquid chromatographic method with fluorescence detection has been developed and validated in a single laboratory for the trace determination of trimetazidine (TMZ) in human plasma. Fluoxetine (FLX) was used as the internal standard. TMZ and FLX were isolated from plasma by protein precipitation with acetonitrile and derivatized by heating with 7-chloro-4-nitrobenzo-2-oxa-1,3-diazole in pH 8 borate buffer at 70C for 30 min. Separations were performed in the isocratic mode on a Nucleosil CN column with the mobile phase acetonitrile10 mM sodium acetate buffer (pH 3.5)methanol (47 + 47 + 6, v/v/v) at a flow rate of 1.0 mL/min. The derivatized samples were excited at 470 nm and monitored at an emission wavelength of 530 nm. Under the optimum chromatographic conditions, a linear relationship with a good correlation coefficient (r 0.9997, n 5) was obtained for the peak area ratio of TMZ to FLX and for TMZ concentrations of 1120 ng/mL. The proposed method has the lowest limits of detection and quantitation reported to date for the determination of TMZ in plasma with values of 0.3 and 0.95 ng/mL, respectively. The values for intra- and interassay precision were satisfactory; the relative standard deviations were 4.04. The accuracy of the method was demonstrated; the recoveries of TMZ from spiked human plasma were 98.13102.83 0.24.04. The method has high throughput because of its simple sample preparation procedure and short run time (<10 min). The results demonstrated that the proposed method would have great value when applied in pharmacokinetic studies for TMZ.

Document Type: Research Article

Affiliations: King Saud University, College of Pharmacy, Department of Pharmaceutical Chemistry, PO Box 2457, Riyadh 11451, Saudi Arabia.

Publication date: September 1, 2008

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  • The Journal of AOAC INTERNATIONAL publishes refereed papers and reviews in the fields of chemical, biological and toxicological analytical chemistry for the purpose of showcasing the most precise, accurate and sensitive methods for analysis of foods, food additives, supplements and contaminants, cosmetics, drugs, toxins, hazardous substances, pesticides, feeds, fertilizers and the environment available at that point in time. The scope of the Journal includes unpublished original research describing new analytical methods, techniques and applications; improved approaches to sampling, both in the field and the laboratory; better methods of preparing samples for analysis; collaborative studies substantiating the performance of a given method; statistical techniques for evaluating data. The Journal will also publish other articles of general interest to its audience, e.g., technical communications; cautionary notes; comments on techniques, apparatus, and reagents.
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