Published:Journal of Chromatographic Science,
ISSN 0021-9665Volume
41, Number 8, September 2003, pp. 405-409
Liposomal Benznidazole: A High-Performance Liquid Chromatographic
Determination for Biodistribution Studies
M.J. Morilla, P.E. Benavidez, M.O. Lopez, and E.L. Romero*
Departamento de Ciencia y Tecnología, Universidad Nacional de Quilmes,
Laboratorio de Diseño de Transportadores de Drogas, Roque Saenz Peña
180, Bernal, B1876BXD, Buenos Aires, Argentina
In this work, an isocratic high-performance
liquid chromatographic method for quantitation of liposomal benznidazole (BNZ)
in biological
tissues is presented. The method comprises protein precipitation together with
an efficient extraction of bulk or liposomal BNZ with acetonitrile–dimethylsulfoxide
(1:1, v/v) at a 2:1 (extraction solvent–tissue matrix, v/v or /vw) ratio;
the process is completed by a final precipitation with trichloroacetic acid.
The resultant supernatants are assayed chromatographically using a Kromasil C18
(25- ¥ 0.4-cm i.d., 100 Å, 5-µm particle size), with an isocratic
mobil phase consisting of acetonitrile–water (40:60, v/v), a flow rate
of 0.9 mL/min, and detected at 324 nm. Bulk BNZ is used as a reference standard
for the analysis of samples containing liposomal BNZ. The assay is linear over
a concentration range of 0.75 (the lowest quantity of analyte determined with
precision and accuracy of ≤ 20%) to 25 µg/mL-g in all liquid and
solid matrices. Within-day precision is better than 6.4% in plasma and 8.6% in
liver, the same for the two assayed concentrations. Between-day precision is
5.4% and 12.3% in plasma and 9% and 6.9% in liver for the two assayed concentrations,
respectively. The absolute recoveries range between 70% and 97%. Therefore, the
method is accurate and precise to be employed for detection of minor quantities
of liposomal BNZ in biological tissues.
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