Development and validation of a high performance liquid 4 chromatography method for oligodeoxynucleotides determination 5 in a novel coagel-based formulation

GABRIELA V. ULLIO GAMBOA A,C, JUAN M. LLABOT A,C, MARÍA F. SANCHEZ-VALLECILLO B,C, BELKYS A. MALETTO B,C, 9 SANTIAGO D. PALMA A,C, DANIEL A. ALLEMANDI

Analytical Chemistry Research

Elsevier B.V.

Año: 2015 p. 20 - 20

2214-1812

he therapeutic benefit of phosphorothioate oligodeoxynucleotides (PS-ODN) containing immune stim- 24 ulatory sequences has been demonstrated in animal models of cancer and infection. Several tools are 25 available for the determination of these oligonucleotides in biological samples and pharmaceutical pre- 26 parations, including UV spectroscopy, dye binding, isotopic tracing, capillary gel electrophoresis (CGE), 27 hybridization-based enzyme-linked immunosorbent assay (ELISA), and chromatography techniques. 28 However, due to inter-assay variability and accuracy problems associated with the afore mentioned 29 methods, we have developed and validated an isocratic high performance liquid chromatographic 30 (HPLC) for analytical determination of PS-ODN containing unmethylated CpG motifs (CpG-ODN). 31 Validation under Food and Drug Administration (FDA) guidelines of the analytical parameters include: 32 linearity (r2 0.9996), LODs (0.86 lg/ml) and LOQ (6.25 lg/ml), infra (0.19?3