High-performance liquid chromatographic analysis for a nonchromophore-containing phosphatidyl inositol analog, 1-[(1-O-octadecyl-2-O-methyl-sn-glcero)-phospho]-1D-3-deoxy-myo-inositol, using indirect UV detection

Phosphatidylinosite-3-kinase (PI3 kinase) is an important constituent of gr owth factor regulation. It is also involved in oncogene signaling pathways. an ether-containing phosphatidyl inositol(PI) analog, OMDPI, 1-[(1-O-octad ecyl-2-O-methyl-sn-glycero)-phospho]-1D-3-deosy-myo-inositol, , is a potent inhibitor of this pathway and may be clinically useful in the treatment of a variety of neoplasms, OMDPI is currently being investigated as an anti-t umor agent by the National Cancer Institute, NIH, OMDPI, a non-chromophore- containing PI analog, is not directly adaptable to the commonly used UV det ection of HPLC. This paper reports the development and validation of an HPL C assay for OMDPI based on indirect UV detection, in which a UV-absorbing i on-pair reagent (the probe), protriptyline, is added to the mobile phase to induce a signal for the compound. The method is sensitive (limit of detect ion <5 <mu>l of 1 mug/ml or 5 ng), precise (R.S.D.<2.5%), linear (r(2)=0.99 95) and accurate (error<0.07%). It is superior to refractive index detectio n and evaporative light scattering detection in either sensitivity or linea rity and does not require special equipment. (C) 2001 Elsevier Science B.V. All rights reserved.