High-performance liquid chromatographic assay for sodium mercaptoundecahydrododecaborate in rat tissues

Mercaptoundecahydrododecaborate (BSH) is an important agent for the boron neutron-capture therapy (BNCT). A sensitive high-performance liquid chromatographic (HPLC) method was developed for measuring BSH concentrations in rat tissues. Various tissue samples containing the drug were homogenized in a 1:1 (g/ml) mixture with phosphate buffered saline. The samples were then deproteinised with 4 volumes of acetonitrile and centrifuged. An aliquot of the supernatant was dried and reconstituted in 200 microliter of Tris-HC1 buffer. The samples were subjected to precolumn derivatization using the thiol reactive monobromobimane (mBB). The drug-mBB adduct was resolved by isocratic elution from a C18 reversed-phase column. The optimized mobile phase was methanol-0.02 M phosphate buffer (43:57, v/v) containing 0.01 M tetrabutylammonium dihydrogen phosphate as the ion-pairing agent with the final pH adjusted to 7.0. The flow-rate was set at 2.0 ml/min. The adduct was monitored by UV absorption at 373 nm. The analysis was completed in less than 15 min. The detection limit was 0.5 microgram/ml (0.25 microgram of boron). The assay method was linear over a concentration range of 0.5 to 50 micrograms/ml. This assay method could be used to evaluate the BSH concentrations in different tissues in studies on the targeted delivery of BSH.