LOC14 to be the most potent PDI inhibitor reported to date( EC50 : 500 nM)

PDI:500 nM(EC50)

LOC14 levels were measured in mouse plasma, striatum and cerebral cortex using Ultra Performance Liquid Chromatography-Tandem Mass Spectrometry (LC-MS/MS).?LOC14 was extracted from mouse plasma by mixing 100 μl of plasma with 900 μl of cold acetonitrile containing 25 ng/ml of cyclocreatine as an internal standard.?After mixing for 5 min. the sample was centrifuged for 10min and the organic layer was transferred to an LC-MS vial and dried under nitrogen.?The sample was reconstituted in 100 μl of 50% acetonitrile and 5 μl was injected onto the LC-MS.?Brain tissue samples were homogenized using a tissue tearor at a concentration of 150 mg/ml in cold LC-MS water followed by extraction with 1.8 ml of cold acetonitrile containing the cyclocreatine internal standard.?The brain samples were mixed and centrifuged and reconstituted the same as for the plasma samples[2].