High-performance liquid chromatographic analysis of physiological amino acids in human brain tumors by pre-column derivatization with phenylisothiocyanate

A reversed-phase high-performance liquid chromatographic technique for the determination of free amino acids in five biopsies of human brain tumors (t wo meningiomas, one glioblastoma and two oligodendrogliomas) is described. The frozen tissues were homogenized, deproteinized with perchloric acid and neutralized with potassium hydroxide. Aliquots of the supernatant containi ng the physiological amino acids are used for pre-column derivatization wit h phenylisothiocyanate. The derivatized PTC-amino acids (phenylthiocarbamyl derivatives) are stable for a five day period if stored as a powder at -20 degrees C in an inert atmosphere and they can be analyzed on a reversed-ph ase column (PicoTag) using a gradient of two eluents with absorption detect ion at a wavelength of 254 nm. Good resolution of several amino acids (>30) is achieved within ca. 60 min. For most amino acids this method is suitabl e for an accurate measurement over a wide range of physiological concentrat ions (50-400 pmol) starting from a very small amount of sample. (C) 1999 Pu blished by Elsevier Science B.V. All rights reserved.