Improved high performance liquid charomatographic method for the determination of tris(hydroxymethyl)aminomethane (THAM) in human plasma, erythrocytes, and whole blood

A sensitive high performance liquid chromatographic (HPLC) assay was developed to quantitate the organic amine proton-acceptor, tris(hydroxymethyl)aminomethane (THAM) in human plamsa, erythrocytes, and whole blood. An aliquot of the sample was heat evaporated (200 degree C for 2 h), derivatized, extracted, and then injected onto a symmetry C sub(8) column. The UV absorbance of the effluent was monitored at 237 nm. 2-Amino-2-methyl-1,3-propanediol was found to be an appropriate internal standard. The method has been applied to quantitate the samples from patients undergoing orthotopic liver transplantation. The assay has also been employed to assess the in vitro time of THAM-uptake into the erythrocytes from blood samples incubated at 37 degree C.