[Role of HPLC in separation analysis in laboratory medicine]

The principle, theory and analytical application of high performance liquid chromatography (HPLC) in laboratory medicine was described. The importance of separation analysis was focussed with respect to accuracy, precision, short-time assay and easy operation. The pretreatment of samples such as deproteinization was also important for the accurate results and the analysis of plasma catecholamines and hemoglobin A1c were described as examples with use of column switching technique, step-wise separation, gradient elution method on TSK gels. We demonstrated the results of plasma free amino acids, gamma-carboxyglutamic acid and catecholamines in the aged which had been engaged in our laboratory as routine test and application with use of HPLC connected to chemiluminescent analyzer. Catecholamine analysis showed the importance of reproducibility and recovery. The reproducibility was within 2% as CV and 93-96% as recovery %. In the case of plasma free amino acid analysis, ASP, GLU, ALA showed decreases in the aged, but CYS did increase with age. Concerning about the drug monitoring of cyclospolin A in the infant whole blood after liver transplantation, the serum levels of cyclospolin A showed the lower concentration than whole blood. The extraction of cyclospolin A was required after deproteinization of whole blood as pretreatment.