ABSTRACT
In the late 1970s the atypical metabolite of ornithine, ornithine-lactam, has been observed in urine samples of patients suffering from hyperornithinemia. However, not least due to insufficient analytical methods, until now there are no data available about ornithine-lactam in human plasma. Here, we describe a new method, which is, for the first time, suitable to identify and quantify ornithine-lactam in human EDTA-plasma. The method was validated according to the requirements of the FDA guidance for bioanalytical method validation. The analytes were extracted on mixed mode cation exchange SPE columns, separated on a silica analytical HPLC column working in the HILIC mode and detected on a tandem mass spectrometer equipped with an ESI ion source. As internal standard newly synthesized stable isotope labeled D(6)-ornithine-lactam was used. The calibration function was linear in the range of 0.1-5 microM. Intra- and inter-day precision and accuracy was better than 14% at all concentration levels. In EDTA-plasma samples from 30 volunteers ornithine-lactam concentrations ranging from 0.136 to 0.653 microM were determined. These concentrations correlated significantly (p<0.001, R(2)=0.784) to those of ornithine in EDTA-plasma.
