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de Boer NK, Reinisch W, Teml A, van Bodegraven AA, Schwab M, Lukas M, et al; Dutch 6-TG working group.
We prepared calibration curves by adding known amounts of 6-TG and 6-MMP to a pool of erythrocyte hemolysates isolated from blood bank samples from healthy blood donors.
Separation of 6-TG and 6-MMP derivative was achieved with an 8-min isocratic elution with 95% mobile phase A-5% mobile phase B at a flow rate of 0.
This was followed by hydrolysis (100 [degrees]C for 45 min) of the 6-TGNs in the separated supernatant to release the 6-TG.
For chromatographic separation of the free 6-TG after both sample preparation procedures described above, we used a modification (7) of the Lennard chromatographic method (21).
The free bases, 6-TG and 6-MP, liberated from the respective nucleoside and nucleotide moiety by acid hydrolysis, were analyzed on an octadecylsilane column after liquid-liquid extraction with dichloromethane.
6-TG and Me6-MP derivative liberated from the nucleotide moiety were analyzed by a reversed-phase HPLC method.