USE OF AN ENZYMATIC MIXTURE AGAINST MET-HB EXCESS DURING OXIMETRY OF DILUTE HB-A SOLUTIONS

: A mixture of NADH, cytochrome-C-reductase and methylene blue was employed to antagonize ferri(met)-Hb formation during oximetries of dilute samples of human Hb-A. Its efficacy is clear at any pH between 6.8, 7.3 and 7.8, in the presence of 100 mM NaCl. In these cases, ferri(met)-Hb decreases of about 2/3, compared with the enzyme-free controls, and p50 increases. On the contrary, when samples are examined at pH 7.3 with 600 mM NaCl and/or 100 mM NaCl plus 1 mM DPG, ferri(met)-Hb of the samples containing the enzymes also decreases, but not more than 1/2 of its initial value, whereas also p50 decreases. Finally, Bohr effect is lower in the samples containing the enzymes. In conclusion, the enzymatic mixture is very useful, but not easy to be handled; so, some observations for its correct use are required.