Red cell pyrimidine 5' nucleotidase deficiency: determination of nucleotidase activity and nucleotide content using HPLC.

The hereditary deficiency of erythrocyte pyrimidine 5' nucleotidase has been investigated using an HPLC anion-exchange procedure designed to measure both red cell nucleotide content and enzymic activity. Red cell nucleotide profiles were determined using a low pH phosphate buffer salt-gradient system in 20 min, whereas a low pH buffer alone permitted the determination of enzymic activity with each of six different nucleotide substrates in less than 4 min. Both the red cell nucleotide profiles and the enzymic activity of haemolysates from two affected brothers and their children agreed well with previously published values. This unified approach should prove useful for detailed studies of deficiencies involving isoenzymes of pyrimidine nucleotidase.