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Thin-layer polyacrylamide electrophoresis and kinetic studies were used to differentiate three different mutant erythrocyte pyruvate kinase (PK) in congenital nonspherocytic hemolytic anemia. In the first family, proband and father had PK which migrated faster than the normal and high Michaelis—Menten constants (Km), whereas mother had normal migration and kinetics, but low PK activity. In the second family, proband and mother had PK which migrated slower than the normal. Proband had markedly high Km and mother intermediately high Km. In the third family, proband and mother had faster migrating PK than the normal. Proband had high Km and mother had intermediately high Km. The parents in the second and third families were consanguineous. Inherited molecular abnormalities of erythrocyte PK were proven to exist by abnormal electrophoretic mobilities, as well as abnormal results of kinetic studies.

We evaluated six deproteinizing methods for determination of uric acid in serum by "high-performance" liquid chromatography with ultraviolet detection: those involving zinc hydroxide, sodium tungstate, trichloroacetic acid, perchloric acid, acetonitrile, and centrifugal ultrafiltration (with Amicon MPS-1 devices). We used a Toyosoda ODS-120A reversed-phase column. The mobile phase was sodium phosphate buffer (40 mmol/L, pH 2.2) containing 20 mL of methanol per liter. Absorbance of the eluate was monitored at 284 nm. The precipitation method with perchloric acid gave high recoveries of uric acid and good precision, and results agreed with those by the uricase-catalase method of Kageyama (Clin Chim Acta 1971;31:421-6).

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