High-performance liquid chromatographic microassay for L-glutamate:glyoxylate aminotransferase activity in human liver. Application in primary hyperoxaluria type 1.
A rapid and sensitive liquid chromatographic technique to determine L-glutamate:glyoxylate and aminotransferase (EC 2.6.1.4) activity in human liver is described. Homogenised tissue was incubated for 60 min in the presence of substrates and the 2-oxoglutarate generated was converted into the corresponding phenylhydrazone which was determined using reversed-phase high-performance liquid chromatography. The procedure allowed the detection of the enzyme activity expressed by 7.5 micrograms of liver protein, it was more sensitive and less time-consuming than the spectrophotometric procedure previously used. No significant differences were found between normal controls and patients with primary hyperoxaluria. In an 8-month-infant with primary hyperoxaluria type 1, the enzyme activity was reduced to 16% of the average control values.
Petrarulo
M
M
Laboratorio Calcolosi Renale, Ospedale Mauriziano Umberto I, Turin, Italy.
Pellegrino
S
S
Marangella
M
M
Cosseddu
D
D
Linari
F
F
D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D006801:Humans; D006960:Hyperoxaluria, Primary / Q000201:enzymology*; D007223:Infant; D008099:Liver / Q000201:enzymology*; D008297:Male; D008875:Middle Aged; D010659:Phenylhydrazines; D012680:Sensitivity and Specificity; D013056:Spectrophotometry, Ultraviolet; D000637:Transaminases / Q000378:metabolism*
D010659:Phenylhydrazines; C030299:phenylhydrazine; D000637:Transaminases; C020649:glutamate-glyoxylate aminotransferase