Rapid determination of p-aminohippuric acid in serum and urine by high-performance liquid chromatography.
We report a simple and rapid HPLC procedure for measuring p-aminohippuric acid in serum and urine. After deproteinization with acetonitrile and addition of p-aminobenzoic acid as an internal standard, the chromatographic run is performed on a C18 column with the absorbance detector set at 275 nm. The separation is carried out in 10 min at a flow-rate of 1.0 ml/min with a mobile phase composed of 7 mmol/l 1-decanesulfonic acid, pH 3.70, and acetonitrile (82:18, v/v). The relationship between p-aminohippuric acid concentration and the p-aminohippuric acid/internal standard peak area is linear up to 100 microg/ml. Within-run precision measured at three different p-aminohippuric acid concentrations ranges from 1.73 to 1.98% in serum and from 0.72 to 1.32% in urine. Between-run precision varies from 1.80 to 4.06% in serum and from 1.05 to 2.66% in urine. Analytical recovery is between 98.26 and 99.44% in serum and from 99.57 to 100.45% in urine. The method is very simple, sensitive, and requires small volumes of sample for the assay (100 microl). Therefore, it could be a useful tool for the analysis of p-aminohippuric acid in the evaluation of renal plasma flow.
Marsilio
R
R
Department of Pediatrics, University of Padua, Italy.
Dall'Amico
R
R
Montini
G
G
Murer
L
L
Ros
M
M
Zacchello
G
G
Zacchello
F
F
D000328:Adult; D002648:Child; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D006801:Humans; D011786:Quality Control; D012680:Sensitivity and Specificity; D010130:p-Aminohippuric Acid / Q000097:blood* / Q000652:urine*
D010130:p-Aminohippuric Acid