TY - JOUR
T1 - Rapid determination of p-aminohippuric acid in serum and urine by high-performance liquid chromatography
AU - Marsilio, R.
AU - Dall'Amico, R.
AU - Montini, G.
AU - Murer, L.
AU - Ros, M.
AU - Zacchello, G.
AU - Zacchello, F.
PY - 1997/12/19
Y1 - 1997/12/19
N2 - 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 l-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 μg/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 3.06% 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 μl). Therefore, it could be a useful tool for the analysis of p-aminohippuric acid in the evaluation of renal plasma flow.
AB - 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 l-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 μg/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 3.06% 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 μl). Therefore, it could be a useful tool for the analysis of p-aminohippuric acid in the evaluation of renal plasma flow.
KW - p-Aminohippuric acid
UR - http://www.scopus.com/inward/record.url?scp=0031578924&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=0031578924&partnerID=8YFLogxK
U2 - 10.1016/S0378-4347(97)00467-2
DO - 10.1016/S0378-4347(97)00467-2
M3 - Article
C2 - 9518171
AN - SCOPUS:0031578924
VL - 704
SP - 359
EP - 364
JO - Journal of Chromatography B: Biomedical Sciences and Applications
JF - Journal of Chromatography B: Biomedical Sciences and Applications
SN - 1387-2273
IS - 1-2
ER -