Simultaneous determination of p-aminohippuric acid, acetyl-p-aminohippuric acid and iothalamate in human plasma and urine by high-performance liquid chromatography.
A sensitive and specific high-performance liquid chromatographic assay was developed for the simultaneous determination of p-aminohippuric acid (PAH), acetyl-p-aminohippuric acid (aPAH), and iothalamate in human plasma and urine. Plasma samples were prepared by protein precipitation with acetonitrile followed by evaporation, reconstitution in mobile phase, and injection onto a C18 reversed-phase column. Urine samples were diluted with 3 volumes of mobile phase prior to injection. Column effluent was monitored by UV detection at 254 nm. The lower limits of quantification in plasma were 0.5 mg/l for PAH and aPAH, and 1.0 mg/l for iothalamate. The within-day and between-day coefficients of variation in plasma and urine were < or =7.8% for all analytes. This method is well suited for renal function studies using iothalamate and PAH, whether administered as a bolus dose or by continuous infusion, to measure glomerular filtration rate and effective renal plasma flow, respectively.
Dowling
T C
TC
Department of Pharmaceutical Sciences, School of Pharmacy, University of Pittsburgh, PA 15261, USA.
Frye
R F
RF
Zemaitis
M A
MA
D000097:Acetonitriles; D000618:Aminohippuric Acids; D001798:Blood Proteins; D011232:Chemical Precipitation; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D003287:Contrast Media; D005919:Glomerular Filtration Rate; D006801:Humans; D007483:Iothalamic Acid / Q000032:analysis*; D011786:Quality Control; D012079:Renal Circulation; D012680:Sensitivity and Specificity; D010130:p-Aminohippuric Acid / Q000032:analysis* / Q000097:blood / Q000652:urine
D000097:Acetonitriles; D000618:Aminohippuric Acids; D001798:Blood Proteins; D003287:Contrast Media; D007483:Iothalamic Acid; C024893:4-acetylaminohippuric acid; D010130:p-Aminohippuric Acid; C032159:acetonitrile