Determination of inulin in plasma and urine by reversed-phase high-performance liquid chromatography.
We report a new HPLC procedure for measuring inulin in plasma and urine. Samples after dilution are boiled in mild acidic conditions and then analyzed on a C18 column. Solvent system A is 3.2 mM HCl, pH 2.5, and B is acetonitrile-3.2 mM HCl (60:40, v/v), pH 2.5. The separation is carried out in 8 min with a flow-rate of 1.0 ml/min and the absorbance monitored at 280 nm. The relationship between inulin and the recorded peak area is linear from 0.2 to 3.2 mg/ml with a correlation coefficient of 0.999 for plasma and 0.999 for urine. Within-run precision, measured at three inulin concentrations, ranged from 0.9 to 1.7% in plasma and from 0.8 to 1.2% in urine. Between-run precision varied in plasma from 2.7 to 3.2% and in urine from 3.0 to 3.3%. Analytical recovery ranged from 102 to 107% in plasma and from 101 to 105% in urine, respectively. The method is sensitive, selective and only 30-microliters samples are required. Therefore, it could be used to evaluate the glomerular filtration rate even in small babies and to perform studies in animals.
Dall'Amico
R
R
Department of Pediatrics, University of Padua, Italy.
Montini
G
G
Pisanello
L
L
Piovesan
G
G
Bottaro
S
S
Cracco
A T
AT
Zacchello
G
G
Zacchello
F
F
D000328:Adult; D002648:Child; D002675:Child, Preschool; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D005919:Glomerular Filtration Rate; D006801:Humans; D006868:Hydrolysis; D007444:Inulin / Q000032:analysis* / Q000097:blood / Q000652:urine; D007674:Kidney Diseases / Q000097:blood / Q000652:urine; D015203:Reproducibility of Results; D013056:Spectrophotometry, Ultraviolet
D007444:Inulin