title: Validation of an analytical methodology to quantify melamine in body fluids using micellar liquid chromatography.
abstract: Measurement of urine and plasma melamine-concentration is helpful in confirming melamine-associated renal diseases. A chromatographic procedure using a C18 column and a micellar mobile phase of sodium dodecyl sulphate (0.2M), buffered at pH 3 and detection set at 210 nm, was reported for the resolution and quantification of melamine in plasma and urine. In this work, direct injection was used, thus avoiding long extraction and experimental procedures. Melamine was eluted in nearly 6.3 min without overlapping the protein band or other endogenous compounds. The optimal mobile phase composition was taken by studying the influence of each chromatographic parameter. Validation was satisfactorily performed following the US Food and Drug Administration (FDA), in terms of: linearity (0.25-25 ppm; r2>0.9995 in both cases), sensitivity, limit of detection (50 ppb), limit of quantification (250 ppb), intra- and inter-day precision (R.S.D. 0.7-10.2% and 1.0-9.1%, respectively) and recovery, calculated as accuracy (85.7-103.8% and 94.8-103.6%, respectively) and robustness (R.S.D.<7.1%). The suggested methodology has been applied to the analysis of real samples of volunteers, and no melamine was found in any of them.
authors.affiliation: Química Bioanalítica, QFA, ESTCE, Universitat Jaume I, 12071 Castelló, Spain.
authors.forename: S
authors.initials: S
authors.lastname: Marco-Peiró
authors.forename: B
authors.initials: B
authors.lastname: Beltrán-Martinavarro
authors.forename: M
authors.initials: M
authors.lastname: Rambla-Alegre
authors.forename: J
authors.initials: J
authors.lastname: Peris-Vicente
authors.forename: J
authors.initials: J
authors.lastname: Esteve-Romero
mesh_terms: D000328:Adult; D001826:Body Fluids / Q000737:chemistry*; D002138:Calibration; D002853:Chromatography, Liquid; D005260:Female; D006801:Humans; D006863:Hydrogen-Ion Concentration; D008297:Male; D008823:Micelles; D008875:Middle Aged; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D012967:Sodium Dodecyl Sulfate / Q000737:chemistry; D014227:Triazines / Q000097:blood* / Q000652:urine*
chemical_list: D008823:Micelles; D014227:Triazines; D012967:Sodium Dodecyl Sulfate; C011907:melamine