title: Direct quantification of pentosidine in urine and serum by HPLC with column switching.
abstract: Concentrations of pentosidine, an advanced glycation end product, are increased in aging, diabetes mellitus, and uremia. Using HPLC with column switching, we developed a direct method of measuring pentosidine in urine and serum. We inject the sample directly onto a gel-filtration precolumn, select ("heart-cut") the eluate fraction containing pentosidine, and introduce this fraction into a reversed-phased column by use of a switching valve. The recovery rate of the complete method was 97.7-99.9%. The intraassay CV was 5.7%, and the interassay CV was 5.8%. The calibration curve showed significant linearity (r = 0.998, P = 0.0001). We examined urinary concentrations of pentosidine in 12 diabetic patients (mean +/- SD, 8.7 +/- 2.3 micromol/mol of creatinine), 32 patients with chronic renal failure (CRF; 36.1 +/- 39.0), 19 osteoporotic patients (7.9 +/- 5.3), and 29 healthy control subjects (5.2 +/- 2.3). In CRF, urinary pentosidine in the patients undergoing hemodialysis was significantly higher than in CRF patients not being treated by hemodialysis (mean, 58.1 vs 18.2; P <0.001). Also, concentrations of urinary and serum pentosidine were significantly correlated (r = 0.797, P = 0.0011). Because this method does not require pretreatment of samples, it is convenient and useful for measuring urinary and serum pentosidine.
authors.affiliation: Department of Orthopedic Surgery, Hamamatsu University School of Medicine, Japan.
authors.forename: M
authors.initials: M
authors.lastname: Takahashi
authors.forename: H
authors.initials: H
authors.lastname: Hoshino
authors.forename: K
authors.initials: K
authors.lastname: Kushida
authors.forename: K
authors.initials: K
authors.lastname: Kawana
authors.forename: T
authors.initials: T
authors.lastname: Inoue
mesh_terms: D000328:Adult; D000368:Aged; D000375:Aging; D001120:Arginine / Q000031:analogs & derivatives* / Q000032:analysis / Q000097:blood / Q000652:urine; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D003920:Diabetes Mellitus / Q000097:blood / Q000652:urine; D006801:Humans; D007676:Kidney Failure, Chronic / Q000097:blood / Q000652:urine; D008239:Lysine / Q000031:analogs & derivatives* / Q000032:analysis / Q000097:blood / Q000652:urine; D008875:Middle Aged; D006435:Renal Dialysis; D013997:Time Factors; D014511:Uremia / Q000097:blood / Q000652:urine
chemical_list: D001120:Arginine; C062187:pentosidine; D008239:Lysine