A very sensitive HPLC method for pentosidine estimation in urine and serum was elaborated. The liquid chromatograph of Shimadzu type CLASS VP version 5.0 including autosampler, quarternary pump, column oven and fluorescence monitor was controlled on line by means of a computer with special software CLASS VP in Windows 98 milieu. As a stationary phase was used CGC glass column Separon SGX C18, 150x3 mm (length x I.D.), filled with 7 m spherical silica, surface modified with octadecylsilyl groups. The mobile phase was 0.02 M heptafluorobutyric acid (HFBA) with 0.01 M (NH4)2SO4 and with a variable concentration of the acetonitrile (ACN). A pentosidine standard was synthesized using the simple polymeranalogical reaction with no require- ment for purification of the reaction product. Variation in the reproducibility of the HPLC estimation was slightly above 1% (RSD), the sensitivity limit was lower than 20 femtomoles, recovery of the whole method including sample hydrolysis and purification with cellulose was 77 ± 3.5%. The method was verified with very good results using a model sample of urine and serum.

        Key words: pentosidine, HPLC, AGE, pentosidine preparation, glycation.