Novel and highly sensitive mixed-polymeric electrokinetic chromatography system for determination of contaminants and impurities of heparin samples

TRIPODI V.,; FLOR S.,; DOBRECKY C.,; CONTIN M.,; LUCANGIOLI, SILVIA

ELECTROPHORESIS

WILEY-V C H VERLAG GMBH

Lugar: WEINHEIM; Año: 2010 vol. 31 p. 3606 - 3612

0173-0835

A mixed-polymeric electrokinetic chromatography system has been developed for the simultaneous determination of a contaminant like oversulfated condroitin sulfate (OSCS) and impurities expressed as dermatan (Der) in heparin (Hep) samples. The EKC system consisted of 0.5 % w/v polymeric β-cyclodextrin, 0.4 % w/v tetronic® 1107 and 400 mM tris-phosphate buffer at pH 3.5. The optimized electrophoretic conditions included the use of an uncoated-silica capillary of 50 cm of total length and 75 µm i.d., an applied voltage of -7 kV, a temperature of 30 ºC and 200 nm UV-detection. The highly sensitive method developed showed low values of LOD, 0.07 % w/w (0.07 µg/ml) (OSCS) and 0.1 % w/w (0.1 µg/ml) (Der), and values of LOQ 0.2 % w/w (0.2 µg/ml) (OSCS) and 0.3 % w/w (0.3 µg/ml) (Der) with a concentration level of heparin sample as low as 0.1 mg/mL. Additional parameters of validation such as specificity, linearity, accuracy, and robustness were evaluated according to international guidelines. Due to its simplicity, high sensitivity and reliability, the proposed method can be an advantageous alternative to the traditional methodologies for the analysis of heparin in raw material and specially in finished products because of the low amounts of heparin sample required.