Acta Pharmaceutica, Vol. 65 No. 1, 2015.
Original scientific paper
https://doi.org/10.1515/acph-2015-0010
Development and validation of a simple and sensitive size-exclusion chromatography method for quantitative determination of heparin in pharmaceuticals
MAJA RADIVOJŠA MATANOVIĆ
; Faculty of Pharmacy, University of Ljubljana, 1000 Ljubljana, Slovenia
IZTOK GRABNAR
; Faculty of Pharmacy, University of Ljubljana, 1000 Ljubljana, Slovenia
PEGI AHLIN GRABNAR
; Faculty of Pharmacy, University of Ljubljana, 1000 Ljubljana, Slovenia
ROBERT ROŠKAR
; Faculty of Pharmacy, University of Ljubljana, 1000 Ljubljana, Slovenia
Abstract
Heparin is widely used as an anticoagulant for the treatment and prevention of various thrombotic diseases. However, due to its high anionic charge, heterogeneity in size distribution and high polarity, its analysis is very challenging. In this paper, a novel method based on size-exclusion chromatography (SEC) for quantitative determination of intact heparin in pharmaceuticals is presented. Analyses were performed on a BioSep-SEC-S 2000 column with L-arginine solution at pH 6.5 as mobile phase and UV detection at 210 nm. The proposed method was found to be selective, linear (R2 > 0.997) over the concentration range of 3.1 to 1222 µg mL–1, with a limit of detection of 1.0 µg mL–1. Intra-day and inter-day precision were below 5.1 % and inaccuracy expressed as bias did not exceed 6.5 %. The reported method is simple, selective, sensitive, and requires no laborious sample preparation, which makes it appropriate for routine quantitative analysis of heparin in pharmaceuticals.
Keywords
heparin; quantitative analysis; size-exclusion chromatography; L-arginine
Hrčak ID:
128744
URI
Publication date:
31.3.2015.
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