Unfractionated Heparin Assay

[tykiser]

I am trying to find an HPLC method for identification of Heparin. Does anyone have an assay method they would be willing to share?
We simply want to test heparin's stability in different solutions and will not be measuring plasma levels. My literature search has been unsuccessful as many of the listed assays are being utilized to find particular functional groups, etc. Any help would be greatly appreciated.

Regards,
Ty

[Uwe Neue]

Size exclusion chromatography might be suitable for your task. I think that you should be able to get methods of suppliers of columns for aqueous SEC.

[Rolandas Plausinaitis]

It is not clear if you mean by assay activity or mass - distribution. For activity measurement method you can reach for USP draft or European Pharmacopeia articles which are quite similar and use enzymatic reaction measurement on UV spectrometer.

For mass profile gel permeation chromatography is used, there is draft USP article, but there is problem - calibration standard is not available. You can seek European Pharmacopeia article, the method has some problems, but standard is available. Also WHO has the standard and citation of the article which is describing measurement method (similar to above).
You can find description of measurement principle using RI/UV detection in: Determination of the Molecular-Weight Distribution of Low-Molecular-Weight Heparins Using High- Performance Gel Permeation Chromatography. Komatsu, Takahata, Tanaka, Ishimitsu, Okada- Biol. Pharm. Bull. 16(12), p. 1189-1193 (1993)

Alternative detection method is to use laser light scattering detector, for example miniDAWN from Wyatt Technology. For this method you don't need the calibration standard. Details are in Analytical Biochemistry, 245, p. 231-241 (1997).