Evaluating mixtures of low molecular weight heparins by chain profiles or chain mapping

A method of producing an enoxaparin preparation, the method comprising the steps of:

• providing an isolated tetrasaccharide fraction of a size fractionated enoxaparin batch;
• evaluating the tetrasaccharide fraction using high performance liquid chromatography (HPLC) to determine the presence of at least one of the chains shown in the following 1st table:
• Peak #
• Assignment
• 1
• ΔUHNS6SI2SHNS6S
• 2
• ΔUHNS6SI2SHNS6S
• ΔUgalHNS6SI2SManNS6S
• 3
• ΔU2SHNS6SI2SHNS (α)
• 4
• ΔU2SHNS6SI2SHNS (β),
• ΔU2SHNS6SI2SManNS (β)
• 5
• ΔU2SHNS6SI2SManNS (α),
• ΔU2SHNSI2SHNS6S (α)
• 6
• ΔU2SHNSI2SHNS6S (β)
• 7
• ΔU2SHNS6SI2SManNS (1,6 anhydro)
• 8
• ΔU2SHNS6SI2SHNS (1,6 anhydro)
• 9
• ΔU2SHNS6SG2SHNS (α)
• 10
• ΔU2SHNS6SGHNS6S (α)
• 11
• ΔU2SHNS6SGManNS6S (α,β)
• 12
• ΔU2SHNS6SI2SHNS6S (α),
• ΔU2SHNS6SI2SManNS6S (β)
• 13
• ΔU2SHNS6SI2SHNS6S (β),
• ΔU2SHNS6SI2SManNS6S (α)
• 14
• ΔU2SHNS6SG2SHNS6S (α)
• 15
• ΔU2SHNS6SG2SHNS6S (β)
• or to determine the presence of a peak in an amount identified in the following 2nd table:
• Chain
• Sequencing
• by SAX-
• HPLC
• Range A
• Distribution
• Area % of
• Range B
• Range C
• by Area %
• HPLC peaks
• Area %
• Area %
• Peak 1
• 0.1
• 0.6
• Present or
• Present or
• Detectable
• Detectable
• Peak 2
• 0.6
• 1.2
• 0.6
• 1.2
• 0.4
• 1.4
• Peak 3
• 0.6
• 1.2
• 0.4
• 1.0
• 0.2
• 1.2
• Peak 4
• 1.8
• 3.6
• 1.7
• 4.1
• 0.9
• 4.9
• Peak 5
• 2.1
• 3.7
• 2.2
• 4.0
• 1.6
• 4.6
• Peak 6
• 0.5
• 1.6
• 0.4
• 1.6
• 0.0
• 2.0
• Peak 7
• 5.9
• 9.0
• 5.6
• 9.2
• 4.4
• 10.4
• Peak 8
• 7.0
• 9.9
• 6.6
• 10.2
• 5.4
• 11.4
• Peak 9
• 0.9
• 1.6
• 0.8
• 1.4
• 0.6
• 1.6
• Peak 10
• 5.0
• 6.2
• 4.9
• 6.7
• 4.3
• 7.3
• Peak 11
• 1.2
• 1.9
• 1.0
• 2.2
• 0.6
• 2.6
• Peak 12
• 39.7
• 48.0
• 38.1
• 48.9
• 34.5
• 52.5
• Peak 13
• 18.1
• 20.7
• 17.8
• 20.8
• 16.8
• 21.8
• Peak 14
• 2.3
• 4.0
• 1.9
• 4.3
• 1.1
• 5.1
• Peak 15
• 0.6
• 1.4
• 0.4
• 1.6
• 0.0
• 2.0
• * Peak numbers referenced in this 2nd Table correlate to the chains associated with said peaks in the 1st Table
• and based upon the determination, further processing the enoxaparin batch by a method including one or more steps selected from the group consisting of classifying, selecting, accepting, discarding, releasing, withholding, processing into a drug product, shipping, moving to a different location, formulating, labeling, packaging, releasing into commerce, selling, and offering for sale the enoxaparin batch.