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

A method of manufacturing an enoxaparin preparation comprising:

• esterification of carboxylate groups on uronic acid residues of a heparin preparation followed by beta-elimination to provide a LMWH preparation;
• providing a high performance liquid chromatography (HPLC) determination of whether a tetrasaccharide structure is present in the LMWH preparation in an amount provided in the following table:
• Range A
• Range B
• Range C
• % Area
• % Area
• % Area
• under
• under
• under
• Peak
• HPLC
• HPLC
• HPLC
• #
• Tetrasaccharide
• curve
• curve
• curve
• 1
• ΔUHNS6SI2SHNS6S
• 0.1
• 0.6
• Present or
• Present or
• Detectable
• Detectable
• 2
• ΔUHNS6SI2SHNS6S
• 0.6
• 1.2
• 0.6
• 1.2
• 0.4
• 1.4
• ΔUgalHNS6SI2SManNS6S
• 3
• ΔU2SHNS6SI2SHNS (α)
• 0.6
• 1.2
• 0.4
• 1.0
• 0.2
• 1.2
• 4
• ΔU2SHNS6SI2SHNS (β),
• 1.8
• 3.6
• 1.7
• 4.1
• 0.9
• 4.9
• ΔU2SHNS6SI2SManNS (β)
• 5
• ΔU2SHNS6SI2SManNS (α),
• 2.1
• 3.7
• 2.2
• 4.0
• 1.6
• 4.6
• ΔU2SHNSI2SHNS6S (α)
• 6
• ΔU2SHNSI2SHNS6S (β)
• 0.5
• 1.6
• 0.4
• 1.6
• 0.0
• 2.0
• 7
• ΔU2SHNS6SI2SManNS
• 5.9
• 9.0
• 5.6
• 9.2
• 4.4
• 10.4
• (1,6 anhydro)
• 8
• ΔU2SHNS6SI2SHNS
• 7.0
• 9.9
• 6.6
• 10.2
• 5.4
• 11.4
• (1,6 anhydro)
• 9
• ΔU2SHNS6SG2SHNS (α)
• 0.9
• 1.6
• 0.8
• 1.4
• 0.6
• 1.6
• 10
• ΔU2SHNS6SGHNS6S (α)
• 5.0
• 6.2
• 4.9
• 6.7
• 4.3
• 7.3
• 11
• ΔU2SHNS6SGManNS6S
• 1.2
• 1.9
• 1.0
• 2.2
• 0.6
• 2.6
• (α, β)
• 12
• ΔU2SHNS6SI2SHNS6S (α),
• 39.7
• 48.0
• 38.1
• 48.9
• 34.5
• 52.5
• ΔU2SHNS6SI2SManNS6S (β)
• 13
• ΔU2SHNS6SI2SHNS6S (β),
• 18.1
• 20.7
• 17.8
• 20.8
• 16.8
• 21.8
• ΔU2SHNS6SI2SManNS6S (α)
• 14
• ΔU2SHNS6SG2SHNS6S (α)
• 2.3
• 4.0
• 1.9
• 4.3
• 1.1
• 5.1
• 15
• ΔU2SHNS6SG2SHNS6S (β)
• 0.6
• 1.4
• 0.4
• 1.6
• 0.0
• 2.0
• and
• if said structures are present in an amount provided in the table, processing said preparation into an enoxaparin drug product to thereby manufacture an enoxaparin preparation.