Enoxaparin Structural Characterization by HILIC-FTMS

Lovenox^® and Clexane^®, the innovator versions of enoxaparin marketed in the U.S. and Europe were purchased from Sanofi-Aventis (Bridgewater, NJ). Generic versions of Lovenox^® were provided by three different manufactures (three current lots of each). Online hydrophilic interaction chromatography (HILIC) Fourier transform mass spectrometry (FTMS) was performed as previous described [4 (link)]. Briefly, enoxaparin injections were diluted into 1 μg/μL and directly injected into a HILIC column (2.0 mm × 150 mm, 200 Å, Phenomenex, Torrance, CA) by an Agilent 1200 autosampler. The LC column was directly connected online to the standard ESI source of LTQ-Orbitrap XL FT-MS (Thermo Fisher Scientific, San-Jose, CA). The enoxaparin intact chain compositions were analyzed under the negative mode. Following raw data acquisition, charge deconvolution was auto-processed using DeconTools [14 –15 (link)] software. Enoxaparin structural assignment was performed by automatic processing with GlycReSoft 1.0 software, developed at Boston University (http://code.google.com/p/glycresoft/downloads/list) [4 (link),12 (link)]. The output on enoxaparin composition from GlycReSoft was then processed using GlycCompSoft to provide automated relative quantification of intact chains present in enoxaparin.

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Wang X., Liu X., Li L., Zhang F., Hu M., Ren F., Chi L, & Linhardt R.J. (2016). GlycCompSoft: Software for Automated Comparison of Low Molecular Weight Heparins Using Top-Down LC/MS Data. PLoS ONE, 11(12), e0167727.

Publication 2016

Lovenox Clexane HILIC column Agilent 1200 autosampler LTQ-Orbitrap XL FT-MS

Chromatography Clexane Enoxaparin Generic Hydrophilic interaction Lovenox Mass spectrometry

Corresponding Organization : Hefei University of Technology

Other organizations : Shandong University, New York State Department of Health, Wadsworth Center, Tokushima University

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Variable analysis

independent variables

Lovenox® and Clexane®, the innovator versions of enoxaparin
Generic versions of Lovenox® from three different manufacturers

dependent variables

Enoxaparin intact chain compositions analyzed under the negative mode

control variables

Enoxaparin injections diluted into 1 μg/μL and directly injected into a HILIC column (2.0 mm × 150 mm, 200 Å, Phenomenex, Torrance, CA) by an Agilent 1200 autosampler
The LC column was directly connected online to the standard ESI source of LTQ-Orbitrap XL FT-MS (Thermo Fisher Scientific, San-Jose, CA)

controls

No positive or negative controls were explicitly mentioned.

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