Quantitative determination of disaccharide content in digested unfragmented heparin and low molecular weight heparin by direct-infusion electrospray mass spectrometry

Characterization of currently marketed heparin products: Analysis of heparin digests by RPIP-UHPLC–QTOF-MS

“…This work Sigma, Iduron II, III 5 mIU each for 60 g, 37 • C overnight Brustkern et al [26] Sigma I, III 10 mIU each for 40 g, 37 • C overnight Korir et al [17] Sigma I, II, III 10 mIU each for 20 g, 37 • C 17 h Zhang et al [18] Home-made I, II, III 10 mIU each for 5 g, 37 • C 10 h Galeotti and Volpi [19] Seikagaku I, II, III 1 IU each for 100 g, 37 • C 8 h Saad and Leary [20] Calbiochem I, II, III 10 mIU each for 20 g, 37 • C overnight Saad et al [21] Sigma, Calbiochem I, II, III 10 mIU each for 100 g, 37 • C 16 h Kinoshita and Sugahara [22] Seikagaku I, II, III 1.7 mIU-I and 0.85 mIU-III a for 1 g, 37 • C 30 min Camara et al [23] Sigma I, II, III 10 mIU each for 10 g, 37 • C 24 h Ruiz-Calero et al [24] Sigma I, II, III 13 mIU-I, 1.7 mIU-II, 1.5 mIU-III for 100 g, 37 • C 24 h Militsopoulou et al [25] Sigma, Seikagaku I, II, III 300 mIU each for 1 mg, 37 • C 90 min a Heparin lyase III contains a small proportion of lyase II. of 20 mM ammonium acetate and 2 mM calcium acetate (pH 7.4) and subjected to enzymatic digestion by adding different combinations of heparin lyase I, II or III dissolved in same buffer.…”

“…Clearly, it is difficult to establish absolute values for the heparin disaccharide composition across other literature studies [17][18][19][20][21][22][23][24][25][26] (Table 3) because of the variation in heparin source, instrumentation and experimental protocol used in the digestions. However, because measurements were performed on the same lots of heparin sodium with two different sample preparation protocols and instrument systems data from this work and the work of Brustkern et al can be compared and contrasted more directly than other literature reports.…”

“…The catalytic mechanism and substrate specificities of these heparin lyases have been studied [11][12][13][14][15]. For most heparin depolymerization studies, heparin lyase I, II and III have been used in combination to achieve complete digestion [16][17][18][19][20][21][22][23][24][25].…”

“…In Kinoshita et al [22], Ruiz-Calero et al [24] and Militsopoulou et al [25] replicates were not specified. In Camara et al [23], average ± error values came from two measurements. b Based on average values of seven samples reported in [18].…”

Isolation and Characterization of Low Molecular Weight Glycosaminoglycans from Marine Mollusc Amussium pleuronectus (Linne) using Chromatography

“…Heparin and low molecular weight heparin (LMWH) are heterogeneous glycosaminoglycans (GAGs) prescribed as anticoagulants [1]. Heparin has molecular mass range between 3,000 and 40,000, which is biosynthesized as a proteoglycan consisting of a small core protein to which polysaccharide side chains are attached.…”