Qualification of HSQC methods for quantitative composition of heparin and low molecular weight heparins

Structural Characterization of the Low-Molecular-Weight Heparin Dalteparin by Combining Different Analytical Strategies

“…The mono/disaccharide percentage content of eleven different batches of Fragmin ® was determined by applying a recently validated method [ 19 ]. The HSQC spectrum reported in Figure 1 shows all major and minor signals used for integration.…”

“…Heparin/heparan sulfate fragment mapping by enzymatic digestion either with either heparinase I, II and III or using a mixture of them is a very common strategy reported in several studies and different analytical methods to run the depolymerized solutions and record interesting information in function to the technique used are often described [ 14 , 15 , 16 , 17 , 18 ]. Moreover, it was recently described that the quantitative NMR-HSQC analysis, applied to determine the mono- and disaccharide composition of heparin and LMWHs, can be used for detailed structural comparison studies between generic or biosimilar drugs with the reference product [ 19 ].…”

Multivariate analysis applied to complex biological medicines

“…While the 13 C-1 H HSQC NMR spectrum of heparin provides a great deal of information, the analysis performed here is mainly qualitative, although through the integration of a number of signals in the 13 C-1 H HSQC NMR spectra of heparin samples, the average disaccharide composition of the polysaccharide can be determined. 29,30 These approaches are much quicker than other traditional methods, such as digestion followed by HPLC or HPLC-MS, and require much less preparation time. Such approaches are highly valuable to the quality control of the heparin pharmaceutical product; the NMR experiment, spectral processing and subsequent multivariate analysis could all be performed within one working day, with the only barrier being the exchange of the sample into D 2 O.…”

Chondroitin, Dermatan, Heparan, and Keratan Sulfate: Structure and Functions

“…The peaks of a 2D-NMR experiment such as HSQC or DEPT-HSQC can be integrated, if specific constraints are satisfied, in order to furnish a quantitative estimation of the GAG sulfation pattern in terms of the relative proportion of disaccharide subunit in the polysaccharide (Guerrini et al 2005;Gargiulo et al 2009;Mauri et al 2017). It is worth noting that this is a nondestructive method for the quali-quantitative determination of sulfation pattern on GAG polysaccharide chains ( Fig.…”