A New Approach for Heparin Standardization: Combination of Scanning UV Spectroscopy, Nuclear Magnetic Resonance and Principal Component Analysis

Abstract

The year 2007 was marked by widespread adverse clinical responses to heparin use, leading to a global recall of potentially affected heparin batches in 2008. Several analytical methods have since been developed to detect impurities in heparin preparations; however, many are costly and dependent on instrumentation with only limited accessibility. A method based on a simple UV-scanning assay, combined with principal component analysis (PCA), was developed to detect impurities, such as glycosaminoglycans, other complex polysaccharides and aromatic compounds, in heparin preparations. Results were confirmed by NMR spectroscopy. This approach provides an additional, sensitive tool to determine heparin purity and safety, even when NMR spectroscopy failed, requiring only standard laboratory equipment and computing facilities.

Detection of a Non-animal Source of Glycosaminoglycans from Edible Mushrooms in Northern Thailand

“…Since DMMB dye reacts with all types of sulfated GAG chains, this assay was not able to distinguish the types of GAGs present in the mushroom extracts. Although our spectrophotometric analysis confirmed the presence of GAG chains at peaks around 190-210 nm (Lima et al, 2011), we also found additional broad peaks around 240-260 nm that were related to other aromatic compounds (de Micalizzi et al, 1998). Some ions, such as calcium or sodium, or the sulfation pattern of GAGs or other contaminants can shift the peaks (McEwen et al, 2009).…”

“…GAGs levels measured by the DMMB dye-binding assay from the top two mushroom extracts (log white fungi and bolete) were highly correlated with the highest absorption peak around 190-210 nm from spectrophotometric analysis; this tentatively represented the carboxylate chromophore of iduronate and glucuronate present in the GAGs chains (Lima et al, 2011), as shown in Figure 3 (L for log white fungi; and I for bolete). Although hygroscopic earthstar contained the highest levels of GAGs according to the DMMB dye-binding assay, its absorption (G) peak at around 190-210 nm was shifted toward the absorption peaks of the shitake mushroom (E), milk white russula (H), rosy russula (D), and sajor-caju mushroom (F) extracts.…”

“…The absorption pattern in the mushroom extracts resembled the chondroitin sulfate standard, with two peaks around 190-300 nm in most extracts. Since GAGs polymers are composed of either iduronic acid or glucuronic acid and N-acetyl chromophores (Kjellén and Lindahl, 1991;Varki et al, 2009), the highest peaks around 190-210 nm, arising from the carboxylate chromophore of iduronate and glucuronate present in the GAGs chains, were responsible for the majority of the absorption pattern (Lima et al, 2011).…”

Low molecular weight heparins: Structural differentiation by spectroscopic and multivariate approaches

“…Multivariate analysis was performed as previously described (Lima et al, 2011) using the software R: A Language and Environment for Statistical Computing (R Foundation for Statistical Computing, Viena, Austria. http://cran.r-project.org/), with prior mean centering.…”

Méretvariált magnetit nanorészecskék előállítása és felületmódosítása polianionos védőrétegekkel

“…Ennek az lehet az oka, hogy a természetes eredetű, marha légcsőből előállított CSA a kinyerési eljárás következtében szennyezésként cetil-piridínium-kloridot (CPC) tartalmaz (Garnjanagoonchorn et al, 2007;Lima et al, 2011), és ez adszorbeálódva a magnetit részecskéken azok egy részét hidrofobizálja, destabilizálja. UV méréssel tudtam igazolni a szennyező jelenlétét, ugyanis a CPC-nak jellemző elnyelése van 260 nm-nél (Lima et al, 2011;Roach et al, 2013;Song et al, 2007). Ez a csúcs megjelent a CSA oldatában mért UV spektrumon, azonban az adszorpció után az egyensúlyi felülúszó már nem tartalmazta ezt a csúcsot, még nagy feleslegben alkalmazott CSA esetében sem.…”