Estimation of the composition of heparin mixtures from various origins using proton nuclear magnetic resonance and multivariate calibration methods

Ruiz-Calero, Victoria; Saurina, Javier; Galcerán, T.; Hernández-Cassou, Santiago; Puignou, L.

doi:10.1007/s00216-002-1315-x

Cited by 21 publications

(10 citation statements)

References 28 publications

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“…11,24 Regulatory concerns about the substitution of one heparin for another or the blending of heparins coming from two different organisms or tissues have lead to the development of assays to assess the provenance of a given heparin. 6,[24][25][26][27][28][29] Crude porcine heparins (the unbleached intermediate used to prepare heparin active pharmaceutical ingredient (API)) are currently assessed by quantitative PCR qPCR, which can sensitively detect the presence of ruminant DNA, suggesting blending of porcine heparin with ovine or bovine heparin. 6,[25][26][27] When using this qPCR method, heparinase is required to degrade the heparin present in the sample to ensure accurate detection, as the heparin-mediated inhibition of PCR was previously described.…”

Section: Introductionmentioning

confidence: 99%

Stable Isotopic Analysis of Porcine, Bovine, and Ovine Heparins

Jasper

Zhang

Poe³

et al. 2015

Journal of Pharmaceutical Sciences

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The assessment of provenance of heparin is becoming a major concern for the pharmaceutical industry and its regulatory bodies. Batch-specific [carbon (δ13C), nitrogen (δ15N), oxygen (δ18O), sulfur (δ34S), and hydrogen (δD)] stable-isotopic compositions of five different animal-derived heparins were performed. Measurements readily allowed their differentiation into groups and/or subgroups based on their isotopic provenance. Principle component analysis showed that a bivariate plot of δ13C and δ18O is the best single, bivariate plot that results in the maximum discrimination ability when only two stable isotopes are used to describe the variation in the data set. Stable-isotopic analyses revealed that (i) stable-isotope measurements on these highly-sulfated polysaccharide (MW ~15 kDa) natural products (“biologics”) were feasible; (ii) in bivariate plots, the δ13C versus δ18O plot reveals a well-defined relationship for source differentiation of hogs raised in the US from hogs raised in Europe and China; (iii) the δD versus δ18O plot revealed the most well-defined relationship for source differentiation based on the hydrologic-environmental isotopes of water (D/H and 18O/16O), and (iv) the δ15N versus δ18O and δ34S versus δ18O relationships are both very similar, possibly reflecting the food sources used by the different heparin producers.

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Section: Introductionmentioning

confidence: 99%

Stable Isotopic Analysis of Porcine, Bovine, and Ovine Heparins

Jasper

Zhang

Poe³

et al. 2015

Journal of Pharmaceutical Sciences

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“…Structural characterization of heparin and heparan sulfate has been aided by the development of more efficient separation techniques and the use of spectroscopic methods such as NMR, − which can also provide information about degree and patterns of sulfation. However, NMR requires large (milligram) amounts of sample material.…”

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confidence: 99%

Compositional Analysis and Quantification of Heparin and Heparan Sulfate by Electrospray Ionization Ion Trap Mass Spectrometry

2003

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A new method using a combination of electrospray ionization mass spectrometry (ESI-MS) and tandem mass spectrometry (MSn) was developed for the identification and quantitative analysis of eight heparan sulfate (HS)- and heparin-derived delta-disaccharides obtained by enzymatic depolymerization. The compositional analysis of nonisomeric disaccharide constituents of heparin/HS was achieved from full-scan MS1 spectra using an internal standard and a calculated response factor for each disaccharide. Diagnostic product ions from MSn spectra of isomeric disaccharides were used for the quantitative analysis of the relative amounts of each of the isomers in mixtures. The protocol was validated using several quality control samples and showed satisfactory accuracy and precision. The analysis is rapid, accurate, and uses no purification or separation steps prior to analysis by MS, thus reducing sample consumption and analysis time of traditional methods. Using this quantitative analysis procedure, percentages of disaccharide compositions for heparins from porcine and bovine intestinal mucosa and heparan sulfate from bovine kidney were determined.

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“…The main differences between porcine and bovine heparin have been previously described on the basis of data obtained by HPLC, MS, 14 or NMR. 23,24 The intra-species compared with inter-species variability for porcine, bovine, and ovine heparin, and the percentage of contamination that can be detected was investigated. SAX-HPLC is the simplest method as it only requires conventional HPLC equipment and provides very good resolution.…”

Section: Discussionmentioning

confidence: 99%

Quantitative PCR and Disaccharide Profiling to Characterize the Animal Origin of Low-Molecular-Weight Heparins

Houiste

Auguste

Macrez

et al. 2007

Clin Appl Thromb Hemost

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Low-molecular-weight heparins (LMWHs) are widely used in the management of thrombosis and acute coronary syndromes. They are obtained by the enzymatic or chemical depolymerization of porcine intestinal heparin. Enoxaparin sodium, a widely used LMWH, has a unique and reproducible oligosaccharide profile which is determined by the origin of the starting material and a tightly controlled manufacturing process. Although other enoxaparin-like LMWHs do exist, specific release criteria including the origin of the crude heparin utilized for their production, have not been established. A quantitative polymerase chain reaction method has been developed to ensure the purity of the porcine origin of crude heparin, with a DNA detection limit as low as 1 ppm for bovine, or 10 ppm for ovine contaminants. This method is routinely used as the release acceptance criterion during enoxaparin sodium manufacturing. Furthermore, when the process removes DNA, other analytical techniques can be used to assess any contamination. Disaccharide profiling after exhaustive depolymerization can determine the presence of at least 10% bovine or 20% ovine material; multivariate analysis is useful to perform the data analysis. Consistent with the availability of newer technology, these methods should be required as acceptance criteria for crude heparins used in the manufacture of LMWHs to ensure their safety, quality, and immunologic profile.

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Estimation of the composition of heparin mixtures from various origins using proton nuclear magnetic resonance and multivariate calibration methods

Cited by 21 publications

References 28 publications

Stable Isotopic Analysis of Porcine, Bovine, and Ovine Heparins

Stable Isotopic Analysis of Porcine, Bovine, and Ovine Heparins

Compositional Analysis and Quantification of Heparin and Heparan Sulfate by Electrospray Ionization Ion Trap Mass Spectrometry

Quantitative PCR and Disaccharide Profiling to Characterize the Animal Origin of Low-Molecular-Weight Heparins

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