Determination of dermatan sulfate and chondroitin sulfate as related substances in heparin by capillary electrophoresis

Bendazzoli, Claudia; Liverani, Lino; Spelta, Franco; Prandi, Massimo; Fiori, Jessica; Gotti, Roberto

doi:10.1016/j.jpba.2010.07.006

Cited by 11 publications

(10 citation statements)

References 20 publications

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“…Some heparin oligosaccharides have been derivatized to incorporate fluorescent tags by reductive amination to improve CE detection [ 31 ]. However, the structural elucidation of heparins may still be problematic due to the high charges and the possible occurrence of numerous isomers such as alternate disaccharide sequences of GlcA and GlcN residues with further O -sulfation, N -sulfation, and N -acetylation [ 32 ].…”

Section: Resultsmentioning

confidence: 99%

Application of 2,3-Naphthalenediamine in Labeling Natural Carbohydrates for Capillary Electrophoresis

et al. 2012

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Neutral and acidic monosaccharide components in Ganoderma lucidum polysaccharide are readily labeled with 2,3-naphthalenediamine, and the resulting saccharide-naphthimidazole (NAIM) derivatives are quantified by capillary electrophoresis (CE) in borate buffer. Using sulfated-α-cyclodextrin as the chiral selector, enantiomers of monosaccharide-NAIMs are resolved on CE in phosphate buffer, allowing a simultaneous determination of the absolute configuration and sugar composition in the mucilage polysaccharide of a medicinal herb Dendrobium huoshanense. Together with the specific enzymatic reactions of various glycoside hydrolases on the NAIM derivatives of glycans, the structures of natural glycans can be deduced from the digestion products identified by CE analysis. Though heparin dissachrides could be successfully derived with the NAIM-labeling method, the heparin derivatives with the same degree of sulfation could not be separated by CE.

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Section: Resultsmentioning

confidence: 99%

Application of 2,3-Naphthalenediamine in Labeling Natural Carbohydrates for Capillary Electrophoresis

et al. 2012

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“…The voltage of -30 kV was chosen for determinations, because it gave rapid migration and good peak shapes and also acceptable separation of degradation products. The reported pH for determination of heparin was between 2.00 to 5.00 (21)(22)(23) and the best separation was achieved at pH of 3.50. The effect of electrolyte concentration was investigated and the results showed that by the enhancement in the concentration, the resolution and the separation were increased, but in concentration of 100 mM peak shapes were degraded extremely, therefore the electrolyte with morality of 72 mM was applied for analyses, also in this concentration force degradation products could be completely separated at 257 nm.…”

Section: Optimization Of the Ce Methodsmentioning

confidence: 99%

“…In the other work, a CE method was used for determination of intact heparin in infusion solutions and plasma with contactless conductivity detection [20]. CE was also applied for determination of impurities, contaminants and related compounds of heparin, for instance; Wielgos and coworkers [21] reported a CE method for determination of impurities, Volpi et al [22] developed a CE method for determination of related substances, Bendazzoli et al [23] presented a CE method for determination of a contaminant in heparin preparations and Liu and co workers [20] developed a rapid and simple CE method for simultaneously separation of heparin, chondroitin sulfate and hyaluronic acid in pharmaceutical preparations and synovial fluid.…”

Section: Introductionmentioning

confidence: 99%

Development and Validation of a Simple and Rapid Capillary Zone Electrophoresis Method for Quantification of Heparin in a Pharmaceutical Product and Stability Studies

Tamizi¹,

Jouyban²

2012

Pharm Anal Acta

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A simple and rapid capillary zone electrophoresis method has been developed and validated for determination of heparin in a pharmaceutical product and evaluation of heparin stability under different stress conditions. The best separation was achieved by a bare fused silica capillary (50 μm i.d.; 50 cm total and 41.5 cm effective length), phosphate buffer (pH=3.50, 72 mM) at 35°C, hydrodynamic injection at 50 mbar for 40 seconds and applied voltage of-30 kV. Heparin and force degradation products were detected by a photo diode array detector at 200 nm and 257 nm, respectively. The proposed method was validated in terms of linearity, accuracy, precision, limit of quantification (LOQ) and limit of detection (LOD). For evaluation of heparin stability, heparin solutions were subjected to thermal (90 ± 1°C), acidic (pH=2.00, 70 ± 1°C) and basic (pH=12.00, 70 ± 1°C) stress conditions, also sun light exposure (in the lab). The results indicated that, the method was linear in the range of 0.312 to 15.0 mg/ml with LOD of 0.078 mg/ ml and LOQ of 0.312 mg/ml, accurate (between 97.27% and 101.0%) and precise (intra-day precision of 0.28 to 1.8 and inter-day precision of 0.78 to 3.2%). The migration time of heparin under optimized conditions, was 2.39 ± 0.03 minutes and force degradation products were separated within 7 minutes. Heparin content in the pharmaceutical product quantified by the method was 99.58 ± 0.70% of the label claim. Meanwhile the method could show the changes in the heparin electropherogram, also formation of degradation products under different stress conditions. Therefore the proposed method could be applied as a fast and suitable technique for determination of heparin in pharmaceutical dosage forms and stability studies.

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“…Thus, much lower sample concentrations were required in this method as compared to published HPLC and other CE assays. A method for the analysis of chondroitin sulfate and dermatan sulfate as impurities of heparin based on the digestion with chondroitinase enzymes followed by CE analysis with detection limits as low as 0.01% has also been published . It is worth to note that genotoxic impurities are typically less often determined by CE, since here particularly sensitive techniques such as GC and LC combined with capable MS are required .…”

Section: Separation and Analysis Of Small Molecules And Nonbiotechnolmentioning

confidence: 99%

Recent advances in capillary electrophoretic migration techniques for pharmaceutical analysis

et al. 2014

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Since the introduction about 30 years ago, CE techniques have gained a significant impact in pharmaceutical analysis. The present review covers recent advances and applications of CE for the analysis of pharmaceuticals. Both small molecules and biomolecules such as proteins are considered. The applications range from the determination of drug-related substances to the analysis of counterions and the determination of physicochemical parameters. Furthermore, general considerations of CE methods in pharmaceutical analysis are described.

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Determination of dermatan sulfate and chondroitin sulfate as related substances in heparin by capillary electrophoresis

Cited by 11 publications

References 20 publications

Application of 2,3-Naphthalenediamine in Labeling Natural Carbohydrates for Capillary Electrophoresis

Application of 2,3-Naphthalenediamine in Labeling Natural Carbohydrates for Capillary Electrophoresis

Development and Validation of a Simple and Rapid Capillary Zone Electrophoresis Method for Quantification of Heparin in a Pharmaceutical Product and Stability Studies

Recent advances in capillary electrophoretic migration techniques for pharmaceutical analysis

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