Separation of a complex mixture of heparin-derived oligosaccharides using reversed-phase high-performance liquid chromatography

Thanawiroon, Charuwan; Linhardt, Robert J.

doi:10.1016/s0021-9673(03)00779-9

Cited by 70 publications

(55 citation statements)

References 25 publications

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“…Extensive experiments were undertaken to optimize the HPLC separation of both small (dp2-dp6) and large (dp10 -dp14) heparin-derived oligosaccharides (48). RPIP-HPLC used an MS-friendly mobile phase including: volatile ion-pairing reagents, tributylamine; volatile inorganic salt, ammonium acetate; and an organic modifier, acetonitrile.…”

Section: Resultsmentioning

confidence: 99%

Liquid Chromatography/Mass Spectrometry Sequencing Approach for Highly Sulfated Heparin-derived Oligosaccharides

Thanawiroon

Rice

Toida

et al. 2004

Journal of Biological Chemistry

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146

152

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Liquid chromatography/mass spectrometry (LC/MS)is applied to the analysis of complex mixtures of oligosaccharides obtained through the controlled, heparinase-catalyzed depolymerization of heparin. Reversedphase ion-pairing chromatography, utilizing a volatile mobile phase, results in the high resolution separation of highly sulfated, heparin-derived oligosaccharides. Simultaneous detection by UV absorbance and electrospray ionization-mass spectrometry (ESI-MS) provides important structural information on the oligosaccharide components of this mixture. Highly sensitive and easily interpretable spectra were obtained through post-column addition of tributylamine in acetonitrile. High resolution mass spectrometry afforded elemental composition of many known and previously unknown heparin-derived oligosaccharides. UV in combination with MS detection led to the identification of oligosaccharides arising from the original non-reducing end (NRE) of the heparin chain. The structural identification of these oligosaccharides provided sequence from a reading frame that begins at the non-reducing terminus of the heparin chain. Interestingly, 16 NRE oligosaccharides are observed, having both an even and an odd number of saccharide residues, most of which are not predicted based on biosynthesis or known pathways of heparin catabolism. Quantification of these NRE oligosaccharides afforded a number-averaged molecular weight consistent with that expected for the pharmaceutical heparin used in this analysis. Molecular ions could be assigned for oligosaccharides as large as a tetradecasaccharide, having a mass of 4625 Da and a net charge of ؊32. Furthermore, MS detection was demonstrated for oligosaccharides with up to 30 saccharide units having a mass of >10,000 Da and a net charge of ؊60.The structural elucidation of complex carbohydrates remains one of the most difficult challenges for chemists, often requiring the application of multiple analytical approaches (1-5). Glycosaminoglycans (GAGs), 1 and heparin in particular, have proven to be extremely difficult to analyze because of high negative charge, polydispersity, and sequence heterogeneity (6, 7). Heparin and low molecular weight heparins, prepared through the controlled chemical or enzymatic fragmentation of heparin (8), are widely used as clinical anticoagulants. Despite their medical importance, these drugs are relatively uncharacterized in terms of their chemical structure. Moreover, heparin and the structurally related heparan sulfate exhibit many additional biological activities, making them of great interest in new drug discovery (9). Numerous challenges can arise from the structural investigation of biologically active heparin oligosaccharides, particularly those in recognition systems involving specific protein-carbohydrate interactions (10, 11). Such biologically important oligosaccharides often contain rare sequences (12, 13) and are present only in minute, often picomole quantities. New derivatization methods (14, 15), chromatography (15, 16), electrophoresis-ba...

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Section: Resultsmentioning

confidence: 99%

Liquid Chromatography/Mass Spectrometry Sequencing Approach for Highly Sulfated Heparin-derived Oligosaccharides

Thanawiroon

Rice

Toida

et al. 2004

Journal of Biological Chemistry

Self Cite

146

152

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“…In general, HPLC can be used to analyze almost all the compounds in the herbal medicines. Thus, over the past decades, HPLC has received the most extensive application in the analysis of herbal medicines [63][64][65][66][67][68][69][70][71][72][73][74][75][76]. Reversed-phase (RP) columns may be the most popular columns used in the analytical separation of herbal medicines.…”

Section: High-performance Liquid Chromatographymentioning

confidence: 99%

Quality control of herbal medicines

Liang¹,

Xie²,

Chan³

2004

Journal of Chromatography B

500

401

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Different chromatographic and electrophoretic techniques commonly used in the instrumental inspection of herbal medicines (HM) are first comprehensively reviewed. Chemical fingerprints obtained by chromatographic and electrophoretic techniques, especially by hyphenated chromatographies, are strongly recommended for the purpose of quality control of herbal medicines, since they might represent appropriately the "chemical integrities" of the herbal medicines and therefore be used for authentication and identification of the herbal products. Based on the conception of phytoequivalence, the chromatographic fingerprints of herbal medicines could be utilized for addressing the problem of quality control of herbal medicines. Several novel chemometric methods for evaluating the fingerprints of herbal products, such as the method based on information theory, similarity estimation, chemical pattern recognition, spectral correlative chromatogram (SCC), multivariate resolution, etc. are discussed in detail with examples, which showed that the combination of chromatographic fingerprints of herbal medicines and the chemometric evaluation might be a powerful tool for quality control of herbal products.

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“…A técnica de CLAE apresenta-se com relativa facilidade de manipulação e permite ampla análise de diferentes compostos. A análise por CLAE deve considerar a necessidade de condições ótimas de separação, o que depende da fase móvel utilizada, pH, pressão da bomba, preparo da amostra etc (Sanyal et al, 2003;Thanawiroon & Linhardt, 2003;Liang et al, 2004).…”

Section: Procedimentosunclassified

O Brasil no contexto de controle de qualidade de plantas medicinais

Souza-Moreira¹,

Salgado²,

Pietro³

2010

Rev. bras. farmacogn.

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RESUMO:Atualmente, as plantas medicinais movem altos valores financeiros em todo o mundo e representam o tipo de tratamento mais acessível para cerca de 80% da população, principalmente entre os países em desenvolvimento. Entretanto, existe ainda uma falta de conhecimento sobre propriedades químicas, farmacológicas e toxicológicas a fim de assegurar a eficácia e segurança das plantas medicinais. Os critérios de eficácia e segurança de plantas medicinais estão relacionados a qualidade, isto é, as plantas necessitam ser corretamente identificadas, cultivadas e coletadas, devem estar livres de material estranho, partes de outras plantas e contaminações inorgânicas e/ ou microbianas. O objetivo deste estudo foi o de elucidar os diferentes processos e padronizações sobre o controle de drogas vegetais, principalmente no Brasil.Unitermos: Plantas medicinais, fitoquímica, controle de qualidade.ABSTRACT: "Brazil in the context of plants and derivates quality control". Nowadays medicinal plants move high financial values at the world and represent a type of treatment more accessible for about 80% of people, mainly among developing countries. However, there is a lack of knowledge about chemical, pharmacological and toxicological properties towards ensure their efficacy and safety. Efficacy and safety criteria of medicinal plants are related to quality, that is, the plants need to be correctly identified, cultivated and harvested, they must be free of strange material, part of other plants and inorganic and/or microorganisms contamination. The aim of this study was to elucidate different process and standardizations about quality control of vegetal drugs mainly in Brazil. ISSN 0102-695X Keywords INTRODUÇÃOPor longos períodos na história, o uso de plantas medicinais foi presente não só devido ao seu caráter alimentar, mas também às suas propriedades de cura, sejam elas reais ou ritualísticas. Entretanto, até a metade do século XX, o uso de medicamentos de origem sintética tornou-se mais amplo que o uso de plantas medicinais, principalmente na sociedade ocidental, com destaque para os países desenvolvidos visto que os considerados países em desenvolvimento permaneceram fazendo uso das propriedades curativas das plantas, certamente devido às difíceis condições econômicas de aquisição dos medicamentos sintéticos

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Separation of a complex mixture of heparin-derived oligosaccharides using reversed-phase high-performance liquid chromatography

Cited by 70 publications

References 25 publications

Liquid Chromatography/Mass Spectrometry Sequencing Approach for Highly Sulfated Heparin-derived Oligosaccharides

Liquid Chromatography/Mass Spectrometry Sequencing Approach for Highly Sulfated Heparin-derived Oligosaccharides

Quality control of herbal medicines

O Brasil no contexto de controle de qualidade de plantas medicinais

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