Heparins are linear sulfated polysaccharides widely used as anticoagulant drugs. Their nonreducing-end (NRE) has been little investigated due to challenges in their characterization, but is known to be partly generated by enzymatic cleavage with heparanases, resulting in N-sulfated glucosamines at the NRE. Uronic NRE (specifically glucuronic acids) have been isolated from porcine heparin, with GlcA-GlcNS,3S,6S identified as a porcine-specific NRE marker. To further characterize NRE in heparinoids, a building block analysis involving exhaustive heparinase digestion and subsequent reductive amination with sulfanilic acid was performed. This study describes a new method for identifying heparin classical building blocks and novel NRE building blocks using strong anion exchange chromatography on AS11 columns for the assay, and ion-pair liquid chromatography-mass spectrometry for building block identification. Porcine, ovine, and bovine intestine heparins were analyzed. Generally, NRE on these three heparins are highly sulfated moieties, particularly with 3-O sulfates, and the observed composition of the NRE is highly dependent on heparin origin. At the highest level of specificity, the isolated marker was only detected in porcine heparin. However, the proportion of glucosamines in the NRE and the proportion of glucuronic/iduronic configurations in the NRE uronic moieties greatly varied between heparin types.

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来自不同动物来源的肝素中的特定非还原性末端：使用磺胺酸还原胺化标记的构建块分析

肝素是广泛用作抗凝药物的线性硫酸化多糖。由于表征方面的挑战，它们的非还原末端 (NRE) 很少被研究，但已知部分是由乙酰肝素酶的酶促裂解产生的，从而在 NRE 处产生 N-硫酸化葡糖胺。已从猪肝素中分离出糖醛 NRE（特别是葡萄糖醛酸），其中 GlcA-GlcNS、3S、6S 被鉴定为猪特异性 NRE 标记物。为了进一步表征类肝素中的 NRE，进行了涉及彻底肝素酶消化和随后用磺胺酸还原胺化的构建块分析。本研究描述了一种使用 AS11 柱上的强阴离子交换色谱来鉴定肝素经典构件和新型 NRE 构件的新方法。和离子对液相色谱-质谱法用于构建块鉴定。分析了猪、绵羊和牛肠肝素。通常，这三种肝素上的 NRE 是高度硫酸化的部分，尤其是 3-O 硫酸盐，并且观察到的 NRE 组成高度依赖于肝素来源。在最高水平的特异性下，仅在猪肝素中检测到分离的标记物。然而，NRE 中葡糖胺的比例和 NRE 糖醛酸部分中葡糖醛酸/艾杜糖醛酸构型的比例在肝素类型之间差异很大。并且观察到的 NRE 的组成高度依赖于肝素来源。在最高水平的特异性下，仅在猪肝素中检测到分离的标记物。然而，NRE 中葡糖胺的比例和 NRE 糖醛酸部分中葡糖醛酸/艾杜糖醛酸构型的比例在肝素类型之间差异很大。并且观察到的 NRE 的组成高度依赖于肝素来源。在最高水平的特异性下，仅在猪肝素中检测到分离的标记物。然而，NRE 中葡糖胺的比例和 NRE 糖醛酸部分中葡糖醛酸/艾杜糖醛酸构型的比例在肝素类型之间差异很大。