Quantitative determination of the glycosaminoglycan Δ-disaccharide composition of serum, platelets and granulocytes by reversed-phase high-performance liquid chromatography
Quantitative determination of the glycosaminoglycan Δ-disaccharide composition of serum, platelets and granulocytes by reversed-phase high-performance liquid chromatography
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DOI：
10.1016/j.chroma.2008.06.007
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发表时间：
2008-08-01
影响因子：
4.1
通讯作者：
Goetting, Christian
中科院分区：
文献类型：
--
作者：
Ambrosius, Michael;Kleesiek, Knut;Goetting, Christian
用荧光团2-氨基吖啶酮（AMAQ）衍生来自硫酸乙酰肝素/肝素（HS/H）的七种δ-二糖标准品和来自软骨素/硫酸皮肤素（CS/DS）和透明质酸（HA）的九种A-二糖标准品，并通过反相HPLC在两次运行中分离，每次运行具有基线分离和非常短的运行时间。这种新的方法有利于从CS/DS/HA和HS/H分离最大数量的δ-二糖，在一个柱和缓冲系统后，荧光标记在目前的两个运行。第一次从CS/DS/HA中分离出9种糖胺聚糖（GAG）δ-二糖，荧光团标记后一次运行。CS/DS/HA和HS/H Delta-二糖的定量限（LOQ）低于0.2 pmol。我们证明了我们的方法对生物样品的适用性。此外，第一次测定了血小板和粒细胞的GAG A-二糖组成的正常范围。(c)2008 Elsevier B. V.保留所有权利。
Seven Delta-disaccharicle standards from heparan sulfate/heparin (HS/H) and nine A-disaccharide standards from chondroitin/dermatan sulfate (CS/DS) and hyaluronic acid (HA) were derivatized with the fluorophore 2-aminoacridone (AMAQ and separated in two runs each by reversed-phase HPLC with baseline separation and very short run times. This novel method facilitates the separation of the largest number of Delta-disaccharides from both CS/DS/HA and HS/H with one column and buffer system after fluorophore labeling in two runs at present. For the first time nine glycosaminoglycan (GAG) Delta-disaccharides from CS/DS/HA were separated after fluorophore labeling in one run. The limits of quantification (LOQs) were below 0.2 pmol for CS/DS/HA and HS/H Delta-disaccharides. We demonstrated applicability of our method for biological samples. Furthermore, normal ranges of the GAG A-disaccharide compositions from platelets and granulocytes were determined for the first time. (c) 2008 Elsevier B.V. All rights reserved.