Quantitative analysis of plasma neutral glycosphingolipids by high performance liquid chromatography of their perbenzoyl derivatives.
Quantitative analysis of plasma neutral glycosphingolipids by high performance liquid chromatography of their perbenzoyl derivatives.
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通过高效液相色谱对其过苯甲酰衍生物进行血浆中性糖鞘脂的定量分析。
DOI：
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发表时间：
1977
影响因子：
6.5
通讯作者：
R. McCluer
中科院分区：
文献类型：
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作者：
M. David;Ullman;R. McCluer
建立了中性糖基神经酰胺及其过氧苯甲酰衍生物的高效液相色谱定量分析方法。在37 ℃下，用10%苯甲酰氯的吡啶溶液将单-、二-、三-和四-糖基神经酰胺中每一种均超过2.5 nmol的样品苯甲酰化16小时。通过溶剂分配将产物与过量试剂分离，并注入到薄膜硅胶（Zipax）柱（2.1 mm × 50 cm）上。用2 - 17%乙酸乙酯的己烷溶液以2ml/min的10分钟线性梯度洗脱衍生物，并记录280 nm处的吸光度。检测器响应与所用样品的重量（2 - 30 nmol）成比例，检测下限为约70 pmol。该方法已用于红细胞和血浆糖脂的定量分析。可使用少至0.5 ml的血浆进行分析。重复分析的相对标准偏差范围为2.0%（葡萄糖基神经酰胺）至5.4%（半乳糖基神经酰胺）。
A quantitative high performance liquid chromatography method for the analysis of neutral glycosylceramides as their perbenzoyl derivatives has been devised. Samples containing more than 2.5 nmol each of mono-, di-, tri-, and tetraglycosylceramide are benzoylated with 10% benzoyl chloride in pyridine at 37degrees C for 16 hr. The products are separated from excess reagents by solvent distribution and injected onto a pellicllar silica gel (Zipax) column (2.1 mm X 50 cm). The derivatives are eluted with a 10 min linear gradient of 2-17% ethyl acetate in hexane at 2 ml/min and absorbance at 280 nm is recorded. The detector response was proportional to the weight of sample used (2-30 nmol) and the lower limit of detection was about 70 pmol. The procedure has been applied to the quantitative analysis of erythrocyte and plasma glycolipids. As little as 0.5 ml of plasma can be used for analysis. The relative standard deviation of repetitive analyses ranged between 2.0% for glucosylceramide to 5.4% for galactosyllactosylceramide.