Analysis of sialyllactoses in blood and urine by high-performance liquid chromatography
Analysis of sialyllactoses in blood and urine by high-performance liquid chromatography
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DOI：
10.1006/abio.1998.3021
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发表时间：
1999-04-10
影响因子：
2.9
通讯作者：
Zopf, D
中科院分区：
文献类型：
--
作者：
Fu, DT;Zopf, D
建立了一种高灵敏度、高选择性的高效液相色谱（HPLC）分析生物体液中寡糖的方法。在这种方法中，生物流体样品，如血清或尿液，通过10，000分子量截留过滤器滤芯过滤，以去除大分子，如蛋白质和脂质。然后如前所述用1-苯基-3-甲基-5-吡唑啉酮（PMP）将滤液中的碳水化合物衍生化[Anal. 180，351-357，（1989）]。通过反相HPLC分离衍生的碳水化合物，并通过245 nm处的UV吸光度监测。碳水化合物的定量分析可以基于其相对于标准校准曲线的积分值来实现。由于中性和酸性碳水化合物可以通过使用Dowex 1-X8阴离子交换树脂分离，因此该方法可以专门用于分析生物流体中的中性、酸性和总碳水化合物。由于PMP特异性地与还原性醛糖反应，因此基本上消除了生物体液中存在的非碳水化合物组分的干扰。该方法已被证明是高度灵敏的，需要低至5 pmol的分析物进行可靠的分析。它也已成功地用于人体血液和尿液样品中碳水化合物药物的药代动力学分析。(C)北京：科学出版社.
A sensitive and highly selective high-performance liquid chromatography (HPLC)-based method has been developed for the analysis of oligosaccharides in biological fluids. In this method, a sample of biological fluid, such as blood serum or urine, is filtered through a 10,000 molecular weight cutoff filter cartridge to remove large molecules such as proteins and lipids. The carbohydrates in the filtrate are then derivatized with 1-phenyl-3-methyl-5-pyrazolone (PMP) as described previously [Anal. Biochem. 180, 351-357, (1989)]. The derivatized carbohydrates are separated by reverse-phase HPLC and monitored by UV absorbance at 245 nm. Quantitative analysis of the carbohydrates can be achieved based on their integration values relative to a standard calibration curve. Since neutral and acidic carbohydrates can be separated by using Dowex 1-X8 anion exchange resin, this method can be used specifically to analyze neutral, acidic, and total carbohydrates in the biological fluids. Because PMP specifically reacts with reducing aldoses, interference from noncarbohydrate components present in the biological fluids is essentially eliminated. This method has proven to be highly sensitive, requiring as little as 5 pmol of analyte for reliable analysis. It has also been used successfully for pharmacokinetic analysis of carbohydrate drugs in human blood and urine samples. (C) 1999 Academic Press.