高效液相色谱法测定白细胞udp半乳糖和udp葡萄糖浓度。

Enzyme & protein Pub Date : 1993-01-01 DOI:10.1159/000468666
M J Palmieri, R A Reynolds, J B Gibson, G T Berry, S Segal
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引用次数: 2

摘要

采用高效液相色谱法分离和定量人白细胞(wbc)中udp半乳糖(UDPGal)和udp葡萄糖(UDPGlu)。三氯乙酸处理的无蛋白滤液在阴离子交换柱(Dionex CarboPac PA-1)上使用梯度为20-40%的磷酸钾缓冲液(pH 4.5)进行层析。UDPGal和UDPGlu的回收率为93% ~ 106%,该方法在较宽的WBC蛋白浓度范围内呈线性。低至2.5 ml (2.2 mg白细胞蛋白)的血容量可用于糖核苷酸的定量回收。33例年龄在1 ~ 65岁的正常人中,UDPGal和UDPGlu的平均值和标准差分别为12.4 +/- 4.2和31.5 +/- 9.3 μ mol/100 g蛋白。UDPGal和UDPGlu值在儿童和成人之间无统计学差异。UDPGal和UDPGlu的浓度与WBC总数或库尔特计数器分析获得的淋巴细胞数量之间没有相关性。从同一血样中制备的白细胞和红细胞中UDPGal和UDPGlu的浓度无相关性。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
Concentration of white blood cell UDPgalactose and UDPglucose determined by high performance liquid chromatography.

We have applied a HPLC method to separate and quantitate UDPgalactose (UDPGal) and UDPglucose (UDPGlu) in human white blood cells (WBCs). Trichloroacetic acid-treated, protein-free filtrates were chromatographed on an anion-exchange column (Dionex CarboPac PA-1) using a gradient of 20-40% potassium phosphate buffer (pH 4.5). Recoveries of UDPGal and UDPGlu ranged from 93 to 106%, and the method was linear over a wide range of WBC protein concentrations. Volumes of blood as low as 2.5 ml (2.2 mg WBC protein) could be used to achieve quantitative recovery of the sugar nucleotides. The mean values and standard deviations of UDPGal and UDPGlu in 33 normal individuals ranging in age from 1 day to 65 years were 12.4 +/- 4.2 and 31.5 +/- 9.3 mumol/100 g protein, respectively. No statistical differences in UDPGal and UDPGlu values were observed between children and adults. No correlation was established between the concentrations of UDPGal and UDPGlu and either total WBC count or the number of lymphocytes obtained from Coulter counter analysis. There was no relationship between the concentrations of UDPGal and UDPGlu in WBCs and RBCs which were prepared from the same blood specimen.

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