Bryson Y J, Sweetman L, Connor J D
Antimicrob Agents Chemother. 1978 Dec;14(6):909-15. doi: 10.1128/AAC.14.6.909.
Previous methods using low- or high-pressure liquid chromatography and UV absorbance for quantitation of arabinosides in plasma can practically detect only >/=200 ng of adenine arabinoside and >/=100 ng of hypoxanthine arabinoside per ml, and they require expensive equipment and expert technical assistance. We describe in this report a simple quantitative microbioassay for arabinosides in human plasma based on their ability to inhibit the cytopathic effect of vaccinia virus in an adenosine deaminase-free cell culture system. Using prior separation of nucleosides in plasma by thin-layer chromatography, followed by the microbioassay, we quantitated adenine arabinoside with a sensitivity of approximately 4 ng/ml and hypoxanthine arabinoside at approximately 625 ng/ml. This assay method is simple, sensitive, and reproducible, requires small plasma samples, measures biological activity, and is adaptable to routine use. It is an important tool for evaluating the pharmacology of adenine arabinoside and Ara-AMP in patients in current clinical trials.
以往采用低压或高压液相色谱及紫外吸光度法对血浆中阿拉伯糖苷进行定量的方法，实际上每毫升只能检测到≥200 ng的阿糖腺苷和≥100 ng的阿糖次黄嘌呤，而且它们需要昂贵的设备和专业的技术支持。在本报告中，我们描述了一种基于人血浆中阿拉伯糖苷抑制无腺苷脱氨酶细胞培养系统中痘苗病毒细胞病变效应能力的简单定量微生物测定法。通过先利用薄层色谱法分离血浆中的核苷，然后进行微生物测定，我们定量测定阿糖腺苷的灵敏度约为4 ng/ml，阿糖次黄嘌呤的灵敏度约为625 ng/ml。该测定方法简单、灵敏、可重复，所需血浆样本量小，可测量生物活性，且适用于常规使用。它是评估当前临床试验中患者阿糖腺苷和阿糖腺苷单磷酸药理学的重要工具。
