Determination of the diol content of chromatographic supports by capillary electrophoresis
Determination of the diol content of chromatographic supports by capillary electrophoresis
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DOI：
10.1016/s0021-9673(96)00742-x
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发表时间：
1997-01-17
影响因子：
4.1
通讯作者：
Hage, DS
中科院分区：
文献类型：
--
作者：
Chattopadhyay, A;Hage, DS
开发了一种毛细管电泳 (CE) 方法，用于测定高性能亲和或尺寸排阻色谱中使用的支持物的二醇含量。该方法涉及用高碘酸盐氧化二醇键合的载体，然后使用 CE 来分离和定量该反应产生的碘酸盐。在优化该测定时考虑了氧化和分离条件。最终方法是通过将已知量的载体与 20 倍过量的高碘酸盐在 pH 4.0、0.5 M 乙酸盐缓冲液中反应来进行，并使用均苯四酸作为内标。氧化5-10分钟后，过滤混合物并将滤液注射到57cmx50μm I.D.上。熔融石英毛细管在 25 kV 下运行，含有 pH 4.0、0.5 M 乙酸盐作为运行缓冲液。每次运行的总分离时间为 5 分钟，对于 1-ml 反应混合物注射 6-nl，检测限为 0.1 mM 碘酸盐（或 0.1 μmol 二醇基团）。通过改变所分析的支持物的量，该方法可用于多孔或无孔支持物。该技术与碘滴定法显示出良好的相关性，但需要更少的样品和时间来执行。
A capillary electrophoresis (CE) method was developed for determining the diol content of supports used in high-performance affinity or size exclusion chromatography. This method involved oxidizing the diol-bonded support with periodate, followed by the use of CE to separate and quantitate the iodate produced by this reaction. Both the oxidation and separation conditions were considered in optimizing this assay. The final method was performed by reacting a known amount of support with a 20-fold excess of periodate in pH 4.0, 0.5 M acetate buffer, with pyromellitic acid being used as an internal standard. After allowing 5-10 min for oxidation, the mixture was filtered and the filtrate was injected onto a 57 cmx50 mu m I.D. fused-silica capillary operated at 25 kV and containing pH 4.0, 0.5 M acetate as the running buffer. The total separation time was 5 min per run and gave a detection limit of 0.1 mM iodate (or 0.1 mu mol diol groups) for a 6-nl injection of a 1-ml reaction mixture. By varying the amount the support that was assayed, this method could be used with either porous or non-porous supports. This technique showed good correlation with an iodometric titration but required much less sample and time to perform.