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首页> 外文期刊>Journal of chromatography, A: Including electrophoresis and other separation methods >A study of the parameters affecting the accuracy of the total pore blocking method
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A study of the parameters affecting the accuracy of the total pore blocking method

机译：影响总孔隙堵塞法精度的参数研究

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We report on a study wherein we investigate the different factors affecting the accuracy of the total pore blocking method to determine the interstitial volume of reversed-phase packed bed columns. Octane, nonane, decane and dodecane were all found to be suitable blocking agents, whereas heptane already dissolves too well in the applied fully aqueous buffers. The method of moments needs to be used to accurately determine the elution times, and a proper correction for the frit volume is needed. Failing to do so can lead to errors on the observed interstitial volume of the order of 2 or more. It has also been shown that the application of a high flow rate or a high pressure does not force the blocking agent out of the mesopores of the particles. The only potential source of loss of blocking agent is dissolution into the mobile phase (even though this is a buffered fully aqueous solution). This effect however only becomes significant after the elution of 400 geometrical column volumes, i.e., orders more than needed for a regular total pore blocking experiment.
机译：我们报告了一项研究，其中我们研究了影响总孔隙封闭方法准确性的不同因素，以确定反相填充床柱的间隙体积。辛烷、壬烷、癸烷和十二烷都被发现是合适的封闭剂，而庚烷在所施加的全水性缓冲液中已经溶解得太好了。需要使用矩法来准确确定洗脱时间，并且需要对熔块体积进行适当的校正。如果不这样做，可能会导致观察到的间质体积出现 2% 或更多的误差。还表明，施加高流速或高压不会迫使阻断剂从颗粒的介孔中排出。封闭剂损失的唯一潜在来源是溶解到流动相中（即使这是一种缓冲的全水溶液）。然而，这种效应只有在洗脱 400 个几何色谱柱体积后才会变得显著，即比常规总孔封闭实验所需的数量级多。

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