کد مقاله کد نشریه سال انتشار مقاله انگلیسی نسخه تمام متن
1199819 1493575 2014 8 صفحه PDF دانلود رایگان
عنوان انگلیسی مقاله ISI
Determination of pore size distributions in capillary-channeled polymer fiber stationary phases by inverse size-exclusion chromatography and implications for fast protein separations
ترجمه فارسی عنوان
تعیین توزیع اندازه ذره در فاز ثابت فیبرهای کانال فیبر مویرگ با استفاده از کروماتوگرافی معکوس اندازه معکوس و پیامدهای جداسازی پروتئین سریع
موضوعات مرتبط
مهندسی و علوم پایه شیمی شیمی آنالیزی یا شیمی تجزیه
چکیده انگلیسی


• iSEC is applied to determine the intra-fiber pore size distributions of C-CP fibers.
• The average pore size of ∼4 nm practically excludes peptides/proteins of >∼10,000 Da.
• van Deemter and Knox plots reveal a virtual absence of mass transfer limitations.
• Characteristic particle diameters (dp) from Knox equation fitting are ∼ 50 μm.

Capillary-channeled polymer (C-CP) fibers have been utilized as liquid chromatography stationary phases, primarily for biomacromolecule separations on the analytical and preparative scales. The collinear packing of the eight-channeled C-CP fibers provides for very efficient flow, allowing operation at high linear velocity (u > 100 mm s−1) and low backpressure (<2000 psi) in analytical-scale separations. To take advantage of these fluid transport properties, there must not be mass transfer limitations as would be imposed by having an appreciably porous phase, wherein solute diffusion limits the overall mass transport rates. To better understand the physical nano-/micro- structure of C-CP fibers, inverse size exclusion chromatography (iSEC) has been employed to determine the pore size distribution (PSD) within C-CP fibers. A diversity of test species (from metal ions to large proteins) was used as probes under non-retaining conditions to obtain a response curve reflecting the apparent partition coefficient (Kd) versus hydrodynamic radii (rm). A mean pore radius (rp) of 4.2 nm with standard deviation (sp) of ±1.1 nm was calculated by fitting the Kd versus rm data to model equations with a Gaussian pore size distribution, and a pore radius of 4.0 ± 0.1 nm was calculated based on a log-normal distribution. The derived mean pore radius is much smaller than traditional support materials, with the standard deviation showing a relatively uniform pore distribution. van Deemter plots were analyzed to provide practical confirmation of the structural implications. Large molecules (e.g., proteins) that are fully excluded from pores have no significant C-terms in the van Deemter plots whereas small molecules that can access the pore volumes display appreciable C-terms, as expected. Fitting of retention data to the Knox equation suggests that the columns operate with a characteristic particle diameter (dp) of ∼53 μm.

ناشر
Database: Elsevier - ScienceDirect (ساینس دایرکت)
Journal: Journal of Chromatography A - Volume 1351, 18 July 2014, Pages 82–89
نویسندگان
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