کد مقاله کد نشریه سال انتشار مقاله انگلیسی نسخه تمام متن
5135241 1493428 2017 9 صفحه PDF دانلود رایگان
عنوان انگلیسی مقاله ISI
Self-packed core shell nano liquid chromatography columns and silica-based monolithic trap columns for targeted proteomics
موضوعات مرتبط
مهندسی و علوم پایه شیمی شیمی آنالیزی یا شیمی تجزیه
پیش نمایش صفحه اول مقاله
Self-packed core shell nano liquid chromatography columns and silica-based monolithic trap columns for targeted proteomics
چکیده انگلیسی


- Nano LC columns fast and simply packed in-house with core shell particles.
- Silica monolith trap columns enhanced total chromatographic performance.
- Targeted detection of CYP27A1 in cancer cells.

Self-preparation of nano liquid chromatography (nLC) columns has advantages regarding cost and flexibility. For targeted proteomics, we evaluated several approaches for particle-packing nLC columns and manufacturing fritless silica-based monolithic trap columns (50 μm inner diameter). Our preferred approach for nLC column preparation was to magnetically stir Accucore core shell particles (C18 stationary phase) in ACN/water (80/20, v/v) suspensions during pressure-driven filling of polymer-fritted standard fused silica capillaries. The columns were ready for use about one hour after preparation had begun. They had comparable peak capacities (peptides) to commercial columns, and satisfactory within/between-column retention time repeatability, suited for targeted proteomics. Packing with commercial capillary housings/nanospray emitters did not improve performance compared to packing with in-house fritted stock fused silica capillary tubing.For trap columns, several recipes for narrow bore silica-based monolithic columns were evaluated, and we found the recipe by Zou et al. (2005) to be reproducible. Compared to the standard C18 trap column for Accucore nLC columns, monolith trap columns (C8 stationary phase) significantly reduced peak widths. The readily prepared in-house columns were used for targeted detection of the enzyme CYP27A1 in cancer cells, which is associated with proliferation and metastasis of breast cancer.

ناشر
Database: Elsevier - ScienceDirect (ساینس دایرکت)
Journal: Journal of Chromatography A - Volume 1498, 19 May 2017, Pages 111-119
نویسندگان
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