These methods have their own advantages, such as sensitivity (real-time monitoring using molecular beacons) or using very little sample (capillary electrophoresis).
All measurements were performed using a capillary cell (sample volume 10 μl) and automated filling at 25 °C.
4. Hemotological measures will be assessed using a 35 μl capillary blood sample drawn from a finger prick blood sample for immediate analysis via a Cholestech LDX system (Cholestech LDX, Cholestech, USA).
With all these considerations in mind, development of a dissolution DNP system suitable for use in analytical chemistry should be able to successfully polarize and quantitatively dissolve extremely small samples, preferably using a capillary-based system for sample delivery.
A small volume of blood is quantitatively taken up using a capillary and transferred to a sample cartridge, which is subsequently inserted into the MS system for automatic analysis.
Briefly, the tryptic peptides (2 μg in each sample) were analyzed using a RP C18 capillary LC column (100 μm × 150 mm, 3 μm) from Michrom Bioresources (Michrom BioResources, Westford, MA, USA).
Peptide samples were separated using a capillary reversed-phase LC column (Agilent Poroshell 120 EC-C18; 2.1 mm ×100 mm, particle size 2.7 μm) and an Agilent 1290 Infinity HPLC system.
Samples were run using a high-resolution capillary gel electrophoresis (CGE) protocols (deltaDOT, UK) under reducing conditions.
When we used a glass capillary as sample, we filled the sample chamber with glycerol to avoid refractive index mismatch on the glass surface which otherwise adds aberrations to the speckle spots.
Fully automated online pre-concentration and separation of the tryptically digested samples was performed using a set of capillary- and nanoHPLC instruments of the 1100 Series (Agilent, Waldbronn, Germany) operated in series.
