Rapid determination of globin chains in red blood cells by capillary electrophoresis using INSTCoated fused‐silica capillary

Abstract

A laboratory-made INSTCoated fused-silica capillary has been newly used for CE separation of four mixtures of proteins in sodium phosphate BGEs at pH 3.0 and 2.5, respectively. The obtained separation efficiencies range from 145,000 theoretical plates per meter for myoglobin to 1,216,000 m(-1) for lysozyme. A total of 49-89% of the number of theoretical plates was obtained in a commercial polyvinyl alcohol coated capillary compared to the INSTCoated capillary under the same experimental conditions, 0-86% was obtained in a laboratory polyacrylamide-coated capillary, and only 0-6% was obtained in an uncoated fused-silica capillary. The RSD values for the intraday repeatability for an INSTCoated capillary were 0.1-1.0% (migration time) and 0.3-2.4% (peak area); RSD values for the interday repeatability in the same capillary are 0.6-1.4% (migration time) and 2.4-5.5% (peak area); RSD values for interday repeatability between different capillaries equaled 1.7-2.1% (migration time) and 2.8-10.9% (peak area). The INSTCoated capillary has been further used for rapid determination of globin chains isolated from red blood cells. A separation of α and β chains prepared from adult blood has been completed in 3 min with a peak resolution of 1.3, and the separation of α and (G)γ chains prepared from newborn blood took 3 min with a peak resolution of 3.6.

Capillary electrophoresis separation of aminoalkanol derivatives of 1,7‐dimethyl‐8,9‐diphenyl‐4‐azatricyclo[5.2.1.02,6]dec‐8‐ene‐3,5,10‐trione as potential anticancer drugs

“…Its potential for automated, rapid, high-efficiency separations makes it appealing as a replacement for some of the more labor-intensive assays carried out in electrophoretic gels and as a complement to companion techniques such as HPLC. Among the many attractive characteristics of this technology is its versatility for analyses of a diverse spectrum of analytes, ranging from small organic ions to macromolecular protein complexes or DNA [10][11][12][13][14][15][16][17][18][19][20][21]. Thus, CE has a significant role in drug discovery and manufacturing processes and has a potential to grow further, due to new developments that can provide highly sensitive and high throughput analysis.…”

Rapid methods for the separation of natural mixtures of beauverolides, cholesterol acyltransferase inhibitors, isolated from the fungus Isaria fumosorosea

“…We used two different experimental conditions: We characterized the cyclic BVDs using non‐aqueous CE with methanol as solvent. Detailed experimental conditions: coated fused silica capillary, inner diameter 25 µm, outer diameter 360 µm, total length 32 cm, length to C 4 D 18 cm; inner capillary surface deactivated by INST‐Coating solution to prevent adsorption of BVDs to capillary surface ; BGE 500 mM acetic acid (Sigma–Aldrich, St. Louis, MO): KOH (Lachema, Neratovice, Czech Republic) (pH 5.5) dissolved in methanol (Sigma–Aldrich); hydrodynamic sample injection by 50 mbar pressure for 4 s; separation voltage +15 kV with simultaneous application of hydrodynamic pressure of 50 mbar to decrease the migration time of BVDs ; corresponding electric current 3.0 µA. We used the C 4 D contactless conductivity detection equipped with sinus‐wave alternating signal of frequency 1 MHz and amplitude 100 V for sensitive detection of BVDs, which yields good results in capillaries of small internal diameter .…”

Electrophoretic large volume sample stacking for sensitive determination of the anti‐microbial agent pentamidine in rat plasma for pharmacological studies

“…Additionally, INST coating solution (Biotaq, Gaithersburg, USA) , was added to background electrolyte (BGE) in an amount 1% v/v to improve preventing EOF creation in capillaries. 1 min washing with the BGE was used between individual analyses; this is completely sufficient for short capillaries with 50 μm id.…”