Performance of a sheathless porous tip sprayer for capillary electrophoresis-electrospray ionization-mass spectrometry of intact proteins.

The performance of a prototype porous tip sprayer for sheathless capillary electrophoresis-mass spectrometry (CE-MS) of intact proteins was studied. Capillaries with a porous tip were inserted in a stainless steel needle filled with static conductive liquid and installed in a conventional electrospray ionization (ESI) source. Using a BGE of 100 mM acetic acid (pH 3.1) and a positively charged capillary coating, a highly reproducible and efficient separation of four model proteins (insulin, carbonic anhydrase II, ribonuclease A and lysozyme) was obtained. The protein mass spectra were of good quality allowing reliable mass determination of the proteins and some of their impurities. Sheath-liquid CE-MS using the same porous tip capillary and an isopropanol-water-acetic acid sheath liquid showed slightly lower to similar analyte responses. However, as noise levels increased with sheath-liquid CE-MS, detection limits were improved by a factor 6.5-20 with sheathless CE-MS. The analyte response in sheathless CE-MS could be enhanced using a nanoESI source and adding 5% isopropanol to the BGE, leading to improved detection limits by 50-fold to 140-fold as compared to sheath liquid interfacing using the same capillary - equivalent to sub-nM detection limits for three out of four proteins. Clearly, the sheathless porous tip sprayer provides high sensitivity CE-MS of intact proteins.

Electrochemical properties of columns in capillary electrochromatography. I. Ohm's law, resistivity and field strength.

The most commonly used type of column in capillary electrochromatography (CEC) consists of a packed segment and an open (but buffer-filled) segment. The two segments differ importantly in two respects: firstly, their electrical resistivity; and secondly, their zeta potentials at a multitude of solid-liquid interfaces. Determination of the magnitude of these properties for each segment cannot be made using only results from the column as a whole. Instead, measurements of resistivity and zeta potentials of an entirely open, unpacked column can be used in conjunction with those of the CEC column to determine the electrochemical nature of both segments. This review of basic electrochemical properties will describe simple procedures that can be used to determine resistance, resistivity, conductivity, conductance, and field strength in each segment. In addition, it will be shown how the properties of each segment add together to give the same properties of the CEC column as a whole. The equations so derived will be applied to data from the literature and conclusions drawn from the results.