Details

Autor(en) / Beteiligte

• Haselberg, Rob
• Ratnayake, Chitra K.
• de Jong, Gerhardus J.
• Somsen, Govert W.

Titel

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

Ist Teil von

• Journal of Chromatography A, 2010-11, Vol.1217 (48), p.7605-7611

Ort / Verlag

Amsterdam: Elsevier B.V

Erscheinungsjahr

2010

Quelle

MEDLINE

Beschreibungen/Notizen

• 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.