Stability Enhancement for Peptide Analysis by Electrospray Using the Triple Quadrupole Mass Spectrometer |
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| Authors: | Houle Wang Kheng B. Lim Ross F. Lawrence William N. Howald J.Alex Taylor Lowell H. Ericsson Kenneth A. Walsh Murray Hackett |
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| Affiliation: | aDepartment of Medicinal Chemistry, University of Washington, Seattle, Washington, 98195;bDepartment of Biochemistry, University of Washington, Seattle, Washington, 98195 |
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| Abstract: | Electrospray ionization sources, used with triple quadrupole mass spectrometers from PE/Sciex (API III+), Micromass (Quattro II), and Finnigan (TSQ 7000), were modified with a 35-gauge stainless steel needle. The dimensions of the needle were 63 μm i.d. by 145 μm o.d. with variable length, depending on the specific instrument. This modification led to enhanced signal stability, improved signal/noise ratios, and lowered sample consumption for a wide range of peptides. Stable baselines were observed with flow rates in the range of 50 nL/min to 5 μL/min. An alternative design, based on a metal wire housed within a fused silica capillary, led to the most stable signals of all during infusion, but caused excessive peak broadening with capillary chromatography. The Finnigan interface was further modified with an external postcolumn addition tee, used in conjunction with capillary liquid chromatography columns of 30 and 50 μm internal diameter. The best results with the modified Finnigan interface were acquired using the 50-μm column at a flow rate of 150 to 200 nL/min. |
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