Detection limits for matrix-assisted laser desorption of polypeptides with an external ion source Fourier-transform mass spectrometer.
Sensitivity in the low-femtomole range with mass resolution greater than 20,000 is demonstrated for several polypeptides analyzed by a mass spectrometer that pairs matrix-assisted laser desorption/ionization (MALDI) and Fourier-transform mass spectrometry (FTMS). The compounds investigated were substance P, renin substrate, melittin, the B-chain of bovine insulin, and bovine insulin. Standard solutions of the polypeptides were prepared with 30% acetonitrile+water, and micropipettes were used to transfer small amounts (1-20 fmol) to a sample probe. The samples were embedded in a large excess of matrix material (2,5-dihydroxybenzoic acid) and ionized by a pulse from an excimer laser. The FTMS instrument used for these experiments has the MALDI source in a separate chamber outside the magnetic field. Ions are extracted from the source and transported by an RF-only quadrupole ion guide to an FTMS analyzer cell mounted in the homogeneous region of a 6.5 T superconducting magnet. The high sensitivity of MALDI-FTMS is due, in part, to the high transfer efficiency of the ion guide, even for ions with a wide range of kinetic energies. The ion guide is easy to use because there are only two adjustments (RF amplitude and DC offset voltage), and unlike electrostatic ion transport means, alignment of it with the axis of the magnetic field is not critical. The mass resolution and sensitivity of MALDI-FTMS is compared with that of MALDI done with time-of-flight, magnetic sector, and quadrupole ion-trap mass spectrometers.
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Department of Chemistry, University of California, Irvine 92717.
McIver
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D000595:Amino Acid Sequence; D000808:Angiotensinogen / Q000737:chemistry; D000818:Animals; D002417:Cattle; D005583:Fourier Analysis; D006801:Humans; D007328:Insulin / Q000737:chemistry; D007834:Lasers; D013058:Mass Spectrometry; D008555:Melitten / Q000737:chemistry; D008969:Molecular Sequence Data; D010455:Peptides / Q000737:chemistry*; D013373:Substance P / Q000737:chemistry
D007328:Insulin; D010455:Peptides; D000808:Angiotensinogen; D008555:Melitten; D013373:Substance P