title: Rapid Online Non-Enzymatic Protein Digestion Analysis with High Pressure Superheated ESI-MS.
abstract: Recently, we reported a new ESI ion source that could electrospray the super-heated aqueous solution with liquid temperature much higher than the normal boiling point (J. Am. Soc. Mass Spectrom. 25, 1862-1869). The boiling of liquid was prevented by pressurizing the ion source to a pressure greater than atmospheric pressure. The maximum operating pressure in our previous prototype was 11 atm, and the highest achievable temperature was 180°C. In this paper, a more compact prototype that can operate up to 27 atm and 250°C liquid temperatures is constructed, and reproducible MS acquisition can be extended to electrospray temperatures that have never before been tested. Here, we apply this super-heated ESI source to the rapid online protein digestion MS. The sample solution is rapidly heated when flowing through a heated ESI capillary, and the digestion products are ionized by ESI in situ when the solution emerges from the tip of the heated capillary. With weak acid such as formic acid as solution, the thermally accelerated digestion (acid hydrolysis) has the selective cleavage at the aspartate (Asp, D) residue sites. The residence time of liquid within the active heating region is about 20 s. The online operation eliminates the need to transfer the sample from the digestion reactor, and the output of the digestive reaction can be monitored and manipulated by the solution flow rate and heater temperature in a near real-time basis.
authors.affiliation: Graduate School, Department of Interdisciplinary Research, University of Yamanashi, Kofu, Yamanashi, 400-8511, Japan, leechuin@yamanashi.ac.jp.
authors.forename: Lee Chuin
authors.initials: LC
authors.lastname: Chen
authors.forename: Masato
authors.initials: M
authors.lastname: Kinoshita
authors.forename: Masato
authors.initials: M
authors.lastname: Noda
authors.forename: Satoshi
authors.initials: S
authors.lastname: Ninomiya
authors.forename: Kenzo
authors.initials: K
authors.lastname: Hiraoka
mesh_terms: D000595:Amino Acid Sequence; D004867:Equipment Design; D006358:Hot Temperature; D008969:Molecular Sequence Data; D010446:Peptide Fragments / Q000032:analysis / Q000737:chemistry*; D011312:Pressure; D011506:Proteins / Q000032:analysis / Q000737:chemistry*; D059748:Proteolysis; D021241:Spectrometry, Mass, Electrospray Ionization / Q000295:instrumentation / Q000379:methods*
chemical_list: D010446:Peptide Fragments; D011506:Proteins
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