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Poster Presentations

Day 2, June 23（Mon.）　

Room P (Maesato East, Foyer, Ocean Wing)

• 2P-AM-25
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Optimization of probe electrospray ionization-mass spectrometry (PESI-MS) for the analysis of an in vitro enzyme reaction system

(¹Grad. Sch. IST, Osaka Univ., ²OTRI, Osaka Univ., ³ICBiotech., Osaka Univ., ⁴JSPS, ⁵Shimadzu)
^oNobuyuki Okahashi^1,2, Takeo Taniguchi¹, Takuma Suzuki^3,4, Takanari Hattori⁵, Hidenori Takahashi⁵, Kohsuke Honda^2,3, Fumio Matsuda^1,2

Probe electrospray ionization (PESI) ionizes metabolites by applying a voltage to a probe with a sample attached. Since PESI introduces only a small volume of solution, it minimizes contamination of the mass spectrometer, making it suitable for high-throughput analysis with minimal sample preparation. This study applied PESI-MS to measure metabolite levels in an enzyme reaction system in vitro containing proteins, buffers, and salts. An engineered glycolysis enzyme mixture was used for the reaction. PESI-QTOF/MS analysis showed that ethanol produced stable signal intensity, and ammonium acetate or ammonium formate improved ATP and NADPH ionization. Time-course analysis of eight metabolites in non-oxidative glycolysis showed expected trends, such as increasing ATP and decreasing ADP. The results were consistent with ion-pair LC-ESI-MS, confirming the quantitative capability of PESI-QTOF/MS.

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