US 11,913,059 B1
Mass spectrometry ionization based-assay for the detection of enzyme activity and/or presence
Trevor G Glaros, North East, MD (US); Elizabeth S Dhummakupt, Abingdon, MD (US); Phillip M Mach, Edgewood, MD (US); and Daniel O Carmany, Belcamp, MD (US)
Assigned to The United States of America as Represented by the Secretary of the Army, Washington, DC (US)
Filed by U.S. Army Combat Capabilities Development Command, Chemical Biological Center, APG, MD (US)
Filed on Oct. 26, 2021, as Appl. No. 17/511,033.
Application 17/511,033 is a division of application No. 15/976,120, filed on May 10, 2018, granted, now 11,162,128.
Claims priority of provisional application 62/509,956, filed on May 23, 2017.
Int. Cl. C12Q 1/46 (2006.01); H01J 49/04 (2006.01); H01J 49/16 (2006.01); G01N 33/68 (2006.01)
CPC C12Q 1/46 (2013.01) [G01N 33/6848 (2013.01); H01J 49/0445 (2013.01); H01J 49/168 (2013.01); C12Y 301/01007 (2013.01); C12Y 301/01008 (2013.01); G01N 2560/00 (2013.01)] 9 Claims
 
1. A method for detecting an analyte of interest using paper spray mass spectrometry, the method comprising:
providing a spray material;
applying a sample that may contain one or more analytes of interest that inhibit the enzymatic activity of acetylcholinesterase (AChE) and/or butyrylcholinesterase (BChE) and wherein said sample includes AChE and BChE enzymes, said sample applied to and disposed on said spray material;
hydrating the sample disposed on the spray material with a solvent to promote enzymatic cleavage, and extract the one or more analytes of interest contained within the sample, said solvent applied to and disposed on said spray material;
applying an enzyme substrate specific to AChE and BChE and comprising 1,1-dimethyl-4-acetylthiomethylpiperidinium iodide (MATP+) and butyrylthiocholine, wherein said enzyme substrate may be applied to the spray material or included in the solvent applied to spray material;
applying a voltage to the spray material creating charged droplets of the one or more analytes of interest contained within the sample; and
using a mass spectrometer to perform mass spectrometry on the droplets to identify the one or more analytes of interest contained in the sample and to determine a presence of the AChE and BChE enzymes contained in the sample.