US 12,117,457 B2
Method for simultaneous analysis of neurotransmitters and their metabolites based on derivatization
Junghyun Son, Seoul (KR); Yoeseph Cho, Seoul (KR); Oh-Seung Kwon, Seoul (KR); Hana Park, Seoul (KR); Jin Woo Park, Seoul (KR); Yejin Lee, Seoul (KR); and Seongeun Jeon, Seoul (KR)
Assigned to Korea Institute of Science and Technology, Seoul (KR)
Filed by KOREA INSTITUTE OF SCIENCE AND TECHNOLOGY, Seoul (KR)
Filed on Mar. 31, 2021, as Appl. No. 17/219,023.
Claims priority of application No. 10-2020-0067399 (KR), filed on Jun. 4, 2020.
Prior Publication US 2021/0389336 A1, Dec. 16, 2021
Int. Cl. G01N 33/94 (2006.01)
CPC G01N 33/9406 (2013.01) [G01N 33/9413 (2013.01); G01N 33/942 (2013.01); G01N 33/9426 (2013.01); G01N 33/944 (2013.01)] 15 Claims
OG exemplary drawing
 
1. A method for simultaneous analysis of neurotransmitters and/or their metabolites, comprising (a) separating analytes comprising a plurality of neurotransmitters and/or their metabolites from a sample selected from body tissues, body fluids, secretions, and excretions, (b) derivatizing the analytes with ethyl chloroformate to obtain derivatives of the plurality of neurotransmitters and/or their metabolites, (c) separating the derivatives of the plurality of neurotransmitters and/or their metabolites by liquid chromatography, and (d) subjecting the separated derivatives of the neurotransmitters or their metabolites to multiple reaction monitoring (MRM) using a mass spectrometer, wherein the neurotransmitters and/or their metabolites have a limit of detection (LOD) of <0.1 ng/ml; and
wherein step (b) comprises (b-1) adding a mixed solution of ethanol and pyrimidine and ethyl chloroformate to the separated analytes to derivatize the analytes and (b-2) adding an organic solvent to the derivatized analytes, extracting the derivatives of the neurotransmitters and/or their metabolites by liquid-liquid extraction, and concentrating the derivatives.