TY - JOUR
T1 - Measurement of Intracellular Nitric Oxide with a Quantitative Mass Spectrometry Probe Approach
AU - Zhong, Zhu Jun
AU - Yao, Zhong Ping
AU - Shi, Zi Qi
AU - Liu, Yang Dan
AU - Liu, Li Fang
AU - Xin, Gui Zhong
N1 - Funding Information:
This work was supported by the National Natural Science Foundation of China (Nos. 81973577, 81773872, and 81874306) and Jiangsu Provincial Natural Science Foundation of China (Grant No. BK20201330). This research was also supported by Talents Planning of Six Summit Fields of Jiangsu Province (No. YY-057).
Publisher Copyright:
© 2021 American Chemical Society
PY - 2021/6/22
Y1 - 2021/6/22
N2 - Nitric oxide (NO) is a molecule of physiological importance, and the function of NO depends on its concentration in biological systems, particularly in cells. Concentration-based analysis of intracellular NO can provide insight into its precise role in health and disease. However, current methods for detecting intracellular NO are still inadequate for quantitative analysis. In this study, we report a quantitative mass spectrometry probe approach to measure NO levels in cells. The probe, Amlodipine (AML), comprises a Hantzsch ester group that reacts with NO to form a pyridine, Dehydro Amlodipine (DAM). Quantification of DAM by ultraperformance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) allows specific measurement of intracellular NO levels. Notably, the AML/NO reaction proceeds rapidly (within 1 s), which is favorable for NO detection considering its large diffusivity and short half-life. Meanwhile, studies under simulated physiological conditions revealed that the AML response to NO is proportional and selective. The presented UPLC-MS/MS method showed high sensitivity (LLOQ = 0.24 nM) and low matrix interference (less than 15%) in DAM quantification. Furthermore, the mass spectrometry probe approach was demonstrated by enabling the measurement of endogenous and exogenous NO in cells. Hence, the quantitative UPLC-MS/MS method developed using AML as a probe is expected to be a new method for intracellular NO analysis.
AB - Nitric oxide (NO) is a molecule of physiological importance, and the function of NO depends on its concentration in biological systems, particularly in cells. Concentration-based analysis of intracellular NO can provide insight into its precise role in health and disease. However, current methods for detecting intracellular NO are still inadequate for quantitative analysis. In this study, we report a quantitative mass spectrometry probe approach to measure NO levels in cells. The probe, Amlodipine (AML), comprises a Hantzsch ester group that reacts with NO to form a pyridine, Dehydro Amlodipine (DAM). Quantification of DAM by ultraperformance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) allows specific measurement of intracellular NO levels. Notably, the AML/NO reaction proceeds rapidly (within 1 s), which is favorable for NO detection considering its large diffusivity and short half-life. Meanwhile, studies under simulated physiological conditions revealed that the AML response to NO is proportional and selective. The presented UPLC-MS/MS method showed high sensitivity (LLOQ = 0.24 nM) and low matrix interference (less than 15%) in DAM quantification. Furthermore, the mass spectrometry probe approach was demonstrated by enabling the measurement of endogenous and exogenous NO in cells. Hence, the quantitative UPLC-MS/MS method developed using AML as a probe is expected to be a new method for intracellular NO analysis.
UR - http://www.scopus.com/inward/record.url?scp=85108725635&partnerID=8YFLogxK
U2 - 10.1021/acs.analchem.1c01259
DO - 10.1021/acs.analchem.1c01259
M3 - Journal article
AN - SCOPUS:85108725635
SN - 0003-2700
VL - 93
SP - 8536
EP - 8543
JO - Analytical Chemistry
JF - Analytical Chemistry
IS - 24
ER -