scholarly journals Peer Review #3 of "Development of a novel nanoflow liquid chromatography-parallel reaction monitoring mass spectrometry-based method for quantification of angiotensin peptides in HUVEC cultures (v0.1)"

2020 ◽  
Keyword(s):  
Mass Spectrometry ◽  
Liquid Chromatography ◽  
Peer Review ◽  
Reaction Monitoring ◽  
Parallel Reaction ◽  
Angiotensin Peptides ◽  
Parallel Reaction Monitoring ◽  
Nanoflow Liquid Chromatography

scholarly journals Development of a novel nanoflow liquid chromatography-parallel reaction monitoring mass spectrometry-based method for quantification of angiotensin peptides in HUVEC cultures

PeerJ ◽  
2020 ◽  
Vol 8 ◽  
pp. e9941
Author(s):  
Chuan He ◽  
Simiao Hu ◽  
Wanxing Zhou
Keyword(s):  
Mass Spectrometry ◽  
Liquid Chromatography ◽  
Culture Supernatant ◽  
Reaction Monitoring ◽  
Ang Ii ◽  
Parallel Reaction ◽  
Parallel Reaction Monitoring ◽  
Relative Standard ◽  
Ang Iv ◽  
Nanoflow Liquid Chromatography

Background This study aimed to develop an analytical method using liquid chromatography tandem mass spectrometry (LC-MS/MS) for the determination of angiotensin (Ang) I, Ang (1-9), Ang II, Ang (1-7), Ang (1-5), Ang III, Ang IV in human umbilical vein endothelial cell (HUVEC) culture supernatant. Methods HUVEC culture supernatant was added with gradient concentrations (0.05–1,000 ng/ml) of standard solutions of the Ang peptides. These samples underwent C18 solid-phase extraction and separation using a preconcentration nano-liquid chromatography mass spectrometry system. The target peptides were detected by a Q Exactive quadrupole orbitrap high-resolution mass spectrometer in the parallel reaction monitoring mode. Ang converting enzyme (ACE) in HUVECs was silenced to examine Ang I metabolism. Results The limit of detection was 0.1 pg for Ang II and Ang III, and 0.5 pg for Ang (1-9), Ang (1-7), and Ang (1-5). The linear detection range was 0.1–2,000 pg (0.05–1,000 ng/ml) for Ang II and Ang III, and 0.5–2,000 pg (0.25–1,000 ng/ml) for Ang (1-9) and Ang (1-5). Intra-day and inter-day precisions (relative standard deviation) were <10%. Ang II, Ang III, Ang IV, and Ang (1-5) were positively correlated with ACE expression by HUVECs, while Ang I, Ang (1-7), and Ang (1-9) were negatively correlated. Conclusion The nanoflow liquid chromatography-parallel reaction monitoring mass spectrometry-based methodology established in this study can evaluate the Ang peptides simultaneously in HUVEC culture supernatant.

scholarly journals Identification of Metabolites of Aurantio-Obtusin in Rats Using Ultra-High-Performance Liquid Chromatography-Q-Exactive Orbitrap Mass Spectrometry with Parallel Reaction Monitoring

2021 ◽  
Vol 2021 ◽  
pp. 1-8
Author(s):  
Shi-han Qin ◽  
Yuan Xu ◽  
Kai-lin Li ◽  
Kai-yan Gong ◽  
Jie Peng ◽  
...  
Keyword(s):  
Mass Spectrometry ◽  
High Performance Liquid Chromatography ◽  
Liquid Chromatography ◽  
High Performance ◽  
Good Effect ◽  
Reaction Monitoring ◽  
Parallel Reaction ◽  
Parallel Reaction Monitoring ◽  
Orbitrap Mass Spectrometry ◽  
Q Exactive

Aurantio-obtusin (AO) is a major anthraquinone compound isolated from Cassiae Semen or Duhaldea nervosa, which possesses diverse pharmacological effects. Previous studies have shown that it has a good effect on lowering blood lipids and treating various diseases. A few studies have also reported about its metabolites. A rapid and reliable method using ultra-high-performance liquid chromatography-Q-Exactive Orbitrap mass spectrometry and multiple data-processing technologies was established to investigate the metabolites of AO in the plasma and various tissues of rats, including the heart, liver, spleen, lung, kidneys, and brain. Finally, a total of 36 metabolites were identified in the plasma of rats, which could be very beneficial for understanding the effective form of AO metabolites leading to new drug discovery. The result demonstrated that this strategy, especially parallel reaction monitoring, has shown a wide range of applications in the identification of metabolites.

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