scholarly journals Development of a Parallel Reaction Monitoring-MS Method To Quantify IGF Proteins in Dogs and a Case of Nonislet Cell Tumor Hypoglycemia

2018 ◽  
Vol 18 (1) ◽  
pp. 18-29 ◽  
Author(s):  
Alberto Valdés ◽  
Moira Lewitt ◽  
Erica Wiss ◽  
Margareta Ramström ◽  
Emma M. Strage
Keyword(s):  
Cell Tumor ◽  
Reaction Monitoring ◽  
Parallel Reaction ◽  
Parallel Reaction Monitoring ◽  
Tumor Hypoglycemia

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.

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