title: Development of a sensitive, accurate and robust liquid chromatography/mass spectrometric method for profiling of angiotensin peptides in plasma and its application for atherosclerotic mice.
abstract: Quantification of angiotensin (Ang) peptides in biological matrices is a challenge due to their low picomolar (pM) concentration and poor analytical performance of current methods. This work aimed to select an optimal strategy for liquid chromatography/mass spectrometry (LC/MS) quantification of major angiotensins in plasma of wild type and atherosclerotic mice. Optimal LC/MS set-up for Ang quantification was chosen, based on analytical performance, from: nanoflow/orbitrap, nanoflow/triple quadrupole and preconcentration nanoflow/triple quadrupole. The best LC/MS configuration (preconcentration nanoflow/triple quadrupole) was validated and used for measurement of angiotensins (Ang I, II, III, IV and (1-7)) in plasma of 6-month-old atherosclerotic apolipoprotein E/LDL receptor double knock-outs (ApoE/LDLR (--/--)) and wild type C57BL/6J (WT) mice. The method established for Ang quantification was selective, accurate and highly sensitive with LLOQ of 5pgmL(-1). The peak area intra-day precisions for Ang II and Ang-(1-7) were in the range 3.0-5.1 and 3.5-5.8, respectively, with corresponding accuracy of 95.4-103.5% and 95.6-106.3%. Plasma angiotensin profile was substantially modified in ApoE/LDLR knock-out mice with increase in concentration of Ang II from 37.6±21.3pgmL(-1) in WT to 200.2±47.6pgmL(-1). Concentrations of Ang I, III and IV were also increased 3-10 fold in ApoE/LDLR (--/--) mice while that of Ang-(1-7) was unchanged. We conclude that the method developed could be effectively used for accurate, comprehensive profiling of angiotensin peptides in mouse plasma. We identified substantial changes in renin-angiotensin system in a genetic mouse model of atherosclerosis consistent with the overactivation of angiotensin converting enzyme (ACE) and the impairment of ACE2.
authors: Department of Biochemistry, Medical University of Gdansk, Debinki 1, 80-211 Gdansk, Poland; Department of Biotechnology and Food Microbiology, Poznan University of Life Sciences, Wojska Polskiego 48, 60-627 Poznan, Poland. Electronic address: m.olkowicz@gumed.edu.pl.
authors: Mariola
authors: M
authors: Olkowicz
authors: Department of Biochemistry, Medical University of Gdansk, Debinki 1, 80-211 Gdansk, Poland.
authors: Adrianna
authors: A
authors: Radulska
authors: Department of Pharmacokinetics and Physical Pharmacy, Jagiellonian University Medical College, Medyczna 9, 30-688 Krakow, Poland; Jagiellonian Centre for Experimental Therapeutics (JCET), Jagiellonian University, Bobrzynskiego 14, 30-348 Krakow, Poland.
authors: Joanna
authors: J
authors: Suraj
authors: Jagiellonian Centre for Experimental Therapeutics (JCET), Jagiellonian University, Bobrzynskiego 14, 30-348 Krakow, Poland.
authors: Agnieszka
authors: A
authors: Kij
authors: Department of Pharmacokinetics and Physical Pharmacy, Jagiellonian University Medical College, Medyczna 9, 30-688 Krakow, Poland; Jagiellonian Centre for Experimental Therapeutics (JCET), Jagiellonian University, Bobrzynskiego 14, 30-348 Krakow, Poland.
authors: Maria
authors: M
authors: Walczak
authors: Jagiellonian Centre for Experimental Therapeutics (JCET), Jagiellonian University, Bobrzynskiego 14, 30-348 Krakow, Poland; Department of Experimental Pharmacology, Jagiellonian University Medical College, Grzegorzecka 16, 31-531 Krakow, Poland.
authors: Stefan
authors: S
authors: Chlopicki
authors: Department of Biochemistry, Medical University of Gdansk, Debinki 1, 80-211 Gdansk, Poland.
authors: Ryszard T
authors: RT
authors: Smolenski
mesh_terms: D000803:Angiotensin I / Q000097:blood; D000804:Angiotensin II / Q000097:blood; D000805:Angiotensin III / Q000097:blood; D000809:Angiotensins / Q000097:blood*; D000818:Animals; D001057:Apolipoproteins E / Q000235:genetics; D050197:Atherosclerosis / Q000097:blood* / Q000235:genetics; D002853:Chromatography, Liquid / Q000379:methods; D013058:Mass Spectrometry / Q000379:methods; D008810:Mice, Inbred C57BL; D018345:Mice, Knockout; D010446:Peptide Fragments / Q000097:blood; D011973:Receptors, LDL / Q000235:genetics
chemical_list: D000809:Angiotensins; D001057:Apolipoproteins E; D010446:Peptide Fragments; D011973:Receptors, LDL; D000804:Angiotensin II; D000805:Angiotensin III; D000803:Angiotensin I; C118790:angiotensin I (1-7)
keywords: Angiotensins; Atherosclerotic mice; High-resolution mass spectrometry; Renin–angiotensin system; Targeted quantitative proteomics