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

Biochemical evaluation of the renin-angiotensin system: the good, bad, and absolute?

“…ANG II and its COOH-terminal intact metabolites were elevated 3-to 10-fold in the apolipoprotein E Ϫ/Ϫ mice, but ANG IV remained the major peptide (550 pM); plasma levels of ANG-(1-7) were unchanged (102). Although the ANG II and ANG-(1-7) values were comparable with RIAbased studies, the higher ANG III and ANG IV content may again reflect inadequate inhibition of aminopeptidases by addition of a standard protease inhibitor cocktail in contrast to extraction methods specifically designed to inhibit peptide processing (102). Finally, Ali et al (7) quantified plasma ANG II and ANG-(1-7) content at 20,000 and 40,000 pM, respectively, in WT mice using UPLC-quardruple (Q)TOF.…”

“…Plasma levels of ANG II, ANG III, and ANG-(1-7) were 36, 32, and 63 pM, respectively; however, ANG IV (158 pM) was the predominant peptide in WT mice (102). ANG II and its COOH-terminal intact metabolites were elevated 3-to 10-fold in the apolipoprotein E Ϫ/Ϫ mice, but ANG IV remained the major peptide (550 pM); plasma levels of ANG-(1-7) were unchanged (102). Although the ANG II and ANG-(1-7) values were comparable with RIAbased studies, the higher ANG III and ANG IV content may again reflect inadequate inhibition of aminopeptidases by addition of a standard protease inhibitor cocktail in contrast to extraction methods specifically designed to inhibit peptide processing (102).…”

“…Indeed, their additional analysis of plasma peptides from female WT mice anesthetized with ketamine revealed far lower plasma ANG III, although the levels of ANG III were twofold higher than ANG II (160 vs. 74 pM) (147). Olkowicz et al (102) quantified plasma angiotensin by MS in male WT mice and in apolipoprotein E Ϫ/Ϫ mice that exhibit a stimulated RAS. Plasma levels of ANG II, ANG III, and ANG-(1-7) were 36, 32, and 63 pM, respectively; however, ANG IV (158 pM) was the predominant peptide in WT mice (102).…”

“…Olkowicz et al (102) quantified plasma angiotensin by MS in male WT mice and in apolipoprotein E Ϫ/Ϫ mice that exhibit a stimulated RAS. Plasma levels of ANG II, ANG III, and ANG-(1-7) were 36, 32, and 63 pM, respectively; however, ANG IV (158 pM) was the predominant peptide in WT mice (102). ANG II and its COOH-terminal intact metabolites were elevated 3-to 10-fold in the apolipoprotein E Ϫ/Ϫ mice, but ANG IV remained the major peptide (550 pM); plasma levels of ANG-(1-7) were unchanged (102).…”

The vasoprotective axes of the renin-angiotensin system: Physiological relevance and therapeutic implications in cardiovascular, hypertensive and kidney diseases

“…Using Liquid chromatography–tandem mass spectrometry (LC-MS/MS), Wysocki et al reported that the level of plasma ANG III was remarkably higher than that of ANG II in wild-type mice on the C57BL6 genetic background (1,047±178 vs. 7.8±2.0 pg/ml), whereas the kidney ANG III level was much lower than that of plasma (172±12 pg/g) [81]. Also using LC-MS/MS, Olkowicz et al reported the plasma ANG III level nearly 35-fold lower at 30.3 ± 3.8 pg/ml in wild-type mice [82]. More reliable ANG II levels in the plasma and kidney may be obtained by the gold-standard high-performance liquid chromatography (HPLC)-based RIA or ELISA approach [78–80].…”