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Titolo:
Specific and non-specific measurements of tissue angiotensin II cascade members
Autore:
Naik, GOA; Moe, GW; Armstrong, PW;
Indirizzi:
St Michaels Hosp, Toronto, ON M5B 1W8, Canada St Michaels Hosp Toronto ONCanada M5B 1W8 p, Toronto, ON M5B 1W8, Canada Univ Toronto, Dept Med, Div Cardiol, Toronto, ON M5B 1W8, Canada Univ Toronto Toronto ON Canada M5B 1W8 rdiol, Toronto, ON M5B 1W8, Canada Univ Alberta, Dept Med, Edmonton, AB, Canada Univ Alberta Edmonton AB Canada Alberta, Dept Med, Edmonton, AB, Canada
Titolo Testata:
JOURNAL OF PHARMACEUTICAL AND BIOMEDICAL ANALYSIS
fascicolo: 5-6, volume: 24, anno: 2001,
pagine: 947 - 955
SICI:
0731-7085(200103)24:5-6<947:SANMOT>2.0.ZU;2-3
Fonte:
ISI
Lingua:
ENG
Soggetto:
PERFORMANCE LIQUID-CHROMATOGRAPHY; CONGESTIVE HEART-FAILURE; RECEPTOR SUBTYPES; PEPTIDES; RADIOIMMUNOASSAY; PLASMA; HPLC; SYSTEM; SEPARATION; ANTISERA;
Keywords:
angiotensins; immunoreactive angiotensins; tissue extraction; solid-phase extraction; triethylammonium formate; ion-pair reverse phase high-performance liquid chromatography; radioimmunoassay;
Tipo documento:
Article
Natura:
Periodico
Settore Disciplinare:
Life Sciences
Citazioni:
42
Recensione:
Indirizzi per estratti:
Indirizzo: Naik, GOA St Michaels Hosp, 8F 30 Bond St, Toronto, ON M5B 1W8, Canada St Michaels Hosp 8F 30 Bond St Toronto ON Canada M5B 1W8 Canada
Citazione:
G.O.A. Naik et al., "Specific and non-specific measurements of tissue angiotensin II cascade members", J PHARM B, 24(5-6), 2001, pp. 947-955

Abstract

Although angiotensin II (ANG II) has been the focal regulatory peptide of the renin-angiotensin system, its proteolytic fragments have recently been demonstrated to have biological effects. Conventional measurement of angiotensins involves radioimmunoassay (RIA), which is a sensitive binding technique capable of measuring low physiological concentrations. However, ANG II antibody cross-reacts with ANG II and its fragments (ANG II cascade), rendering RIA measurement alone to be a non-specific measure of immunoreactive ANG II (ir-ANG II). On the other hand, high-performance liquid chromatography (HPLC) is capable of separating immunoreactive ANG II cascade members, but may not be sensitive enough to detect these low peptide concentrations often present in biological samples. Consequently, a reverse-phase HPLC method, with triethylammoniun formate as an ion-pair reagent, was developed to separate ANG II and its fragments, ANG III, ANG IV and ANG V. This HPLC separation was applied to extracts from normal canine hearts and ANG II cascadeimmunoreactive fractions were collected. Collected fractions were quantified by RIA, with the use of separate standard curves. The isocratic HPLC separation of ANG II, ANG III, ANG IV and ANG V was achieved in less than 5 min with adjacent peaks having baseline resolution. Measured cardiac left ventricle ANG III, ANG IV and ANG V concentrations (mean +/- SD) were 5.3 +/- 2.2, 4.0 +/- 1.0 and 3.1 +/- 1.0 fmol/g (n = 9), respectively. There was a significant difference (P = 0.003, n = 9) between left ventricular imlnunoreactive ANG II and 'true' ANG II, corrected for recovery rates of 86.2 +/- 22.5 and 53.5 +/- 16.2 fmol/g, respectively. We conclude that the combination of HPLC with RIA ensures the specific measurement of the ANG II cascade family members while non-chromatographic processing of tissue renders ANG II measurement non-specific. In addition, the use of triethylammonium formate as mobile phase additive is superior in the HPLC separation of the angiotensins. (C) 2001 Elsevier Science B.V. All rights reserved.

ASDD Area Sistemi Dipartimentali e Documentali, Università di Bologna, Catalogo delle riviste ed altri periodici
Documento generato il 28/03/20 alle ore 14:05:26