title: UHPLC-MS/MS method with protein precipitation extraction for the simultaneous quantification of ten antihypertensive drugs in human plasma from resistant hypertensive patients.
abstract: Today the management of resistant hypertension is a critical health problem: the main difficulty on this field is the discrimination of cases of poor therapeutic adherence from cases of real resistance. This gives rise to the need of high throughput and reliable quantification methods for the Therapeutic Drug Monitoring (TDM) of antihypertensive drugs. The aim of this work was the development and validation of a UHPLC-Tandem mass spectrometry assay for this application and its use in plasma from patients with resistant hypertension. The novelty of this method resides in the ability to simultaneously quantify a wide panel of antihypertensive drugs: amlodipine, atenolol, clonidine, chlortalidone, doxazosin, hydrochlorothiazide, nifedipine, olmesartan, ramipril and telmisartan. Moreover, this method stands out for its simplicity and cheapness, resulting feasible for clinical routine. Both standards and quality controls were prepared in human plasma. After the addition of internal standard, each sample underwent protein precipitation with acetonitrile and was then dried. Extracts were resuspended in water:acetonitrile 90:10 (0.05% formic acid) and then injected into the chromatographic system. Chromatographic separation was performed on an Acquity(®) UPLC HSS T3 1.8μm 2.1×150mm column, with a gradient of water and acetonitrile, both added with 0.05% formic acid. Accuracy, intra-day and inter-day precision fitted FDA guidelines for all analytes, while matrix effects and recoveries resulted stable between samples for each analyte. Finally, we tested this method by monitoring plasma concentrations in 22 hypertensive patients with good results. This simple analytical method could represent a useful tool for the management of antihypertensive therapy.
authors: Unit of Infectious Diseases(2), University of Turin, Department of Medical Sciences, Amedeo di Savoia Hospital, Turin, Italy. Electronic address: amedeo.denicolo@unito.it.
authors: Amedeo
authors: A
authors: De Nicolò
authors: Unit of Infectious Diseases(2), University of Turin, Department of Medical Sciences, Amedeo di Savoia Hospital, Turin, Italy.
authors: Valeria
authors: V
authors: Avataneo
authors: Division of Internal Medicine and Hypertension Unit, University of Turin, Department of Medical Sciences, AOU "Città della Salute e della Scienza", Turin, Italy.
authors: Franco
authors: F
authors: Rabbia
authors: Unit of Infectious Diseases(2), University of Turin, Department of Medical Sciences, Amedeo di Savoia Hospital, Turin, Italy.
authors: Gabriele
authors: G
authors: Bonifacio
authors: Unit of Infectious Diseases(2), University of Turin, Department of Medical Sciences, Amedeo di Savoia Hospital, Turin, Italy.
authors: Jessica
authors: J
authors: Cusato
authors: Unit of Infectious Diseases(2), University of Turin, Department of Medical Sciences, Amedeo di Savoia Hospital, Turin, Italy.
authors: Cristina
authors: C
authors: Tomasello
authors: Division of Internal Medicine and Hypertension Unit, University of Turin, Department of Medical Sciences, AOU "Città della Salute e della Scienza", Turin, Italy.
authors: Elisa
authors: E
authors: Perlo
authors: Division of Internal Medicine and Hypertension Unit, University of Turin, Department of Medical Sciences, AOU "Città della Salute e della Scienza", Turin, Italy.
authors: Paolo
authors: P
authors: Mulatero
authors: Division of Internal Medicine and Hypertension Unit, University of Turin, Department of Medical Sciences, AOU "Città della Salute e della Scienza", Turin, Italy.
authors: Franco
authors: F
authors: Veglio
authors: Division of Internal Medicine and Hypertension Unit, University of Turin, Department of Medical Sciences, AOU "Città della Salute e della Scienza", Turin, Italy.
authors: Giovanni
authors: G
authors: Di Perri
authors: Unit of Infectious Diseases(2), University of Turin, Department of Medical Sciences, Amedeo di Savoia Hospital, Turin, Italy.
authors: Antonio
authors: A
authors: D'Avolio
mesh_terms: D000959:Antihypertensive Agents / Q000097:blood* / Q000737:chemistry* / Q000627:therapeutic use; D002851:Chromatography, High Pressure Liquid / Q000379:methods; D016903:Drug Monitoring / Q000379:methods; D006801:Humans; D006973:Hypertension / Q000188:drug therapy; D010949:Plasma / Q000737:chemistry*; D015203:Reproducibility of Results; D053719:Tandem Mass Spectrometry / Q000379:methods
chemical_list: D000959:Antihypertensive Agents
keywords: Antihypertensive drugs; Hypertension; Liquid chromatography; TDM; Tandem mass spectrometry; Therapeutic drug monitoring