Development of UHPLC-MS/MS methods to quantify 25 antihypertensive drugs in serum in a cohort of patients treated for hypertension

Christian W Thorstensen; Per-Erik Clasen; Stine Rognstad; Renate Haldsrud; Siri Føreid; Trine Helstrøm; Ola Undrum Bergland; Lene Vernås Halvorsen; Arleen Aune; Eirik Olsen; Karl Marius Brobak; Aud Høieggen; Ingebjørg Gustavsen; Anne Cecilie K Larstorp; Camilla Lund Søraas; Mimi Stokke Opdal

doi:10.1016/j.jpba.2022.114908

Development of UHPLC-MS/MS methods to quantify 25 antihypertensive drugs in serum in a cohort of patients treated for hypertension

J Pharm Biomed Anal. 2022 Sep 20:219:114908. doi: 10.1016/j.jpba.2022.114908. Epub 2022 Jun 23.

Authors

Christian W Thorstensen¹, Per-Erik Clasen¹, Stine Rognstad², Renate Haldsrud¹, Siri Føreid¹, Trine Helstrøm¹, Ola Undrum Bergland³, Lene Vernås Halvorsen⁴, Arleen Aune⁵, Eirik Olsen⁶, Karl Marius Brobak⁷, Aud Høieggen⁴, Ingebjørg Gustavsen¹, Anne Cecilie K Larstorp⁸, Camilla Lund Søraas⁹, Mimi Stokke Opdal¹⁰

Affiliations

¹ Department of Pharmacology, Oslo University Hospital, Ullevål, Oslo, Norway.
² Department of Pharmacology, Oslo University Hospital, Ullevål, Oslo, Norway; Section of Cardiovascular and Renal Research, Oslo University Hospital, Ullevål, Oslo, Norway; Institute of Clinical Medicine, Medical Faculty, University of Oslo, Oslo, Norway. Electronic address: stirog@ous-hf.no.
³ Section of Cardiovascular and Renal Research, Oslo University Hospital, Ullevål, Oslo, Norway; Institute of Clinical Medicine, Medical Faculty, University of Oslo, Oslo, Norway.
⁴ Section of Cardiovascular and Renal Research, Oslo University Hospital, Ullevål, Oslo, Norway; Institute of Clinical Medicine, Medical Faculty, University of Oslo, Oslo, Norway; Department of Nephrology, Oslo University Hospital, Ullevål, Oslo, Norway.
⁵ Department of Clinical Science, University of Bergen, Bergen, Norway; Department of Heart Disease, Haukeland University Hospital, Bergen, Norway.
⁶ Department of Cardiology, St Olav University Hospital, Trondheim, Norway; Norwegian University of Science and Technology, Trondheim, Norway.
⁷ Section of Nephrology, University Hospital of North Norway, Tromsø, Norway; Metabolic and Renal Research Group, The Arctic University of Norway, Tromsø, Norway.
⁸ Section of Cardiovascular and Renal Research, Oslo University Hospital, Ullevål, Oslo, Norway; Institute of Clinical Medicine, Medical Faculty, University of Oslo, Oslo, Norway; Department of Medical Biochemistry, Oslo University Hospital, Ullevål, Oslo, Norway.
⁹ Section of Cardiovascular and Renal Research, Oslo University Hospital, Ullevål, Oslo, Norway; Section for Environmental and Occupational Medicine, Oslo University Hospital, Ullevål, Oslo, Norway.
¹⁰ Department of Pharmacology, Oslo University Hospital, Ullevål, Oslo, Norway; Institute of Clinical Medicine, Medical Faculty, University of Oslo, Oslo, Norway.

PMID: 35803015
DOI: 10.1016/j.jpba.2022.114908

Abstract

We developed three ultra-high pressure liquid chromatography coupled to mass spectrometry detection (UHPLC-MS/MS) methods to quantify 25 antihypertensive drugs in serum samples. Patient-reported drug lists were collected, and drug concentrations were analysed in samples from 547 patients, half with uncontrolled hypertension, and all treated with ≥ 2 antihypertensive drugs. For sample preparation, serum was mixed with deuterated internal standards and acetonitrile and precipitated. Aliquots of the supernatant were injected on UHPLC-MSMS with a C18 reversed phase column. The mobile phase was 0.1 % HCOOH (formic acid) in water and 0.1 % HCOOH in acetonitrile (except in methanol for spironolactone/canrenone) at a flow rate of 0.4 mL/min. The calibrators and internal controls were prepared in Autonorm™. The calibration ranges were wide, and the models were linear or quadratic with squared correlation coefficients ≥ 0.97. The limits of detection and quantification, specificity, carry-over, and matrix effects were acceptable. The accuracy of the internal controls was in the range 85-121 %, and the intermediate precision for all drugs was 4-28 %. The patient-reported antihypertensive drug use and the detected serum drug concentrations were in accordance with that most frequently prescribed nationally. The percent non-detectable level was 5-10 % for bendroflumethiazide, doxazosin, nifedipine, and ramipril. Often the drug dose chosen was lower than the recommended maximum daily dose. We report the maximum (C_max) and minimum (C_min) drug concentrations after drug intake. The inter-individual pharmacokinetic variability at C_min was 18-fold for hydrochlorothiazide, 22-fold for losartan carboxyl acid, 26-fold for amlodipine, 44-fold for candesartan, and 50-fold for valsartan. Our methods are suitable for measuring antihypertensive drugs in patient serum for therapy control.

Keywords: Antihypertensive drugs; Drug non-adherence; Serum drug concentration; Therapy control; Ultra-high pressure liquid chromatography coupled to mass spectrometry detection (UHPLC-MS/MS).

MeSH terms

Acetonitriles
Antihypertensive Agents*
Chromatography, High Pressure Liquid / methods
Humans
Hypertension* / drug therapy
Tandem Mass Spectrometry / methods

Substances

Acetonitriles
Antihypertensive Agents