Simultaneous Determination of Carvedilol, Enalaprilat, and Perindoprilat in Human Plasma Using LC–MS/MS and Its Application to a Pharmacokinetic Pilot Study
A method for the extraction and quantification of carvedilol, enalaprilat, and perindoprilat in 50 µL human plasma, using high-performance liquid chromatography with tandem mass spectrometry (LC–MS/MS) detection was developed and validated. Samples were prepared via protein precipitation with chroma...
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Published in | Chromatographia Vol. 85; no. 5; pp. 455 - 468 |
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Main Authors | , , , , , , , , , |
Format | Journal Article |
Language | English |
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01.05.2022
Springer Nature B.V |
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Abstract | A method for the extraction and quantification of carvedilol, enalaprilat, and perindoprilat in 50 µL human plasma, using high-performance liquid chromatography with tandem mass spectrometry (LC–MS/MS) detection was developed and validated. Samples were prepared via protein precipitation with chromatographic separation on a Restek Ultra II Biphenyl column using gradient elution at a corresponding flowrate of 300 µL/min. Electrospray ionisation with mass detection at unit resolution in the multiple reaction monitoring (MRM) mode on an AB Sciex API 5500 mass spectrometer was used. Accuracy, precision, selectivity, sensitivity, matrix effects, recovery, process efficiency, and stability were assessed over the validation period. The assay was validated over the calibration range 0.2–200 ng/mL for all three analytes. The inter- and intra-day precision expressed as the coefficient of variation (CV) and accuracy (%Nom) all fell within acceptable limits. The overall recovery was calculated as 72.9%, 77.1%, and 77.0% for carvedilol, enalaprilat, and perindoprilat respectively, with the recovery being shown to be reproducible at the low, medium and high end of the calibration range for all three analytes. The method proved to be specific for all three analytes with no significant matrix effects observed. The validated method facilitated the analysis of carvedilol, enalaprilat, and perindoprilat in human plasma collected from adults as part of a pilot pharmacokinetic study. This validated analytical method lays the foundation for determining adherence in heart failure patients prescribed with carvedilol, enalapril and perindopril. |
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AbstractList | A method for the extraction and quantification of carvedilol, enalaprilat, and perindoprilat in 50 µL human plasma, using high-performance liquid chromatography with tandem mass spectrometry (LC–MS/MS) detection was developed and validated. Samples were prepared via protein precipitation with chromatographic separation on a Restek Ultra II Biphenyl column using gradient elution at a corresponding flowrate of 300 µL/min. Electrospray ionisation with mass detection at unit resolution in the multiple reaction monitoring (MRM) mode on an AB Sciex API 5500 mass spectrometer was used. Accuracy, precision, selectivity, sensitivity, matrix effects, recovery, process efficiency, and stability were assessed over the validation period. The assay was validated over the calibration range 0.2–200 ng/mL for all three analytes. The inter- and intra-day precision expressed as the coefficient of variation (CV) and accuracy (%Nom) all fell within acceptable limits. The overall recovery was calculated as 72.9%, 77.1%, and 77.0% for carvedilol, enalaprilat, and perindoprilat respectively, with the recovery being shown to be reproducible at the low, medium and high end of the calibration range for all three analytes. The method proved to be specific for all three analytes with no significant matrix effects observed. The validated method facilitated the analysis of carvedilol, enalaprilat, and perindoprilat in human plasma collected from adults as part of a pilot pharmacokinetic study. This validated analytical method lays the foundation for determining adherence in heart failure patients prescribed with carvedilol, enalapril and perindopril. A method for the extraction and quantification of carvedilol, enalaprilat, and perindoprilat in 50 µL human plasma, using high-performance liquid chromatography with tandem mass spectrometry (LC–MS/MS) detection was developed and validated. Samples were prepared via protein precipitation with chromatographic separation on a Restek Ultra II Biphenyl column using gradient elution at a corresponding flowrate of 300 µL/min. Electrospray ionisation with mass detection at unit resolution in the multiple reaction monitoring (MRM) mode on an AB Sciex API 5500 mass spectrometer was used. Accuracy, precision, selectivity, sensitivity, matrix effects, recovery, process efficiency, and stability were assessed over the validation period. The assay was validated over the calibration range 0.2–200 ng/mL for all three analytes. The inter- and intra-day precision expressed as the coefficient of variation (CV) and accuracy (%Nom) all fell within acceptable limits. The overall recovery was calculated as 72.9%, 77.1%, and 77.0% for carvedilol, enalaprilat, and perindoprilat respectively, with the recovery being shown to be reproducible at the low, medium and high end of the calibration range for all three analytes. The method proved to be specific for all three analytes with no significant matrix effects observed. The validated method facilitated the analysis of carvedilol, enalaprilat, and perindoprilat in human plasma collected from adults as part of a pilot pharmacokinetic study. This validated analytical method lays the foundation for determining adherence in heart failure patients prescribed with carvedilol, enalapril and perindopril. |
Author | van der Merwe, Marthinus Maartens, Gary Castel, Sandra Kellermann, Tracy Joubert, Anton Norman, Jennifer Sliwa, Karen Joubert, Andre Sinxadi, Phumla Wiesner, Lubbe |
Author_xml | – sequence: 1 givenname: Andre surname: Joubert fullname: Joubert, Andre organization: Division of Clinical Pharmacology, Department of Medicine, University of Cape Town – sequence: 2 givenname: Tracy surname: Kellermann fullname: Kellermann, Tracy organization: Division of Clinical Pharmacology, Department of Medicine, University of Cape Town, Division of Clinical Pharmacology, Department of Medicine, Stellenbosch University – sequence: 3 givenname: Anton surname: Joubert fullname: Joubert, Anton organization: Division of Clinical Pharmacology, Department of Medicine, University of Cape Town – sequence: 4 givenname: Marthinus surname: van der Merwe fullname: van der Merwe, Marthinus organization: Division of Clinical Pharmacology, Department of Medicine, University of Cape Town – sequence: 5 givenname: Jennifer surname: Norman fullname: Norman, Jennifer organization: Division of Clinical Pharmacology, Department of Medicine, University of Cape Town – sequence: 6 givenname: Sandra surname: Castel fullname: Castel, Sandra organization: Division of Clinical Pharmacology, Department of Medicine, University of Cape Town – sequence: 7 givenname: Karen surname: Sliwa fullname: Sliwa, Karen organization: Department of Medicine, Faculty of Health Sciences, Cape Heart Institute, University of Cape Town, Division of Cardiology, Department of Medicine, Groote Schuur Hospital, University of Cape Town – sequence: 8 givenname: Gary surname: Maartens fullname: Maartens, Gary organization: Division of Clinical Pharmacology, Department of Medicine, University of Cape Town – sequence: 9 givenname: Phumla surname: Sinxadi fullname: Sinxadi, Phumla organization: Division of Clinical Pharmacology, Department of Medicine, University of Cape Town – sequence: 10 givenname: Lubbe orcidid: 0000-0002-9070-8699 surname: Wiesner fullname: Wiesner, Lubbe email: lubbe.wiesner@uct.ac.za organization: Division of Clinical Pharmacology, Department of Medicine, University of Cape Town |
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Cites_doi | 10.1007/s00216-017-0552-y 10.1093/bjaceaccp/mkh042 10.1002/phar.2107 10.2165/11318100-000000000-00000 10.1365/s10337-008-0848-3 10.1111/j.1365-2125.1992.tb04006.x 10.1161/JAHA.115.002606 10.1016/j.amjcard.2006.07.015 10.1016/j.gheart.2019.07.004 10.1097/00005344-199209000-00024 10.1016/j.jnma.2020.02.003 10.1111/j.1365-2710.2005.00646.x 10.1055/b-0034-75318 10.1111/j.1365-2125.1995.tb04463.x 10.1023/B:CARD.0000025756.32499.6f 10.1016/j.jchromb.2005.06.012 10.3810/pgm.2013.07.2687 10.1016/j.drudis.2020.08.005 10.1093/eurheartj/ehab368 10.1016/j.jpba.2006.12.017 10.1021/ac020361s 10.1016/j.jchromb.2017.06.029 10.2147/rrcc.s50658 10.1016/S0011-393X(04)90003-3 10.1177/00912700022009576 |
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Keywords | Adherence Perindoprilat LC–MS/MS Carvedilol Heart failure (HF) Enalaprilat |
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SubjectTerms | Analytical Chemistry Blood plasma Calibration Chemistry Chemistry and Materials Science Chromatography Coefficient of variation High performance liquid chromatography Human performance Ions Laboratory Medicine Mass spectrometry Mathematical analysis Original Pharmacokinetics Pharmacology Pharmacy Proteomics Recovery Selectivity Stability analysis |
Title | Simultaneous Determination of Carvedilol, Enalaprilat, and Perindoprilat in Human Plasma Using LC–MS/MS and Its Application to a Pharmacokinetic Pilot Study |
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