Development and validation of an LC‒MS/MS method for the determination of cyclocreatine phosphate and its related endogenous biomolecules in rat heart tissues.

Autor: Abo-Elmagd IF; Bioanalysis Research Group, School of Pharmacy, New Giza University, Giza, 12256, Egypt., Mahmoud AM; Analytical Chemistry Department, Faculty of Pharmacy, Cairo University, Cairo, 11562, Egypt. amr.bekhet@pharma.cu.edu.eg.; Department of Chemistry, School of Pharmacy, Newgiza University (NGU), New Giza, Km 22 Cairo-Alex Road, Cairo, Egypt. amr.bekhet@pharma.cu.edu.eg., Al-Ghobashy MA; Bioanalysis Research Group, School of Pharmacy, New Giza University, Giza, 12256, Egypt.; Analytical Chemistry Department, Faculty of Pharmacy, Cairo University, Cairo, 11562, Egypt., Nebsen M; Analytical Chemistry Department, Faculty of Pharmacy, Cairo University, Cairo, 11562, Egypt., Rabie MA; Department of Pharmacology and Toxicology, Faculty of Pharmacy, Cairo University, Cairo, 11562, Egypt.; Faculty of Pharmacy and Drug Technology, Egyptian Chinese University (ECU), Cairo, 19346, Egypt., Mohamed AF; Department of Pharmacology and Toxicology, Faculty of Pharmacy, Cairo University, Cairo, 11562, Egypt.; Faculty of Pharmacy, King Salman International University (KSIU), South Sinai Government, Ras Sedr city, 46612, Egypt., Ahmed LA; Department of Pharmacology and Toxicology, Faculty of Pharmacy, Cairo University, Cairo, 11562, Egypt., El Sayed NS; Department of Pharmacology and Toxicology, Faculty of Pharmacy, Cairo University, Cairo, 11562, Egypt., Arafa RK; Biomedical Sciences Program & Drug Design and Discovery Lab, Zewail City of Science and Technology, Cairo, 12578, Egypt., Todd R; ProChem International, LLC, Sheboygan, Wisconsin, USA., Elgebaly SA; Nour Heart, Inc., Vienna, Virginia, USA.; University of Connecticut, Faculty of Medicine, Farmington, Connecticut, USA.
Jazyk: angličtina
Zdroj: BMC chemistry [BMC Chem] 2024 Nov 04; Vol. 18 (1), pp. 214. Date of Electronic Publication: 2024 Nov 04.
DOI: 10.1186/s13065-024-01304-1
Abstrakt: The cardioprotective drug cyclocreatine phosphate has been awarded Food and Drug Administration-orphan drug designation for the prevention of ischemic injury to enhance cardiac graft recovery and survival in heart transplantation. Cyclocreatine phosphate is the water-soluble derivative of cyclocreatine. Estimating the levels of Cyclocreatine phosphate, Adenosine triphosphate, Creatine Phosphate, Creatine and Cyclocreatine helps us in understanding the energy state as well as evaluating the heart cells' function. The quantification of endogenous compounds imposes a challenging task for analysts because of the absence of a true blank matrix, whose use is required according to international guidelines. Recently, the International Council for Harmonization issued a new guideline that contains guidance on the validation of methods used to quantify endogenous components, such as the background subtraction approach that was employed in our current study. Specifically, we developed and validated a sensitive, reliable and accurate liquid chromatography-tandem mass spectrometry assay to determine simultaneously the levels of mentioned endogenous compounds in rat heart tissue. Tissue samples were prepared by protein precipitation extraction using water: methanol (1:1). Using Ultra Performance Liquid Chromatography, Chromatographic separation was achieved with ZORBAX Eclipse Plus C18 4.6 × 100 mm,3.5 μm column and conditions as following: ammonium acetate (pH 8.5): acetonitrile, 70:30 mobile phase, 0.7 mL/min flow rate and 25 °C temperature. Electrospray ionization mass detector with Multiple reaction monitoring mode was then employed, using both positive and negative modes, Analysis was carried out using 5.00-2000.00 ng/mL linear concentration range within 2 min for each analyte. According to Food and Drug Administration guidelines for bioanalytical methods, validation was carried out. We investigated the matrix effect, recovery efficiency and process efficiency for the analyte in neat solvent, postextraction matrix and tissue. The results stated mean percentage recoveries higher than 99%, accuracy 93.32-111.99%, and Relative Standard Deviation (RSD) below 15% within the concentration range of our study which indicated that target analytes' stability in their real matrix is sufficient under the employed experimental conditions.
(© 2024. The Author(s).)
Databáze: MEDLINE
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