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LC-MS/MS method for the simultaneous determination of clarithromycin, rifampicin and their main metabolites in horse plasma, epithelial lining fluid and broncho-alveolar cells.
Abstract: Clarithromycin (CLA) is a well established macrolide antibiotic which is frequently used in therapy of airway diseases in foals. It is extensively metabolized by CYP3A4 resulting in the antimicrobial active metabolite 14-hydroxyclarithromycin (OH-CLA). Rifampicin (RIF) is often comedicated to prevent resistance and augment therapy. RIF is a known inducer for metabolizing enzymes and transporter proteins. Therefore, comedication might bare the risks of pharmacokinetic drug interactions which were investigated in a clinical trial. As no adequate method to determine CLA, RIF and their main metabolites OH-CLA and 25-O-desacetylrifampicin (DAc-RIF) were described so far, we developed a selective and sensitive assay to measure concentrations of all four substances simultaneously in plasma, epithelial lining fluid (ELF) and broncho-alveolar cells (BAC) of foals. Drugs were measured after extraction with methyl tert-butyl ether using roxithromycin as internal standard and liquid chromatography coupled with tandem mass spectrometry (LC-MS/MS) for detection. The chromatography was done isocratically using 25mM ammonium acetate buffer (pH 4)/acetonitrile (45%/55%, flow rate 200μl/min). The MS/MS analysis was performed in the positive ion mode (m/z transitions: CLA, 748.5-590.1; OH-CLA, 764.1-606.1; RIF, 823.1-791.2; DAc-RIF, 781.1-749.1 and 837.3-679.2 for the internal standard). The method was validated according to selectivity, linearity, accuracy, precision, recovery, matrix effects and stability. The validation ranges for all substances were 2.5-25 for the low and 25-250ng/ml for the high validation range. The described assay was shown to be valid and successfully applied to measure disposition of CLA, OH-CLA, RIF and DAc-RIF in plasma, ELF and BAC of foals in a clinical trial.
Copyright © 2011 Elsevier B.V. All rights reserved.
Publication Date: 2011-01-22 PubMed ID: 21310577DOI: 10.1016/j.jpba.2011.01.019Google Scholar: Lookup The Equine Research Bank provides access to a large database of publicly available scientific literature. Inclusion in the Research Bank does not imply endorsement of study methods or findings by Mad Barn.
- Clinical Trial
- Journal Article
- Research Support
- Non-U.S. Gov't
- Validation Study
Summary
This research summary has been generated with artificial intelligence and may contain errors and omissions. Refer to the original study to confirm details provided. Submit correction.
This study explores the development of a new LC-MS/MS method for simultaneous detection of two key antibiotics (Clarithromycin and Rifampicin) and their metabolites in horse plasma, epithelial lining fluid, and broncho-alveolar cells.
Introduction and Background
- This study pertains to the development of a specific chemical assay to measure the concentration of two antibiotics, Clarithromycin (CLA) and Rifampicin (RIF), and their main metabolic byproducts (14-hydroxyclarithromycin and 25-O-desacetylrifampicin) in various samples drawn from foals, including plasma, broncho-alveolar cells (BAC), and epithelial lining fluid (ELF).
- CLA, a popular macrolide antibiotic, is typically used in treating airway diseases in young horses. While it gets considerably metabolized into the antimicrobial agent, 14-hydroxyclarithromycin, RIF is usually co-administered to enhance the effectiveness of the therapy and mitigate possible drug resistance.
- Nevertheless, the combined administration of CLA and RIF runs the accompanying risk of unfavorable pharmacokinetic drug interactions, which the researchers assessed within a clinical trial.
Methods and Techniques
- Initially, the scientists devised a novel sensitive assay, as none of the previous techniques were adequate for the simultaneous determination of CLA, RIF, and their key metabolites.
- In achieving this, the drugs were initially extracted with methyl tert-butyl ether and roxithromycin was used as the internal standard.
- Detection was carried out via liquid chromatography combined with tandem mass spectrometry (LC-MS/MS).
- Detailed settings for the chromatography and MS/MS analysis are also provided, including the specifics around the isocratic method, buffer system, flow rate, and positive ion mode for the MS/MS process.
Validation and Application
- The developed method was validated based on parameters like selectivity, accuracy, precision, linearity, recovery, matrix effects, and stability.
- The validation ranges for all substances were set between specific units for both low and high validation intervals.
- Upon validation, the devised process turned out to be precise and successfully used for measuring the disposition of CLA, OH-CLA, RIF, and DAc-RIF within plasma, BAC, and ELF of foals as a part of a clinical experiment.
Cite This Article
APA
Oswald S, Peters J, Venner M, Siegmund W.
(2011).
LC-MS/MS method for the simultaneous determination of clarithromycin, rifampicin and their main metabolites in horse plasma, epithelial lining fluid and broncho-alveolar cells.
J Pharm Biomed Anal, 55(1), 194-201.
https://doi.org/10.1016/j.jpba.2011.01.019 Publication
Researcher Affiliations
- Department of Clinical Pharmacology, Ernst-Moritz-Arndt University Greifswald, Greifswald, Germany. stefan.oswald@uni-greifswald.de
MeSH Terms
- Animals
- Anti-Bacterial Agents / blood
- Anti-Bacterial Agents / metabolism
- Anti-Bacterial Agents / pharmacokinetics
- Biotransformation
- Bronchioles / cytology
- Bronchioles / metabolism
- Bronchoalveolar Lavage Fluid / chemistry
- Bronchoalveolar Lavage Fluid / cytology
- Chromatography, High Pressure Liquid / veterinary
- Clarithromycin / analogs & derivatives
- Clarithromycin / blood
- Clarithromycin / metabolism
- Clarithromycin / pharmacokinetics
- Drug Interactions
- Drug Stability
- Horse Diseases / blood
- Horse Diseases / drug therapy
- Horses
- Limit of Detection
- Pulmonary Alveoli / cytology
- Pulmonary Alveoli / metabolism
- Reproducibility of Results
- Respiratory Tract Infections / blood
- Respiratory Tract Infections / drug therapy
- Respiratory Tract Infections / veterinary
- Rifampin / analogs & derivatives
- Rifampin / blood
- Rifampin / metabolism
- Rifampin / pharmacokinetics
- Spectrometry, Mass, Electrospray Ionization / veterinary
- Tandem Mass Spectrometry / veterinary
Citations
This article has been cited 9 times.- Liu L, Zhang L, Zheng X, Liu X, Liu W, Wu J. LC-MS/MS-based multiplex antibacterial platform for therapeutic drug monitoring in intensive care unit patients. Front Pharmacol 2023;14:1116071.
- Kozak J, Tyszczuk-Rotko K, Wójciak M, Sowa I, Rotko M. First Screen-Printed Sensor (Electrochemically Activated Screen-Printed Boron-Doped Diamond Electrode) for Quantitative Determination of Rifampicin by Adsorptive Stripping Voltammetry. Materials (Basel) 2021 Jul 29;14(15).
- Kobuchi S, Aoki M, Inoue C, Murakami H, Kuwahara A, Nakamura T, Yasui H, Ito Y, Takada K, Sakaeda T. Transport of Azithromycin into Extravascular Space in Rats. Antimicrob Agents Chemother 2016 Nov;60(11):6823-6827.
- Shimomura H, Andachi S, Aono T, Kigure A, Yamamoto Y, Miyajima A, Hirota T, Imanaka K, Majima T, Masuyama H, Tatsumi K, Aoyama T. Serum concentrations of clarithromycin and rifampicin in pulmonary Mycobacterium avium complex disease: long-term changes due to drug interactions and their association with clinical outcomes. J Pharm Health Care Sci 2015;1:32.
- Patil JS, Suresh S, Sureshbabu AR, Rajesh MS. Development and validation of liquid chromatography-mass spectrometry method for the estimation of rifampicin in plasma. Indian J Pharm Sci 2011 Sep;73(5):558-63.
- Zouari M, Cetinkaya A, Ozkan SA. Comparative analysis of LC-MS/MS and electrochemical immunosensor for clarithromycin detection in plasma. Mikrochim Acta 2025 Oct 28;192(11):758.
- Mostafa EA, Azim MA, ElZaher AA, ElKady EF, Fouad MA, Ghazy FH, Radi EA, El Makarim Saleh MA, El Kerdawy AM. Correlating physico-chemical properties of analytes with Hansen solubility parameters of solvents using machine learning algorithm for predicting suitable extraction solvent. Sci Rep 2024 Aug 13;14(1):18741.
- Du Y, Yu L, Deng B, Li Q, Hu J, Li L, Xu Y, Song L, Xie F, Wang Y, Chen Y, Liu C, Zhai X, Lu Y. Pharmacokinetic Interactions Between Tegoprazan and the Combination of Clarithromycin, Amoxicillin and Bismuth in Healthy Chinese Subjects: An Open-Label, Single-Center, Multiple-Dosage, Self-Controlled, Phase I Trial. Clin Drug Investig 2024 May;44(5):343-355.
- Shen L, Liao K, Yang E, Yang F, Lin W, Wang J, Fan S, Huang X, Chen L, Shen H, Jin H, Ruan Y, Liu X, Zeng G, Xu JF, Pi J. Macrophage targeted iron oxide nanodecoys augment innate immunological and drug killings for more effective Mycobacterium Tuberculosis clearance. J Nanobiotechnology 2023 Oct 10;21(1):369.
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