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Multiplex Therapeutic Drug Monitoring by Isotope-dilution HPLC-MS/MS of Antibiotics in Critical Illnesses
Published on: August 30, 2018
Analytical method for the determination of usually prescribed antibiotics in human nails using UHPLC-MS/MS.
María Del Carmen Gómez-Regalado1, Lydia Espín-Moreno1, Laura Martín-Pozo2
1Department of Analytical Chemistry, University of Granada, E-18071 Granada, Spain.
Abstract:
Antibiotics are a group of drugs used for the treatment of bacterial diseases. They are used in both human and veterinary medicine and, although they are not permitted, they are sometimes used as growth promoters. The present research compares two extraction techniques: ultrasound-assisted extraction (UAE) and microwave-assisted extraction (MAE) in order to evaluate their efficiency in the determination of 17 usually prescribed antibiotics in human nails. The extraction parameters were optimized using multivariate techniques. Once both techniques were compared, MAE was selected as optimal due to its greater experimental practicability together with the better extraction efficiencies it provides. Target analytes were detected and quantified by ultra-high performance liquid chromatography with tandem mass spectrometry detection (UHPLC-MS/MS). The run time was 20 min. The methodology was then successfully validated, obtaining acceptable analytical parameters according to the guide used. Limits of detection were between 0.3 and 3 ng g-1 and limits of quantification were in the range from 1.0 to 4.0 ng g-1. Recovery percentages ranged from 87.5% to 114.2%, and precision (in terms of standard deviation) was less than 15% in all cases. Finally, the optimized method was applied to nails taken from 10 volunteers and the results revealed the presence of one or more antibiotics in all the samples examined. The most commonly found antibiotic was sulfamethoxazole, followed by danofloxacin and levofloxacin. The results demonstrated, on the one hand, the presence of these compounds in the human body and, on the other hand, the suitability of nails as a non-invasive biomarker of exposure.
Insights
Human nails can detect common antibiotics like sulfamethoxazole, danofloxacin, and levofloxacin. This study found nails are a suitable non-invasive biomarker for antibiotic exposure monitoring.
Area of Science:
- Analytical Chemistry
- Environmental Science
- Forensic Toxicology
Background:
- Antibiotics are crucial for treating bacterial infections in humans and animals.
- Non-permitted use of antibiotics as growth promoters raises public health concerns.
- Human nails offer a potential non-invasive matrix for detecting drug exposure.
Purpose of the Study:
- To compare ultrasound-assisted extraction (UAE) and microwave-assisted extraction (MAE) for antibiotic determination in human nails.
- To optimize extraction parameters using multivariate techniques.
- To validate a method for quantifying 17 prescribed antibiotics in human nail samples.
Main Methods:
- Microwave-assisted extraction (MAE) was optimized and selected over UAE for its efficiency and practicality.
- Antibiotics were quantified using ultra-high performance liquid chromatography with tandem mass spectrometry (UHPLC-MS/MS).
- Method validation included assessment of limits of detection (LOD), quantification (LOQ), recovery, and precision.
Main Results:
- MAE provided better extraction efficiencies and experimental practicability compared to UAE.
- The validated UHPLC-MS/MS method achieved LODs between 0.3-3 ng/g and LOQs of 1.0-4.0 ng/g.
- All 10 volunteer samples contained one or more antibiotics, with sulfamethoxazole being the most prevalent.
Conclusions:
- Human nails serve as a suitable non-invasive biomarker for assessing exposure to commonly prescribed antibiotics.
- The developed MAE coupled with UHPLC-MS/MS method is effective for antibiotic determination in nail samples.
- Widespread presence of antibiotics in the human body highlights the importance of monitoring exposure through accessible biomarkers.
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