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From a Natural Product to Its Biosynthetic Gene Cluster: A Demonstration Using Polyketomycin from Streptomyces diastatochromogenes Tü6028
Published on: January 13, 2017
Electromembrane extraction of streptomycin from biological fluids
Frederik André Hansen1, Stig Pedersen-Bjergaard2
1Department of Pharmacy, University of Oslo, P.O. Box 1068 Blindern, 0316 Oslo, Norway.
Abstract:
In this fundamental study, streptomycin was extracted successfully from urine and plasma using electromembrane extraction (EME). Streptomycin is an aminoglycoside with log P -7.6 and was selected as an extremely polar model analyte. EME is a microextraction technique, where charged analytes are extracted under the influence of an electrical field, from sample, through a supported liquid membrane (SLM), and into an acceptor solution. The SLM comprised 2-nitrophenyl pentyl ether (NPPE) mixed with bis(2-ethylhexyl) phosphate (DEHP). DEHP served as ionic carrier and facilitated transfer of streptomycin across the SLM. For EME from urine and protein precipitated plasma, the optimal DEHP content in the SLM was 45-50% w/w. From untreated plasma, the content of DEHP was increased to 75% w/w in order to suppress interference from plasma proteins. Most endogenous substances with UV absorbance were not extracted into the acceptor. Proteins and phospholipids were also discriminated, with <0.6% of proteins and <0.02% of phospholipids found in the acceptor after EME. Thus, despite the fact that the SLM was permeable to more polar molecules, the EME still provided very efficient sample cleanup. Extraction process efficiencies of 98% and 61% were achieved from urine and plasma, respectively, with linear calibration (R2 > 0.9929), absence of significant matrix effects (94-112%), accuracy of 94-125%, and RSD ≤ 15% except at LLOQ. The average current during extractions was 67 µA or less. The findings of this paper demonstrated that EME is feasible for extraction of basic analytes of extreme polarity.
Insights
Electromembrane extraction (EME) successfully isolated streptomycin, a highly polar antibiotic, from biological samples like urine and plasma. This microextraction technique offers efficient cleanup and accurate quantification for polar analytes.
Area of Science:
- Analytical Chemistry
- Separation Science
- Biomedical Analysis
Background:
- Streptomycin, an aminoglycoside antibiotic, is an extremely polar analyte (log P -7.6).
- Efficient extraction of highly polar compounds from complex biological matrices like urine and plasma remains challenging.
- Electromembrane extraction (EME) is a microextraction technique utilizing an electrical field for analyte transfer.
Purpose of the Study:
- To investigate the feasibility of electromembrane extraction (EME) for isolating the highly polar analyte streptomycin.
- To optimize EME conditions for extracting streptomycin from urine and plasma samples.
- To evaluate the cleanup efficiency and analytical performance of EME for streptomycin determination.
Main Methods:
- Streptomycin extraction from urine and plasma using electromembrane extraction (EME).
- Supported liquid membrane (SLM) composed of 2-nitrophenyl pentyl ether (NPPE) and bis(2-ethylhexyl) phosphate (DEHP) as an ionic carrier.
- Optimization of DEHP content in the SLM for different sample types (urine, precipitated plasma, untreated plasma) to enhance extraction efficiency and minimize matrix interference.
Main Results:
- Successful extraction of streptomycin from urine and plasma with high process efficiencies (98% for urine, 61% for plasma).
- Effective discrimination against proteins (<0.6%) and phospholipids (<0.02%), indicating significant sample cleanup.
- Achieved linear calibration (R² > 0.9929), minimal matrix effects (94-112%), good accuracy (94-125%), and acceptable precision (RSD ≤ 15% except at LLOQ).
Conclusions:
- Electromembrane extraction (EME) is a viable and efficient technique for the extraction of extremely polar basic analytes like streptomycin from biological fluids.
- The optimized EME method provides effective sample cleanup, minimizing interference from complex biological matrices.
- EME demonstrates potential for the quantitative analysis of highly polar pharmaceuticals and other relevant compounds in clinical and forensic settings.
