Abstract
An electrically-assisted microextraction method called electromembrane extraction, followed by a simple high performance liquid chromatography and ultraviolet detection was developed and validated for determining phenobarbital in biological samples. The major parameters influencing the electromembrane extraction procedure including solvent composition, voltage, pH of acceptor and donor solutions, salt effect, and time of extraction were evaluated and optimized. The drug was extracted from the donor aqueous sample solution (pH 9) to the acceptor aqueous solution (pH 13). The donor and acceptor phases were separated by a hollow fiber dipped in 1-octanol as a supported liquid membrane. A voltage of 40 V during 20 minutes was applied as the driving force. The enrichment factor was obtained >51 which enhanced the sensitivity of the instrument. Limit of detection and limit of quantitation were 7.5 and 25 ng/ mL, respectively. The method was linear over the range of 25-1000 ng/mL for phenobarbital (R2 >0.9998) with repeatability (%RSD) between 0.4% and 6.8% (n = 3). The proposed method was successfully applied to human plasma and urine samples with relative recovery of 70-80% and %RSD < 6.8%.
Highlights
Application of new microextraction methods for analytical purposes has become an interesting alternative to conventional sample preparation methods in pharmaceutical analysis
The major parameters associated with the efficiency of electromembrane extraction (EME) were further evaluated and optimized
In order to ensure the maximum extraction efficiency, the parameters influencing on the EME procedure including membrane composition, pH of donor and acceptor solutions, applied voltage, and extraction time were evaluated, and whereby suitable conditions for maximum extraction were obtained
Summary
Application of new microextraction methods for analytical purposes has become an interesting alternative to conventional sample preparation methods in pharmaceutical analysis. Electrically-assisted microextraction or electromembrane extraction (EME) has recently been introduced as a new miniaturized sample preparation technique for pharmaceutical and biomedical analysis (Pedersen-Bjergaard, Rasmussen, 2006; Balchen et al, 2007). In this method, the charged analyte is extracted from donor solution into the acceptor solution. Considering the advantages of electromembrane extraction technique, in the present research for the first time, a rapid and simple EME-HPLC-UV method was developed and validated for quantification of phenobarbital in human plasma and urine samples. The major parameters associated with the efficiency of EME were further evaluated and optimized
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