On-line coupling of Micro-Extraction by Packed Sorbent with Sequential Injection Chromatography system for direct extraction and determination of betaxolol in human urine
Abstract
A novel approach for automation of Micro-Extraction by Packed Sorbent (MEPS), a solid phase extraction technique, is presented, enabling precise and repeatable liquid handling due to the employment of sequential injection technique. The developed system was used for human urine sample clean-up and pre-concentration of betaxolol before its separation and determination.
A commercial MEPS C-18 cartridge Was integrated into an SIChrom (TM) system. The chromatographic separation was performed on a monolithic High Resolution C18 (50 x 4.6 mm) column which was coupled on-line in the system with Micro-Extraction using an additional selection valve.
A mixture of acetonitrile and aqueous solution of 0.5% triethylamine with acetic acid, pH adjusted to 4.5 in ratio 30:70 was used as a mobile phase for elution of betaxolol from MEPS directly onto the monolithic column where the separation took place. Betaxolol was quantified by a fluorescence detector at wavelengths lambda(ex)= 220 nm and lambda(em)=305 nm.
The linear calibration range of 5-400 ng mL(-1), with limit of detection 1.5 ng mL(-1) and limit of quantification 5 ng mL(-1) and correlation r= 0.9998 for both the standard and urine matrix calibration were achieved. The system recovery was 105 +/- 5%; 100 +/- 4%; 108 +/- 1% for three concentration levels of betaxolol in 10 times diluted urine - 5, 20 and 200 ng mL(-1), respectively.