On-line coupling of Micro-Extraction by Packed Sorbent with Sequential Injection Chromatography system for direct extraction and determination of betaxolol in human urine

• Automation of Micro-Extraction by Packet Sorbent in flow manifold.
• On-line hyphenation of MEPS and chromatography in one SIC system.
• Extending of capabilities of the Sequential Injection Chromatography.
• Automated, fast and sensitive determination of betaxolol in human urine.
• Extending of capabilities of MEPS automation in biomedical analysis.

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™ system. The chromatographic separation was performed on a monolithic High Resolution C18 (50×4.6 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 λex=220 nm and λem=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.

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