Biocompatible in-tube solid phase microextraction coupled with liquid chromatography-fluorescence detection for determination of interferon α in plasma samples

• Published in: Journal of Chromatography A Vol. 1218; no. 21; pp. 3376 - 3381
• Main Authors: Chaves, Andréa R., Silva, Bruno J.G., Lanças, Fernando M., Queiroz, Maria Eugênia C.
• Format: Journal Article Conference Proceeding
• Language: English
• Published: Amsterdam Elsevier B.V 27-05-2011; Elsevier
• Subjects: Analysis; Automated; Biocompatibility; Biocompatible Materials - chemistry; Biological and medical sciences; Chromatography; Chromatography, Liquid - methods; Drugs; General pharmacology; Humans; In-tube SPME; Interferon; Interferon alpha 2a; Interferon-gamma - blood; Interferon-gamma - isolation & purification; Linear Models; Liquid chromatography; Macromolecules; Medical sciences; Monitoring; Pharmaceutical Preparations - chemistry; Pharmaceutical Preparations - isolation & purification; Pharmacology. Drug treatments; Plasma sample; Recombinant Proteins; Reproducibility of Results; Restricted access material; Solid Phase Microextraction - instrumentation; Solid Phase Microextraction - methods; Spectrometry, Fluorescence; Interferon alpha 2a; Restricted access material; Plasma sample; In-tube SPME; Trace analysis; Validation; Biological fluid; Automation; Fluorescence detector; Chemical analysis; Restricted access medium column; Alpha interferon; Cytokine; Interferon alpha; HPLC chromatography; Blood; Chemical enrichment; Blood plasma; Solid phase microextraction; Therapeutic drug monitoring; Sample preparation; Biocompatibility; Quantitative analysis
• ISSN: 0021-9673; 1873-3778
• DOI: 10.1016/j.chroma.2010.11.039

The present work demonstrates the successful application of automated biocompatible in-tube solid-phase microextraction coupled with liquid chromatography (in-tube SPME/LC) for determination of interferon alpha
2a (IFN α
2a) in plasma samples for therapeutic drug monitoring. A restricted access material (RAM, protein-coated silica) was employed for preparation of a lab-made biocompatible in-tube SPME capillary that enables the direct injection of biological fluids as well as the simultaneous exclusion of macromolecules by chemical diffusion barrier and drug pre-concentration. The in-tube SPME variables, such as sample volume, draw/eject volume, number of draw–eject cycles, and desorption mode were optimized, to improve the sensitivity of the proposed method. The IFN α
2a analyses in plasma sample were carried out within 25
min (sample preparation and LC analyses). The response of the proposed method was linear over a dynamic range, from 0.06 to 3.0
MIU
mL
−1, with correlation coefficient equal to 0.998. The interday precision of the method presented coefficient of variation lower than 8%. The proposed automated method has adequate analytical sensitivity and selectivity for determination of IFN α
2a in plasma samples for therapeutic drug monitoring.