In vivo solid phase microextraction sampling of human saliva for non-invasive and on-site monitoring

• 5 min invivo SPME sampling of saliva offers unbiased on-site monitoring.
• TF-SPME–LC–MS provides fast determination of prohibited substances in saliva.
• TF-SPME provides very good sample clean-up, preventing matrix effect.
• The TF-SPME–LC–MS method offers enough sensitivity to detect steroids in saliva.

On-site sample preparation is an analytical approach based on direct sampling from the system under investigation. It has the advantage of combining sampling and sample preparation into a single step, thus generally is fast, minimizes the potential sources of error and eliminates the risks for analytes instability. For such analysis solid phase microextraction in thin film geometry (TF-SPME) can provide robust and convenient in vivo sampling, offering in the same time faster analysis and higher extraction recovery (i.e., better sensitivity) due to large surface to volume ratio.In this study, TF-SPME in coated blade and membrane formats with a single extraction phase were used for in vivo and ex vivo saliva extraction and separation by LC and GC, respectively. Due to applicability for wide range of polarity of analytes as well as thermal and solvent stability during the desorption, hydrophilic lipophilic balanced particles (HLB) were chosen as extraction phase and used for fast (5 min) in vivo and ex vivo sampling. The results of metabolomic profiling of the saliva are indicating that even 5 min in vivo sampling using TF-SPME followed by GC and LC analyses provides complementary coverage of wide range of analytes with different physical and chemical properties. To demonstrate the applicability of the method for doping analyses, the SPME–LC–MS/MS method was validated for simultaneous quantification of 49 prohibited substances with limit of quantification (LOQ) ranging between 0.004 and 0.98 ng mL−1. Moreover, the method was also validated and successfully applied for determination of endogenous steroids in saliva where the concentrations of the analytes are substantially low. The developed assay offers fast and reliable multiresidue analysis of saliva as an attractive alternative to the standard analysis methods.

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