科研之友
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摘要
Background: Already available sample preparation technologies for liquid chromatography-tandem mass spectrometry have substantial shortcomings with respect to automation. A novel approach is based on gel-like polymeric material with defined absorption chemistry, which is immobilized in micro-plate wells. It is referred to as Tecan Immobilized Coating Extraction (TM) (TICE (TM)) technology and it enables easy automation on liquid handling systems. We aimed to study the performance of Tecan AC Extraction Plate (TM) based on this principle by addressing 25-hydroxyvitamin D (25OHD) as an exemplary analyte. Methods: A protocol for extraction of 25OHD from serum samples based on TICE (TM) technology was implemented on a robotic liquid handling system Freedom EVO (R) (Tecan). An isotope-dilution ultra-performance liquid chromatography-tandem mass spectrometry method was used for quantification. Performance was tested according to a comprehensive protocol. Results: Linearity was found over a range from 4.3 to 65.8 ng/mL for 25OHD(3). The coefficients of variation for the intra-day and inter-day precision were <6% and accuracy ranged between 96.9% and 99.8% for 25OHD(3). Recovery was 84% and efficient control of matrix effects was verified. High sample throughput could be observed with 96 samples prepared in <60 min. Close agreement of results was found for clinical samples analyzed with a second tandem mass spectrometry method based on protein precipitation and two-dimensional ultra-performance liquid chromatography for sample preparation (r = 0.988, n = 73). Conclusions: The new TICE (TM) technology was found to be a useful process for sample preparation in clinical mass spectrometry. Full automation suited for routine analysis was achieved.
- 出版日期2014-3
