15
Table 2: Method accuracy and precision results for FNN and NFNN extracted from mouse brain for three extracted batches using three different lots of synthetic SLE plates. Six replicates were extracted for each QC concentration level.
compared to diatomaceous earth SLE, the synthetic SLE sorbent provided higher overall analyte recoveries, improved assay reproducibility and greater phospholipid depletion for brain and cerebellum extracts. The developed SLE assay in the 96-well plate format is amenable to fast and automated sample preparation in high throughput laboratories.
References
1. S. Wang, M. Cyronak, E. Yang, J. Pharm. Biomed. Anal., 43, 2007, 701. 2. P.J. Taylor, Clin. Biochem., 38, 2005, 328.
Table 3: Method accuracy and precision results for three batches extracted from mouse cerebellum. Six replicates were extracted for each QC concentration level. Quality control samples were spiked in cerebellum homogenate, and then quantified against a calibration curve derived from brain homogenate.
3. M. Lahaie, J.N. Mess, M. Furtado, F. Garofolo, Bioanalysis, 2, 2010, 1011.
4. R. Bonfi glio, R.C. King, T.V. Olah, K. Merkle, Rapid Commun. Mass Spectrom., 13, 1999, 1175.
5. H. Jiang, H. Cao, Y. Zhang, D.M. Fast, J. Chromatogr. B., 891-892, 2012, 71.
6. L. Zhao, Agilent Technologies Publication, 5994-0949EN.
7. D. Lucas, Agilent Technologies Publication, 5994-0950EN.
For Research Use Only. Not for use in diagnostic procedures.
Table 4: Matrix effect evaluation from eight lots of brain homogenate. Three replicates were extracted at each QC concentration level.
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Conclusions
A sample preparation method using a synthetic SLE sorbent (96-well plate format, 200 mg) was developed and validated for the quantitative determination of fenfl uramine and norfenfl uramine in mouse brain, and cross-validated in mouse cerebellum. The SLE method was optimised for elution solvent, elution volume, number of elution aliquots, and sample equilibrium
time based on analyte recovery, method reproducibility and matrix effects. The developed SLE method using synthetic SLE sorbent was subsequently validated for method selectivity and sensitivity, calibration curve linearity, accuracy and precision, and matrix effect in multiple donor lots. All acceptance criteria were met for calibration curve linearity and intra- and inter-day accuracy and precision, the latter derived from three synthetic SLE sorbent lots. When
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