| Description |
This thesis presents the validation results on a modified heavy metal panel assay known as HYMET4 Blood Panel. It lays out the importance of metal testing in the clinical setting, and presents the underlying motivations that compelled the improvement of the current assay. We validated the measurement of arsenic, cadmium, lead, and mercury in whole blood using an Agilent Isocratic HPLC system and autosampler as the sample introduction system, herein referred to as Isocratic Pump Direct Injection (IPDI). The autosampler accommodates two 45 vial holders that increases sample throughputs. Sample preparation and introduction have been modified as well as data analysis parameters. Validation studies conducted were imprecision, sensitivity, accuracy, Analytical Measurement Range (AMR) or linearity, recovery, and carryover. EDTA, gold, DMSA, and DMPS have been used to study mercury stability in solution. The data from the validation studies of all four metals in the panel are analyzed. The results of the imprecision, sensitivity, accuracy, AMR, recovery, and carryover studies are as follows and are well promising. Of the four chelators used for the mercury stability study, DMPS gave the best results. The validation results suggest that the modifications made to the HYMET4 Blood Panel assay have substantially improved the assay. The successful validation of the modified assay also suggests that the improved assay will increase the laboratory throughput with the use of the two 45 vial holders. It will increase sensitivity of the results using the newer analytical system. It will also reduce current sample volume by one fifth, which leads to laboratory cost saving and patient specimen volume reduction. It has been proposed based on comparison data that the use of Cetac autosampler will give a more robust touch to the improvement process of the assay. Cetac autosampler is engineered uniquely for use in trace element testing as opposed to the IPDI system engineered for biochemical analysis. |
