Matrix effects initial testing

The diligent analyst would develop a robust method with rigorous matrix effect tests on multiple lots, including hemolyzed and lipidemic samples. An initial test would be a spike-recovery evaluation on at least six individual lots. Samples should be spiked at or near the LLOQ, and at a high level near the ULOQ. If matrix interference were indicated by unacceptable relative error (RE) percentage in certain lots, the spiked sample of the unacceptable lots should be diluted with the standard calibrator matrix to estimate the minimum dilution requirement (MDR) at and above which the spike-recovery is acceptable. The spike-recovery test should then be repeated with the test samples diluted at the MDR. Note that this approach will increase the LLOQ for a less sensitive assay. If sensitivity is an issue, then other venues will be required to address the matrix effect problem. For example, the method can be modified to include sample clean-up, antibodies and/or assay conditions may be changed, or the study purpose may be tolerable to acknowledge that the method may not be selective for a few patients (whose data may require special interpretation). 

Initial Test of Matrix Effects Assay matrices are typically the most troublesome component in LBA. There should be careful consideration of this variable during method validation. Two types of tests are used to address the two major concerns surrounding matrix effects. These are performed in consideration of (1) whether there is a matrix difference between the standards and anticipated study samples that impacts the relative accuracy of an assay and (2) whether there are inter-individual or disease-specific differences in matrix in the target patient population. Two types of tests are used to evaluate such matrix effects spike recovery, where known amounts of analyte are mixed ( spiked ) into characterized matrix, and parallelism in patient samples. However, limited availability of patient samples may prevent the latter testing during the method feasibility phase. 

Alternatively (or initially) the mixture is treated as a whole and tested in its crude state. The advantage of this strategy includes the relevancy of the tested sample to its environmental counterpart, decreased potential for artefact formation, and inclusion of combined effects of chemicals in the mixture. Moreover if the mixture is representative of others in its class (e.g., diesel emissions from different sources would share certain characteristics), it may be possible to extrapolate results across samples. This method also circumvents the labor-intensive process of individual testing of multiple chemicals. But sometimes a complex mixture is too cytotoxic to be tested directly in a bioassay. Furthermore, it may be incompatible with the test system because of the physical matrix. Other disadvantages include the inability to specify the constituent of the mixture responsible for the toxicity, as well as potential masking effects (e.g., the masking of mutagenicity by cytotoxicity). 

While the estimates of the autocorrelation coefficients for the Cg time series (lower rows in 1 to ordy change slightly, the estimates the autocorrelation coefficients for the Benzene time series (upper rows in to 3) are clearly affected since three parameters are dropped from the model. The remaining coefficients are affected, too. In particular, the lagged cross-correlations to the Cg time series change from 1.67 to 2.51 and from -2.91 to -2.67 (right upper entries in 1 and This confirms the serious effect of even unobtrusive outliers in multivariate times series analysis. By incorporating the outliers effects, the model s AIC decreases from -4.22 to -4.72. Similarly, SIC decreases from -4.05 to -4.17. The analyses of residuals. show a similar pattern as for the initial model and reveal no serious hints for cross- or auto-correlation. i Now, the multivariate Jarque-Bera test does not reject the hypothesis of multivariate normally distributed variables (at a 5% level). The residuals empirical covariance matrix is finally estimated as... 

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