## 3 Results
### Metabolite Variance across Multiwell Fluidic System to Ascertain Sites of Generated Cell Byproducts
#### 3.8.1 Trending Relevance  
Study outcomes of discrete, continuous, and trending models are displayed in Figures 10-12; LDH (Figure 10), CYP3A4 (Figure 11), and GSH (Figure 12). In figures, vertical columns are categorized (a1-a3) HepaRGs exposed to ASA, (b1-b3) rat hepatocytes exposed to ASA, (c1-c3) HepaRGs exposed to NEF, and (d1-d3) rat hepatocytes exposed to NEF. Within figures, horizontal rows are demarcated by a1-\(\upmu\)d1, a2-\(\upmu\)d2, and a3-\(\upmu\)d3 as defined in Table 5. From the same SOP, trending effects of DMSO vehicle, additional exposure compounds, and additional assays are corroborated in Appendix A with Figure A1 and in the supplemental section as illustrated in supplemental Figure S1. LDH ASSAY--Functional Indicator (Studies 1-4)  
LDH is found in cells. LDH discharge is used as a biomarker of cell cytotoxicity as release can increase mitochondrial damage [19]. **ASA exposure on HepaRG cells (Study 1)** is displayed in Figure 10(a1). LDH outcomes, for DMSO vs. ASA, are non-significant (ns) as revealed in wells 3-11. **ASA exposure on rat hepatocytes (Study 2)** is displayed in Figure 10(b1). ASA induces LDH release in upstream well 3 (*) and downstream wells 10 (*) and 11 (*). **NEF exposure on HepaRG cells (Study 3)** is displayed in Figure 10(c1). NEF stimulates LDH in upstream wells 3 (***), 4 (***), 5 (*), and 6 (***); thereafter, wells \begin{table}
\begin{tabular}{p{42.7 pt} p{113.8 pt} p{113.8 pt}} \hline \hline \multicolumn{1}{c}{In Brief (Figure 9(b1–b3)):} & \multicolumn{1}{c}{3\(\upsigma\) UCL is minimum at 24 h and increases over time to indicate that cell-generated APAP-GSH has the highest variance at later APAP exposure periods with both upstream sites and prolonged exposure times augmenting downstream device sites with conditioned media; Figure 9b3 wells 9 and 10.} \\ \hline
**APAP-GLU** & Highest MR @ 72 h & Most well–well variations are 68.1 \(\upmu\)M \& 48.6 \(\upmu\)M occurring at 72 h in downstream wells 9 and 10 (Figure 9(c3)). \\ \hline Mean MR @ 72 h & Uniform well–well \(\upmu\)s are present in upstream wells 4–8 (0.91–\(\upmu\)21.24 \(\upmu\)M) and downstream well 11 (3.9 \(\upmu\)M) (Figure 9c3). \\ \hline Negligeable MR @ 24 h & Limited well–well \(\upmu\) found in upstream wells 4 and 5 (–0.0 \(\upmu\)M) (Figure 9(c1)). \\ \hline MR Compilations @ 24, 48, 72 h & 24 h data mean is the lowest (9.0 \(\upmu\)M) with UCL at the lowest distinction (30.0 \(\upmu\)M) to drive least variation as across device culture wells; Figure 9(c1). \\  & 48 h data mean is midrange (21.0 \(\upmu\)M) with UCL also a midrange distinction (79.0 \(\upmu\)M); Figure 9(c2). MRs also display notably high variations in culture wells 8 and 9 (63.5 \(\upmu\)M and 34.7 \(\upmu\)M). \\  & 72 h mean (23.0 \(\upmu\)M) and the UCL (92.0 \(\upmu\)M) are the highest when compared alongside G-24 h and Glu-48 h; Figure 9(c1–c3). \\ \hline In Brief (Figure 9(c1–c3)): & 3\(\upsigma\) UCL is minimum at 24 h and increases over time to indicate that cell-generated APAP-Glu has high variance at both intermediate and late APAP exposure periods (Figure 9(c2) wells 8–9 and Figure 9(c3) wells 9–10). \\ \hline \hline \end{tabular}
\end{table} 
Table 4: _Cont._7-11 register as non-significant cell function activities. **NEF exposure on rat hepatocytes (Study 4)** is displayed in Figure 10(d1). NEF stimulates LDH in upstream well 3 (***), midstream well 7 (***), and downstream wells 10 (**) and 11 (*). Interior wells 4-6 and 8-9 register as non-significant. For Studies 1-4, LDH trending responses are further summarized in Table 6.  
MR control charts) to display functional fold-changes amid adjacent wells. (**a3--HepARG line graph**) ASA does not induce a trending variation. (**b3--rat hepatocyte line graph**) ASA stimulates a process shift at upstream well 3 and special cause in downstream wells 10 and 11. (**c3--HepARG line graph**) NEF induces a trending process shift at upstream wells 4-7.