*3.9. Statistical Analyses*

The method was optimized and validated with a statistical treatment to increase the AF recovery and to save time and reagent waste. AFs peaks were simultaneously separated with no interfering. Significance (*p* < 0.05) of the data was analyzed by using a Student's *t*-test. The relative standard deviation (RSD) was calculated using Equation (1).

$$\text{RSD} = \text{Si} \times 100 \text{\AA} \tag{1}$$

S = standard deviation, and x = mean of the data.

**Author Contributions:** Designed the experiments, A.F.A. and J.-H.Y.; Performed the whole experiments and drafted the paper, A.F.A.; Supervised the experiments and revised the draft, J.-H.Y. All authors have read and agreed to the published version of the manuscript.

**Funding:** We thank Wendy Bedale of the Food Research Institute at the University of Wisconsin-Madison for critically reviewing the manuscript. Support for this research was provided by Food Research Institute and the University of Wisconsin-Madison Office of the Vice Chancellor for Research and Graduate Education (OVCRGE) with funding from the Wisconsin Alumni Research Foundation.

**Conflicts of Interest:** The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.
