{"id":11,"date":"2024-04-04T00:39:46","date_gmt":"2024-04-03T21:39:46","guid":{"rendered":"https:\/\/sisu.ut.ee\/lcms_method_validation\/104-experimental-design\/"},"modified":"2024-04-04T00:41:47","modified_gmt":"2024-04-03T21:41:47","slug":"104-experimental-design","status":"publish","type":"page","link":"https:\/\/sisu.ut.ee\/lcms_method_validation\/104-experimental-design\/","title":{"rendered":"10.4 Experimental design"},"content":{"rendered":"<p>\r\n\tAdditionally to the One factor at a Time approach (OFAT, explained in <a data-url=\"https:\/\/sisu.ut.ee\/lcms_method_validation\/103-different-ways-evaluate-robustness\" href=\"https:\/\/sisu.ut.ee\/lcms_method_validation\/103-different-ways-evaluate-robustness\" target=\"_blank\" title=\"\" rel=\"noopener\">10.3<\/a>), robustness can also <span><span><span style=\"color:#2f2f2f\">be estimated by varying multiple method parameters (factors) at the same time. Such approaches are strongly recommended if it is expected that different method parameters have <i>interactions<\/i>: meaning that the value of one parameter influences another value of a different parameter. In chromatography, the parameter interactions between some of the parameters are rather the rule than the <\/span><\/span><\/span><span><span>exception <\/span><\/span><span><span>[<\/span><\/span><span><span>ref 65]. Therefore, using OFAT can lead to getting only a partial and\/or flawed picture of the method robustness<span style=\"color:#2f2f2f\">. For example, the retention behaviour of the acidic compounds in RP is influenced by the mobile phase pH. Additionally, changing the additive concentration can at the same time strongly influence pH and, in turn, changing pH can influence the effect of the additive. These interactions will remain unnoticed with One factor at a Time approaches.<\/span><\/span><\/span>\r\n<\/p>\r\n\r\n<p>\r\n\t<span><span><span style=\"color:#2f2f2f\">A statistical approach to validation procedures is the Design of Experiment (DoE). This tool has been broadly applied for many years in method optimization but is also relevant for the method <\/span><\/span><\/span><span><span>validation process. Even though a DoE is not yet prominently illustrated in the validation guidelines for analytical method development, publications in recent years have shown [ref 66 and ref 67] that it is on its way to becoming a valuable mainstream tool for evaluating robustness. Additionally, a DoE chapter has been added to the 10th version of <span style=\"color:#2f2f2f\">the European Pharmacopoeia (Ph. Eur.) and will certainly influence the future of validation guidelines.<\/span><\/span><\/span>\r\n<\/p>\r\n\r\n<h3 style=\"margin-bottom: 7.5pt;text-align: justify\">\r\n\t<span style=\"background:white\"><span style=\"line-height:normal\"><span><span style=\",serif\"><span style=\"color:#2f2f2f\">How to perform a DoE<\/span><\/span><\/span><\/span><\/span>\r\n<\/h3>\r\n\r\n<h4 style=\"text-align: center\">\r\n\t<div class=\"ratio ratio-16x9 mb-3\"><div class=\"video-placeholder-wrapper video-placeholder-wrapper--16x9\">\n\t\t\t    <div class=\"video-placeholder d-flex justify-content-center align-items-center\">\n\t\t\t        <div class=\"overlay text-white p-2 w-100 text-center d-block justify-content-center align-items-center\">\n\t\t\t            <div>To view third-party content, please accept cookies.<\/div>\n\t\t\t            <button class=\"btn btn-secondary btn-sm mt-1 consent-change\">Change consent<\/button>\n\t\t\t        <\/div>\n\t\t\t    <\/div>\n\t\t\t<\/div>\n<\/div><br>Design of Experiment<br><a data-url=\"https:\/\/www.uttv.ee\/naita?id=32135\" href=\"https:\/\/www.uttv.ee\/naita?id=32135\" target=\"_blank\" title=\"\" rel=\"noopener\">https:\/\/www.uttv.ee\/naita?id=32135<\/a>\r\n<\/h4>\r\n\r\n<h4 style=\"text-align: center\">\r\n\t<a data-url=\"https:\/\/youtu.be\/D71f9uFqHDA\" href=\"https:\/\/youtu.be\/D71f9uFqHDA\" target=\"_blank\" title=\"\" rel=\"noopener\">https:\/\/youtu.be\/D71f9uFqHDA<\/a>\r\n<\/h4>\r\n\r\n<p style=\"margin-bottom:7.5pt;text-align:justify\">\r\n\t<span><span><span style=\"color:#2f2f2f\">The most basic way to study the robustness of a method is called <i>full factorial<\/i> design, carried out as in the following example for a robustness evaluation:<\/span><\/span><\/span>\r\n<\/p>\r\n\r\n<p>\r\n\t<span><span><span style=\"color:#2f2f2f\">a) Brainstorm and list all factors which are likely to influence the robustness of a newly developed chromatographic method. The DoE approach is needed to determine if the factors like the eluent pH (let\u2019s call it factor A), the additive concentration of the eluent (B) and the column temperature (C) have a critical impact on the retention time (also called response variable). The three factors (A, B and C) are set on two coded levels, a high (+) and a low level (-), which are depicted in the table below. We are using one level above and one below our optimal factor set to evaluate the robustness in both directions. The factor levels themselves are chosen by the chemist in charge of the validation and should be placed in a reasonable distance from the optimal factor setting. In practice, this means considering errors in laboratory work that appear while creating e.g. a new eluent solution and then predicting based on these thoughts \u0394pH or \u0394Additive conc. values, which can be used as upper and lower levels in DoE. <\/span><\/span><\/span><br><span><span><span style=\"color:#2f2f2f\">To calculate the number of experiments at different factor combinations, a simple formula can be used:<\/span><\/span><\/span>\r\n<\/p>\r\n\r\n<h6 style=\"text-align: center\">\r\n\t<img loading=\"lazy\" decoding=\"async\" width=\"280\" height=\"112\" class=\"alignnone wp-image-77\" style=\"width: 58px;height: 20px\" src=\"https:\/\/sisu.ut.ee\/wp-content\/uploads\/sites\/130\/doe_eq1_01.png\" title=\"doe_eq1_01.png\" alt=\"DoE_Eq1\"><span style=\"background:white\"><span style=\"line-height:normal\"><span><span style=\",serif\"><span style=\"color:#2f2f2f\"><i>\u00a0 \u00a0<\/i><\/span><\/span><\/span><\/span><\/span>(Eq 1)\r\n<\/h6>\r\n\r\n<p>\r\n\t<span style=\"background:white\"><span style=\"line-height:normal\"><span style=\"color:#2f2f2f\">w<\/span><\/span><\/span>h<span style=\"background:white\"><span style=\"line-height:normal\"><span style=\"color:#2f2f2f\">ere\u00a0\u00a0 \u00a0<i>n<\/i> \u2013 number of experiments performed at different combinations<\/span><\/span><\/span><br><span style=\"background:white\"><span style=\"line-height:normal\"><i><span style=\"color:#2f2f2f\">\u00a0 \u00a0 \u00a0 \u00a0 \u00a0 \u00a0 \u00a0 k<\/span><\/i><span style=\"color:#2f2f2f\"> \u2013 number of factors.<\/span><\/span><\/span>\r\n<\/p>\r\n\r\n<p style=\"margin-bottom:7.5pt;text-align:justify\">\r\n\t<span><span><span style=\"color:#2f2f2f\">In the current example <i>k<\/i> = 3: pH, additive concentration, column temperature. Thus, we need to perform 8 experiments in total to determine the robustness of the response variables at two levels.<\/span><\/span><\/span>\r\n<\/p>\r\n\r\n<p style=\"margin-bottom:7.5pt;text-align:justify\">\r\n\t<span><span><span style=\"color:#2f2f2f\">b) Construct a measurement plan that would contain all possible combinations of the factor levels for one substance in the mixture.<\/span><\/span><\/span>\r\n<\/p>\r\n\r\n<p style=\"margin-bottom:7.5pt;text-align:justify\">\r\n\t<i><span><span><span style=\"color:#2f2f2f\">Note 1: We are monitoring the retention times of all analyte substances, so by performing the experiments later, we can create tables like the following ones for all substances which are planned to be separated in the mixture and calculate the specific effects for every substance individually.<\/span><\/span><\/span><\/i>\r\n<\/p>\r\n\r\n<p style=\"margin-bottom:7.5pt;text-align:justify\">\r\n\t<i><span><span><span style=\"color:#2f2f2f\">Not<\/span><\/span><\/span><\/i><em>e 2: If repeatability is a concern to you then it is recommended to use a central point in DoE with only \u201coptimal\u201d setting and repeating this point multiple t<\/em><i><span><span>i<span style=\"color:#2f2f2f\">mes at random in the experimental plan instead of repeating the whole design itself.<\/span><\/span><\/span><\/i>\r\n<\/p>\r\n\r\n<p style=\"margin-bottom:7.5pt;text-align:justify\">\r\n\t<span><span><span style=\"color:#2f2f2f\">In a step-by-step manner:<\/span><\/span><\/span>\r\n<\/p>\r\n\r\n<p style=\"text-align:justify\">\r\n\t<span><span><span style=\"color:#2f2f2f\">First, notice that in the table, we first add + signs to half of the experiments of the first factor column (pH).<\/span><\/span><\/span>\r\n<\/p>\r\n\r\n<table class=\"table table-hover\" align=\"center\" border=\"1\" width=\"402\">\r\n\t<thead>\r\n\t\t<tr style=\"height:24.0pt!msorm\">\r\n\t\t\t<th nowrap scope=\"col\" style=\"width: 61.35pt\" width=\"123\">\r\n\t\t\t\t<p align=\"center\" style=\"margin-bottom:.0001pt;text-align:center;padding:0cm5.4pt0cm5.4pt!msorm\">\r\n\t\t\t\t\t<span style=\"line-height:normal\"><span style=\"height:24.0pt\"><span style=\"color:black\">Experiment number<\/span><\/span><\/span>\r\n\t\t\t\t<\/p>\r\n\t\t\t<\/th>\r\n\t\t\t<th nowrap scope=\"col\" style=\"width: 30.55pt\" width=\"61\">\r\n\t\t\t\t<p align=\"center\" style=\"margin-bottom:.0001pt;text-align:center;padding:0cm5.4pt0cm5.4pt!msorm\">\r\n\t\t\t\t\t<span style=\"line-height:normal\"><span style=\"height:24.0pt\"><span style=\"color:black\">A<\/span><\/span><\/span>\r\n\t\t\t\t<\/p>\r\n\r\n\t\t\t\t<p align=\"center\" style=\"margin-bottom:.0001pt;text-align:center;padding:0cm5.4pt0cm5.4pt!msorm\">\r\n\t\t\t\t\t<br><span style=\"line-height:normal\"><span style=\"height:24.0pt\"><span style=\"color:black\">pH <\/span><\/span><\/span>\r\n\t\t\t\t<\/p>\r\n\t\t\t<\/th>\r\n\t\t\t<th scope=\"col\" style=\"width: 63.8pt\" width=\"128\">\r\n\t\t\t\t<p align=\"center\" style=\"margin-bottom:.0001pt;text-align:center;padding:0cm5.4pt0cm5.4pt!msorm\">\r\n\t\t\t\t\t<span style=\"line-height:normal\"><span style=\"height:24.0pt\"><span style=\"color:black\">B<\/span><\/span><\/span>\r\n\t\t\t\t<\/p>\r\n\r\n\t\t\t\t<p align=\"center\" style=\"margin-bottom:.0001pt;text-align:center;padding:0cm5.4pt0cm5.4pt!msorm\">\r\n\t\t\t\t\t<span style=\"line-height:normal\"><span style=\"height:24.0pt\"><span style=\"color:black\">Additive conc.<br>[mmol\/L]<\/span><\/span><\/span>\r\n\t\t\t\t<\/p>\r\n\t\t\t<\/th>\r\n\t\t\t<th scope=\"col\" style=\"width: 45.3pt\" width=\"91\">\r\n\t\t\t\t<p align=\"center\" style=\"margin-bottom:.0001pt;text-align:center;padding:0cm5.4pt0cm5.4pt!msorm\">\r\n\t\t\t\t\t<span style=\"line-height:normal\"><span style=\"height:24.0pt\"><span style=\"color:black\">C<\/span><\/span><\/span>\r\n\t\t\t\t<\/p>\r\n\r\n\t\t\t\t<p align=\"center\" style=\"margin-bottom:.0001pt;text-align:center;padding:0cm5.4pt0cm5.4pt!msorm\">\r\n\t\t\t\t\t<span style=\"line-height:normal\"><span style=\"height:24.0pt\"><span style=\"color:black\">Column Temp.<br>[\u00b0C]<\/span><\/span><\/span>\r\n\t\t\t\t<\/p>\r\n\t\t\t<\/th>\r\n\t\t<\/tr>\r\n\t<\/thead>\r\n\t<tbody>\r\n\t\t<tr style=\"height:12.0pt!msorm\">\r\n\t\t\t<td nowrap style=\"width: 61.35pt\" width=\"123\">\r\n\t\t\t\t<p align=\"center\" style=\"margin-bottom:.0001pt;text-align:center;padding:0cm5.4pt0cm5.4pt!msorm\">\r\n\t\t\t\t\t<span style=\"line-height:normal\"><span style=\"height:12.0pt\"><span style=\"color:black\">1<\/span><\/span><\/span>\r\n\t\t\t\t<\/p>\r\n\t\t\t<\/td>\r\n\t\t\t<td nowrap style=\"width: 30.55pt\" width=\"61\">\r\n\t\t\t\t<p align=\"center\" style=\"margin-bottom:.0001pt;text-align:center;padding:0cm5.4pt0cm5.4pt!msorm\">\r\n\t\t\t\t\t<span style=\"line-height:normal\"><span style=\"height:12.0pt\"><b><span style=\"color:black\">+<\/span><\/b><\/span><\/span>\r\n\t\t\t\t<\/p>\r\n\t\t\t<\/td>\r\n\t\t\t<td nowrap style=\"width: 63.8pt\" width=\"128\">\r\n\t\t\t\t<p align=\"center\" style=\"margin-bottom:.0001pt;text-align:center;padding:0cm5.4pt0cm5.4pt!msorm\">\r\n\t\t\t\t\t\u00a0\r\n\t\t\t\t<\/p>\r\n\t\t\t<\/td>\r\n\t\t\t<td nowrap style=\"width: 45.3pt\" width=\"91\">\r\n\t\t\t\t<p align=\"center\" style=\"margin-bottom:.0001pt;text-align:center;padding:0cm5.4pt0cm5.4pt!msorm\">\r\n\t\t\t\t\t\u00a0\r\n\t\t\t\t<\/p>\r\n\t\t\t<\/td>\r\n\t\t<\/tr>\r\n\t\t<tr style=\"height:12.0pt!msorm\">\r\n\t\t\t<td nowrap style=\"width: 61.35pt\" width=\"123\">\r\n\t\t\t\t<p align=\"center\" style=\"margin-bottom:.0001pt;text-align:center;padding:0cm5.4pt0cm5.4pt!msorm\">\r\n\t\t\t\t\t<span style=\"line-height:normal\"><span style=\"height:12.0pt\"><span style=\"color:black\">2<\/span><\/span><\/span>\r\n\t\t\t\t<\/p>\r\n\t\t\t<\/td>\r\n\t\t\t<td nowrap style=\"width: 30.55pt\" width=\"61\">\r\n\t\t\t\t<p align=\"center\" style=\"margin-bottom:.0001pt;text-align:center;padding:0cm5.4pt0cm5.4pt!msorm\">\r\n\t\t\t\t\t<span style=\"line-height:normal\"><span style=\"height:12.0pt\"><b><span style=\"color:black\">+<\/span><\/b><\/span><\/span>\r\n\t\t\t\t<\/p>\r\n\t\t\t<\/td>\r\n\t\t\t<td nowrap style=\"width: 63.8pt\" width=\"128\">\r\n\t\t\t\t<p align=\"center\" style=\"margin-bottom:.0001pt;text-align:center;padding:0cm5.4pt0cm5.4pt!msorm\">\r\n\t\t\t\t\t\u00a0\r\n\t\t\t\t<\/p>\r\n\t\t\t<\/td>\r\n\t\t\t<td nowrap style=\"width: 45.3pt\" width=\"91\">\r\n\t\t\t\t<p align=\"center\" style=\"margin-bottom:.0001pt;text-align:center;padding:0cm5.4pt0cm5.4pt!msorm\">\r\n\t\t\t\t\t\u00a0\r\n\t\t\t\t<\/p>\r\n\t\t\t<\/td>\r\n\t\t<\/tr>\r\n\t\t<tr style=\"height:12.0pt!msorm\">\r\n\t\t\t<td nowrap style=\"width: 61.35pt\" width=\"123\">\r\n\t\t\t\t<p align=\"center\" style=\"margin-bottom:.0001pt;text-align:center;padding:0cm5.4pt0cm5.4pt!msorm\">\r\n\t\t\t\t\t<span style=\"line-height:normal\"><span style=\"height:12.0pt\"><span style=\"color:black\">3<\/span><\/span><\/span>\r\n\t\t\t\t<\/p>\r\n\t\t\t<\/td>\r\n\t\t\t<td nowrap style=\"width: 30.55pt\" width=\"61\">\r\n\t\t\t\t<p align=\"center\" style=\"margin-bottom:.0001pt;text-align:center;padding:0cm5.4pt0cm5.4pt!msorm\">\r\n\t\t\t\t\t<span style=\"line-height:normal\"><span style=\"height:12.0pt\"><b><span style=\"color:black\">+<\/span><\/b><\/span><\/span>\r\n\t\t\t\t<\/p>\r\n\t\t\t<\/td>\r\n\t\t\t<td nowrap style=\"width: 63.8pt\" width=\"128\">\r\n\t\t\t\t<p align=\"center\" style=\"margin-bottom:.0001pt;text-align:center;padding:0cm5.4pt0cm5.4pt!msorm\">\r\n\t\t\t\t\t\u00a0\r\n\t\t\t\t<\/p>\r\n\t\t\t<\/td>\r\n\t\t\t<td nowrap style=\"width: 45.3pt\" width=\"91\">\r\n\t\t\t\t<p align=\"center\" style=\"margin-bottom:.0001pt;text-align:center;padding:0cm5.4pt0cm5.4pt!msorm\">\r\n\t\t\t\t\t\u00a0\r\n\t\t\t\t<\/p>\r\n\t\t\t<\/td>\r\n\t\t<\/tr>\r\n\t\t<tr style=\"height:12.0pt!msorm\">\r\n\t\t\t<td nowrap style=\"width: 61.35pt\" width=\"123\">\r\n\t\t\t\t<p align=\"center\" style=\"margin-bottom:.0001pt;text-align:center;padding:0cm5.4pt0cm5.4pt!msorm\">\r\n\t\t\t\t\t<span style=\"line-height:normal\"><span style=\"height:12.0pt\"><span style=\"color:black\">4<\/span><\/span><\/span>\r\n\t\t\t\t<\/p>\r\n\t\t\t<\/td>\r\n\t\t\t<td nowrap style=\"width: 30.55pt\" width=\"61\">\r\n\t\t\t\t<p align=\"center\" style=\"margin-bottom:.0001pt;text-align:center;padding:0cm5.4pt0cm5.4pt!msorm\">\r\n\t\t\t\t\t<span style=\"line-height:normal\"><span style=\"height:12.0pt\"><b><span style=\"color:black\">+<\/span><\/b><\/span><\/span>\r\n\t\t\t\t<\/p>\r\n\t\t\t<\/td>\r\n\t\t\t<td nowrap style=\"width: 63.8pt\" width=\"128\">\r\n\t\t\t\t<p align=\"center\" style=\"margin-bottom:.0001pt;text-align:center;padding:0cm5.4pt0cm5.4pt!msorm\">\r\n\t\t\t\t\t\u00a0\r\n\t\t\t\t<\/p>\r\n\t\t\t<\/td>\r\n\t\t\t<td nowrap style=\"width: 45.3pt\" width=\"91\">\r\n\t\t\t\t<p align=\"center\" style=\"margin-bottom:.0001pt;text-align:center;padding:0cm5.4pt0cm5.4pt!msorm\">\r\n\t\t\t\t\t\u00a0\r\n\t\t\t\t<\/p>\r\n\t\t\t<\/td>\r\n\t\t<\/tr>\r\n\t\t<tr style=\"height:12.0pt!msorm\">\r\n\t\t\t<td nowrap style=\"width: 61.35pt\" width=\"123\">\r\n\t\t\t\t<p align=\"center\" style=\"margin-bottom:.0001pt;text-align:center;padding:0cm5.4pt0cm5.4pt!msorm\">\r\n\t\t\t\t\t<span style=\"line-height:normal\"><span style=\"height:12.0pt\"><span style=\"color:black\">5<\/span><\/span><\/span>\r\n\t\t\t\t<\/p>\r\n\t\t\t<\/td>\r\n\t\t\t<td nowrap style=\"width: 30.55pt\" width=\"61\">\r\n\t\t\t\t<p align=\"center\" style=\"margin-bottom:.0001pt;text-align:center;padding:0cm5.4pt0cm5.4pt!msorm\">\r\n\t\t\t\t\t<span style=\"line-height:normal\"><span style=\"height:12.0pt\"><b><span style=\"color:black\">\u2013<\/span><\/b><\/span><\/span>\r\n\t\t\t\t<\/p>\r\n\t\t\t<\/td>\r\n\t\t\t<td nowrap style=\"width: 63.8pt\" width=\"128\">\r\n\t\t\t\t<p align=\"center\" style=\"margin-bottom:.0001pt;text-align:center;padding:0cm5.4pt0cm5.4pt!msorm\">\r\n\t\t\t\t\t\u00a0\r\n\t\t\t\t<\/p>\r\n\t\t\t<\/td>\r\n\t\t\t<td nowrap style=\"width: 45.3pt\" width=\"91\">\r\n\t\t\t\t<p align=\"center\" style=\"margin-bottom:.0001pt;text-align:center;padding:0cm5.4pt0cm5.4pt!msorm\">\r\n\t\t\t\t\t\u00a0\r\n\t\t\t\t<\/p>\r\n\t\t\t<\/td>\r\n\t\t<\/tr>\r\n\t\t<tr style=\"height:12.0pt!msorm\">\r\n\t\t\t<td nowrap style=\"width: 61.35pt\" width=\"123\">\r\n\t\t\t\t<p align=\"center\" style=\"margin-bottom:.0001pt;text-align:center;padding:0cm5.4pt0cm5.4pt!msorm\">\r\n\t\t\t\t\t<span style=\"line-height:normal\"><span style=\"height:12.0pt\"><span style=\"color:black\">6<\/span><\/span><\/span>\r\n\t\t\t\t<\/p>\r\n\t\t\t<\/td>\r\n\t\t\t<td nowrap style=\"width: 30.55pt\" width=\"61\">\r\n\t\t\t\t<p align=\"center\" style=\"margin-bottom:.0001pt;text-align:center;padding:0cm5.4pt0cm5.4pt!msorm\">\r\n\t\t\t\t\t<span style=\"line-height:normal\"><span style=\"height:12.0pt\"><b><span style=\"color:black\">\u2013<\/span><\/b><\/span><\/span>\r\n\t\t\t\t<\/p>\r\n\t\t\t<\/td>\r\n\t\t\t<td nowrap style=\"width: 63.8pt\" width=\"128\">\r\n\t\t\t\t<p align=\"center\" style=\"margin-bottom:.0001pt;text-align:center;padding:0cm5.4pt0cm5.4pt!msorm\">\r\n\t\t\t\t\t\u00a0\r\n\t\t\t\t<\/p>\r\n\t\t\t<\/td>\r\n\t\t\t<td nowrap style=\"width: 45.3pt\" width=\"91\">\r\n\t\t\t\t<p align=\"center\" style=\"margin-bottom:.0001pt;text-align:center;padding:0cm5.4pt0cm5.4pt!msorm\">\r\n\t\t\t\t\t\u00a0\r\n\t\t\t\t<\/p>\r\n\t\t\t<\/td>\r\n\t\t<\/tr>\r\n\t\t<tr style=\"height:12.0pt!msorm\">\r\n\t\t\t<td nowrap style=\"width: 61.35pt\" width=\"123\">\r\n\t\t\t\t<p align=\"center\" style=\"margin-bottom:.0001pt;text-align:center;padding:0cm5.4pt0cm5.4pt!msorm\">\r\n\t\t\t\t\t<span style=\"line-height:normal\"><span style=\"height:12.0pt\"><span style=\"color:black\">7<\/span><\/span><\/span>\r\n\t\t\t\t<\/p>\r\n\t\t\t<\/td>\r\n\t\t\t<td nowrap style=\"width: 30.55pt\" width=\"61\">\r\n\t\t\t\t<p align=\"center\" style=\"margin-bottom:.0001pt;text-align:center;padding:0cm5.4pt0cm5.4pt!msorm\">\r\n\t\t\t\t\t<span style=\"line-height:normal\"><span style=\"height:12.0pt\"><b><span style=\"color:black\">\u2013<\/span><\/b><\/span><\/span>\r\n\t\t\t\t<\/p>\r\n\t\t\t<\/td>\r\n\t\t\t<td nowrap style=\"width: 63.8pt\" width=\"128\">\r\n\t\t\t\t<p align=\"center\" style=\"margin-bottom:.0001pt;text-align:center;padding:0cm5.4pt0cm5.4pt!msorm\">\r\n\t\t\t\t\t\u00a0\r\n\t\t\t\t<\/p>\r\n\t\t\t<\/td>\r\n\t\t\t<td nowrap style=\"width: 45.3pt\" width=\"91\">\r\n\t\t\t\t<p align=\"center\" style=\"margin-bottom:.0001pt;text-align:center;padding:0cm5.4pt0cm5.4pt!msorm\">\r\n\t\t\t\t\t\u00a0\r\n\t\t\t\t<\/p>\r\n\t\t\t<\/td>\r\n\t\t<\/tr>\r\n\t\t<tr style=\"height:12.4pt!msorm\">\r\n\t\t\t<td nowrap style=\"width: 61.35pt\" width=\"123\">\r\n\t\t\t\t<p align=\"center\" style=\"margin-bottom:.0001pt;text-align:center;padding:0cm5.4pt0cm5.4pt!msorm\">\r\n\t\t\t\t\t<span style=\"line-height:normal\"><span style=\"height:12.4pt\"><span style=\"color:black\">8<\/span><\/span><\/span>\r\n\t\t\t\t<\/p>\r\n\t\t\t<\/td>\r\n\t\t\t<td nowrap style=\"width: 30.55pt\" width=\"61\">\r\n\t\t\t\t<p align=\"center\" style=\"margin-bottom:.0001pt;text-align:center;padding:0cm5.4pt0cm5.4pt!msorm\">\r\n\t\t\t\t\t<span style=\"line-height:normal\"><span style=\"height:12.4pt\"><b><span style=\"color:black\">\u2013<\/span><\/b><\/span><\/span>\r\n\t\t\t\t<\/p>\r\n\t\t\t<\/td>\r\n\t\t\t<td nowrap style=\"width: 63.8pt\" width=\"128\">\r\n\t\t\t\t<p align=\"center\" style=\"margin-bottom:.0001pt;text-align:center;padding:0cm5.4pt0cm5.4pt!msorm\">\r\n\t\t\t\t\t\u00a0\r\n\t\t\t\t<\/p>\r\n\t\t\t<\/td>\r\n\t\t\t<td nowrap style=\"width: 45.3pt\" width=\"91\">\r\n\t\t\t\t<p align=\"center\" style=\"margin-bottom:.0001pt;text-align:center;padding:0cm5.4pt0cm5.4pt!msorm\">\r\n\t\t\t\t\t\u00a0\r\n\t\t\t\t<\/p>\r\n\t\t\t<\/td>\r\n\t\t<\/tr>\r\n\t<\/tbody>\r\n<\/table>\r\n\r\n<p>\r\n\t<span style=\"background:white\"><span style=\"line-height:normal\"><span style=\"color:#2f2f2f\">Then we fill in the next factor<\/span><span style=\"color:#2f2f2f\"> by splitting the first <\/span><span style=\"color:#2f2f2f\">factors + signs in half.<\/span><\/span><\/span>\r\n<\/p>\r\n\r\n<table class=\"table table-hover\" align=\"center\" border=\"1\" width=\"402\">\r\n\t<thead>\r\n\t\t<tr style=\"height:24.0pt!msorm\">\r\n\t\t\t<th nowrap scope=\"col\" style=\"width: 61.35pt\" width=\"123\">\r\n\t\t\t\t<p align=\"center\" style=\"margin-bottom:.0001pt;text-align:center;padding:0cm5.4pt0cm5.4pt!msorm\">\r\n\t\t\t\t\t<span style=\"line-height:normal\"><span style=\"height:24.0pt\"><span><span style=\",serif\"><span style=\"color:black\">Experiment number<\/span><\/span><\/span><\/span><\/span>\r\n\t\t\t\t<\/p>\r\n\t\t\t<\/th>\r\n\t\t\t<th nowrap scope=\"col\" style=\"width: 30.55pt\" width=\"61\">\r\n\t\t\t\t<p align=\"center\" style=\"margin-bottom:.0001pt;text-align:center;padding:0cm5.4pt0cm5.4pt!msorm\">\r\n\t\t\t\t\t<span style=\"line-height:normal\"><span style=\"height:24.0pt\"><span lang=\"DE\"><span style=\",serif\"><span style=\"color:black\">A<\/span><\/span><\/span><\/span><\/span>\r\n\t\t\t\t<\/p>\r\n\r\n\t\t\t\t<p align=\"center\" style=\"margin-bottom:.0001pt;text-align:center;padding:0cm5.4pt0cm5.4pt!msorm\">\r\n\t\t\t\t\t<br><span style=\"line-height:normal\"><span style=\"height:24.0pt\"><span><span style=\",serif\"><span style=\"color:black\">pH <\/span><\/span><\/span><\/span><\/span>\r\n\t\t\t\t<\/p>\r\n\t\t\t<\/th>\r\n\t\t\t<th scope=\"col\" style=\"width: 63.8pt\" width=\"128\">\r\n\t\t\t\t<p align=\"center\" style=\"margin-bottom:.0001pt;text-align:center;padding:0cm5.4pt0cm5.4pt!msorm\">\r\n\t\t\t\t\t<span style=\"line-height:normal\"><span style=\"height:24.0pt\"><span><span style=\",serif\"><span style=\"color:black\">B<\/span><\/span><\/span><\/span><\/span>\r\n\t\t\t\t<\/p>\r\n\r\n\t\t\t\t<p align=\"center\" style=\"margin-bottom:.0001pt;text-align:center;padding:0cm5.4pt0cm5.4pt!msorm\">\r\n\t\t\t\t\t<span style=\"line-height:normal\"><span style=\"height:24.0pt\"><span><span style=\",serif\"><span style=\"color:black\">Additive conc.<br>[mmol\/L]<\/span><\/span><\/span><\/span><\/span>\r\n\t\t\t\t<\/p>\r\n\t\t\t<\/th>\r\n\t\t\t<th scope=\"col\" style=\"width: 45.3pt\" width=\"91\">\r\n\t\t\t\t<p align=\"center\" style=\"margin-bottom:.0001pt;text-align:center;padding:0cm5.4pt0cm5.4pt!msorm\">\r\n\t\t\t\t\t<span style=\"line-height:normal\"><span style=\"height:24.0pt\"><span lang=\"DE\"><span style=\",serif\"><span style=\"color:black\">C<\/span><\/span><\/span><\/span><\/span>\r\n\t\t\t\t<\/p>\r\n\r\n\t\t\t\t<p align=\"center\" style=\"margin-bottom:.0001pt;text-align:center;padding:0cm5.4pt0cm5.4pt!msorm\">\r\n\t\t\t\t\t<span style=\"line-height:normal\"><span style=\"height:24.0pt\"><span><span style=\",serif\"><span style=\"color:black\">Column Temp.<br>[\u00b0C]<\/span><\/span><\/span><\/span><\/span>\r\n\t\t\t\t<\/p>\r\n\t\t\t<\/th>\r\n\t\t<\/tr>\r\n\t<\/thead>\r\n\t<tbody>\r\n\t\t<tr style=\"height:12.0pt!msorm\">\r\n\t\t\t<td nowrap style=\"width: 61.35pt\" width=\"123\">\r\n\t\t\t\t<p align=\"center\" style=\"margin-bottom:.0001pt;text-align:center;padding:0cm5.4pt0cm5.4pt!msorm\">\r\n\t\t\t\t\t<span style=\"line-height:normal\"><span style=\"height:12.0pt\"><span><span style=\",serif\"><span style=\"color:black\">1<\/span><\/span><\/span><\/span><\/span>\r\n\t\t\t\t<\/p>\r\n\t\t\t<\/td>\r\n\t\t\t<td nowrap style=\"width: 30.55pt\" width=\"61\">\r\n\t\t\t\t<p align=\"center\" style=\"margin-bottom:.0001pt;text-align:center;padding:0cm5.4pt0cm5.4pt!msorm\">\r\n\t\t\t\t\t<span style=\"line-height:normal\"><span style=\"height:12.0pt\"><b><span><span style=\",serif\"><span style=\"color:black\">+<\/span><\/span><\/span><\/b><\/span><\/span>\r\n\t\t\t\t<\/p>\r\n\t\t\t<\/td>\r\n\t\t\t<td nowrap style=\"width: 63.8pt\" width=\"128\">\r\n\t\t\t\t<p align=\"center\" style=\"margin-bottom:.0001pt;text-align:center;padding:0cm5.4pt0cm5.4pt!msorm\">\r\n\t\t\t\t\t<span style=\"line-height:normal\"><span style=\"height:12.0pt\"><b><span><span style=\",serif\"><span style=\"color:black\">+<\/span><\/span><\/span><\/b><\/span><\/span>\r\n\t\t\t\t<\/p>\r\n\t\t\t<\/td>\r\n\t\t\t<td nowrap style=\"width: 45.3pt\" width=\"91\">\r\n\t\t\t\t<p align=\"center\" style=\"margin-bottom:.0001pt;text-align:center;padding:0cm5.4pt0cm5.4pt!msorm\">\r\n\t\t\t\t\t\u00a0\r\n\t\t\t\t<\/p>\r\n\t\t\t<\/td>\r\n\t\t<\/tr>\r\n\t\t<tr style=\"height:12.0pt!msorm\">\r\n\t\t\t<td nowrap style=\"width: 61.35pt\" width=\"123\">\r\n\t\t\t\t<p align=\"center\" style=\"margin-bottom:.0001pt;text-align:center;padding:0cm5.4pt0cm5.4pt!msorm\">\r\n\t\t\t\t\t<span style=\"line-height:normal\"><span style=\"height:12.0pt\"><span><span style=\",serif\"><span style=\"color:black\">2<\/span><\/span><\/span><\/span><\/span>\r\n\t\t\t\t<\/p>\r\n\t\t\t<\/td>\r\n\t\t\t<td nowrap style=\"width: 30.55pt\" width=\"61\">\r\n\t\t\t\t<p align=\"center\" style=\"margin-bottom:.0001pt;text-align:center;padding:0cm5.4pt0cm5.4pt!msorm\">\r\n\t\t\t\t\t<span style=\"line-height:normal\"><span style=\"height:12.0pt\"><b><span><span style=\",serif\"><span style=\"color:black\">+<\/span><\/span><\/span><\/b><\/span><\/span>\r\n\t\t\t\t<\/p>\r\n\t\t\t<\/td>\r\n\t\t\t<td nowrap style=\"width: 63.8pt\" width=\"128\">\r\n\t\t\t\t<p align=\"center\" style=\"margin-bottom:.0001pt;text-align:center;padding:0cm5.4pt0cm5.4pt!msorm\">\r\n\t\t\t\t\t<span style=\"line-height:normal\"><span style=\"height:12.0pt\"><b><span><span style=\",serif\"><span style=\"color:black\">+<\/span><\/span><\/span><\/b><\/span><\/span>\r\n\t\t\t\t<\/p>\r\n\t\t\t<\/td>\r\n\t\t\t<td nowrap style=\"width: 45.3pt\" width=\"91\">\r\n\t\t\t\t<p align=\"center\" style=\"margin-bottom:.0001pt;text-align:center;padding:0cm5.4pt0cm5.4pt!msorm\">\r\n\t\t\t\t\t\u00a0\r\n\t\t\t\t<\/p>\r\n\t\t\t<\/td>\r\n\t\t<\/tr>\r\n\t\t<tr style=\"height:12.0pt!msorm\">\r\n\t\t\t<td nowrap style=\"width: 61.35pt\" width=\"123\">\r\n\t\t\t\t<p align=\"center\" style=\"margin-bottom:.0001pt;text-align:center;padding:0cm5.4pt0cm5.4pt!msorm\">\r\n\t\t\t\t\t<span style=\"line-height:normal\"><span style=\"height:12.0pt\"><span><span style=\",serif\"><span style=\"color:black\">3<\/span><\/span><\/span><\/span><\/span>\r\n\t\t\t\t<\/p>\r\n\t\t\t<\/td>\r\n\t\t\t<td nowrap style=\"width: 30.55pt\" width=\"61\">\r\n\t\t\t\t<p align=\"center\" style=\"margin-bottom:.0001pt;text-align:center;padding:0cm5.4pt0cm5.4pt!msorm\">\r\n\t\t\t\t\t<span style=\"line-height:normal\"><span style=\"height:12.0pt\"><b><span><span style=\",serif\"><span style=\"color:black\">+<\/span><\/span><\/span><\/b><\/span><\/span>\r\n\t\t\t\t<\/p>\r\n\t\t\t<\/td>\r\n\t\t\t<td nowrap style=\"width: 63.8pt\" width=\"128\">\r\n\t\t\t\t<p align=\"center\" style=\"margin-bottom:.0001pt;text-align:center;padding:0cm5.4pt0cm5.4pt!msorm\">\r\n\t\t\t\t\t<span style=\"line-height:normal\"><span style=\"height:12.0pt\"><b><span><span style=\",serif\"><span style=\"color:black\">\u2013<\/span><\/span><\/span><\/b><\/span><\/span>\r\n\t\t\t\t<\/p>\r\n\t\t\t<\/td>\r\n\t\t\t<td nowrap style=\"width: 45.3pt\" width=\"91\">\r\n\t\t\t\t<p align=\"center\" style=\"margin-bottom:.0001pt;text-align:center;padding:0cm5.4pt0cm5.4pt!msorm\">\r\n\t\t\t\t\t\u00a0\r\n\t\t\t\t<\/p>\r\n\t\t\t<\/td>\r\n\t\t<\/tr>\r\n\t\t<tr style=\"height:12.0pt!msorm\">\r\n\t\t\t<td nowrap style=\"width: 61.35pt\" width=\"123\">\r\n\t\t\t\t<p align=\"center\" style=\"margin-bottom:.0001pt;text-align:center;padding:0cm5.4pt0cm5.4pt!msorm\">\r\n\t\t\t\t\t<span style=\"line-height:normal\"><span style=\"height:12.0pt\"><span><span style=\",serif\"><span style=\"color:black\">4<\/span><\/span><\/span><\/span><\/span>\r\n\t\t\t\t<\/p>\r\n\t\t\t<\/td>\r\n\t\t\t<td nowrap style=\"width: 30.55pt\" width=\"61\">\r\n\t\t\t\t<p align=\"center\" style=\"margin-bottom:.0001pt;text-align:center;padding:0cm5.4pt0cm5.4pt!msorm\">\r\n\t\t\t\t\t<span style=\"line-height:normal\"><span style=\"height:12.0pt\"><b><span><span style=\",serif\"><span style=\"color:black\">+<\/span><\/span><\/span><\/b><\/span><\/span>\r\n\t\t\t\t<\/p>\r\n\t\t\t<\/td>\r\n\t\t\t<td nowrap style=\"width: 63.8pt\" width=\"128\">\r\n\t\t\t\t<p align=\"center\" style=\"margin-bottom:.0001pt;text-align:center;padding:0cm5.4pt0cm5.4pt!msorm\">\r\n\t\t\t\t\t<span style=\"line-height:normal\"><span style=\"height:12.0pt\"><b><span><span style=\",serif\"><span style=\"color:black\">\u2013<\/span><\/span><\/span><\/b><\/span><\/span>\r\n\t\t\t\t<\/p>\r\n\t\t\t<\/td>\r\n\t\t\t<td nowrap style=\"width: 45.3pt\" width=\"91\">\r\n\t\t\t\t<p align=\"center\" style=\"margin-bottom:.0001pt;text-align:center;padding:0cm5.4pt0cm5.4pt!msorm\">\r\n\t\t\t\t\t\u00a0\r\n\t\t\t\t<\/p>\r\n\t\t\t<\/td>\r\n\t\t<\/tr>\r\n\t\t<tr style=\"height:12.0pt!msorm\">\r\n\t\t\t<td nowrap style=\"width: 61.35pt\" width=\"123\">\r\n\t\t\t\t<p align=\"center\" style=\"margin-bottom:.0001pt;text-align:center;padding:0cm5.4pt0cm5.4pt!msorm\">\r\n\t\t\t\t\t<span style=\"line-height:normal\"><span style=\"height:12.0pt\"><span><span style=\",serif\"><span style=\"color:black\">5<\/span><\/span><\/span><\/span><\/span>\r\n\t\t\t\t<\/p>\r\n\t\t\t<\/td>\r\n\t\t\t<td nowrap style=\"width: 30.55pt\" width=\"61\">\r\n\t\t\t\t<p align=\"center\" style=\"margin-bottom:.0001pt;text-align:center;padding:0cm5.4pt0cm5.4pt!msorm\">\r\n\t\t\t\t\t<span style=\"line-height:normal\"><span style=\"height:12.0pt\"><b><span><span style=\",serif\"><span style=\"color:black\">\u2013<\/span><\/span><\/span><\/b><\/span><\/span>\r\n\t\t\t\t<\/p>\r\n\t\t\t<\/td>\r\n\t\t\t<td nowrap style=\"width: 63.8pt\" width=\"128\">\r\n\t\t\t\t<p align=\"center\" style=\"margin-bottom:.0001pt;text-align:center;padding:0cm5.4pt0cm5.4pt!msorm\">\r\n\t\t\t\t\t<span style=\"line-height:normal\"><span style=\"height:12.0pt\"><b><span><span style=\",serif\"><span style=\"color:black\">+<\/span><\/span><\/span><\/b><\/span><\/span>\r\n\t\t\t\t<\/p>\r\n\t\t\t<\/td>\r\n\t\t\t<td nowrap style=\"width: 45.3pt\" width=\"91\">\r\n\t\t\t\t<p align=\"center\" style=\"margin-bottom:.0001pt;text-align:center;padding:0cm5.4pt0cm5.4pt!msorm\">\r\n\t\t\t\t\t\u00a0\r\n\t\t\t\t<\/p>\r\n\t\t\t<\/td>\r\n\t\t<\/tr>\r\n\t\t<tr style=\"height:12.0pt!msorm\">\r\n\t\t\t<td nowrap style=\"width: 61.35pt\" width=\"123\">\r\n\t\t\t\t<p align=\"center\" style=\"margin-bottom:.0001pt;text-align:center;padding:0cm5.4pt0cm5.4pt!msorm\">\r\n\t\t\t\t\t<span style=\"line-height:normal\"><span style=\"height:12.0pt\"><span><span style=\",serif\"><span style=\"color:black\">6<\/span><\/span><\/span><\/span><\/span>\r\n\t\t\t\t<\/p>\r\n\t\t\t<\/td>\r\n\t\t\t<td nowrap style=\"width: 30.55pt\" width=\"61\">\r\n\t\t\t\t<p align=\"center\" style=\"margin-bottom:.0001pt;text-align:center;padding:0cm5.4pt0cm5.4pt!msorm\">\r\n\t\t\t\t\t<span style=\"line-height:normal\"><span style=\"height:12.0pt\"><b><span><span style=\",serif\"><span style=\"color:black\">\u2013<\/span><\/span><\/span><\/b><\/span><\/span>\r\n\t\t\t\t<\/p>\r\n\t\t\t<\/td>\r\n\t\t\t<td nowrap style=\"width: 63.8pt\" width=\"128\">\r\n\t\t\t\t<p align=\"center\" style=\"margin-bottom:.0001pt;text-align:center;padding:0cm5.4pt0cm5.4pt!msorm\">\r\n\t\t\t\t\t<span style=\"line-height:normal\"><span style=\"height:12.0pt\"><b><span><span style=\",serif\"><span style=\"color:black\">+<\/span><\/span><\/span><\/b><\/span><\/span>\r\n\t\t\t\t<\/p>\r\n\t\t\t<\/td>\r\n\t\t\t<td nowrap style=\"width: 45.3pt\" width=\"91\">\r\n\t\t\t\t<p align=\"center\" style=\"margin-bottom:.0001pt;text-align:center;padding:0cm5.4pt0cm5.4pt!msorm\">\r\n\t\t\t\t\t\u00a0\r\n\t\t\t\t<\/p>\r\n\t\t\t<\/td>\r\n\t\t<\/tr>\r\n\t\t<tr style=\"height:12.0pt!msorm\">\r\n\t\t\t<td nowrap style=\"width: 61.35pt\" width=\"123\">\r\n\t\t\t\t<p align=\"center\" style=\"margin-bottom:.0001pt;text-align:center;padding:0cm5.4pt0cm5.4pt!msorm\">\r\n\t\t\t\t\t<span style=\"line-height:normal\"><span style=\"height:12.0pt\"><span><span style=\",serif\"><span style=\"color:black\">7<\/span><\/span><\/span><\/span><\/span>\r\n\t\t\t\t<\/p>\r\n\t\t\t<\/td>\r\n\t\t\t<td nowrap style=\"width: 30.55pt\" width=\"61\">\r\n\t\t\t\t<p align=\"center\" style=\"margin-bottom:.0001pt;text-align:center;padding:0cm5.4pt0cm5.4pt!msorm\">\r\n\t\t\t\t\t<span style=\"line-height:normal\"><span style=\"height:12.0pt\"><b><span><span style=\",serif\"><span style=\"color:black\">\u2013<\/span><\/span><\/span><\/b><\/span><\/span>\r\n\t\t\t\t<\/p>\r\n\t\t\t<\/td>\r\n\t\t\t<td nowrap style=\"width: 63.8pt\" width=\"128\">\r\n\t\t\t\t<p align=\"center\" style=\"margin-bottom:.0001pt;text-align:center;padding:0cm5.4pt0cm5.4pt!msorm\">\r\n\t\t\t\t\t<span style=\"line-height:normal\"><span style=\"height:12.0pt\"><b><span><span style=\",serif\"><span style=\"color:black\">\u2013<\/span><\/span><\/span><\/b><\/span><\/span>\r\n\t\t\t\t<\/p>\r\n\t\t\t<\/td>\r\n\t\t\t<td nowrap style=\"width: 45.3pt\" width=\"91\">\r\n\t\t\t\t<p align=\"center\" style=\"margin-bottom:.0001pt;text-align:center;padding:0cm5.4pt0cm5.4pt!msorm\">\r\n\t\t\t\t\t\u00a0\r\n\t\t\t\t<\/p>\r\n\t\t\t<\/td>\r\n\t\t<\/tr>\r\n\t\t<tr style=\"height:12.4pt!msorm\">\r\n\t\t\t<td nowrap style=\"width: 61.35pt\" width=\"123\">\r\n\t\t\t\t<p align=\"center\" style=\"margin-bottom:.0001pt;text-align:center;padding:0cm5.4pt0cm5.4pt!msorm\">\r\n\t\t\t\t\t<span style=\"line-height:normal\"><span style=\"height:12.4pt\"><span><span style=\",serif\"><span style=\"color:black\">8<\/span><\/span><\/span><\/span><\/span>\r\n\t\t\t\t<\/p>\r\n\t\t\t<\/td>\r\n\t\t\t<td nowrap style=\"width: 30.55pt\" width=\"61\">\r\n\t\t\t\t<p align=\"center\" style=\"margin-bottom:.0001pt;text-align:center;padding:0cm5.4pt0cm5.4pt!msorm\">\r\n\t\t\t\t\t<span style=\"line-height:normal\"><span style=\"height:12.4pt\"><b><span><span style=\",serif\"><span style=\"color:black\">\u2013<\/span><\/span><\/span><\/b><\/span><\/span>\r\n\t\t\t\t<\/p>\r\n\t\t\t<\/td>\r\n\t\t\t<td nowrap style=\"width: 63.8pt\" width=\"128\">\r\n\t\t\t\t<p align=\"center\" style=\"margin-bottom:.0001pt;text-align:center;padding:0cm5.4pt0cm5.4pt!msorm\">\r\n\t\t\t\t\t<span style=\"line-height:normal\"><span style=\"height:12.4pt\"><b><span><span style=\",serif\"><span style=\"color:black\">\u2013<\/span><\/span><\/span><\/b><\/span><\/span>\r\n\t\t\t\t<\/p>\r\n\t\t\t<\/td>\r\n\t\t\t<td nowrap style=\"width: 45.3pt\" width=\"91\">\r\n\t\t\t\t<p align=\"center\" style=\"margin-bottom:.0001pt;text-align:center;padding:0cm5.4pt0cm5.4pt!msorm\">\r\n\t\t\t\t\t\u00a0\r\n\t\t\t\t<\/p>\r\n\t\t\t<\/td>\r\n\t\t<\/tr>\r\n\t<\/tbody>\r\n<\/table>\r\n\r\n<p>\r\n\t<span style=\"background:white\"><span style=\"line-height:normal\"><span style=\"color:#2f2f2f\">And we do the same for the third (all other columns that remain).<\/span><\/span><\/span>\r\n<\/p>\r\n\r\n<table class=\"table table-hover\" align=\"center\" border=\"1\" width=\"402\">\r\n\t<thead>\r\n\t\t<tr style=\"height:24.0pt!msorm\">\r\n\t\t\t<th nowrap scope=\"col\" style=\"width: 61.35pt\" width=\"123\">\r\n\t\t\t\t<p align=\"center\" style=\"margin-bottom:.0001pt;text-align:center;padding:0cm5.4pt0cm5.4pt!msorm\">\r\n\t\t\t\t\t<span style=\"line-height:normal\"><span style=\"height:24.0pt\"><span><span style=\",serif\"><span style=\"color:black\">Experiment number<\/span><\/span><\/span><\/span><\/span>\r\n\t\t\t\t<\/p>\r\n\t\t\t<\/th>\r\n\t\t\t<th nowrap scope=\"col\" style=\"width: 30.55pt\" width=\"61\">\r\n\t\t\t\t<p align=\"center\" style=\"margin-bottom:.0001pt;text-align:center;padding:0cm5.4pt0cm5.4pt!msorm\">\r\n\t\t\t\t\t<span style=\"line-height:normal\"><span style=\"height:24.0pt\"><span lang=\"DE\"><span style=\",serif\"><span style=\"color:black\">A<\/span><\/span><\/span><\/span><\/span>\r\n\t\t\t\t<\/p>\r\n\r\n\t\t\t\t<p align=\"center\" style=\"margin-bottom:.0001pt;text-align:center;padding:0cm5.4pt0cm5.4pt!msorm\">\r\n\t\t\t\t\t<br><span style=\"line-height:normal\"><span style=\"height:24.0pt\"><span><span style=\",serif\"><span style=\"color:black\">pH <\/span><\/span><\/span><\/span><\/span>\r\n\t\t\t\t<\/p>\r\n\t\t\t<\/th>\r\n\t\t\t<th scope=\"col\" style=\"width: 63.8pt\" width=\"128\">\r\n\t\t\t\t<p align=\"center\" style=\"margin-bottom:.0001pt;text-align:center;padding:0cm5.4pt0cm5.4pt!msorm\">\r\n\t\t\t\t\t<span style=\"line-height:normal\"><span style=\"height:24.0pt\"><span><span style=\",serif\"><span style=\"color:black\">B<\/span><\/span><\/span><\/span><\/span>\r\n\t\t\t\t<\/p>\r\n\r\n\t\t\t\t<p align=\"center\" style=\"margin-bottom:.0001pt;text-align:center;padding:0cm5.4pt0cm5.4pt!msorm\">\r\n\t\t\t\t\t<span style=\"line-height:normal\"><span style=\"height:24.0pt\"><span><span style=\",serif\"><span style=\"color:black\">Additive conc.<br>[mmol\/L]<\/span><\/span><\/span><\/span><\/span>\r\n\t\t\t\t<\/p>\r\n\t\t\t<\/th>\r\n\t\t\t<th scope=\"col\" style=\"width: 45.3pt\" width=\"91\">\r\n\t\t\t\t<p align=\"center\" style=\"margin-bottom:.0001pt;text-align:center;padding:0cm5.4pt0cm5.4pt!msorm\">\r\n\t\t\t\t\t<span style=\"line-height:normal\"><span style=\"height:24.0pt\"><span lang=\"DE\"><span style=\",serif\"><span style=\"color:black\">C<\/span><\/span><\/span><\/span><\/span>\r\n\t\t\t\t<\/p>\r\n\r\n\t\t\t\t<p align=\"center\" style=\"margin-bottom:.0001pt;text-align:center;padding:0cm5.4pt0cm5.4pt!msorm\">\r\n\t\t\t\t\t<span style=\"line-height:normal\"><span style=\"height:24.0pt\"><span><span style=\",serif\"><span style=\"color:black\">Column Temp.<br>[\u00b0C]<\/span><\/span><\/span><\/span><\/span>\r\n\t\t\t\t<\/p>\r\n\t\t\t<\/th>\r\n\t\t<\/tr>\r\n\t<\/thead>\r\n\t<tbody>\r\n\t\t<tr style=\"height:12.0pt!msorm\">\r\n\t\t\t<td nowrap style=\"width: 61.35pt\" width=\"123\">\r\n\t\t\t\t<p align=\"center\" style=\"margin-bottom:.0001pt;text-align:center;padding:0cm5.4pt0cm5.4pt!msorm\">\r\n\t\t\t\t\t<span style=\"line-height:normal\"><span style=\"height:12.0pt\"><span><span style=\",serif\"><span style=\"color:black\">1<\/span><\/span><\/span><\/span><\/span>\r\n\t\t\t\t<\/p>\r\n\t\t\t<\/td>\r\n\t\t\t<td nowrap style=\"width: 30.55pt\" width=\"61\">\r\n\t\t\t\t<p align=\"center\" style=\"margin-bottom:.0001pt;text-align:center;padding:0cm5.4pt0cm5.4pt!msorm\">\r\n\t\t\t\t\t<span style=\"line-height:normal\"><span style=\"height:12.0pt\"><b><span><span style=\",serif\"><span style=\"color:black\">+<\/span><\/span><\/span><\/b><\/span><\/span>\r\n\t\t\t\t<\/p>\r\n\t\t\t<\/td>\r\n\t\t\t<td nowrap style=\"width: 63.8pt\" width=\"128\">\r\n\t\t\t\t<p align=\"center\" style=\"margin-bottom:.0001pt;text-align:center;padding:0cm5.4pt0cm5.4pt!msorm\">\r\n\t\t\t\t\t<span style=\"line-height:normal\"><span style=\"height:12.0pt\"><b><span><span style=\",serif\"><span style=\"color:black\">+<\/span><\/span><\/span><\/b><\/span><\/span>\r\n\t\t\t\t<\/p>\r\n\t\t\t<\/td>\r\n\t\t\t<td nowrap style=\"width: 45.3pt\" width=\"91\">\r\n\t\t\t\t<p align=\"center\" style=\"margin-bottom:.0001pt;text-align:center;padding:0cm5.4pt0cm5.4pt!msorm\">\r\n\t\t\t\t\t<span style=\"line-height:normal\"><span style=\"height:12.0pt\"><b><span><span style=\",serif\"><span style=\"color:black\">+<\/span><\/span><\/span><\/b><\/span><\/span>\r\n\t\t\t\t<\/p>\r\n\t\t\t<\/td>\r\n\t\t<\/tr>\r\n\t\t<tr style=\"height:12.0pt!msorm\">\r\n\t\t\t<td nowrap style=\"width: 61.35pt\" width=\"123\">\r\n\t\t\t\t<p align=\"center\" style=\"margin-bottom:.0001pt;text-align:center;padding:0cm5.4pt0cm5.4pt!msorm\">\r\n\t\t\t\t\t<span style=\"line-height:normal\"><span style=\"height:12.0pt\"><span><span style=\",serif\"><span style=\"color:black\">2<\/span><\/span><\/span><\/span><\/span>\r\n\t\t\t\t<\/p>\r\n\t\t\t<\/td>\r\n\t\t\t<td nowrap style=\"width: 30.55pt\" width=\"61\">\r\n\t\t\t\t<p align=\"center\" style=\"margin-bottom:.0001pt;text-align:center;padding:0cm5.4pt0cm5.4pt!msorm\">\r\n\t\t\t\t\t<span style=\"line-height:normal\"><span style=\"height:12.0pt\"><b><span><span style=\",serif\"><span style=\"color:black\">+<\/span><\/span><\/span><\/b><\/span><\/span>\r\n\t\t\t\t<\/p>\r\n\t\t\t<\/td>\r\n\t\t\t<td nowrap style=\"width: 63.8pt\" width=\"128\">\r\n\t\t\t\t<p align=\"center\" style=\"margin-bottom:.0001pt;text-align:center;padding:0cm5.4pt0cm5.4pt!msorm\">\r\n\t\t\t\t\t<span style=\"line-height:normal\"><span style=\"height:12.0pt\"><b><span><span style=\",serif\"><span style=\"color:black\">+<\/span><\/span><\/span><\/b><\/span><\/span>\r\n\t\t\t\t<\/p>\r\n\t\t\t<\/td>\r\n\t\t\t<td nowrap style=\"width: 45.3pt\" width=\"91\">\r\n\t\t\t\t<p align=\"center\" style=\"margin-bottom:.0001pt;text-align:center;padding:0cm5.4pt0cm5.4pt!msorm\">\r\n\t\t\t\t\t<span style=\"line-height:normal\"><span style=\"height:12.0pt\"><b><span><span style=\",serif\"><span style=\"color:black\">\u2013<\/span><\/span><\/span><\/b><\/span><\/span>\r\n\t\t\t\t<\/p>\r\n\t\t\t<\/td>\r\n\t\t<\/tr>\r\n\t\t<tr style=\"height:12.0pt!msorm\">\r\n\t\t\t<td nowrap style=\"width: 61.35pt\" width=\"123\">\r\n\t\t\t\t<p align=\"center\" style=\"margin-bottom:.0001pt;text-align:center;padding:0cm5.4pt0cm5.4pt!msorm\">\r\n\t\t\t\t\t<span style=\"line-height:normal\"><span style=\"height:12.0pt\"><span><span style=\",serif\"><span style=\"color:black\">3<\/span><\/span><\/span><\/span><\/span>\r\n\t\t\t\t<\/p>\r\n\t\t\t<\/td>\r\n\t\t\t<td nowrap style=\"width: 30.55pt\" width=\"61\">\r\n\t\t\t\t<p align=\"center\" style=\"margin-bottom:.0001pt;text-align:center;padding:0cm5.4pt0cm5.4pt!msorm\">\r\n\t\t\t\t\t<span style=\"line-height:normal\"><span style=\"height:12.0pt\"><b><span><span style=\",serif\"><span style=\"color:black\">+<\/span><\/span><\/span><\/b><\/span><\/span>\r\n\t\t\t\t<\/p>\r\n\t\t\t<\/td>\r\n\t\t\t<td nowrap style=\"width: 63.8pt\" width=\"128\">\r\n\t\t\t\t<p align=\"center\" style=\"margin-bottom:.0001pt;text-align:center;padding:0cm5.4pt0cm5.4pt!msorm\">\r\n\t\t\t\t\t<span style=\"line-height:normal\"><span style=\"height:12.0pt\"><b><span><span style=\",serif\"><span style=\"color:black\">\u2013<\/span><\/span><\/span><\/b><\/span><\/span>\r\n\t\t\t\t<\/p>\r\n\t\t\t<\/td>\r\n\t\t\t<td nowrap style=\"width: 45.3pt\" width=\"91\">\r\n\t\t\t\t<p align=\"center\" style=\"margin-bottom:.0001pt;text-align:center;padding:0cm5.4pt0cm5.4pt!msorm\">\r\n\t\t\t\t\t<span style=\"line-height:normal\"><span style=\"height:12.0pt\"><b><span><span style=\",serif\"><span style=\"color:black\">+<\/span><\/span><\/span><\/b><\/span><\/span>\r\n\t\t\t\t<\/p>\r\n\t\t\t<\/td>\r\n\t\t<\/tr>\r\n\t\t<tr style=\"height:12.0pt!msorm\">\r\n\t\t\t<td nowrap style=\"width: 61.35pt\" width=\"123\">\r\n\t\t\t\t<p align=\"center\" style=\"margin-bottom:.0001pt;text-align:center;padding:0cm5.4pt0cm5.4pt!msorm\">\r\n\t\t\t\t\t<span style=\"line-height:normal\"><span style=\"height:12.0pt\"><span><span style=\",serif\"><span style=\"color:black\">4<\/span><\/span><\/span><\/span><\/span>\r\n\t\t\t\t<\/p>\r\n\t\t\t<\/td>\r\n\t\t\t<td nowrap style=\"width: 30.55pt\" width=\"61\">\r\n\t\t\t\t<p align=\"center\" style=\"margin-bottom:.0001pt;text-align:center;padding:0cm5.4pt0cm5.4pt!msorm\">\r\n\t\t\t\t\t<span style=\"line-height:normal\"><span style=\"height:12.0pt\"><b><span><span style=\",serif\"><span style=\"color:black\">+<\/span><\/span><\/span><\/b><\/span><\/span>\r\n\t\t\t\t<\/p>\r\n\t\t\t<\/td>\r\n\t\t\t<td nowrap style=\"width: 63.8pt\" width=\"128\">\r\n\t\t\t\t<p align=\"center\" style=\"margin-bottom:.0001pt;text-align:center;padding:0cm5.4pt0cm5.4pt!msorm\">\r\n\t\t\t\t\t<span style=\"line-height:normal\"><span style=\"height:12.0pt\"><b><span><span style=\",serif\"><span style=\"color:black\">\u2013<\/span><\/span><\/span><\/b><\/span><\/span>\r\n\t\t\t\t<\/p>\r\n\t\t\t<\/td>\r\n\t\t\t<td nowrap style=\"width: 45.3pt\" width=\"91\">\r\n\t\t\t\t<p align=\"center\" style=\"margin-bottom:.0001pt;text-align:center;padding:0cm5.4pt0cm5.4pt!msorm\">\r\n\t\t\t\t\t<span style=\"line-height:normal\"><span style=\"height:12.0pt\"><b><span><span style=\",serif\"><span style=\"color:black\">\u2013<\/span><\/span><\/span><\/b><\/span><\/span>\r\n\t\t\t\t<\/p>\r\n\t\t\t<\/td>\r\n\t\t<\/tr>\r\n\t\t<tr style=\"height:12.0pt!msorm\">\r\n\t\t\t<td nowrap style=\"width: 61.35pt\" width=\"123\">\r\n\t\t\t\t<p align=\"center\" style=\"margin-bottom:.0001pt;text-align:center;padding:0cm5.4pt0cm5.4pt!msorm\">\r\n\t\t\t\t\t<span style=\"line-height:normal\"><span style=\"height:12.0pt\"><span><span style=\",serif\"><span style=\"color:black\">5<\/span><\/span><\/span><\/span><\/span>\r\n\t\t\t\t<\/p>\r\n\t\t\t<\/td>\r\n\t\t\t<td nowrap style=\"width: 30.55pt\" width=\"61\">\r\n\t\t\t\t<p align=\"center\" style=\"margin-bottom:.0001pt;text-align:center;padding:0cm5.4pt0cm5.4pt!msorm\">\r\n\t\t\t\t\t<span style=\"line-height:normal\"><span style=\"height:12.0pt\"><b><span><span style=\",serif\"><span style=\"color:black\">\u2013<\/span><\/span><\/span><\/b><\/span><\/span>\r\n\t\t\t\t<\/p>\r\n\t\t\t<\/td>\r\n\t\t\t<td nowrap style=\"width: 63.8pt\" width=\"128\">\r\n\t\t\t\t<p align=\"center\" style=\"margin-bottom:.0001pt;text-align:center;padding:0cm5.4pt0cm5.4pt!msorm\">\r\n\t\t\t\t\t<span style=\"line-height:normal\"><span style=\"height:12.0pt\"><b><span><span style=\",serif\"><span style=\"color:black\">+<\/span><\/span><\/span><\/b><\/span><\/span>\r\n\t\t\t\t<\/p>\r\n\t\t\t<\/td>\r\n\t\t\t<td nowrap style=\"width: 45.3pt\" width=\"91\">\r\n\t\t\t\t<p align=\"center\" style=\"margin-bottom:.0001pt;text-align:center;padding:0cm5.4pt0cm5.4pt!msorm\">\r\n\t\t\t\t\t<span style=\"line-height:normal\"><span style=\"height:12.0pt\"><b><span><span style=\",serif\"><span style=\"color:black\">+<\/span><\/span><\/span><\/b><\/span><\/span>\r\n\t\t\t\t<\/p>\r\n\t\t\t<\/td>\r\n\t\t<\/tr>\r\n\t\t<tr style=\"height:12.0pt!msorm\">\r\n\t\t\t<td nowrap style=\"width: 61.35pt\" width=\"123\">\r\n\t\t\t\t<p align=\"center\" style=\"margin-bottom:.0001pt;text-align:center;padding:0cm5.4pt0cm5.4pt!msorm\">\r\n\t\t\t\t\t<span style=\"line-height:normal\"><span style=\"height:12.0pt\"><span><span style=\",serif\"><span style=\"color:black\">6<\/span><\/span><\/span><\/span><\/span>\r\n\t\t\t\t<\/p>\r\n\t\t\t<\/td>\r\n\t\t\t<td nowrap style=\"width: 30.55pt\" width=\"61\">\r\n\t\t\t\t<p align=\"center\" style=\"margin-bottom:.0001pt;text-align:center;padding:0cm5.4pt0cm5.4pt!msorm\">\r\n\t\t\t\t\t<span style=\"line-height:normal\"><span style=\"height:12.0pt\"><b><span><span style=\",serif\"><span style=\"color:black\">\u2013<\/span><\/span><\/span><\/b><\/span><\/span>\r\n\t\t\t\t<\/p>\r\n\t\t\t<\/td>\r\n\t\t\t<td nowrap style=\"width: 63.8pt\" width=\"128\">\r\n\t\t\t\t<p align=\"center\" style=\"margin-bottom:.0001pt;text-align:center;padding:0cm5.4pt0cm5.4pt!msorm\">\r\n\t\t\t\t\t<span style=\"line-height:normal\"><span style=\"height:12.0pt\"><b><span><span style=\",serif\"><span style=\"color:black\">+<\/span><\/span><\/span><\/b><\/span><\/span>\r\n\t\t\t\t<\/p>\r\n\t\t\t<\/td>\r\n\t\t\t<td nowrap style=\"width: 45.3pt\" width=\"91\">\r\n\t\t\t\t<p align=\"center\" style=\"margin-bottom:.0001pt;text-align:center;padding:0cm5.4pt0cm5.4pt!msorm\">\r\n\t\t\t\t\t<span style=\"line-height:normal\"><span style=\"height:12.0pt\"><b><span><span style=\",serif\"><span style=\"color:black\">\u2013<\/span><\/span><\/span><\/b><\/span><\/span>\r\n\t\t\t\t<\/p>\r\n\t\t\t<\/td>\r\n\t\t<\/tr>\r\n\t\t<tr style=\"height:12.0pt!msorm\">\r\n\t\t\t<td nowrap style=\"width: 61.35pt\" width=\"123\">\r\n\t\t\t\t<p align=\"center\" style=\"margin-bottom:.0001pt;text-align:center;padding:0cm5.4pt0cm5.4pt!msorm\">\r\n\t\t\t\t\t<span style=\"line-height:normal\"><span style=\"height:12.0pt\"><span><span style=\",serif\"><span style=\"color:black\">7<\/span><\/span><\/span><\/span><\/span>\r\n\t\t\t\t<\/p>\r\n\t\t\t<\/td>\r\n\t\t\t<td nowrap style=\"width: 30.55pt\" width=\"61\">\r\n\t\t\t\t<p align=\"center\" style=\"margin-bottom:.0001pt;text-align:center;padding:0cm5.4pt0cm5.4pt!msorm\">\r\n\t\t\t\t\t<span style=\"line-height:normal\"><span style=\"height:12.0pt\"><b><span><span style=\",serif\"><span style=\"color:black\">\u2013<\/span><\/span><\/span><\/b><\/span><\/span>\r\n\t\t\t\t<\/p>\r\n\t\t\t<\/td>\r\n\t\t\t<td nowrap style=\"width: 63.8pt\" width=\"128\">\r\n\t\t\t\t<p align=\"center\" style=\"margin-bottom:.0001pt;text-align:center;padding:0cm5.4pt0cm5.4pt!msorm\">\r\n\t\t\t\t\t<span style=\"line-height:normal\"><span style=\"height:12.0pt\"><b><span><span style=\",serif\"><span style=\"color:black\">\u2013<\/span><\/span><\/span><\/b><\/span><\/span>\r\n\t\t\t\t<\/p>\r\n\t\t\t<\/td>\r\n\t\t\t<td nowrap style=\"width: 45.3pt\" width=\"91\">\r\n\t\t\t\t<p align=\"center\" style=\"margin-bottom:.0001pt;text-align:center;padding:0cm5.4pt0cm5.4pt!msorm\">\r\n\t\t\t\t\t<span style=\"line-height:normal\"><span style=\"height:12.0pt\"><b><span><span style=\",serif\"><span style=\"color:black\">+<\/span><\/span><\/span><\/b><\/span><\/span>\r\n\t\t\t\t<\/p>\r\n\t\t\t<\/td>\r\n\t\t<\/tr>\r\n\t\t<tr style=\"height:12.4pt!msorm\">\r\n\t\t\t<td nowrap style=\"width: 61.35pt\" width=\"123\">\r\n\t\t\t\t<p align=\"center\" style=\"margin-bottom:.0001pt;text-align:center;padding:0cm5.4pt0cm5.4pt!msorm\">\r\n\t\t\t\t\t<span style=\"line-height:normal\"><span style=\"height:12.4pt\"><span><span style=\",serif\"><span style=\"color:black\">8<\/span><\/span><\/span><\/span><\/span>\r\n\t\t\t\t<\/p>\r\n\t\t\t<\/td>\r\n\t\t\t<td nowrap style=\"width: 30.55pt\" width=\"61\">\r\n\t\t\t\t<p align=\"center\" style=\"margin-bottom:.0001pt;text-align:center;padding:0cm5.4pt0cm5.4pt!msorm\">\r\n\t\t\t\t\t<span style=\"line-height:normal\"><span style=\"height:12.4pt\"><b><span><span style=\",serif\"><span style=\"color:black\">\u2013<\/span><\/span><\/span><\/b><\/span><\/span>\r\n\t\t\t\t<\/p>\r\n\t\t\t<\/td>\r\n\t\t\t<td nowrap style=\"width: 63.8pt\" width=\"128\">\r\n\t\t\t\t<p align=\"center\" style=\"margin-bottom:.0001pt;text-align:center;padding:0cm5.4pt0cm5.4pt!msorm\">\r\n\t\t\t\t\t<span style=\"line-height:normal\"><span style=\"height:12.4pt\"><b><span><span style=\",serif\"><span style=\"color:black\">\u2013<\/span><\/span><\/span><\/b><\/span><\/span>\r\n\t\t\t\t<\/p>\r\n\t\t\t<\/td>\r\n\t\t\t<td nowrap style=\"width: 45.3pt\" width=\"91\">\r\n\t\t\t\t<p align=\"center\" style=\"margin-bottom:.0001pt;text-align:center;padding:0cm5.4pt0cm5.4pt!msorm\">\r\n\t\t\t\t\t<span style=\"line-height:normal\"><span style=\"height:12.4pt\"><b><span><span style=\",serif\"><span style=\"color:black\">\u2013<\/span><\/span><\/span><\/b><\/span><\/span>\r\n\t\t\t\t<\/p>\r\n\t\t\t<\/td>\r\n\t\t<\/tr>\r\n\t<\/tbody>\r\n<\/table>\r\n\r\n<p>\r\n\t\u00a0\r\n<\/p>\r\n\r\n<p style=\"margin-bottom:7.5pt;text-align:justify\">\r\n\t<span><span><span style=\"color:#2f2f2f\">c) Behind all the + and \u2013 signs are factor levels that are used to carry out the experiments. The decoded factors for this Full Factorial Design can be seen in the table below. Practically this means in our example to perform 8 LC<\/span><\/span><\/span> runs with 4 different eluent solutions and varying temperatures in the column compartment according to the decoded values in the table below. All experiments need to be carried out in a random order to prevent a bias on the response variable. The<span><span><span style=\"color:#2f2f2f\"> reason for this is that unknown factors might be aligned with each other if the experiments are not performed in random order. From those 8 runs we collect all the retention times of all substances in the mixture in order to determine the robustness individually for every analyte. It is always possible to use more response variables than just the one (retention time) depicted here in this example.<\/span><\/span><\/span>\r\n<\/p>\r\n\r\n<p style=\"margin-bottom:7.5pt;text-align:justify\">\r\n\t<span><span><span style=\"color:#2f2f2f\">Now in this step, we perform all the experiments and transfer the results, in our case, the retention time, to the table.<\/span><\/span><\/span>\r\n<\/p>\r\n\r\n<p>\r\n\t\u00a0\r\n<\/p>\r\n\r\n<div align=\"center\">\r\n\t<table class=\"table table-hover\" align=\"center\" border=\"1\" style=\"border-collapse: collapse\" width=\"698\">\r\n\t\t<thead>\r\n\t\t\t<tr style=\"height:28.55pt\">\r\n\t\t\t\t<th nowrap scope=\"col\" style=\"width: 52.8pt\" width=\"106\">\r\n\t\t\t\t\t<p align=\"center\" style=\"margin-bottom:.0001pt;text-align:center;padding:0cm5.4pt0cm5.4pt\">\r\n\t\t\t\t\t\t<span style=\"line-height:normal\"><span style=\"height:28.55pt\"><span style=\"color:black\">Experiment number<\/span><\/span><\/span>\r\n\t\t\t\t\t<\/p>\r\n\t\t\t\t<\/th>\r\n\t\t\t\t<th scope=\"col\" style=\"width: 67.45pt\" width=\"135\">\r\n\t\t\t\t\t<p align=\"center\" style=\"margin-bottom:.0001pt;text-align:center;padding:0cm5.4pt0cm5.4pt\">\r\n\t\t\t\t\t\t<span style=\"line-height:normal\"><span style=\"height:28.55pt\"><span style=\"color:black\">Actual order of measurement<\/span><\/span><\/span>\r\n\t\t\t\t\t<\/p>\r\n\t\t\t\t<\/th>\r\n\t\t\t\t<th nowrap scope=\"col\" style=\"width: 23.1pt\" width=\"46\">\r\n\t\t\t\t\t<p align=\"center\" style=\"margin-bottom:.0001pt;text-align:center;padding:0cm5.4pt0cm5.4pt\">\r\n\t\t\t\t\t\t<span style=\"line-height:normal\"><span style=\"height:28.55pt\"><b><span style=\"color:black\">A<\/span><\/b><\/span><\/span>\r\n\t\t\t\t\t<\/p>\r\n\r\n\t\t\t\t\t<p align=\"center\" style=\"margin-bottom:.0001pt;text-align:center;padding:0cm5.4pt0cm5.4pt\">\r\n\t\t\t\t\t\t\u00a0\r\n\t\t\t\t\t<\/p>\r\n\r\n\t\t\t\t\t<p align=\"center\" style=\"margin-bottom:.0001pt;text-align:center;padding:0cm5.4pt0cm5.4pt\">\r\n\t\t\t\t\t\t<span style=\"line-height:normal\"><span style=\"height:28.55pt\"><span style=\"color:black\">pH <\/span><\/span><\/span>\r\n\t\t\t\t\t<\/p>\r\n\t\t\t\t<\/th>\r\n\t\t\t\t<th scope=\"col\" style=\"width: 63.8pt\" width=\"128\">\r\n\t\t\t\t\t<p align=\"center\" style=\"margin-bottom:.0001pt;text-align:center;padding:0cm5.4pt0cm5.4pt\">\r\n\t\t\t\t\t\t<span style=\"line-height:normal\"><span style=\"height:28.55pt\"><b><span style=\"color:black\">B<\/span><\/b><\/span><\/span>\r\n\t\t\t\t\t<\/p>\r\n\r\n\t\t\t\t\t<p align=\"center\" style=\"margin-bottom:.0001pt;text-align:center;padding:0cm5.4pt0cm5.4pt\">\r\n\t\t\t\t\t\t<span style=\"line-height:normal\"><span style=\"height:28.55pt\"><span style=\"color:black\">Additive conc.<br>[mmol\/L]<\/span><\/span><\/span>\r\n\t\t\t\t\t<\/p>\r\n\t\t\t\t<\/th>\r\n\t\t\t\t<th scope=\"col\" style=\"width: 70.9pt\" width=\"142\">\r\n\t\t\t\t\t<p align=\"center\" style=\"margin-bottom:.0001pt;text-align:center;padding:0cm5.4pt0cm5.4pt\">\r\n\t\t\t\t\t\t<span style=\"line-height:normal\"><span style=\"height:28.55pt\"><b><span style=\"color:black\">C<\/span><\/b><\/span><\/span>\r\n\t\t\t\t\t<\/p>\r\n\r\n\t\t\t\t\t<p align=\"center\" style=\"margin-bottom:.0001pt;text-align:center;padding:0cm5.4pt0cm5.4pt\">\r\n\t\t\t\t\t\t<span style=\"line-height:normal\"><span style=\"height:28.55pt\"><span style=\"color:black\">Column Temp.<br>[\u00b0C]<\/span><\/span><\/span>\r\n\t\t\t\t\t<\/p>\r\n\t\t\t\t<\/th>\r\n\t\t\t\t<th scope=\"col\" style=\"width: 70.85pt\" width=\"142\">\r\n\t\t\t\t\t<p align=\"center\" style=\"margin-bottom:.0001pt;text-align:center;padding:0cm5.4pt0cm5.4pt\">\r\n\t\t\t\t\t\t<span style=\"line-height:normal\"><span style=\"height:28.55pt\"><b><span style=\"color:black\">Response 1<\/span><\/b><\/span><\/span>\r\n\t\t\t\t\t<\/p>\r\n\r\n\t\t\t\t\t<p align=\"center\" style=\"margin-bottom:.0001pt;text-align:center;padding:0cm5.4pt0cm5.4pt\">\r\n\t\t\t\t\t\t<span style=\"line-height:normal\"><span style=\"height:28.55pt\"><span style=\"color:black\">Ret. Time<br>[min]<\/span><\/span><\/span>\r\n\t\t\t\t\t<\/p>\r\n\t\t\t\t<\/th>\r\n\t\t\t<\/tr>\r\n\t\t<\/thead>\r\n\t\t<tbody>\r\n\t\t\t<tr style=\"height:13.9pt\">\r\n\t\t\t\t<td nowrap style=\"width: 52.8pt\" width=\"106\">\r\n\t\t\t\t\t<p align=\"center\" style=\"margin-bottom:.0001pt;text-align:center;padding:0cm5.4pt0cm5.4pt\">\r\n\t\t\t\t\t\t<span style=\"line-height:normal\"><span style=\"height:13.9pt\"><b><span style=\"color:black\">1<\/span><\/b><\/span><\/span>\r\n\t\t\t\t\t<\/p>\r\n\t\t\t\t<\/td>\r\n\t\t\t\t<td style=\"width: 67.45pt\" width=\"135\">\r\n\t\t\t\t\t<p align=\"center\" style=\"margin-bottom:.0001pt;text-align:center;padding:0cm5.4pt0cm5.4pt\">\r\n\t\t\t\t\t\t<span style=\"line-height:normal\"><span style=\"height:13.9pt\"><span style=\"color:black\">3<\/span><\/span><\/span>\r\n\t\t\t\t\t<\/p>\r\n\t\t\t\t<\/td>\r\n\t\t\t\t<td nowrap>\r\n\t\t\t\t\t<p style=\"text-align: center\">\r\n\t\t\t\t\t\t9.8\r\n\t\t\t\t\t<\/p>\r\n\t\t\t\t<\/td>\r\n\t\t\t\t<td nowrap>\r\n\t\t\t\t\t<p style=\"text-align: center\">\r\n\t\t\t\t\t\t5.2\r\n\t\t\t\t\t<\/p>\r\n\t\t\t\t<\/td>\r\n\t\t\t\t<td nowrap>\r\n\t\t\t\t\t<p style=\"text-align: center\">\r\n\t\t\t\t\t\t31\r\n\t\t\t\t\t<\/p>\r\n\t\t\t\t<\/td>\r\n\t\t\t\t<td nowrap style=\"width: 70.85pt\" width=\"142\">\r\n\t\t\t\t\t<p align=\"center\" style=\"margin-bottom:.0001pt;text-align:center;padding:0cm5.4pt0cm5.4pt\">\r\n\t\t\t\t\t\t<span style=\"line-height:normal\"><span style=\"height:13.9pt\"><span style=\"color:black\">8.31<\/span><\/span><\/span>\r\n\t\t\t\t\t<\/p>\r\n\t\t\t\t<\/td>\r\n\t\t\t<\/tr>\r\n\t\t\t<tr style=\"height:13.9pt\">\r\n\t\t\t\t<td nowrap style=\"width: 52.8pt\" width=\"106\">\r\n\t\t\t\t\t<p align=\"center\" style=\"margin-bottom:.0001pt;text-align:center;padding:0cm5.4pt0cm5.4pt\">\r\n\t\t\t\t\t\t<span style=\"line-height:normal\"><span style=\"height:13.9pt\"><b><span style=\"color:black\">2<\/span><\/b><\/span><\/span>\r\n\t\t\t\t\t<\/p>\r\n\t\t\t\t<\/td>\r\n\t\t\t\t<td style=\"width: 67.45pt\" width=\"135\">\r\n\t\t\t\t\t<p align=\"center\" style=\"margin-bottom:.0001pt;text-align:center;padding:0cm5.4pt0cm5.4pt\">\r\n\t\t\t\t\t\t<span style=\"line-height:normal\"><span style=\"height:13.9pt\"><span style=\"color:black\">4<\/span><\/span><\/span>\r\n\t\t\t\t\t<\/p>\r\n\t\t\t\t<\/td>\r\n\t\t\t\t<td nowrap>\r\n\t\t\t\t\t<p style=\"text-align: center\">\r\n\t\t\t\t\t\t9.8\r\n\t\t\t\t\t<\/p>\r\n\t\t\t\t<\/td>\r\n\t\t\t\t<td nowrap>\r\n\t\t\t\t\t<p style=\"text-align: center\">\r\n\t\t\t\t\t\t5.2\r\n\t\t\t\t\t<\/p>\r\n\t\t\t\t<\/td>\r\n\t\t\t\t<td nowrap>\r\n\t\t\t\t\t<p style=\"text-align: center\">\r\n\t\t\t\t\t\t29\r\n\t\t\t\t\t<\/p>\r\n\t\t\t\t<\/td>\r\n\t\t\t\t<td nowrap style=\"width: 70.85pt\" width=\"142\">\r\n\t\t\t\t\t<p align=\"center\" style=\"margin-bottom:.0001pt;text-align:center;padding:0cm5.4pt0cm5.4pt\">\r\n\t\t\t\t\t\t<span style=\"line-height:normal\"><span style=\"height:13.9pt\"><span style=\"color:black\">8.1<\/span><span style=\"color:black\">0<\/span><\/span><\/span>\r\n\t\t\t\t\t<\/p>\r\n\t\t\t\t<\/td>\r\n\t\t\t<\/tr>\r\n\t\t\t<tr style=\"height:13.9pt\">\r\n\t\t\t\t<td nowrap style=\"width: 52.8pt\" width=\"106\">\r\n\t\t\t\t\t<p align=\"center\" style=\"margin-bottom:.0001pt;text-align:center;padding:0cm5.4pt0cm5.4pt\">\r\n\t\t\t\t\t\t<span style=\"line-height:normal\"><span style=\"height:13.9pt\"><b><span style=\"color:black\">3<\/span><\/b><\/span><\/span>\r\n\t\t\t\t\t<\/p>\r\n\t\t\t\t<\/td>\r\n\t\t\t\t<td style=\"width: 67.45pt\" width=\"135\">\r\n\t\t\t\t\t<p align=\"center\" style=\"margin-bottom:.0001pt;text-align:center;padding:0cm5.4pt0cm5.4pt\">\r\n\t\t\t\t\t\t<span style=\"line-height:normal\"><span style=\"height:13.9pt\"><span style=\"color:black\">2<\/span><\/span><\/span>\r\n\t\t\t\t\t<\/p>\r\n\t\t\t\t<\/td>\r\n\t\t\t\t<td nowrap style=\"text-align: center\">\r\n\t\t\t\t\t<p>\r\n\t\t\t\t\t\t9.8\r\n\t\t\t\t\t<\/p>\r\n\t\t\t\t<\/td>\r\n\t\t\t\t<td nowrap>\r\n\t\t\t\t\t<p style=\"text-align: center\">\r\n\t\t\t\t\t\t4.8\r\n\t\t\t\t\t<\/p>\r\n\t\t\t\t<\/td>\r\n\t\t\t\t<td nowrap>\r\n\t\t\t\t\t<p style=\"text-align: center\">\r\n\t\t\t\t\t\t31\r\n\t\t\t\t\t<\/p>\r\n\t\t\t\t<\/td>\r\n\t\t\t\t<td nowrap style=\"width: 70.85pt\" width=\"142\">\r\n\t\t\t\t\t<p align=\"center\" style=\"margin-bottom:.0001pt;text-align:center;padding:0cm5.4pt0cm5.4pt\">\r\n\t\t\t\t\t\t<span style=\"line-height:normal\"><span style=\"height:13.9pt\"><span style=\"color:black\">7.24<\/span><\/span><\/span>\r\n\t\t\t\t\t<\/p>\r\n\t\t\t\t<\/td>\r\n\t\t\t<\/tr>\r\n\t\t\t<tr style=\"height:13.9pt\">\r\n\t\t\t\t<td nowrap style=\"width: 52.8pt\" width=\"106\">\r\n\t\t\t\t\t<p align=\"center\" style=\"margin-bottom:.0001pt;text-align:center;padding:0cm5.4pt0cm5.4pt\">\r\n\t\t\t\t\t\t<span style=\"line-height:normal\"><span style=\"height:13.9pt\"><b><span style=\"color:black\">4<\/span><\/b><\/span><\/span>\r\n\t\t\t\t\t<\/p>\r\n\t\t\t\t<\/td>\r\n\t\t\t\t<td style=\"width: 67.45pt\" width=\"135\">\r\n\t\t\t\t\t<p align=\"center\" style=\"margin-bottom:.0001pt;text-align:center;padding:0cm5.4pt0cm5.4pt\">\r\n\t\t\t\t\t\t<span style=\"line-height:normal\"><span style=\"height:13.9pt\"><span style=\"color:black\">5<\/span><\/span><\/span>\r\n\t\t\t\t\t<\/p>\r\n\t\t\t\t<\/td>\r\n\t\t\t\t<td nowrap>\r\n\t\t\t\t\t<p style=\"text-align: center\">\r\n\t\t\t\t\t\t9.8\r\n\t\t\t\t\t<\/p>\r\n\t\t\t\t<\/td>\r\n\t\t\t\t<td nowrap>\r\n\t\t\t\t\t<p style=\"text-align: center\">\r\n\t\t\t\t\t\t4.8\r\n\t\t\t\t\t<\/p>\r\n\t\t\t\t<\/td>\r\n\t\t\t\t<td nowrap>\r\n\t\t\t\t\t<p style=\"text-align: center\">\r\n\t\t\t\t\t\t29\r\n\t\t\t\t\t<\/p>\r\n\t\t\t\t<\/td>\r\n\t\t\t\t<td nowrap style=\"width: 70.85pt\" width=\"142\">\r\n\t\t\t\t\t<p align=\"center\" style=\"margin-bottom:.0001pt;text-align:center;padding:0cm5.4pt0cm5.4pt\">\r\n\t\t\t\t\t\t<span style=\"line-height:normal\"><span style=\"height:13.9pt\"><span style=\"color:black\">7.43<\/span><\/span><\/span>\r\n\t\t\t\t\t<\/p>\r\n\t\t\t\t<\/td>\r\n\t\t\t<\/tr>\r\n\t\t\t<tr style=\"height:13.9pt\">\r\n\t\t\t\t<td nowrap style=\"width: 52.8pt\" width=\"106\">\r\n\t\t\t\t\t<p align=\"center\" style=\"margin-bottom:.0001pt;text-align:center;padding:0cm5.4pt0cm5.4pt\">\r\n\t\t\t\t\t\t<span style=\"line-height:normal\"><span style=\"height:13.9pt\"><b><span style=\"color:black\">5<\/span><\/b><\/span><\/span>\r\n\t\t\t\t\t<\/p>\r\n\t\t\t\t<\/td>\r\n\t\t\t\t<td style=\"width: 67.45pt\" width=\"135\">\r\n\t\t\t\t\t<p align=\"center\" style=\"margin-bottom:.0001pt;text-align:center;padding:0cm5.4pt0cm5.4pt\">\r\n\t\t\t\t\t\t<span style=\"line-height:normal\"><span style=\"height:13.9pt\"><span style=\"color:black\">6<\/span><\/span><\/span>\r\n\t\t\t\t\t<\/p>\r\n\t\t\t\t<\/td>\r\n\t\t\t\t<td nowrap>\r\n\t\t\t\t\t<p style=\"text-align: center\">\r\n\t\t\t\t\t\t9.4\r\n\t\t\t\t\t<\/p>\r\n\t\t\t\t<\/td>\r\n\t\t\t\t<td nowrap>\r\n\t\t\t\t\t<p style=\"text-align: center\">\r\n\t\t\t\t\t\t5.2\r\n\t\t\t\t\t<\/p>\r\n\t\t\t\t<\/td>\r\n\t\t\t\t<td nowrap>\r\n\t\t\t\t\t<p style=\"text-align: center\">\r\n\t\t\t\t\t\t31\r\n\t\t\t\t\t<\/p>\r\n\t\t\t\t<\/td>\r\n\t\t\t\t<td nowrap style=\"width: 70.85pt\" width=\"142\">\r\n\t\t\t\t\t<p align=\"center\" style=\"margin-bottom:.0001pt;text-align:center;padding:0cm5.4pt0cm5.4pt\">\r\n\t\t\t\t\t\t<span style=\"line-height:normal\"><span style=\"height:13.9pt\"><span style=\"color:black\">8.32<\/span><\/span><\/span>\r\n\t\t\t\t\t<\/p>\r\n\t\t\t\t<\/td>\r\n\t\t\t<\/tr>\r\n\t\t\t<tr style=\"height:13.9pt\">\r\n\t\t\t\t<td nowrap style=\"width: 52.8pt\" width=\"106\">\r\n\t\t\t\t\t<p align=\"center\" style=\"margin-bottom:.0001pt;text-align:center;padding:0cm5.4pt0cm5.4pt\">\r\n\t\t\t\t\t\t<span style=\"line-height:normal\"><span style=\"height:13.9pt\"><b><span style=\"color:black\">6<\/span><\/b><\/span><\/span>\r\n\t\t\t\t\t<\/p>\r\n\t\t\t\t<\/td>\r\n\t\t\t\t<td style=\"width: 67.45pt\" width=\"135\">\r\n\t\t\t\t\t<p align=\"center\" style=\"margin-bottom:.0001pt;text-align:center;padding:0cm5.4pt0cm5.4pt\">\r\n\t\t\t\t\t\t<span style=\"line-height:normal\"><span style=\"height:13.9pt\"><span style=\"color:black\">7<\/span><\/span><\/span>\r\n\t\t\t\t\t<\/p>\r\n\t\t\t\t<\/td>\r\n\t\t\t\t<td nowrap>\r\n\t\t\t\t\t<p style=\"text-align: center\">\r\n\t\t\t\t\t\t9.4\r\n\t\t\t\t\t<\/p>\r\n\t\t\t\t<\/td>\r\n\t\t\t\t<td nowrap>\r\n\t\t\t\t\t<p style=\"text-align: center\">\r\n\t\t\t\t\t\t5.2\r\n\t\t\t\t\t<\/p>\r\n\t\t\t\t<\/td>\r\n\t\t\t\t<td nowrap>\r\n\t\t\t\t\t<p style=\"text-align: center\">\r\n\t\t\t\t\t\t29\r\n\t\t\t\t\t<\/p>\r\n\t\t\t\t<\/td>\r\n\t\t\t\t<td nowrap style=\"width: 70.85pt\" width=\"142\">\r\n\t\t\t\t\t<p align=\"center\" style=\"margin-bottom:.0001pt;text-align:center;padding:0cm5.4pt0cm5.4pt\">\r\n\t\t\t\t\t\t<span style=\"line-height:normal\"><span style=\"height:13.9pt\"><span style=\"color:black\">8.92<\/span><\/span><\/span>\r\n\t\t\t\t\t<\/p>\r\n\t\t\t\t<\/td>\r\n\t\t\t<\/tr>\r\n\t\t\t<tr style=\"height:13.9pt\">\r\n\t\t\t\t<td nowrap style=\"width: 52.8pt\" width=\"106\">\r\n\t\t\t\t\t<p align=\"center\" style=\"margin-bottom:.0001pt;text-align:center;padding:0cm5.4pt0cm5.4pt\">\r\n\t\t\t\t\t\t<span style=\"line-height:normal\"><span style=\"height:13.9pt\"><b><span style=\"color:black\">7<\/span><\/b><\/span><\/span>\r\n\t\t\t\t\t<\/p>\r\n\t\t\t\t<\/td>\r\n\t\t\t\t<td style=\"width: 67.45pt\" width=\"135\">\r\n\t\t\t\t\t<p align=\"center\" style=\"margin-bottom:.0001pt;text-align:center;padding:0cm5.4pt0cm5.4pt\">\r\n\t\t\t\t\t\t<span style=\"line-height:normal\"><span style=\"height:13.9pt\"><span style=\"color:black\">1<\/span><\/span><\/span>\r\n\t\t\t\t\t<\/p>\r\n\t\t\t\t<\/td>\r\n\t\t\t\t<td nowrap>\r\n\t\t\t\t\t<p style=\"text-align: center\">\r\n\t\t\t\t\t\t9.4\r\n\t\t\t\t\t<\/p>\r\n\t\t\t\t<\/td>\r\n\t\t\t\t<td nowrap>\r\n\t\t\t\t\t<p style=\"text-align: center\">\r\n\t\t\t\t\t\t4.8\r\n\t\t\t\t\t<\/p>\r\n\t\t\t\t<\/td>\r\n\t\t\t\t<td nowrap>\r\n\t\t\t\t\t<p style=\"text-align: center\">\r\n\t\t\t\t\t\t31\r\n\t\t\t\t\t<\/p>\r\n\t\t\t\t<\/td>\r\n\t\t\t\t<td nowrap style=\"width: 70.85pt\" width=\"142\">\r\n\t\t\t\t\t<p align=\"center\" style=\"margin-bottom:.0001pt;text-align:center;padding:0cm5.4pt0cm5.4pt\">\r\n\t\t\t\t\t\t<span style=\"line-height:normal\"><span style=\"height:13.9pt\"><span style=\"color:black\">9.84<\/span><\/span><\/span>\r\n\t\t\t\t\t<\/p>\r\n\t\t\t\t<\/td>\r\n\t\t\t<\/tr>\r\n\t\t\t<tr style=\"height:14.5pt\">\r\n\t\t\t\t<td nowrap style=\"width: 52.8pt\" width=\"106\">\r\n\t\t\t\t\t<p align=\"center\" style=\"margin-bottom:.0001pt;text-align:center;padding:0cm5.4pt0cm5.4pt\">\r\n\t\t\t\t\t\t<span style=\"line-height:normal\"><span style=\"height:14.5pt\"><b><span style=\"color:black\">8<\/span><\/b><\/span><\/span>\r\n\t\t\t\t\t<\/p>\r\n\t\t\t\t<\/td>\r\n\t\t\t\t<td style=\"width: 67.45pt\" width=\"135\">\r\n\t\t\t\t\t<p align=\"center\" style=\"margin-bottom:.0001pt;text-align:center;padding:0cm5.4pt0cm5.4pt\">\r\n\t\t\t\t\t\t<span style=\"line-height:normal\"><span style=\"height:14.5pt\"><span style=\"color:black\">8<\/span><\/span><\/span>\r\n\t\t\t\t\t<\/p>\r\n\t\t\t\t<\/td>\r\n\t\t\t\t<td nowrap>\r\n\t\t\t\t\t<p style=\"text-align: center\">\r\n\t\t\t\t\t\t9.4\r\n\t\t\t\t\t<\/p>\r\n\t\t\t\t<\/td>\r\n\t\t\t\t<td nowrap>\r\n\t\t\t\t\t<p style=\"text-align: center\">\r\n\t\t\t\t\t\t4.8\r\n\t\t\t\t\t<\/p>\r\n\t\t\t\t<\/td>\r\n\t\t\t\t<td nowrap>\r\n\t\t\t\t\t<p style=\"text-align: center\">\r\n\t\t\t\t\t\t29\r\n\t\t\t\t\t<\/p>\r\n\t\t\t\t<\/td>\r\n\t\t\t\t<td nowrap style=\"width: 70.85pt\" width=\"142\">\r\n\t\t\t\t\t<p align=\"center\" style=\"margin-bottom:.0001pt;text-align:center;padding:0cm5.4pt0cm5.4pt\">\r\n\t\t\t\t\t\t<span style=\"line-height:normal\"><span style=\"height:14.5pt\"><span style=\"color:black\">9.93<\/span><\/span><\/span>\r\n\t\t\t\t\t<\/p>\r\n\t\t\t\t<\/td>\r\n\t\t\t<\/tr>\r\n\t\t<\/tbody>\r\n\t<\/table>\r\n\r\n\t<p style=\"text-align: left\">\r\n\t\t\u00a0\r\n\t<\/p>\r\n\r\n\t<p style=\"margin-bottom: 7.5pt;text-align: justify\">\r\n\t\t<span><span><span style=\"color:#2f2f2f\">d) Now it is time to calculate the actual effects of the three factors on the response variables. This is done by finding the average for the response at a high value (\u201c+\u201d) and subtracting the average of the response at the low value (\u201c-\u201d). In case of a retention time, we would have:<\/span><\/span><\/span>\r\n\t<\/p>\r\n\r\n\t<p style=\"text-align: left\">\r\n\t\t\u00a0\r\n\t<\/p>\r\n<\/div>\r\n\r\n<h6 style=\"text-align: center\">\r\n\t<img loading=\"lazy\" decoding=\"async\" width=\"1116\" height=\"130\" class=\"alignnone wp-image-78\" style=\"width: 420px;height: 45px\" src=\"https:\/\/sisu.ut.ee\/wp-content\/uploads\/sites\/130\/factora1.png\" title=\"factora1.png\" alt=\"FA1\" srcset=\"https:\/\/sisu.ut.ee\/wp-content\/uploads\/sites\/130\/factora1.png 1116w, https:\/\/sisu.ut.ee\/wp-content\/uploads\/sites\/130\/factora1-300x35.png 300w, https:\/\/sisu.ut.ee\/wp-content\/uploads\/sites\/130\/factora1-1024x119.png 1024w, https:\/\/sisu.ut.ee\/wp-content\/uploads\/sites\/130\/factora1-768x89.png 768w\" sizes=\"auto, (max-width: 1116px) 100vw, 1116px\">\u00a0 \u00a0 \u00a0 \u00a0(Eq 2)\r\n<\/h6>\r\n\r\n<p>\r\n\t\u00a0\r\n<\/p>\r\n\r\n<p style=\"margin-bottom:7.5pt;text-align:justify\">\r\n\t<span><span><span style=\"color:#2f2f2f\">Effect of the factor A on the response would be calculated as follows:<\/span><\/span><\/span>\r\n<\/p>\r\n\r\n<p>\r\n\t\u00a0\r\n<\/p>\r\n\r\n<h6 style=\"margin-bottom: 7.5pt;text-align: center\">\r\n\t<img loading=\"lazy\" decoding=\"async\" width=\"1343\" height=\"100\" class=\"alignnone wp-image-79\" style=\"width: 470px;height: 35px\" src=\"https:\/\/sisu.ut.ee\/wp-content\/uploads\/sites\/130\/factora2.png\" title=\"factora2.png\" alt=\"FA2\" srcset=\"https:\/\/sisu.ut.ee\/wp-content\/uploads\/sites\/130\/factora2.png 1343w, https:\/\/sisu.ut.ee\/wp-content\/uploads\/sites\/130\/factora2-300x22.png 300w, https:\/\/sisu.ut.ee\/wp-content\/uploads\/sites\/130\/factora2-1024x76.png 1024w, https:\/\/sisu.ut.ee\/wp-content\/uploads\/sites\/130\/factora2-768x57.png 768w\" sizes=\"auto, (max-width: 1343px) 100vw, 1343px\">(Eq 3)\r\n<\/h6>\r\n\r\n<p>\r\n\t<span><span><span style=\"color:#2f2f2f\">This is now the effect that the change in pH has on the response variable retention time. This calculation can be performed in a similar manner for all factors A, B, C, and all interactions.<\/span><\/span><\/span>\r\n<\/p>\r\n\r\n<div align=\"center\">\r\n\t<table class=\"table table-hover\" align=\"center\" border=\"1\" style=\"border-collapse: collapse\" width=\"368\">\r\n\t\t<thead>\r\n\t\t\t<tr style=\"height:16.75pt\">\r\n\t\t\t\t<th colspan=\"3\" nowrap scope=\"col\" style=\"width: 183.8pt\" width=\"368\">\r\n\t\t\t\t\t<p align=\"center\" style=\"margin-bottom:.0001pt;text-align:center;padding:0cm5.4pt0cm5.4pt\">\r\n\t\t\t\t\t\t<span style=\"line-height:normal\"><span style=\"height:16.75pt\"><span style=\"color:black\">Results retention time [<\/span><span style=\"color:black\">min]<\/span><\/span><\/span>\r\n\t\t\t\t\t<\/p>\r\n\t\t\t\t<\/th>\r\n\t\t\t<\/tr>\r\n\t\t<\/thead>\r\n\t\t<tbody>\r\n\t\t\t<tr style=\"height:16.75pt\">\r\n\t\t\t\t<td nowrap style=\"width: 50.95pt\" width=\"102\">\r\n\t\t\t\t\t<p align=\"center\" style=\"margin-bottom:.0001pt;text-align:center;padding:0cm5.4pt0cm5.4pt\">\r\n\t\t\t\t\t\t<span style=\"line-height:normal\"><span style=\"height:16.75pt\"><span style=\"color:black\">A<\/span><\/span><\/span>\r\n\t\t\t\t\t<\/p>\r\n\t\t\t\t<\/td>\r\n\t\t\t\t<td nowrap style=\"width: 69.05pt\" width=\"138\">\r\n\t\t\t\t\t<p align=\"center\" style=\"margin-bottom:.0001pt;text-align:center;padding:0cm5.4pt0cm5.4pt\">\r\n\t\t\t\t\t\t<span style=\"line-height:normal\"><span style=\"height:16.75pt\"><span style=\"color:black\">B<\/span><\/span><\/span>\r\n\t\t\t\t\t<\/p>\r\n\t\t\t\t<\/td>\r\n\t\t\t\t<td nowrap style=\"width: 63.8pt\" width=\"128\">\r\n\t\t\t\t\t<p align=\"center\" style=\"margin-bottom:.0001pt;text-align:center;padding:0cm5.4pt0cm5.4pt\">\r\n\t\t\t\t\t\t<span style=\"line-height:normal\"><span style=\"height:16.75pt\"><span style=\"color:black\">C<\/span><\/span><\/span>\r\n\t\t\t\t\t<\/p>\r\n\t\t\t\t<\/td>\r\n\t\t\t<\/tr>\r\n\t\t\t<tr style=\"height:16.75pt\">\r\n\t\t\t\t<td nowrap style=\"width: 50.95pt\" width=\"102\">\r\n\t\t\t\t\t<p align=\"center\" style=\"margin-bottom:.0001pt;text-align:center;padding:0cm5.4pt0cm5.4pt\">\r\n\t\t\t\t\t\t<span style=\"line-height:normal\"><span style=\"height:16.75pt\"><span style=\"color:black\">pH<\/span><\/span><\/span>\r\n\t\t\t\t\t<\/p>\r\n\t\t\t\t<\/td>\r\n\t\t\t\t<td nowrap style=\"width: 69.05pt\" width=\"138\">\r\n\t\t\t\t\t<p align=\"center\" style=\"margin-bottom:.0001pt;text-align:center;padding:0cm5.4pt0cm5.4pt\">\r\n\t\t\t\t\t\t<span style=\"line-height:normal\"><span style=\"height:16.75pt\"><span style=\"color:black\">Additive conc.<\/span><\/span><\/span>\r\n\t\t\t\t\t<\/p>\r\n\t\t\t\t<\/td>\r\n\t\t\t\t<td nowrap style=\"width: 63.8pt\" width=\"128\">\r\n\t\t\t\t\t<p align=\"center\" style=\"margin-bottom:.0001pt;text-align:center;padding:0cm5.4pt0cm5.4pt\">\r\n\t\t\t\t\t\t<span style=\"line-height:normal\"><span style=\"height:16.75pt\"><span style=\"color:black\">Column Temp<\/span><\/span><\/span>\r\n\t\t\t\t\t<\/p>\r\n\t\t\t\t<\/td>\r\n\t\t\t<\/tr>\r\n\t\t\t<tr>\r\n\t\t\t\t<td nowrap>\r\n\t\t\t\t\t<p style=\"text-align: center\">\r\n\t\t\t\t\t\t-1.48\r\n\t\t\t\t\t<\/p>\r\n\t\t\t\t<\/td>\r\n\t\t\t\t<td nowrap style=\"width: 69.05pt\" width=\"138\">\r\n\t\t\t\t\t<p align=\"center\" style=\"margin-bottom:.0001pt;text-align:center;padding:0cm5.4pt0cm5.4pt\">\r\n\t\t\t\t\t\t<span style=\"line-height:normal\"><span style=\"height:17.4pt\"><span style=\"color:black\">-0.20<\/span><\/span><\/span>\r\n\t\t\t\t\t<\/p>\r\n\t\t\t\t<\/td>\r\n\t\t\t\t<td nowrap style=\"width: 63.8pt\" width=\"128\">\r\n\t\t\t\t\t<p align=\"center\" style=\"margin-bottom:.0001pt;text-align:center;padding:0cm5.4pt0cm5.4pt\">\r\n\t\t\t\t\t\t<span style=\"line-height:normal\"><span style=\"height:17.4pt\"><span style=\"color:black\">-0.17<\/span><\/span><\/span>\r\n\t\t\t\t\t<\/p>\r\n\t\t\t\t<\/td>\r\n\t\t\t<\/tr>\r\n\t\t<\/tbody>\r\n\t<\/table>\r\n<\/div>\r\n\r\n<p style=\"margin-bottom:7.5pt;text-align:justify\">\r\n\t\u00a0\r\n<\/p>\r\n\r\n<p style=\"margin-bottom:7.5pt;text-align:justify\">\r\n\t<span><span><span style=\"color:#2f2f2f\">From this depiction, it is clear that the pH value especially has the highest influence on the retention time, in the way that a decrease in pH increases the retention time of the analyte, which would imply that the method is not robust regarding a pH change of \u00b1 0.2 pH units.<\/span><\/span><\/span>\r\n<\/p>\r\n\r\n<p style=\"margin-bottom:7.5pt;text-align:justify\">\r\n\t<span><span><span style=\"color:#2f2f2f\">This can also be visualized in the following plot. By the slope, it is easily deductible that the impact of the pH on the response variable retention time is greater than the effect of the additive concentration in the eluent. In general, the steeper the slope is, the more impact a factor has on the response variable. The effect size describes the numeric influence of the factor on the response variable in the set design region and can be used to distinguish the importance of effects from the less important ones.<\/span><\/span><\/span>\r\n<\/p>\r\n\r\n<p style=\"margin-bottom:7.5pt;text-align:justify\">\r\n\t\u00a0\r\n<\/p>\r\n\r\n<p style=\"margin-bottom:7.5pt;text-align:center\">\r\n\t<img loading=\"lazy\" decoding=\"async\" width=\"1584\" height=\"1075\" class=\"alignnone wp-image-80\" style=\"height: 300px;width: 442px\" src=\"https:\/\/sisu.ut.ee\/wp-content\/uploads\/sites\/130\/graph_effectsize_retention_time.png\" title=\"graph_effectsize_retention_time.png\" alt=\"gr_effs_ret_t\" srcset=\"https:\/\/sisu.ut.ee\/wp-content\/uploads\/sites\/130\/graph_effectsize_retention_time.png 1584w, https:\/\/sisu.ut.ee\/wp-content\/uploads\/sites\/130\/graph_effectsize_retention_time-300x204.png 300w, https:\/\/sisu.ut.ee\/wp-content\/uploads\/sites\/130\/graph_effectsize_retention_time-1024x695.png 1024w, https:\/\/sisu.ut.ee\/wp-content\/uploads\/sites\/130\/graph_effectsize_retention_time-768x521.png 768w, https:\/\/sisu.ut.ee\/wp-content\/uploads\/sites\/130\/graph_effectsize_retention_time-1536x1042.png 1536w\" sizes=\"auto, (max-width: 1584px) 100vw, 1584px\">\r\n<\/p>\r\n\r\n<h4 style=\"text-align:center\">\r\n\tFigure 1 Main effect plot of pH, Additive conc. and Column temp. on the Retention time\r\n<\/h4>\r\n\r\n<p style=\"margin-bottom:7.5pt;text-align:justify\">\r\n\t<span><span><span style=\"color:#2f2f2f\">The influence of the additive concentration in our chosen design region is less impactful on the retention time than pH change in our chosen design region on the retention time. Thereby, we might assume that the method is robust towards changes in additive concentration. But, as will be shown in the following section, this assumption would be wrong! In that\u2019s section we will show the importance and benefits of also monitoring factor interactions instead of only the individual effects of the factors.<\/span><\/span><\/span>\r\n<\/p>\r\n\r\n<p style=\"margin-bottom:7.5pt;text-align:justify\">\r\n\t<span><span><span style=\"color:#2f2f2f\">e) <u>Interactions are important<\/u><\/span><\/span><\/span>\r\n<\/p>\r\n\r\n<p style=\"margin-bottom:7.5pt;text-align:justify\">\r\n\t<span><span><span style=\"color:#2f2f2f\">One of the major advantages of a DoE approach is the determination of the effects of interactions on the response variable. To calculate these interactions, we proceed the same way as with the normal factors A, B and C. The coded value for an interaction is deducted by the multiplication of the single effects meaning that AB= A*B which might look like this<\/span><\/span><\/span><br><span><span><span style=\"color:#2f2f2f\">+ * + = + or this + * \u2013 = -.<\/span><\/span><\/span>\r\n<\/p>\r\n\r\n<p style=\"margin-bottom:7.5pt;text-align:justify\">\r\n\t<span><span><span style=\"color:#2f2f2f\">So, for the interaction of AB (pH and additive conc.), we get coded values like this:<\/span><\/span><\/span>\r\n<\/p>\r\n\r\n<p>\r\n\t<span><span><span style=\"color:#2f2f2f\">If we do the multiplication for all the interaction combinations, it creates the following table with the interactions.<\/span><\/span><\/span>\r\n<\/p>\r\n\r\n<p style=\"text-align: justify\">\r\n\t\u00a0\r\n<\/p>\r\n\r\n<div align=\"center\">\r\n\t<table class=\"table table-hover\" align=\"center\" border=\"1\" style=\"border-collapse: collapse\" width=\"688\">\r\n\t\t<thead>\r\n\t\t\t<tr style=\"height:20.8pt\">\r\n\t\t\t\t<th scope=\"col\" style=\"width: 3cm\" width=\"170\">\r\n\t\t\t\t\t<p align=\"center\" style=\"margin-bottom:.0001pt;text-align:center;padding:3.75pt3.75pt3.75pt3.75pt\">\r\n\t\t\t\t\t\t<span style=\"height:20.8pt\"><span><span style=\",serif\"><span style=\"color:black\">Experiment<\/span><\/span><\/span><\/span>\r\n\t\t\t\t\t<\/p>\r\n\t\t\t\t<\/th>\r\n\t\t\t\t<th scope=\"col\" style=\"width: 3cm\" width=\"170\">\r\n\t\t\t\t\t<p align=\"center\" style=\"margin-bottom:.0001pt;text-align:center;padding:3.75pt3.75pt3.75pt3.75pt\">\r\n\t\t\t\t\t\t<span style=\"height:20.8pt\"><b><span><span style=\",serif\"><span style=\"color:black\">AB<\/span><\/span><\/span><\/b><\/span>\r\n\t\t\t\t\t<\/p>\r\n\t\t\t\t<\/th>\r\n\t\t\t\t<th scope=\"col\" style=\"width: 49.1pt\" width=\"98\">\r\n\t\t\t\t\t<p align=\"center\" style=\"margin-bottom:.0001pt;text-align:center;padding:3.75pt3.75pt3.75pt3.75pt\">\r\n\t\t\t\t\t\t<span style=\"height:20.8pt\"><b><span><span style=\",serif\"><span style=\"color:black\">BC<\/span><\/span><\/span><\/b><\/span>\r\n\t\t\t\t\t<\/p>\r\n\t\t\t\t<\/th>\r\n\t\t\t\t<th scope=\"col\" style=\"width: 45.3pt\" width=\"91\">\r\n\t\t\t\t\t<p align=\"center\" style=\"margin-bottom:.0001pt;text-align:center;padding:3.75pt3.75pt3.75pt3.75pt\">\r\n\t\t\t\t\t\t<span style=\"height:20.8pt\"><b><span><span style=\",serif\"><span style=\"color:black\">AC<\/span><\/span><\/span><\/b><\/span>\r\n\t\t\t\t\t<\/p>\r\n\t\t\t\t<\/th>\r\n\t\t\t\t<th scope=\"col\" style=\"width: 39.75pt\" width=\"80\">\r\n\t\t\t\t\t<p align=\"center\" style=\"margin-bottom:.0001pt;text-align:center;padding:3.75pt3.75pt3.75pt3.75pt\">\r\n\t\t\t\t\t\t<span style=\"height:20.8pt\"><b><span><span style=\",serif\"><span style=\"color:black\">ABC<\/span><\/span><\/span><\/b><\/span>\r\n\t\t\t\t\t<\/p>\r\n\t\t\t\t<\/th>\r\n\t\t\t\t<th scope=\"col\" style=\"width: 39.65pt\" width=\"79\">\r\n\t\t\t\t\t<p align=\"center\" style=\"margin-bottom:.0001pt;text-align:center;padding:3.75pt3.75pt3.75pt3.75pt\">\r\n\t\t\t\t\t\t<span style=\"height:20.8pt\"><b><span><span style=\",serif\"><span style=\"color:black\">Ret. Time<br>[min]<\/span><\/span><\/span><\/b><\/span>\r\n\t\t\t\t\t<\/p>\r\n\t\t\t\t<\/th>\r\n\t\t\t<\/tr>\r\n\t\t<\/thead>\r\n\t\t<tbody>\r\n\t\t\t<tr style=\"height:12.0pt\">\r\n\t\t\t\t<td style=\"width: 3cm\" width=\"170\">\r\n\t\t\t\t\t<p align=\"center\" style=\"margin-bottom:.0001pt;text-align:center;padding:3.75pt3.75pt3.75pt3.75pt\">\r\n\t\t\t\t\t\t<span style=\"height:12.0pt\"><b><span><span style=\",serif\">1<\/span><\/span><\/b><\/span>\r\n\t\t\t\t\t<\/p>\r\n\t\t\t\t<\/td>\r\n\t\t\t\t<td style=\"width: 3cm\" width=\"170\">\r\n\t\t\t\t\t<p align=\"center\" style=\"margin-bottom:.0001pt;text-align:center;padding:3.75pt3.75pt3.75pt3.75pt\">\r\n\t\t\t\t\t\t<span style=\"height:12.0pt\"><b><span><span style=\",serif\"><span style=\"color:black\">+<\/span><\/span><\/span><\/b><\/span>\r\n\t\t\t\t\t<\/p>\r\n\t\t\t\t<\/td>\r\n\t\t\t\t<td style=\"width: 49.1pt\" width=\"98\">\r\n\t\t\t\t\t<p align=\"center\" style=\"margin-bottom:.0001pt;text-align:center;padding:3.75pt3.75pt3.75pt3.75pt\">\r\n\t\t\t\t\t\t<span style=\"height:12.0pt\"><b><span><span style=\",serif\"><span style=\"color:black\">+<\/span><\/span><\/span><\/b><\/span>\r\n\t\t\t\t\t<\/p>\r\n\t\t\t\t<\/td>\r\n\t\t\t\t<td style=\"width: 45.3pt\" width=\"91\">\r\n\t\t\t\t\t<p align=\"center\" style=\"margin-bottom:.0001pt;text-align:center;padding:3.75pt3.75pt3.75pt3.75pt\">\r\n\t\t\t\t\t\t<span style=\"height:12.0pt\"><b><span><span style=\",serif\"><span style=\"color:black\">+<\/span><\/span><\/span><\/b><\/span>\r\n\t\t\t\t\t<\/p>\r\n\t\t\t\t<\/td>\r\n\t\t\t\t<td style=\"width: 39.75pt\" width=\"80\">\r\n\t\t\t\t\t<p align=\"center\" style=\"margin-bottom:.0001pt;text-align:center;padding:3.75pt3.75pt3.75pt3.75pt\">\r\n\t\t\t\t\t\t<span style=\"height:12.0pt\"><b><span><span style=\",serif\"><span style=\"color:black\">+<\/span><\/span><\/span><\/b><\/span>\r\n\t\t\t\t\t<\/p>\r\n\t\t\t\t<\/td>\r\n\t\t\t\t<td style=\"width: 39.65pt\" width=\"79\">\r\n\t\t\t\t\t<p align=\"center\" style=\"margin-bottom:.0001pt;text-align:center;padding:3.75pt3.75pt3.75pt3.75pt\">\r\n\t\t\t\t\t\t<span style=\"height:12.0pt\"><span><span style=\",serif\">8.31<\/span><\/span><\/span>\r\n\t\t\t\t\t<\/p>\r\n\t\t\t\t<\/td>\r\n\t\t\t<\/tr>\r\n\t\t\t<tr style=\"height:12.0pt\">\r\n\t\t\t\t<td style=\"width: 3cm\" width=\"170\">\r\n\t\t\t\t\t<p align=\"center\" style=\"margin-bottom:.0001pt;text-align:center;padding:3.75pt3.75pt3.75pt3.75pt\">\r\n\t\t\t\t\t\t<span style=\"height:12.0pt\"><b><span><span style=\",serif\">2<\/span><\/span><\/b><\/span>\r\n\t\t\t\t\t<\/p>\r\n\t\t\t\t<\/td>\r\n\t\t\t\t<td style=\"width: 3cm\" width=\"170\">\r\n\t\t\t\t\t<p align=\"center\" style=\"margin-bottom:.0001pt;text-align:center;padding:3.75pt3.75pt3.75pt3.75pt\">\r\n\t\t\t\t\t\t<span style=\"height:12.0pt\"><b><span><span style=\",serif\"><span style=\"color:black\">+<\/span><\/span><\/span><\/b><\/span>\r\n\t\t\t\t\t<\/p>\r\n\t\t\t\t<\/td>\r\n\t\t\t\t<td style=\"width: 49.1pt\" width=\"98\">\r\n\t\t\t\t\t<p align=\"center\" style=\"margin-bottom:.0001pt;text-align:center;padding:3.75pt3.75pt3.75pt3.75pt\">\r\n\t\t\t\t\t\t<span style=\"height:12.0pt\"><b><span><span style=\",serif\"><span style=\"color:#767171\">\u2013<\/span><\/span><\/span><\/b><\/span>\r\n\t\t\t\t\t<\/p>\r\n\t\t\t\t<\/td>\r\n\t\t\t\t<td style=\"width: 45.3pt\" width=\"91\">\r\n\t\t\t\t\t<p align=\"center\" style=\"margin-bottom:.0001pt;text-align:center;padding:3.75pt3.75pt3.75pt3.75pt\">\r\n\t\t\t\t\t\t<span style=\"height:12.0pt\"><b><span><span style=\",serif\"><span style=\"color:#767171\">\u2013<\/span><\/span><\/span><\/b><\/span>\r\n\t\t\t\t\t<\/p>\r\n\t\t\t\t<\/td>\r\n\t\t\t\t<td style=\"width: 39.75pt\" width=\"80\">\r\n\t\t\t\t\t<p align=\"center\" style=\"margin-bottom:.0001pt;text-align:center;padding:3.75pt3.75pt3.75pt3.75pt\">\r\n\t\t\t\t\t\t<span style=\"height:12.0pt\"><b><span><span style=\",serif\"><span style=\"color:#767171\">\u2013<\/span><\/span><\/span><\/b><\/span>\r\n\t\t\t\t\t<\/p>\r\n\t\t\t\t<\/td>\r\n\t\t\t\t<td style=\"width: 39.65pt\" width=\"79\">\r\n\t\t\t\t\t<p align=\"center\" style=\"margin-bottom:.0001pt;text-align:center;padding:3.75pt3.75pt3.75pt3.75pt\">\r\n\t\t\t\t\t\t<span style=\"height:12.0pt\"><span><span style=\",serif\">8.10<\/span><\/span><\/span>\r\n\t\t\t\t\t<\/p>\r\n\t\t\t\t<\/td>\r\n\t\t\t<\/tr>\r\n\t\t\t<tr style=\"height:12.0pt\">\r\n\t\t\t\t<td style=\"width: 3cm\" width=\"170\">\r\n\t\t\t\t\t<p align=\"center\" style=\"margin-bottom:.0001pt;text-align:center;padding:3.75pt3.75pt3.75pt3.75pt\">\r\n\t\t\t\t\t\t<span style=\"height:12.0pt\"><b><span><span style=\",serif\">3<\/span><\/span><\/b><\/span>\r\n\t\t\t\t\t<\/p>\r\n\t\t\t\t<\/td>\r\n\t\t\t\t<td style=\"width: 3cm\" width=\"170\">\r\n\t\t\t\t\t<p align=\"center\" style=\"margin-bottom:.0001pt;text-align:center;padding:3.75pt3.75pt3.75pt3.75pt\">\r\n\t\t\t\t\t\t<span style=\"height:12.0pt\"><b><span><span style=\",serif\"><span style=\"color:black\">\u2013<\/span><\/span><\/span><\/b><\/span>\r\n\t\t\t\t\t<\/p>\r\n\t\t\t\t<\/td>\r\n\t\t\t\t<td style=\"width: 49.1pt\" width=\"98\">\r\n\t\t\t\t\t<p align=\"center\" style=\"margin-bottom:.0001pt;text-align:center;padding:3.75pt3.75pt3.75pt3.75pt\">\r\n\t\t\t\t\t\t<span style=\"height:12.0pt\"><b><span><span style=\",serif\"><span style=\"color:#767171\">\u2013<\/span><\/span><\/span><\/b><\/span>\r\n\t\t\t\t\t<\/p>\r\n\t\t\t\t<\/td>\r\n\t\t\t\t<td style=\"width: 45.3pt\" width=\"91\">\r\n\t\t\t\t\t<p align=\"center\" style=\"margin-bottom:.0001pt;text-align:center;padding:3.75pt3.75pt3.75pt3.75pt\">\r\n\t\t\t\t\t\t<span style=\"height:12.0pt\"><b><span><span style=\",serif\"><span style=\"color:black\">+<\/span><\/span><\/span><\/b><\/span>\r\n\t\t\t\t\t<\/p>\r\n\t\t\t\t<\/td>\r\n\t\t\t\t<td style=\"width: 39.75pt\" width=\"80\">\r\n\t\t\t\t\t<p align=\"center\" style=\"margin-bottom:.0001pt;text-align:center;padding:3.75pt3.75pt3.75pt3.75pt\">\r\n\t\t\t\t\t\t<span style=\"height:12.0pt\"><b><span><span style=\",serif\"><span style=\"color:#767171\">\u2013<\/span><\/span><\/span><\/b><\/span>\r\n\t\t\t\t\t<\/p>\r\n\t\t\t\t<\/td>\r\n\t\t\t\t<td style=\"width: 39.65pt\" width=\"79\">\r\n\t\t\t\t\t<p align=\"center\" style=\"margin-bottom:.0001pt;text-align:center;padding:3.75pt3.75pt3.75pt3.75pt\">\r\n\t\t\t\t\t\t<span style=\"height:12.0pt\"><span><span style=\",serif\">7.24<\/span><\/span><\/span>\r\n\t\t\t\t\t<\/p>\r\n\t\t\t\t<\/td>\r\n\t\t\t<\/tr>\r\n\t\t\t<tr style=\"height:12.0pt\">\r\n\t\t\t\t<td style=\"width: 3cm\" width=\"170\">\r\n\t\t\t\t\t<p align=\"center\" style=\"margin-bottom:.0001pt;text-align:center;padding:3.75pt3.75pt3.75pt3.75pt\">\r\n\t\t\t\t\t\t<span style=\"height:12.0pt\"><b><span><span style=\",serif\">4<\/span><\/span><\/b><\/span>\r\n\t\t\t\t\t<\/p>\r\n\t\t\t\t<\/td>\r\n\t\t\t\t<td style=\"width: 3cm\" width=\"170\">\r\n\t\t\t\t\t<p align=\"center\" style=\"margin-bottom:.0001pt;text-align:center;padding:3.75pt3.75pt3.75pt3.75pt\">\r\n\t\t\t\t\t\t<span style=\"height:12.0pt\"><b><span><span style=\",serif\"><span style=\"color:black\">\u2013<\/span><\/span><\/span><\/b><\/span>\r\n\t\t\t\t\t<\/p>\r\n\t\t\t\t<\/td>\r\n\t\t\t\t<td style=\"width: 49.1pt\" width=\"98\">\r\n\t\t\t\t\t<p align=\"center\" style=\"margin-bottom:.0001pt;text-align:center;padding:3.75pt3.75pt3.75pt3.75pt\">\r\n\t\t\t\t\t\t<span style=\"height:12.0pt\"><b><span><span style=\",serif\"><span style=\"color:black\">+<\/span><\/span><\/span><\/b><\/span>\r\n\t\t\t\t\t<\/p>\r\n\t\t\t\t<\/td>\r\n\t\t\t\t<td style=\"width: 45.3pt\" width=\"91\">\r\n\t\t\t\t\t<p align=\"center\" style=\"margin-bottom:.0001pt;text-align:center;padding:3.75pt3.75pt3.75pt3.75pt\">\r\n\t\t\t\t\t\t<span style=\"height:12.0pt\"><b><span><span style=\",serif\"><span style=\"color:#767171\">\u2013<\/span><\/span><\/span><\/b><\/span>\r\n\t\t\t\t\t<\/p>\r\n\t\t\t\t<\/td>\r\n\t\t\t\t<td style=\"width: 39.75pt\" width=\"80\">\r\n\t\t\t\t\t<p align=\"center\" style=\"margin-bottom:.0001pt;text-align:center;padding:3.75pt3.75pt3.75pt3.75pt\">\r\n\t\t\t\t\t\t<span style=\"height:12.0pt\"><b><span><span style=\",serif\"><span style=\"color:black\">+<\/span><\/span><\/span><\/b><\/span>\r\n\t\t\t\t\t<\/p>\r\n\t\t\t\t<\/td>\r\n\t\t\t\t<td style=\"width: 39.65pt\" width=\"79\">\r\n\t\t\t\t\t<p align=\"center\" style=\"margin-bottom:.0001pt;text-align:center;padding:3.75pt3.75pt3.75pt3.75pt\">\r\n\t\t\t\t\t\t<span style=\"height:12.0pt\"><span><span style=\",serif\">7.43<\/span><\/span><\/span>\r\n\t\t\t\t\t<\/p>\r\n\t\t\t\t<\/td>\r\n\t\t\t<\/tr>\r\n\t\t\t<tr style=\"height:12.0pt\">\r\n\t\t\t\t<td style=\"width: 3cm\" width=\"170\">\r\n\t\t\t\t\t<p align=\"center\" style=\"margin-bottom:.0001pt;text-align:center;padding:3.75pt3.75pt3.75pt3.75pt\">\r\n\t\t\t\t\t\t<span style=\"height:12.0pt\"><b><span><span style=\",serif\">5<\/span><\/span><\/b><\/span>\r\n\t\t\t\t\t<\/p>\r\n\t\t\t\t<\/td>\r\n\t\t\t\t<td style=\"width: 3cm\" width=\"170\">\r\n\t\t\t\t\t<p align=\"center\" style=\"margin-bottom:.0001pt;text-align:center;padding:3.75pt3.75pt3.75pt3.75pt\">\r\n\t\t\t\t\t\t<span style=\"height:12.0pt\"><b><span><span style=\",serif\"><span style=\"color:black\">\u2013<\/span><\/span><\/span><\/b><\/span>\r\n\t\t\t\t\t<\/p>\r\n\t\t\t\t<\/td>\r\n\t\t\t\t<td style=\"width: 49.1pt\" width=\"98\">\r\n\t\t\t\t\t<p align=\"center\" style=\"margin-bottom:.0001pt;text-align:center;padding:3.75pt3.75pt3.75pt3.75pt\">\r\n\t\t\t\t\t\t<span style=\"height:12.0pt\"><b><span><span style=\",serif\"><span style=\"color:black\">+<\/span><\/span><\/span><\/b><\/span>\r\n\t\t\t\t\t<\/p>\r\n\t\t\t\t<\/td>\r\n\t\t\t\t<td style=\"width: 45.3pt\" width=\"91\">\r\n\t\t\t\t\t<p align=\"center\" style=\"margin-bottom:.0001pt;text-align:center;padding:3.75pt3.75pt3.75pt3.75pt\">\r\n\t\t\t\t\t\t<span style=\"height:12.0pt\"><b><span><span style=\",serif\"><span style=\"color:#767171\">\u2013<\/span><\/span><\/span><\/b><\/span>\r\n\t\t\t\t\t<\/p>\r\n\t\t\t\t<\/td>\r\n\t\t\t\t<td style=\"width: 39.75pt\" width=\"80\">\r\n\t\t\t\t\t<p align=\"center\" style=\"margin-bottom:.0001pt;text-align:center;padding:3.75pt3.75pt3.75pt3.75pt\">\r\n\t\t\t\t\t\t<span style=\"height:12.0pt\"><b><span><span style=\",serif\"><span style=\"color:#767171\">\u2013<\/span><\/span><\/span><\/b><\/span>\r\n\t\t\t\t\t<\/p>\r\n\t\t\t\t<\/td>\r\n\t\t\t\t<td style=\"width: 39.65pt\" width=\"79\">\r\n\t\t\t\t\t<p align=\"center\" style=\"margin-bottom:.0001pt;text-align:center;padding:3.75pt3.75pt3.75pt3.75pt\">\r\n\t\t\t\t\t\t<span style=\"height:12.0pt\"><span><span style=\",serif\">8.32<\/span><\/span><\/span>\r\n\t\t\t\t\t<\/p>\r\n\t\t\t\t<\/td>\r\n\t\t\t<\/tr>\r\n\t\t\t<tr style=\"height:12.0pt\">\r\n\t\t\t\t<td style=\"width: 3cm\" width=\"170\">\r\n\t\t\t\t\t<p align=\"center\" style=\"margin-bottom:.0001pt;text-align:center;padding:3.75pt3.75pt3.75pt3.75pt\">\r\n\t\t\t\t\t\t<span style=\"height:12.0pt\"><b><span><span style=\",serif\">6<\/span><\/span><\/b><\/span>\r\n\t\t\t\t\t<\/p>\r\n\t\t\t\t<\/td>\r\n\t\t\t\t<td style=\"width: 3cm\" width=\"170\">\r\n\t\t\t\t\t<p align=\"center\" style=\"margin-bottom:.0001pt;text-align:center;padding:3.75pt3.75pt3.75pt3.75pt\">\r\n\t\t\t\t\t\t<span style=\"height:12.0pt\"><b><span><span style=\",serif\"><span style=\"color:black\">\u2013<\/span><\/span><\/span><\/b><\/span>\r\n\t\t\t\t\t<\/p>\r\n\t\t\t\t<\/td>\r\n\t\t\t\t<td style=\"width: 49.1pt\" width=\"98\">\r\n\t\t\t\t\t<p align=\"center\" style=\"margin-bottom:.0001pt;text-align:center;padding:3.75pt3.75pt3.75pt3.75pt\">\r\n\t\t\t\t\t\t<span style=\"height:12.0pt\"><b><span><span style=\",serif\"><span style=\"color:#767171\">\u2013<\/span><\/span><\/span><\/b><\/span>\r\n\t\t\t\t\t<\/p>\r\n\t\t\t\t<\/td>\r\n\t\t\t\t<td style=\"width: 45.3pt\" width=\"91\">\r\n\t\t\t\t\t<p align=\"center\" style=\"margin-bottom:.0001pt;text-align:center;padding:3.75pt3.75pt3.75pt3.75pt\">\r\n\t\t\t\t\t\t<span style=\"height:12.0pt\"><b><span><span style=\",serif\"><span style=\"color:black\">+<\/span><\/span><\/span><\/b><\/span>\r\n\t\t\t\t\t<\/p>\r\n\t\t\t\t<\/td>\r\n\t\t\t\t<td style=\"width: 39.75pt\" width=\"80\">\r\n\t\t\t\t\t<p align=\"center\" style=\"margin-bottom:.0001pt;text-align:center;padding:3.75pt3.75pt3.75pt3.75pt\">\r\n\t\t\t\t\t\t<span style=\"height:12.0pt\"><b><span><span style=\",serif\"><span style=\"color:black\">+<\/span><\/span><\/span><\/b><\/span>\r\n\t\t\t\t\t<\/p>\r\n\t\t\t\t<\/td>\r\n\t\t\t\t<td style=\"width: 39.65pt\" width=\"79\">\r\n\t\t\t\t\t<p align=\"center\" style=\"margin-bottom:.0001pt;text-align:center;padding:3.75pt3.75pt3.75pt3.75pt\">\r\n\t\t\t\t\t\t<span style=\"height:12.0pt\"><span><span style=\",serif\">8.92<\/span><\/span><\/span>\r\n\t\t\t\t\t<\/p>\r\n\t\t\t\t<\/td>\r\n\t\t\t<\/tr>\r\n\t\t\t<tr style=\"height:12.0pt\">\r\n\t\t\t\t<td style=\"width: 3cm\" width=\"170\">\r\n\t\t\t\t\t<p align=\"center\" style=\"margin-bottom:.0001pt;text-align:center;padding:3.75pt3.75pt3.75pt3.75pt\">\r\n\t\t\t\t\t\t<span style=\"height:12.0pt\"><b><span><span style=\",serif\">7<\/span><\/span><\/b><\/span>\r\n\t\t\t\t\t<\/p>\r\n\t\t\t\t<\/td>\r\n\t\t\t\t<td style=\"width: 3cm\" width=\"170\">\r\n\t\t\t\t\t<p align=\"center\" style=\"margin-bottom:.0001pt;text-align:center;padding:3.75pt3.75pt3.75pt3.75pt\">\r\n\t\t\t\t\t\t<span style=\"height:12.0pt\"><span><span style=\",serif\"><span style=\"color:black\">+<\/span><\/span><\/span><\/span>\r\n\t\t\t\t\t<\/p>\r\n\t\t\t\t<\/td>\r\n\t\t\t\t<td style=\"width: 49.1pt\" width=\"98\">\r\n\t\t\t\t\t<p align=\"center\" style=\"margin-bottom:.0001pt;text-align:center;padding:3.75pt3.75pt3.75pt3.75pt\">\r\n\t\t\t\t\t\t<span style=\"height:12.0pt\"><b><span><span style=\",serif\"><span style=\"color:#767171\">\u2013<\/span><\/span><\/span><\/b><\/span>\r\n\t\t\t\t\t<\/p>\r\n\t\t\t\t<\/td>\r\n\t\t\t\t<td style=\"width: 45.3pt\" width=\"91\">\r\n\t\t\t\t\t<p align=\"center\" style=\"margin-bottom:.0001pt;text-align:center;padding:3.75pt3.75pt3.75pt3.75pt\">\r\n\t\t\t\t\t\t<span style=\"height:12.0pt\"><b><span><span style=\",serif\"><span style=\"color:#767171\">\u2013<\/span><\/span><\/span><\/b><\/span>\r\n\t\t\t\t\t<\/p>\r\n\t\t\t\t<\/td>\r\n\t\t\t\t<td style=\"width: 39.75pt\" width=\"80\">\r\n\t\t\t\t\t<p align=\"center\" style=\"margin-bottom:.0001pt;text-align:center;padding:3.75pt3.75pt3.75pt3.75pt\">\r\n\t\t\t\t\t\t<span style=\"height:12.0pt\"><b><span><span style=\",serif\"><span style=\"color:black\">+<\/span><\/span><\/span><\/b><\/span>\r\n\t\t\t\t\t<\/p>\r\n\t\t\t\t<\/td>\r\n\t\t\t\t<td style=\"width: 39.65pt\" width=\"79\">\r\n\t\t\t\t\t<p align=\"center\" style=\"margin-bottom:.0001pt;text-align:center;padding:3.75pt3.75pt3.75pt3.75pt\">\r\n\t\t\t\t\t\t<span style=\"height:12.0pt\"><span><span style=\",serif\">9.84<\/span><\/span><\/span>\r\n\t\t\t\t\t<\/p>\r\n\t\t\t\t<\/td>\r\n\t\t\t<\/tr>\r\n\t\t\t<tr style=\"height:12.4pt\">\r\n\t\t\t\t<td style=\"width: 3cm\" width=\"170\">\r\n\t\t\t\t\t<p align=\"center\" style=\"margin-bottom:.0001pt;text-align:center;padding:3.75pt3.75pt3.75pt3.75pt\">\r\n\t\t\t\t\t\t<span style=\"height:12.4pt\"><b><span><span style=\",serif\">8<\/span><\/span><\/b><\/span>\r\n\t\t\t\t\t<\/p>\r\n\t\t\t\t<\/td>\r\n\t\t\t\t<td style=\"width: 3cm\" width=\"170\">\r\n\t\t\t\t\t<p align=\"center\" style=\"margin-bottom:.0001pt;text-align:center;padding:3.75pt3.75pt3.75pt3.75pt\">\r\n\t\t\t\t\t\t<span style=\"height:12.4pt\"><span><span style=\",serif\"><span style=\"color:black\">+<\/span><\/span><\/span><\/span>\r\n\t\t\t\t\t<\/p>\r\n\t\t\t\t<\/td>\r\n\t\t\t\t<td style=\"width: 49.1pt\" width=\"98\">\r\n\t\t\t\t\t<p align=\"center\" style=\"margin-bottom:.0001pt;text-align:center;padding:3.75pt3.75pt3.75pt3.75pt\">\r\n\t\t\t\t\t\t<span style=\"height:12.0pt\"><b><span><span style=\",serif\"><span style=\"color:black\">+<\/span><\/span><\/span><\/b><\/span>\r\n\t\t\t\t\t<\/p>\r\n\t\t\t\t<\/td>\r\n\t\t\t\t<td style=\"width: 45.3pt\" width=\"91\">\r\n\t\t\t\t\t<p align=\"center\" style=\"margin-bottom:.0001pt;text-align:center;padding:3.75pt3.75pt3.75pt3.75pt\">\r\n\t\t\t\t\t\t<span style=\"height:12.4pt\"><b><span><span style=\",serif\"><span style=\"color:black\">+<\/span><\/span><\/span><\/b><\/span>\r\n\t\t\t\t\t<\/p>\r\n\t\t\t\t<\/td>\r\n\t\t\t\t<td style=\"width: 39.75pt\" width=\"80\">\r\n\t\t\t\t\t<p align=\"center\" style=\"margin-bottom:.0001pt;text-align:center;padding:3.75pt3.75pt3.75pt3.75pt\">\r\n\t\t\t\t\t\t<span style=\"height:12.4pt\"><b><span><span style=\",serif\"><span style=\"color:#767171\">\u2013<\/span><\/span><\/span><\/b><\/span>\r\n\t\t\t\t\t<\/p>\r\n\t\t\t\t<\/td>\r\n\t\t\t\t<td style=\"width: 39.65pt\" width=\"79\">\r\n\t\t\t\t\t<p align=\"center\" style=\"margin-bottom:.0001pt;text-align:center;padding:3.75pt3.75pt3.75pt3.75pt\">\r\n\t\t\t\t\t\t<span style=\"height:12.4pt\"><span><span style=\",serif\">9.93<\/span><\/span><\/span>\r\n\t\t\t\t\t<\/p>\r\n\t\t\t\t<\/td>\r\n\t\t\t<\/tr>\r\n\t\t<\/tbody>\r\n\t<\/table>\r\n<\/div>\r\n\r\n<p style=\"text-align: justify\">\r\n\t\u00a0\r\n<\/p>\r\n\r\n<p style=\"margin-bottom:7.5pt;text-align:justify\">\r\n\t<span><span><span style=\"color:#2f2f2f\">Now the effects of the interactions can be calculated with the same formula from above.<\/span><\/span><\/span>\r\n<\/p>\r\n\r\n<p style=\"margin-bottom:7.5pt;text-align:justify\">\r\n\t<span><span><span style=\"color:#2f2f2f\">We can calculate the interaction effect the same way we calculated the single factor effect:<\/span><\/span><\/span>\r\n<\/p>\r\n\r\n<h6 style=\"margin-bottom:7.5pt;text-align:justify\">\r\n\t<img loading=\"lazy\" decoding=\"async\" width=\"1465\" height=\"97\" class=\"alignnone wp-image-82\" style=\"width: 604px;height: 40px\" src=\"https:\/\/sisu.ut.ee\/wp-content\/uploads\/sites\/130\/factorab1_02.png\" title=\"factorab1_02.png\" alt=\"FAB1\" srcset=\"https:\/\/sisu.ut.ee\/wp-content\/uploads\/sites\/130\/factorab1_02.png 1465w, https:\/\/sisu.ut.ee\/wp-content\/uploads\/sites\/130\/factorab1_02-300x20.png 300w, https:\/\/sisu.ut.ee\/wp-content\/uploads\/sites\/130\/factorab1_02-1024x68.png 1024w, https:\/\/sisu.ut.ee\/wp-content\/uploads\/sites\/130\/factorab1_02-768x51.png 768w\" sizes=\"auto, (max-width: 1465px) 100vw, 1465px\">(Eq 4)\r\n<\/h6>\r\n\r\n<h6>\r\n\t<img loading=\"lazy\" decoding=\"async\" width=\"1456\" height=\"183\" class=\"alignnone wp-image-81\" style=\"width: 544px;height: 75px\" src=\"https:\/\/sisu.ut.ee\/wp-content\/uploads\/sites\/130\/factorab2.png\" title=\"factorab2.png\" alt=\"FAB2\" srcset=\"https:\/\/sisu.ut.ee\/wp-content\/uploads\/sites\/130\/factorab2.png 1456w, https:\/\/sisu.ut.ee\/wp-content\/uploads\/sites\/130\/factorab2-300x38.png 300w, https:\/\/sisu.ut.ee\/wp-content\/uploads\/sites\/130\/factorab2-1024x129.png 1024w, https:\/\/sisu.ut.ee\/wp-content\/uploads\/sites\/130\/factorab2-768x97.png 768w\" sizes=\"auto, (max-width: 1456px) 100vw, 1456px\">(Eq 5)\r\n<\/h6>\r\n\r\n<p>\r\n\t<span style=\",serif\"><span style=\"color:#2f2f2f\">Now we have the complete picture of all effects on the response variable. <\/span><\/span>\r\n<\/p>\r\n\r\n<table class=\"table table-hover\" align=\"center\" border=\"1\" style=\"border-collapse: collapse;margin-left: 7.5pt;margin-right: 7.5pt\" width=\"1115\">\r\n\t<thead>\r\n\t\t<tr style=\"height:16.2pt\">\r\n\t\t\t<th colspan=\"7\" nowrap scope=\"col\" style=\"width: 557.7pt\" width=\"1115\">\r\n\t\t\t\t<p align=\"center\" style=\"margin-bottom:.0001pt;text-align:center;padding:0cm5.4pt0cm5.4pt\">\r\n\t\t\t\t\t<span style=\"line-height:normal\"><span style=\"height:16.2pt\"><b><span lang=\"EN-GB\"><span style=\",serif\"><span style=\"color:black\">Results <\/span><\/span><\/span><\/b><span lang=\"EN-GB\"><span style=\",serif\"><span style=\"color:black\">(<\/span><\/span><\/span><span><span style=\",serif\"><span style=\"color:black\">factor <\/span><\/span><\/span><span lang=\"EN-GB\"><span style=\",serif\"><span style=\"color:black\">and interaction effect sizes)<\/span><\/span><\/span><\/span><\/span>\r\n\t\t\t\t<\/p>\r\n\r\n\t\t\t\t<p align=\"center\" style=\"margin-bottom:.0001pt;text-align:center;padding:0cm5.4pt0cm5.4pt\">\r\n\t\t\t\t\t<span style=\"line-height:normal\"><span style=\"height:16.2pt\"><b><span lang=\"EN-GB\"><span style=\",serif\"><span style=\"color:black\">For retention time<\/span><\/span><\/span><\/b><\/span><\/span>\r\n\t\t\t\t<\/p>\r\n\t\t\t<\/th>\r\n\t\t<\/tr>\r\n\t<\/thead>\r\n\t<tbody>\r\n\t\t<tr style=\"height:15.55pt\">\r\n\t\t\t<td style=\"width: 35.2pt\" width=\"70\">\r\n\t\t\t\t<p align=\"center\" style=\"margin-bottom:.0001pt;text-align:center;padding:0cm5.4pt0cm5.4pt\">\r\n\t\t\t\t\t<span style=\"line-height:normal\"><span style=\"height:15.55pt\"><b><span><span style=\",serif\"><span style=\"color:black\">A<\/span><\/span><\/span><\/b><\/span><\/span>\r\n\t\t\t\t<\/p>\r\n\t\t\t<\/td>\r\n\t\t\t<td style=\"width: 67.25pt\" width=\"135\">\r\n\t\t\t\t<p align=\"center\" style=\"margin-bottom:.0001pt;text-align:center;padding:0cm5.4pt0cm5.4pt\">\r\n\t\t\t\t\t<span style=\"line-height:normal\"><span style=\"height:15.55pt\"><b><span><span style=\",serif\"><span style=\"color:black\">B<\/span><\/span><\/span><\/b><\/span><\/span>\r\n\t\t\t\t<\/p>\r\n\t\t\t<\/td>\r\n\t\t\t<td style=\"width: 89.6pt\" width=\"179\">\r\n\t\t\t\t<p align=\"center\" style=\"margin-bottom:.0001pt;text-align:center;padding:0cm5.4pt0cm5.4pt\">\r\n\t\t\t\t\t<span style=\"line-height:normal\"><span style=\"height:15.55pt\"><b><span><span style=\",serif\"><span style=\"color:black\">C<\/span><\/span><\/span><\/b><\/span><\/span>\r\n\t\t\t\t<\/p>\r\n\t\t\t<\/td>\r\n\t\t\t<td style=\"width: 53.5pt\" width=\"107\">\r\n\t\t\t\t<p align=\"center\" style=\"margin-bottom:.0001pt;text-align:center;padding:0cm5.4pt0cm5.4pt\">\r\n\t\t\t\t\t<span style=\"line-height:normal\"><span style=\"height:15.55pt\"><b><span><span style=\",serif\"><span style=\"color:black\">AB<\/span><\/span><\/span><\/b><\/span><\/span>\r\n\t\t\t\t<\/p>\r\n\t\t\t<\/td>\r\n\t\t\t<td style=\"width: 100.9pt\" width=\"202\">\r\n\t\t\t\t<p align=\"center\" style=\"margin-bottom:.0001pt;text-align:center;padding:0cm5.4pt0cm5.4pt\">\r\n\t\t\t\t\t<span style=\"line-height:normal\"><span style=\"height:15.55pt\"><b><span><span style=\",serif\"><span style=\"color:black\">BC<\/span><\/span><\/span><\/b><\/span><\/span>\r\n\t\t\t\t<\/p>\r\n\t\t\t<\/td>\r\n\t\t\t<td style=\"width: 85.45pt\" width=\"171\">\r\n\t\t\t\t<p align=\"center\" style=\"margin-bottom:.0001pt;text-align:center;padding:0cm5.4pt0cm5.4pt\">\r\n\t\t\t\t\t<span style=\"line-height:normal\"><span style=\"height:15.55pt\"><b><span><span style=\",serif\"><span style=\"color:black\">AC<\/span><\/span><\/span><\/b><\/span><\/span>\r\n\t\t\t\t<\/p>\r\n\t\t\t<\/td>\r\n\t\t\t<td style=\"width: 125.8pt\" width=\"252\">\r\n\t\t\t\t<p align=\"center\" style=\"margin-bottom:.0001pt;text-align:center;padding:0cm5.4pt0cm5.4pt\">\r\n\t\t\t\t\t<span style=\"line-height:normal\"><span style=\"height:15.55pt\"><b><span><span style=\",serif\"><span style=\"color:black\">ABC<\/span><\/span><\/span><\/b><\/span><\/span>\r\n\t\t\t\t<\/p>\r\n\t\t\t<\/td>\r\n\t\t<\/tr>\r\n\t\t<tr style=\"height:15.55pt\">\r\n\t\t\t<td style=\"width: 35.2pt\" width=\"70\">\r\n\t\t\t\t<p align=\"center\" style=\"margin-bottom:.0001pt;text-align:center;padding:0cm5.4pt0cm5.4pt\">\r\n\t\t\t\t\t<span style=\"line-height:normal\"><span style=\"height:15.55pt\"><b><span><span style=\",serif\"><span style=\"color:black\">pH<\/span><\/span><\/span><\/b><\/span><\/span>\r\n\t\t\t\t<\/p>\r\n\t\t\t<\/td>\r\n\t\t\t<td style=\"width: 67.25pt\" width=\"135\">\r\n\t\t\t\t<p align=\"center\" style=\"margin-bottom:.0001pt;text-align:center;padding:0cm5.4pt0cm5.4pt\">\r\n\t\t\t\t\t<span style=\"line-height:normal\"><span style=\"height:15.55pt\"><span><span style=\",serif\"><span style=\"color:black\">Additive<\/span><\/span><\/span><span lang=\"DE\"><span style=\",serif\"><span style=\"color:black\"> conc.<\/span><\/span><\/span><\/span><\/span>\r\n\t\t\t\t<\/p>\r\n\t\t\t<\/td>\r\n\t\t\t<td style=\"width: 89.6pt\" width=\"179\">\r\n\t\t\t\t<p align=\"center\" style=\"margin-bottom:.0001pt;text-align:center;padding:0cm5.4pt0cm5.4pt\">\r\n\t\t\t\t\t<span style=\"line-height:normal\"><span style=\"height:15.55pt\"><span><span style=\",serif\"><span style=\"color:black\">Column Temp.<\/span><\/span><\/span><\/span><\/span>\r\n\t\t\t\t<\/p>\r\n\t\t\t<\/td>\r\n\t\t\t<td style=\"width: 53.5pt\" width=\"107\">\r\n\t\t\t\t<p align=\"center\" style=\"margin-bottom:.0001pt;text-align:center;padding:0cm5.4pt0cm5.4pt\">\r\n\t\t\t\t\t<span style=\"line-height:normal\"><span style=\"height:15.55pt\"><span><span style=\",serif\"><span style=\"color:black\">pH * Additive<\/span><\/span><\/span><span lang=\"DE\"><span style=\",serif\"><span style=\"color:black\"> conc.<\/span><\/span><\/span><\/span><\/span>\r\n\t\t\t\t<\/p>\r\n\t\t\t<\/td>\r\n\t\t\t<td>\r\n\t\t\t\t<p align=\"center\" style=\"margin-bottom:.0001pt;text-align:center;padding:0cm5.4pt0cm5.4pt\">\r\n\t\t\t\t\t<span style=\"line-height:normal\"><span style=\"height:15.55pt\"><span><span style=\",serif\"><span style=\"color:black\">Additive * Column Temp.<\/span><\/span><\/span><\/span><\/span>\r\n\t\t\t\t<\/p>\r\n\t\t\t<\/td>\r\n\t\t\t<td>\r\n\t\t\t\t<p align=\"center\" style=\"margin-bottom:.0001pt;text-align:center;padding:0cm5.4pt0cm5.4pt\">\r\n\t\t\t\t\t<span style=\"line-height:normal\"><span style=\"height:15.55pt\"><span><span style=\",serif\"><span style=\"color:black\">pH * Column Temp.<\/span><\/span><\/span><\/span><\/span>\r\n\t\t\t\t<\/p>\r\n\t\t\t<\/td>\r\n\t\t\t<td>\r\n\t\t\t\t<p align=\"center\" style=\"margin-bottom:.0001pt;text-align:center;padding:0cm5.4pt0cm5.4pt\">\r\n\t\t\t\t\t<span style=\"line-height:normal\"><span style=\"height:15.55pt\"><span><span style=\",serif\"><span style=\"color:black\">pH * Additive * Column Temp.<\/span><\/span><\/span><\/span><\/span>\r\n\t\t\t\t<\/p>\r\n\t\t\t<\/td>\r\n\t\t<\/tr>\r\n\t\t<tr style=\"height:16.2pt\">\r\n\t\t\t<td>\r\n\t\t\t\t<p style=\"text-align: center\">\r\n\t\t\t\t\t<strong>-1.48<\/strong>\r\n\t\t\t\t<\/p>\r\n\t\t\t<\/td>\r\n\t\t\t<td>\r\n\t\t\t\t<p style=\"text-align: center\">\r\n\t\t\t\t\t<span style=\"line-height:normal\"><span style=\"height:16.2pt\"><span><span style=\",serif\"><span style=\"color:black\">-0.20<\/span><\/span><\/span><\/span><\/span>\r\n\t\t\t\t<\/p>\r\n\t\t\t<\/td>\r\n\t\t\t<td>\r\n\t\t\t\t<p align=\"center\" style=\"margin-bottom:.0001pt;text-align:center;padding:0cm5.4pt0cm5.4pt\">\r\n\t\t\t\t\t<span style=\"line-height:normal\"><span style=\"height:16.2pt\"><span><span style=\",serif\"><span style=\"color:black\">-0.17<\/span><\/span><\/span><\/span><\/span>\r\n\t\t\t\t<\/p>\r\n\t\t\t<\/td>\r\n\t\t\t<td>\r\n\t\t\t\t<p style=\"text-align: center\">\r\n\t\t\t\t\t<strong>1.07<\/strong>\r\n\t\t\t\t<\/p>\r\n\t\t\t<\/td>\r\n\t\t\t<td>\r\n\t\t\t\t<p align=\"center\" style=\"margin-bottom:.0001pt;text-align:center;padding:0cm5.4pt0cm5.4pt\">\r\n\t\t\t\t\t<span style=\"line-height:normal\"><span style=\"height:16.2pt\"><span><span style=\",serif\"><span style=\"color:black\">-0.03<\/span><\/span><\/span><\/span><\/span>\r\n\t\t\t\t<\/p>\r\n\t\t\t<\/td>\r\n\t\t\t<td>\r\n\t\t\t\t<p align=\"center\" style=\"margin-bottom:.0001pt;text-align:center;padding:0cm5.4pt0cm5.4pt\">\r\n\t\t\t\t\t<span style=\"line-height:normal\"><span style=\"height:16.2pt\"><span><span style=\",serif\"><span style=\"color:black\">0.18<\/span><\/span><\/span><\/span><\/span>\r\n\t\t\t\t<\/p>\r\n\t\t\t<\/td>\r\n\t\t\t<td>\r\n\t\t\t\t<p align=\"center\" style=\"margin-bottom:.0001pt;text-align:center;padding:0cm5.4pt0cm5.4pt\">\r\n\t\t\t\t\t<span style=\"line-height:normal\"><span style=\"height:16.2pt\"><span><span style=\",serif\"><span style=\"color:black\">0.23<\/span><\/span><\/span><\/span><\/span>\r\n\t\t\t\t<\/p>\r\n\t\t\t<\/td>\r\n\t\t<\/tr>\r\n\t<\/tbody>\r\n<\/table>\r\n\r\n<p style=\"text-align:justify\">\r\n\t\u00a0\r\n<\/p>\r\n\r\n<p style=\"margin-bottom:7.5pt;text-align:justify\">\r\n\t<span><span><span style=\"color:#2f2f2f\">The two-factor interaction between a pH and an additive concentration has a big influence on the retention time. This implies that our method is not yet robust towards pH and additionally towards additive concentration. In our case, the conclusion comes from the comparison of numeric values of the effect size with each other. There is strong evidence towards the significance of pH and <\/span><\/span><\/span><span><span><span style=\"color:black\">pH*Additive conc<\/span><\/span><\/span><span><span><span style=\"color:#2f2f2f\">. However, easy identification of significant effects is not always given, and in these cases, it is strongly recommended to use an ANOVA approach to determine which effects and factors <\/span><\/span><\/span><span><span>are statistically significant. A good ANOVA description in the context of a DoE can be found in the book [ref 68]. However, all results from a DoE should always be evaluated from a practical scientific standpoint.<\/span><\/span>\r\n<\/p>\r\n\r\n<p style=\"margin-bottom:7.5pt;text-align:justify\">\r\n\t<span><span>In general, it is assumed that the importance of interactions will decrease <span style=\"color:#2f2f2f\">with additional interacting factors, so it is unlikely that there will be very significant (three), four or five-factor interactions.<\/span><\/span><\/span><br><span><span><span style=\"color:#2f2f2f\">Reaching this conclusion is only possible with the help of a DoE. If the user would use the OFAT approach, the effects of the interactions on the response variables would go completely unnoticed. Only with the application of a DoE we were able to see the impact of the two-factor interaction on the response variable.<\/span><\/span><\/span>\r\n<\/p>\r\n\r\n<p style=\"margin-bottom:7.5pt;text-align:justify\">\r\n\t<span><span><span style=\"color:#2f2f2f\">Another way of portraying interactions is with the following plots. In both plots one factor is kept constant while the other factor is compared on its influence on the response variable. The plots firstly show us that there is an interaction between factor A and B because the slope changes visibly when only one factor is changed. Secondly, we can say that the interaction of AB is more influenced by the -1 setting for B than by the +1 setting of B.<\/span><\/span><\/span>\r\n<\/p>\r\n\r\n<p style=\"text-align:justify\">\r\n\t\u00a0\r\n<\/p>\r\n\r\n<div>\r\n\t<div class=\"msocomtxt\" id=\"_com_1\">\r\n\t\t<p class=\"MsoCommentText\" style=\"text-align: center\">\r\n\t\t\t<img loading=\"lazy\" decoding=\"async\" width=\"2197\" height=\"723\" class=\"alignnone wp-image-88\" style=\"width: 669px;height: 220px\" src=\"https:\/\/sisu.ut.ee\/wp-content\/uploads\/sites\/130\/doe_effectb-_01.png\" title=\"doe_effectb-_01.png\" alt=\"doeffectb\" srcset=\"https:\/\/sisu.ut.ee\/wp-content\/uploads\/sites\/130\/doe_effectb-_01.png 2197w, https:\/\/sisu.ut.ee\/wp-content\/uploads\/sites\/130\/doe_effectb-_01-300x99.png 300w, https:\/\/sisu.ut.ee\/wp-content\/uploads\/sites\/130\/doe_effectb-_01-1024x337.png 1024w, https:\/\/sisu.ut.ee\/wp-content\/uploads\/sites\/130\/doe_effectb-_01-768x253.png 768w, https:\/\/sisu.ut.ee\/wp-content\/uploads\/sites\/130\/doe_effectb-_01-1536x505.png 1536w, https:\/\/sisu.ut.ee\/wp-content\/uploads\/sites\/130\/doe_effectb-_01-2048x674.png 2048w, https:\/\/sisu.ut.ee\/wp-content\/uploads\/sites\/130\/doe_effectb-_01-1920x632.png 1920w\" sizes=\"auto, (max-width: 2197px) 100vw, 2197px\">\r\n\t\t<\/p>\r\n\r\n\t\t<h4 class=\"MsoCommentText\" style=\"text-align: center\">\r\n\t\t\tFigure 2 Interaction plots of the factors (A) pH and (B) Additive conc.\u00a0\r\n\t\t<\/h4>\r\n\r\n\t\t<p style=\"margin-bottom:7.5pt;text-align:justify\">\r\n\t\t\t<span><span><span style=\"color:#2f2f2f\">f) contour plots<\/span><\/span><\/span>\r\n\t\t<\/p>\r\n\r\n\t\t<p style=\"margin-bottom:7.5pt;text-align:justify\">\r\n\t\t\t<span><span><span style=\"color:#2f2f2f\">A different\u00a0way for the\u00a0interpretation of the interactions on the response variable are the so-called contour plots, as seen in the figures below. These pots have been calculated and plotted in the \u201cR\u201d software with the packages pid and rsm. The lines in the plot depict the interaction between pH and additive concentration and their effect on the response variable. The black dots in the corners of the contour plot describe the design space where the measurement points have been set. The more the lines are bent, the stronger is the interaction. The examples make this evident. When looking at the figure of pH and additive concentration, we see strongly bent lines in the chosen design space, and therefore we can assume a strong interaction between the two factors. On the contrary, if we look at the interaction between pH and column temperature, then we only see a very weak interaction between those factors.<\/span><\/span><\/span>\r\n\t\t<\/p>\r\n\r\n\t\t<div>\r\n\t\t\t<img loading=\"lazy\" decoding=\"async\" width=\"2835\" height=\"845\" class=\"alignnone wp-image-91\" style=\"width: 839px;height: 250px\" src=\"https:\/\/sisu.ut.ee\/wp-content\/uploads\/sites\/130\/interactions.png\" title=\"interactions.png\" alt=\"interactions\" srcset=\"https:\/\/sisu.ut.ee\/wp-content\/uploads\/sites\/130\/interactions.png 2835w, https:\/\/sisu.ut.ee\/wp-content\/uploads\/sites\/130\/interactions-300x89.png 300w, https:\/\/sisu.ut.ee\/wp-content\/uploads\/sites\/130\/interactions-1024x305.png 1024w, https:\/\/sisu.ut.ee\/wp-content\/uploads\/sites\/130\/interactions-768x229.png 768w, https:\/\/sisu.ut.ee\/wp-content\/uploads\/sites\/130\/interactions-1536x458.png 1536w, https:\/\/sisu.ut.ee\/wp-content\/uploads\/sites\/130\/interactions-2048x610.png 2048w, https:\/\/sisu.ut.ee\/wp-content\/uploads\/sites\/130\/interactions-1920x572.png 1920w\" sizes=\"auto, (max-width: 2835px) 100vw, 2835px\">\r\n\t\t<\/div>\r\n\r\n\t\t<h4 class=\"MsoCommentText\" style=\"text-align: center\">\r\n\t\t\tFigure 3 Contour plots of the interactions\r\n\t\t<\/h4>\r\n\r\n\t\t<p class=\"MsoCommentText\">\r\n\t\t\t\u00a0\r\n\t\t<\/p>\r\n\r\n\t\t<p class=\"MsoCommentText\">\r\n\t\t\t\u00a0\r\n\t\t<\/p>\r\n\r\n\t\t<p class=\"MsoCommentText\">\r\n\t\t\t\u00a0\r\n\t\t<\/p>\r\n\t<\/div>\r\n<\/div>\r\n<br><div class=\"wp-block-group attached-files-group is-layout-constrained wp-block-group-is-layout-constrained\"><div class=\"wp-block-file\"><a href=\"https:\/\/sisu.ut.ee\/wp-content\/uploads\/sites\/130\/10.4_experimental_design.pdf\" target=\"_blank\" rel=\"noreferrer noopener\">10.4_experimental_design.pdf<\/a><\/div><\/div>","protected":false},"excerpt":{"rendered":"<p>Additionally to the One factor at a Time approach (OFAT, explained in 10.3), robustness can also be estimated by varying multiple method parameters (factors) at the same time. Such approaches are strongly recommended if it is expected that different method &#8230;<\/p>\n","protected":false},"author":60,"featured_media":0,"parent":0,"menu_order":0,"comment_status":"closed","ping_status":"closed","template":"","meta":{"_acf_changed":false,"inline_featured_image":false,"footnotes":""},"class_list":["post-11","page","type-page","status-publish","hentry"],"acf":[],"_links":{"self":[{"href":"https:\/\/sisu.ut.ee\/lcms_method_validation\/wp-json\/wp\/v2\/pages\/11","targetHints":{"allow":["GET"]}}],"collection":[{"href":"https:\/\/sisu.ut.ee\/lcms_method_validation\/wp-json\/wp\/v2\/pages"}],"about":[{"href":"https:\/\/sisu.ut.ee\/lcms_method_validation\/wp-json\/wp\/v2\/types\/page"}],"author":[{"embeddable":true,"href":"https:\/\/sisu.ut.ee\/lcms_method_validation\/wp-json\/wp\/v2\/users\/60"}],"replies":[{"embeddable":true,"href":"https:\/\/sisu.ut.ee\/lcms_method_validation\/wp-json\/wp\/v2\/comments?post=11"}],"version-history":[{"count":2,"href":"https:\/\/sisu.ut.ee\/lcms_method_validation\/wp-json\/wp\/v2\/pages\/11\/revisions"}],"predecessor-version":[{"id":904,"href":"https:\/\/sisu.ut.ee\/lcms_method_validation\/wp-json\/wp\/v2\/pages\/11\/revisions\/904"}],"wp:attachment":[{"href":"https:\/\/sisu.ut.ee\/lcms_method_validation\/wp-json\/wp\/v2\/media?parent=11"}],"curies":[{"name":"wp","href":"https:\/\/api.w.org\/{rel}","templated":true}]}}