A novel internet-based reaction monitoring, control and autonomous self-optimization platform for chemical synthesis.

Transcription

1 A novel internet-based reaction monitoring, control and autonomous self-optimization platform for chemical synthesis. Daniel E. Fitzpatrick, * Claudio Battilocchio, Steven V. Ley Innovative Technology Centre, Department of Chemistry, University of Cambridge, Lensfield Road, Cambridge CB2 1EW, U.K. Supporting Information 1.0 Simple reaction automation Experiment Configuration A Vapourtec R2/R4 unit provided the pump, temperature control and switching valve support required for this experiment. The mass spectrometer sampling system consisted of a Rheodyne MRA valve (which was configured with a sampling setting of 34), a Knauer K120 pump (set to 0.1 ml min -1 ) and an Advion Expression MS (Figure 1). Figure S1. The equipment used for the simple automation experiment involving a fixed volume of reaction solution being passed multiple times through a packed column.

2 The Advion MS was configured to monitor peaks at (corresponding to the starting nitrile) and (corresponding to the amide product). The MS unit sent intensity data in an analogue format to an Arduino (Figure 2), which was connected to an RS232 cable. LeyLab queried this Arduino board via RS232 to get MS readings on a per-second basis. Figure S2. The analogue-to-serial converter used as the interface between LeyLab and the Advion MS. A voltage between 0-5 V was applied on the cables from the MS (left side) depending on the intensity of the monitored peaks. The Arduino board converted these to a digital representation (integer) between The MS make-up solvent consisted of an 80:20 ratio of deionized water and acetonitrile, with 0.1% (volumetric basis) formic acid. A 1 M solution of 3-cyanopyridine in water (7.5 g of the nitrile in 75 ml water) was put into flask A. The camera control (see below) threshold for the multi-pass experiment was set to 13% from the bottom of the capture frame. A 10 mm Omnifit column was packed with 2.0 g of MnO 2 and placed into a column jacket, which itself was secured into the R2/R4 reactor system. Machine Vision Liquid Level Detection Initially two mass balances were used to monitor the liquid levels in each flask for this experiment, however we noticed that the balance readings were not consistent over time. To investigate the cause of this problem, we recorded the output from a balance over a few days. During this time nothing was placed or removed from the balance tray. As can be seen in Figure 3, the mass readings changed by almost 20 g over the course of five days. Initially we tested whether the root cause of this change was electromagnetic interference (from nearby electrical equipment, including two mass spectrometers) however this was not found to have any influence. Fluctuations in daily temperature seemed to play a small role in mass readings, so we decided to investigate other weather effects. A very strong inverse

3 correlation was seen tying these changes to the local atmospheric pressure at the time: as pressure rose, the mass reading decreased. Figure S3. Top: the mass balance readings over the course of five days; Bottom: recorded atmospheric pressure of the local Cambridge area over the same time period. To mitigate this problem, we switched to using a webcam-based liquid level solution instead. The operating code to enable this was written in python and ran on a Raspberry Pi computer. The process followed to determine liquid level is shown in Figure 4. Figure S4. The process followed by a python script when determining liquid levels by machine vision.

4 Experiment Code The automation code that was supplied to LeyLab during experiment creation is shown in Appendix A. 2.0 Description of Complex algorithm There are four possible ways for the Complex method to select experimental conditions: Reflection A reflection is the most common method for the system to choose new experiment set points. In this case, the system ranks conditions from best to worst before reflecting the worst iteration through the centroid of a plane connecting the other points. This is shown in a figure in the main article. Extension This method only occurs if the previous iteration was obtained via a reflection. If the last iteration (i.e. the reflection) was the best performing compared to the previous, then the system extends the previous iteration along the path formed between the reflected point and the worst performing iteration. This method is described with a figure in the main article. Retraction A retraction can only occur if the previous iteration was obtained from a reflection. If the reflected iteration was the worst performing in the ranked list, then the system chooses new experimental conditions by moving the reflected point back towards the previously worst point. Again, this has been described with a figure in the main text. Shrinking If the previous iteration was a retraction, and it was still the worst performing of the lot, then a shrinking occurs. In this situation, the system takes the best n + 1 iterations (where n is the number of optimization parameters) and sets the best performing iteration as an anchor. It then chooses n new iterations by moving all the other best performing iterations towards the anchor. Having carried out these new iterations, the system performs a reflection and carries on as normal. For more detail regarding the process followed by LeyLab during optimization following the Complex method, refer to reference 22 in the main article text. 3.0 Three dimensional optimization Experiment Configuration The experiment configuration for the 3D optimization experiment was very similar to that described in section 1. A Vapourtec R2/R4 unit provided pump and temperature control support, a Rheodyne MRA valve allowed for sampling of the product stream, a Knauer K120 pump

5 supplied MS make-up solution to the MS and an Avion Expression MS was used as an analytical tool (Figure 5). A 100 psi back pressure regulator was placed after the MRA sampling valve. Figure S5. The equipment used for the three dimensional optimization experiment. A schematic showing equipment layout can be found in the body of the main article. In this case, the MS make-up solution consisted of a 50:50 H 2 O:MeCN ratio with 0.1 % (vol.) formic acid. The Knauer pump was set to 0.1 ml min -1. The MS was configured to monitor the same peaks as before, and the same Arduino-based analogue to serial converter was used to enable LeyLab to collect MS data. Flask A was filled with 250 ml of 1 M solution of 3-cyanopyridine in H 2 O (26 g in 250 ml), flask B consisted solely of deionized H 2 O. LeyLab was configured to optimize three parameters within the following boundaries: Temperature: o C Residence Time (tau): min Concentration: M LeyLab controlled reaction temperature by adjusting the column set point, residence time was manipulated by changing the overall flowrate through the system (pump A + pump B) and inlet concentration was changed by altering the ratio of the two pump flowrates (Pump A:Pump B). The system was configured to allow 2 column volumes (1 column volume was 3 ml) to pass before sampling 1 column volume. This was so that the system reached steady state before calculating the performance of the iteration. Between each iteration, the system was flushed with deionized H 2 O (1 ml min -1 for 20 minutes) so as to remove any remaining reaction solution from the column.

6 Equations The evaluation function for this experiment is shown in the main article text. Two equations were used to calculate the flowrates of pump A and pump B based on iteration set points. These were found from a series of simple mass balance calculations. + = =. + Where: F A = flowrate of pump A F B = flowrate of pump B V = reactor volume (3 ml) = residence time (adjusted by LeyLab) C = reactor inlet concentration (adjusted by LeyLab) C A = concentration of solution in flask A (1.0 M) Experiment Code The automation code that was supplied to LeyLab during experiment creation is shown in Appendix B. Optimization Set Points and Data Table S1. Experimental set points generated by LeyLab for each iteration with associated evaluation function response. Iteration Number 4.0 Five dimensional optimization Experiment Configuration Temperature (deg. C) Residence Time (min) Inlet Concentration (mol/l) Evaluated response The 5D optimization experiment used a Vapourtec R2/R4 unit (providing two pumps, value support and reactor coil temperature control), two Knauer K120 pumps (for triphenylphosphine and solvent reservoirs) and a Mettler-Toledo FlowIR (for in-line detection). The equipment is shown in Figure 6. A 100 psi back pressure regulator was placed after the FlowIR. A schematic showing equipment layout is in the main article.

7 Figure S6. Equipment used in the five dimensional optimization experiment. A schematic representation of experimental layout can be found in the main article. There were a total of four 250 ml reservoirs in this experiment, containing solutions of carbon tetrabromide (0.5 M in MeCN), α-methylbenzyl alcohol (0.5 M in MeCN), triphenylphosphine (0.25 M in MeCN) and acetonitrile. Each reservoir was connected to the reactor system through its own independent pump. LeyLab was configured to optimize five parameters within the following boundaries: Temperature: o C Residence Time (tau): min Concentration: M Equivalents of CBr 4 : Equivalents of PPh 3 : For each iteration the system was configured to pump 15 ml of reaction mixture before switching to solvent. LeyLab used only one third of the total reaction volume (5 ml) to account for dispersion thus ensuring that steady state conditions had been reached. Between each iteration the reactor coil was flushed with 2.0 ml min -1 for 5.5 minutes. Equations Two evaluation functions were used for this optimization experiment, as shown in the main article text. Four equations were used by LeyLab to calculate equipment parameters based on iteration set points. These were derived from simple mass balances over the system. =. =.. ( + +1) =.

8 Where = ( + +1).. 1 F A = flowrate of pump A F B = flowrate of pump B F C = flowrate of pump C F D = flowrate of pump D V = reactor volume (10 ml) = residence time (adjusted by LeyLab) x = equivalents of CBr 4 (adjusted by LeyLab) y = equivalents of PPh 3 (adjusted by LeyLab) z = overall concentration (adjusted by LeyLab) a = concentration CBr 4 (0.5 M) b = concentration of alcohol (0.5 M) c = concentration PPh 3 (0.25 M) Experiment Code The automation code that was supplied to LeyLab during experiment creation is shown in Appendix C. Optimization Set Points and Data Table S2. Experimental set points generated by LeyLab for each iteration with associated evaluation function response. Temperature (deg. C) Residence Time (min) Equivalents CBr4 Equivalents PPh3 Overall Concentration (mol/l) Evaluated response Iteration Number

9 5.0 General Experimental 1 H-NMR spectra were recorded on a Bruker Avance DPX-600 spectrometer with the residual solvent peak as the internal reference (CDCl 3 = 7.26 ppm, d 6 -DMSO = 2.50 ppm). 1 H resonances are reported to the nearest 0.01 ppm. 13 C-NMR spectra were recorded on the same spectrometers with the central resonance of the solvent peak as the internal reference (CDCl 3 = ppm, d 6 - DMSO = ppm). All 13 C resonances are reported to the nearest 0.1 ppm. DEPT 135, COSY, HMQC, and HMBC experiments were used to aid structural determination and spectral assignment. The multiplicity of 1H signals are indicated as: s = singlet, d = doublet, dd = doublet of doublet, ddd = doublet of doublet of doublet, t = triplet, q = quadruplet, sext = sextet, m = multiplet, br. = broad, or combinations of thereof. Coupling constants (J) are quoted in Hz and reported to the nearest 0.1 Hz. Where appropriate, averages of the signals from peaks displaying multiplicity were used to calculate the value of the coupling constant. Unless stated otherwise, reagents were obtained from commercial sources and used without purification. The removal of solvent under reduced pressure was carried out on a standard rotary evaporator. Synthesis of 3-Pyridine carboxamide. An Omnifit column (10 mm length, 1.0mm i.d.) was packed with 2.0 g manganese dioxide, with small plugs of celite placed at both ends of the column. A 0.28 M solution of 3-cyanopyridine was prepared in distilled water (1.46 g in 50 ml) and placed at the inlet of a Vapourtec R2/R4 pump. A 100 psi back pressure regulator was placed downstream of the column. The column was heated to 81 o C while the system was primed with distilled water. Once the system temperature had stabilized, the solution of 3-cyanopyridine was pumped at ml min -1 through the column. A yield of 82% ( 1 H NMR with internal standard) was obtained after the system had reached steady state. δ H (600 MHz; d6-dmso; 25 C) 7.48 (1 H, dd, J 7.9, J 4.8), 7.62 (1 H, s), 8.18 (1 H, s), 8.21 (1 H, d, J 7.9), 8.69 (1 H, d, J 4.8), 9.04 (1 H, s). δ C (150MHz; d6-dmso; 25 C) , , , , , Characterization data correspond to that previously reported (Battilocchio, C.; Hawkins, J. M.; Ley, S. V. Org. Lett. 2014, 16, ). Synthesis of 1-Bromoethylbenzene. 0.5 M, 0.5 M and 0.25 M solutions of carbon tetrabromide, 1-phenylethanol and triphenylphosphine respectively were prepared in acetonitrile and placed at the feed to three pumps. A fourth pump was fed by a solution containing just acetonitrile. The solutions of CBr 4, 1-phenylethanol and PPh 3 were pumped at ml.min -1, ml.min -1 and ml.min -1 respectively through a 10 ml PTFE coil heated to 111 o C. Acetonitrile was pumped through the system at ml.min -1 in order to dilute the overall concentration of the reaction mixture. The back pressure of the system was set to 100 psi. A yield of 92% ( 1 H NMR with internal standard) was obtained after the system had reached steady state.

10 δ H (600 MHz; CDCl 3 ; 25 C) 2.06 (3 H, d, J 6.9), 5.23 (1 H, q, J 6.9), 7.30 (1 H, t, J 7.3), 7.36 (2 H, t, J 7.3), 7.45 (2 H, d, J 7.3). δ C (150 MHz; CDCl 3 ; 25 C) 26.84, 49.58, , , , Characterization data match that previously reported (Denton, R.M.; An, J.; Adeniran, B.; Blake, A.J.; Lewis, W.; Poulton, A.M. J. Org. Chem. 2011, 76, ).

11 3-Pyridine carboxamide NMR

12 1-Bromoethylbenzene NMR

13 Appendix A Simple Automation Code <?php function experiment_start(){ //Define variables newvariable('currentpass', 0); newvariable('numberofpasses', 20); newvariable('flaskacutoff', 10); newvariable('flaskbcutoff', 10); //Equipment set points executeequipmentfunction('vapourtec', 'setflowrate', ['pumpa', 0]); executeequipmentfunction('vapourtec', 'setflowrate', ['pumpb', 0]); executeequipmentfunction('vapourtec', 'keypress', ['pumpb', 'reagent']); executeequipmentfunction('vapourtec', 'start', []); addlistener("getvariable('currentpass')","greaterthan","getvariable('numberofpasses')", "experiment_shutdown()"); pumpatob(); function experiment_shutdown(){ //This code executes when the experiment ends executeequipmentfunction('vapourtec', 'stop', []); stopexperiment(); function pumpatob(){ updatevariable('currentpass', intval(getvariable('currentpass'))+1); executeequipmentfunction('vapourtec', 'keypress', ['pumpb', 'reagent']); executeequipmentfunction('vapourtec', 'setflowrate', ['pumpa', 1]); executeequipmentfunction('vapourtec', 'setflowrate', ['pumpb', 0]); addlistener("getequipmentdata('camera')['1']['percentfrombase']","lessthanequalto","getva riable('flaskacutoff')", "pumpbtoa()"); function pumpbtoa(){ updatevariable('currentpass', intval(getvariable('currentpass'))+1); executeequipmentfunction('vapourtec', 'keypress', ['pumpb', 'solvent']); executeequipmentfunction('vapourtec', 'setflowrate', ['pumpa', 0]); executeequipmentfunction('vapourtec', 'setflowrate', ['pumpb', 1]);?> addlistener("getequipmentdata('camera')['2']['percentfrombase']","lessthanequalto","getva riable('flaskbcutoff')", "pumpatob()");

14 Appendix B Three Dimensional Optimization Code <?php function experiment_start(){ //Define variables newvariable('flaskacutoff', 12); newvariable('waitingfunction', 0); newvariable('columntemperaturesetpoint', 0); newvariable('currenttau', 0); newvariable('currentconc', 0); //Equipment set points executeequipmentfunction('vapourtec', 'setflowrate', ['pumpa', 0]); executeequipmentfunction('vapourtec', 'setflowrate', ['pumpb', 0]); executeequipmentfunction('vapourtec', 'keypress', ['pumpb', 'reagent']); executeequipmentfunction('vapourtec', 'start', []); //Add flask empty listeners addlistener("getequipmentdata('camera')['1']['percentfrombase']","lessthanequalto","getva riable('flaskacutoff')", "experiment_shutdown()"); //Register simplex simplexadd('evaluationfunction','waitingfunction','betweeniterationsfunction',0); minutes simplexaddvariable('temp', 'gettemp', 'settemp([x])', '100', '30'); simplexaddvariable('tau', 'gettau', 'settau([x])', '20', '2'); simplexaddvariable('conc', 'getconc', 'setconc([x])', '1', '0.05'); //residence time in //Generate setpoints simplexgeneratesetpoints(); //Start simplex simplexnext(); //Simplex functions function settemp($x){ //Set column temperature $temperature = round((float)$x, 0); executeequipmentfunction('vapourtec', 'settemperature', [3, $temperature]); updatevariable('columntemperaturesetpoint', $temperature); function settau($x){ $tau = (float)$x; updatevariable('currenttau', $tau); function setconc($x){ $conc = (float)$x; updatevariable('currentconc', $conc); function gettemp(){ function gettau(){ function getconc(){ function evaluationfunction(){ global $experimentid; global $simplexid; //Find out how long 1 CV is $tau = getvariable('currenttau'); $residencetime = (float)$tau * 60; $twocv = 1 * $residencetime; //only 1 CV so that one CV has been allowed through the column and thus read by MS

15 //Get advion readings $sql = "SELECT ed.* FROM equipmentdata ed INNER JOIN equipmentlist el ON ed.equipmentid=el.id WHERE el.name='advion' AND el.experimentid='".mysql_real_escape_string($experimentid)."' ORDER BY date DESC LIMIT ".(string)round($twocv, 0); $result = mysql_query($sql); $startingreading = 0.0; $productreading = 0.0; while($row = mysql_fetch_array($result, MYSQL_ASSOC)){ $array = JSON_decode($row['data'], true); $startingreading += $array['analogue']['a0']; $productreading += $array['analogue']['a1']; $startingreading = $startingreading/(float)intval($twocv); $productreading = $productreading/(float)intval($twocv); //Get evaluation (ratio of product to starting material) $evaluation = $productreading/$startingreading; return $evaluation; function waitingfunction(){ //Flush column executeequipmentfunction('vapourtec', 'setflowrate', ['pumpa', 0]); executeequipmentfunction('vapourtec', 'setflowrate', ['pumpb', 1]); $time = time() ; //flush for 20 mins addlistener("time()","greaterthanequalto",$time, "updatevariable('waitingfunction', intval(getvariable('waitingfunction'))+1)"); //Wait for column temperature addlistener("round((float)getequipmentdata('vapourtec')['temperatures']['slot3']['tempera ture'],0)","equals","round((float)getvariable('columntemperaturesetpoint'),0)", "updatevariable('waitingfunction', intval(getvariable('waitingfunction'))+1)"); //Add listener for flush and temperature addlistener("getvariable('waitingfunction')","greaterthanequalto","intval(2)", "setpumps()"); function setpumps(){ global $experimentid; global $simplexid; updatevariable('waitingfunction', 0); //Get tau $tau = (float)getvariable('currenttau'); //Get concentration $conc = (float)getvariable('currentconc'); //Calculate flow rates $totalflowrate = 3.0/$tau; $Fa = ($conc*$totalflowrate)/1.0; $Fb = $totalflowrate-$fa; $pumpaflowrate = round((float)$fa, 3); $pumpbflowrate = round((float)$fb, 3); //Set pumps executeequipmentfunction('vapourtec', 'setflowrate', ['pumpa', $pumpaflowrate]); executeequipmentfunction('vapourtec', 'setflowrate', ['pumpb', $pumpbflowrate]); //Wait 3CV + 5 mins (for MS stability) $time = round(time() + ($tau*60*3) + (60*5), 0); addlistener("time()","greaterthanequalto",$time, "simplexnext()");

16 function betweeniterationsfunction(){ //Do nothing simplexnext(); function experiment_shutdown(){ //This code executes when the experiment ends executeequipmentfunction('vapourtec', 'stop', []);?> stopexperiment();

17 Appendix C Five Dimensional Optimization Code <?php function experiment_start(){ //Define variables newvariable('waitingfunction', 0); newvariable('coiltemperaturesetpoint', 0); newvariable('currenttau', 0); newvariable('currentequiva', 0); newvariable('currentequivc', 0); newvariable('currentoverallconc', 0); newvariable('currenttppflowrate', 0); newvariable('currentsolventflowrate', 0); //Equipment set points executeequipmentfunction('vapourtec', 'setflowrate', ['pumpa', 0]); executeequipmentfunction('vapourtec', 'setflowrate', ['pumpb', 0]); executeequipmentfunction('vapourtec', 'keypress', ['pumpa', 'solvent']); executeequipmentfunction('vapourtec', 'keypress', ['pumpb', 'solvent']); executeequipmentfunction('vapourtec', 'start', []); executeequipmentfunction('knauersolvent', 'Stop', []); executeequipmentfunction('knauertpp', 'Stop', []); //Register simplex simplexadd('evaluationfunction','waitingfunction','betweeniterationsfunction',0); simplexaddvariable('temp', 'gettemp', 'settemp([x])', '140', '30'); simplexaddvariable('tau', 'gettau', 'settau([x])', '10', '2'); simplexaddvariable('equiva', 'getequiva', 'setequiva([x])', '1.9', '0.1'); simplexaddvariable('equivc', 'getequivc', 'setequivc([x])', '1.9', '0.1'); simplexaddvariable('overallconc', 'getoverallconc', 'setoverallconc([x])', '0.375', '0.05'); //Generate setpoints simplexgeneratesetpoints(); //Start simplex simplexnext(); //Simplex functions function settemp($x){ //Set column temperature $temperature = round((float)$x, 0); executeequipmentfunction('vapourtec', 'settemperature', [2, $temperature]); updatevariable('coiltemperaturesetpoint', $temperature); function settau($x){ $tau = (float)$x; updatevariable('currenttau', $tau); function setequiva($x){ $equiva = (float)$x; updatevariable('currentequiva', $equiva); function setequivc($x){ $equivc = (float)$x; updatevariable('currentequivc', $equivc); function setoverallconc($x){ $overallconc = (float)$x; updatevariable('currentoverallconc', $overallconc);

18 function gettemp(){ function gettau(){ function getequiva(){ function getequivc(){ function getoverallconc(){ function evaluationfunction(){ global $experimentid; global $simplexid; //Find out how many datapoints in one residence time $tau = getvariable('currenttau'); $residencetime = round((float)$tau * 60 * 0.33, 0); //Only process data for 33% of residence time to account for dispersion //Get IR readings for 0.33 tau (3.3 ml) $sql = "SELECT ed.* FROM equipmentdata ed INNER JOIN equipmentlist el ON ed.equipmentid=el.id WHERE el.name='flowir' AND el.experimentid='".mysql_real_escape_string($experimentid)."' ORDER BY date DESC LIMIT ".(string)round($residencetime, 0); $result = mysql_query($sql); $sma = 0.0; $smb = 0.0; $smc = 0.0; $ptppo1 = 0.0; $ptppo2 = 0.0; while($row = mysql_fetch_array($result, MYSQL_ASSOC)){ $array = JSON_decode($row['data'], true); $sma += $array[0]['value']; $smb += $array[1]['value']; $smc += $array[2]['value']; $ptppo1 += $array[3]['value']; $ptppo2 += $array[4]['value']; $sma = $sma/(float)intval($residencetime); $smb = $smb/(float)intval($residencetime); $smc = $smc/(float)intval($residencetime); $ptppo1 = $ptppo1/(float)intval($residencetime); $ptppo2 = $ptppo2/(float)intval($residencetime); //Get evaluation $tau = (float)getvariable('currenttau'); $z = (float)getvariable('currentoverallconc'); $p = (float)$ptppo1; $b = abs((float)$smb); $x = (float)getvariable('currentequiva'); $y = (float)getvariable('currentequivc'); if($b == 0){ $b = (float) ; $evaluation = 0.01*((1.0/$tau) + $z) + $p*($p/$b) *(1/($x + $y)); return $evaluation; function waitingfunction(){ //Flush coils executeequipmentfunction('vapourtec', 'keypress', ['pumpa', 'solvent']); executeequipmentfunction('vapourtec', 'keypress', ['pumpb', 'solvent']); executeequipmentfunction('vapourtec', 'setflowrate', ['pumpa', 0.75]); executeequipmentfunction('vapourtec', 'setflowrate', ['pumpb', 0.75]); executeequipmentfunction('knauersolvent', 'setflowrate', [0.5]); executeequipmentfunction('knauersolvent', 'Start', []); executeequipmentfunction('knauertpp', 'Stop', []); $time = time() + 330;

19 addlistener("time()","greaterthanequalto",$time, "updatevariable('waitingfunction', intval(getvariable('waitingfunction'))+1)"); //Wait for coil temperature addlistener("round((float)getequipmentdata('vapourtec')['temperatures']['slot2']['tempera ture'],0)","equals","round((float)getvariable('coiltemperaturesetpoint'),0)", "updatevariable('waitingfunction', intval(getvariable('waitingfunction'))+1)"); //Add listener for flush and temperatures addlistener("getvariable('waitingfunction')","greaterthanequalto","intval(2)", "setpumps()"); function setpumps(){ global $experimentid; global $simplexid; $conca = 0.5; $concb = 0.5; $concc = 0.25; updatevariable('waitingfunction', 0); //Get experiment variables $tau = (float)getvariable('currenttau'); $x = (float)getvariable('currentequiva'); $y = (float)getvariable('currentequivc'); $z = (float)getvariable('currentoverallconc'); //Calculate flowrates $flowrateb = ($z * 10)/($tau * $concb * ($x + $y + 1)); $flowratea = (($x * $concb)/$conca) * $flowrateb; $flowratec = (($y * $concb)/$concc) * $flowrateb; $flowrated = $flowrateb * ((($concb * ($x + $y + 1))/$z) - (($x * $concb)/$conca) - (($y * $concb)/$concc) - 1); $flowrateb = round((float)$flowrateb, 3); $flowratea = round((float)$flowratea, 3); $flowratec = round((float)$flowratec, 3); $flowrated = round((float)$flowrated, 3); //Set pumps executeequipmentfunction('vapourtec', 'keypress', ['pumpa', 'reagent']); executeequipmentfunction('vapourtec', 'keypress', ['pumpb', 'reagent']); executeequipmentfunction('vapourtec', 'setflowrate', ['pumpa', $flowratea]); executeequipmentfunction('vapourtec', 'setflowrate', ['pumpb', $flowrateb]); executeequipmentfunction('knauersolvent', 'setflowrate', [$flowrated]); executeequipmentfunction('knauersolvent', 'Start', []); executeequipmentfunction('knauertpp', 'setflowrate', [$flowratec]); executeequipmentfunction('knauertpp', 'Start', []); //Update value for knauer flowrates updatevariable('currentsolventflowrate', $flowrated); updatevariable('currenttppflowrate', $flowratec); //Inject 15mL, then switch to just solvent $time = round(time() + ($tau*60*1.25), 0); addlistener("time()","greaterthanequalto",$time, "switchtosolvent()"); //Add listener for simplex evaluation $time = round(time() + ($tau*60*2.25), 0); addlistener("time()","greaterthanequalto",$time, "simplexnext()"); function switchtosolvent(){

20 global $experimentid; global $simplexid; //Switch valve and set flowrate executeequipmentfunction('vapourtec', 'keypress', ['pumpa', 'solvent']); executeequipmentfunction('vapourtec', 'keypress', ['pumpb', 'solvent']); //Set solvent knauer flowrate $currenttpp = (float)getvariable('currenttppflowrate'); $currentsolvent = (float)getvariable('currentsolventflowrate'); $solventflowrate = round($currenttpp + $currentsolvent, 3); executeequipmentfunction('knauersolvent', 'setflowrate', [$solventflowrate]); executeequipmentfunction('knauersolvent', 'Start', []); executeequipmentfunction('knauertpp', 'Stop', []); function betweeniterationsfunction(){ //Do nothing simplexnext(); function experiment_shutdown(){ //This code executes when the experiment ends executeequipmentfunction('vapourtec', 'stop', []); executeequipmentfunction('knauersolvent', 'Stop', []); executeequipmentfunction('knauertpp', 'Stop', []);?> stopexperiment();