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S to re in -2 0 C fre e z e r M u ltic h a n n e l P ip e ttor M u ltic h a n n e l P ip e ttor
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Figure 17.7. Adopted working process owchart related to the 96-well plate based log D determination.
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rearranged. This capability is needed to combine samples from different plates into one 96-well plate enabling use of the 96-channel liquid handler. When the samples are submitted, a le containing information on the speci c samples, such as the corporate ID, lot number, name of submitter, and plate ID, is also submitted along with the plate (step 1 in Figure 17.8). Molecular weights and Q1 mass are obtained and the log D values calculated (step 3 in Figure 17.8). Depending on c log D values, compounds were divided into two major groups, i.e., in or out of the -2 to 2 range (step 4 in Figure 17.8). If the c log D value is within the -2 to 2 range, 1 mL of stock samples will be used. If the c log D value is out of the range, 10 mL of stock samples will be used. Following the sequence of the sorted c log D values, samples are arranged in 96-well plates for actual analysis. The compounds are arranged in a 96well plate as shown in step 5 of Figure 17.8. This is the plate map for placing samples in the plate. After the samples are arranged, proper amounts of octanol and aqueous buffers are added into the wells and fully mixed to equilibrate. After reaching equilibrium, the samples from both the octanol and aqueous phases are transferred to autosampler plates using EDP3 multi-channel pipettes. If required, the Apricot pipettor would be used to dilute the samples appropriately. General rules for dilution require no dilution for either layer if c log D is <2. If c log D is >2, then the
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Obtained plate map associate with each plate
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THRX # Lot Number Barcode Plate B Plate name THRX-228191 1 THRX-967078 1 1 We THRX-993812 THRX # ll THRX-102134 1 E1 T HRX -228191 44557 * lli_782_78-1 Plate Map 44560 * lli_782_78-1 Plate Map 44562 S ubmitter * lli_782_78-1 Plate Map Lot Plate 44563 Craig H .lli_782_78-1 Plate Map * 1 CH u-673-158
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G1 T HRX -287480 1 Craig H . B1 T HRX -120650 1 Li Li Create a list of all samples submitted Li Li C1 T HRX -991652 1
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Obtained a) MW and Q1 mass and
THRX # THRX-228191 THRX-287480 THRX-120650 THRX-991652 Lot 1 1 1 1 Mol Weight 596.4304 671.5434 594.4579 652.4945 Q1 Mass 482.96 558.07 480.98 539.02
b) c log D using templates and the THRX #
T H RX # cpH 4 3 .8 0 .3 2 .6 2 .8 cp H 7 cp H 9 3 .8 2 .3 2 .6 5 .4 4 .3 3 .9 3 .5 6 .5 T H R X -2 28 1 91 T H R X -2 87 4 80 T H R X -1 20 6 50 T H R X -9 91 6 52
Step 4: Step 5: Arrange samples in 96-well plate
1 2 3
Sort sample by plate map, Q1 mass and clog D
Plate Q1 Mass clog D 496.98 468.93 549.02 1 0 549.02 1 1.1 1.7 1.8 Li Li lli_782_78-1 Plate Map Li Li lli_782_78-1 Plate Map Craig H. CHu-673-183 6 7 8 9 Craig H. CHu-673-183
THRX # THRX-107414 THRX-633481 THRX-342551 4 5 THRX-102826
Lot Submitter 1 1 1 1
H X 25467 T R -1 H X 91052 T R -6 H X 90281 T R -4 H X 62775 T R -2 H X 01405 T R -1 H X 38031 T R -5 H X 63392 T R -5 H X 64857 T R -3 H X 54635 T R -1 H X 17977 T R -1 H X 01335 A T R -1
H X 88179 T R -1 H X 14337 T R -1 H X 78785 T R -1 H X 42189 T R -3 H X 10000 T R -1 H X 33166 T R -1 H X 10600 T R -5 H X 55699 T R -6 H X 06654 T R -6 H X 29445 T R -1 H X 96220 B T R -5
1 2 3 4 5 6 7 8 9 10 11 12 H X 52967 T R -1 H X 94150 T R -1 H X 66465 T R -1 H X 06364 T R -7 H X 89181 T R -2 H X 11775 T R -1 H X 47573 T R -7 H X 41894 T R -2 H X 11281 T R -7 H X 67717 T R -1 H X 27636 C T R -1 A
H X 05398 D T R -5 H X 15587 E T R -1
H X 11946 F T R -2 H X 04730 G T R -1
H X 86256 H T R -1
aq; THRX-- aq; THRX-- aq; THRX-- aq; THRX-- aq; THRX-- aq; THRX-- aq; THRX-- aq; THRX-- aq; THRX-- aq; THRX-- aq; THRX-- aq; THRX-125467 191052 690281 462775 201405 138031 563392 564857 354635 117977 101335 178828 o; THRX-- o; THRX-- o; THRX-- o; THRX-- o; THRX-- o; THRX-- o; THRX-- o; THRX-- o; THRX-- o; THRX-- o; THRX-- o; THRX-T R -1 H X 81415 T R -4 H X 63551 T R -4 H X 38181 T R -1 H X 95074 T R -8 H X 90008 T R -1 H X 01566 T R -3 H X 60257 T R -1 H X 55307 T R -4 H X 13995 T R -8 H X 13176 114337 178785 142189 310000 133166 110600 555699 606654 629445 196220 691154 B 588179 aq; THRX-- aq; THRX-- aq; THRX-- aq; THRX-- aq; THRX-- aq; THRX-- aq; THRX-- aq; THRX-- aq; THRX-- aq; THRX-- aq; THRX-- aq; THRX-T R -2 H X 09279 T R -1 H X 22179 T R -7 H X 99113 T R -1 H X 10378 T R -7 H X 31519 T R -6 H X 98187 T R -9 H X 89399 T R -2 H X 49558 T R -2 H X 02865 T R -1 H X 10506 194150 166465 106364 789181 211775 147573 741894 211281 767717 127636 895326 C 152967 o; THRX-- o; THRX-- o; THRX-- o; THRX-- o; THRX-- o; THRX-- o; THRX-- o; THRX-- o; THRX-- o; THRX-- o; THRX-- o; THRX-181415 463551 438181 195074 890008 101566 360257 155307 413995 813176 352126 D 505398 T R -1 H X 73238 T R -1 H X 65372 T R -8 H X 02054 T R -4 H X 13614 T R -1 H X 56623 T R -1 H X 54238 T R -8 H X 36878 T R -7 H X 52996 T R -1 H X 99890 T R -7 H X 76541 aq; THRX-- aq; THRX-- aq; THRX-- aq; THRX-- aq; THRX-- aq; THRX-- aq; THRX-- aq; THRX-- aq; THRX-- aq; THRX-- aq; THRX-- aq; THRX-209279 122179 799113 110378 731519 698187 989399 249558 202865 110506 322556 E 115587 T R -5 H X 89752 T R -1 H X 94416 T R -1 H X 60362 T R -3 H X 06020 T R -5 H X 39953 T R -9 H X 40280 T R -9 H X 51096 T R -7 H X 80170 T R -2 H X 01752 T R -1 H X 94230 o; THRX-- o; THRX-- o; THRX-- o; THRX-- o; THRX-- o; THRX-- o; THRX-- o; THRX-- o; THRX-- o; THRX-- o; THRX-- o; THRX-Step 6: 802054 413614 156623 154238 836878 752996 199890 776541 802054 173238 165372 F 211946 aq; THRX-- aq; THRX-- aq; THRX-- aq;dilution of samples usingaq; THRX-- aq; THRX-- aq; THRX-- aq; THRX-- aq; THRX-THRX-- aq; aq; THRX-- Perform THRX 15685 THRX--93385 THRXaq; THRX--RX 76800 THRXPersonal Pipettor Apricot -672396 THRX 55179 THRX 19867 T R -5 H X 63109 T R -7 H X 96545 T R -1 H X 40478 -1 T R -1 HX -1 24587 T -4 H -1 -1 589752 194416 160362 306020 539953 940280 951096 780170 201752 194230 104691 G 104730 - Transfer THRX-- o; THRX-- o;aqueous phases using a multichannel o; THRX-octanol and THRX-- o; THRX-- o; THRX-- o; THRX-- o; THRX-- o; THRX-o; THRX-- o; THRX-- o; THRX-- o; note: aq = undiluted 115685 193385 124587 undiluted octanol layer aqueous layer; o = 476800 672396 155179 119867 516239 563109 796545 140478 H 186256 pipette into a new 96-well plate