Davids Project Data

IMCS Time
Sample Name Time Actual Codeine Concentration Calculated Concentration

50% N/A 25 20.28
CO N/A 50 48.66
3x N/A 150 159.21
10x N/A 500 541.64
15x N/A 750 796.39
25x N/A 1250 1363.26
50x N/A 2500 2295.54
LCU N/A 30 27.11
MCU N/A 50 47.61
HCU N/A 150 172.04
2500ng/mL Codeine 15 min 15 min 2500 200.52
Accuracy
81.14
97.32
106.14
108.33
106.19
109.06
91.82
90.36
95.22
114.69 Average % Recovery Standard Deviation %CV
8.021%
8.901% 8.45% 0.44% 5.22%
8.420%
16.076%
16.849% 16.80% 0.69% 4.13%
17.461%
23.950%
24.027% 23.60% 0.68% 2.88%
22.813%
24.466%
24.534% 24.71% 0.37% 1.51%
25.144%
31.234%
29.693% 29.53% 1.79% 6.07%
27.662%
36.394%
38.118% 37.41% 0.90% 2.41%
37.726%
IMCSzyme™ Incubation Time Trials

40.00%
35.00%
30.00%
Percent Recovery
25.00%
20.00%
15.00%
10.00%
5.00%
0.00%
15 mi n 30 mi n 45 mi n 60 mi n 90 mi n 120 mi n
Incubation Time
IMCS Activity (Volume)
Sample Name Enzyme Activity Units Actual Codeine Concentration
50% 0U 25
CO 0U 50
3x 0U 150
10x 0U 500
15x 0U 750
25x 0U 1250
50x 0U 2500
Blank 0U 30
LCU 0U 30
MCU 0U 50
HCU 0U 150
2500ng/mL Codeine 2.5uL B-Gluc 125U 2500
2500ng/mL Codeine 5uL B-Gluc 250U 2500
60.00%
50.00%
Percent Recovery
40.00%
30.00%
20.00%
10.00%
0.00%
125U
Calculated Concentration Accuracy
24.58 98.30
50.15 100.31
153.35 102.23
484.82 96.96
776.72 103.56
1230.44 98.43
2505.15 100.21
<0 N/A
32 106.68
35.22 70.44
154.7 103.14 Average % Recovery Standard Deviation %CV
436.03 17.441%
459.4 18.376% 17.57% 0.75% 4.26%
422.44 16.898%
551.18 22.047%
531.47 21.259% 21.30% 0.73% 3.44%
514.56 20.582%
789.26 31.570%
837.81 33.512% 33.24% 1.55% 4.65%
865.65 34.626%
1182.91 47.316%
1161.73 46.469% 46.92% 0.43% 0.91%
1174.08 46.963%
1242.1 49.684%
1257.11 50.284% 49.52% 0.86% 1.74%
1214.55 48.582%
IMCSzyme™ Activity Units Trials

60.00%
50.00%
Percent Recovery
40.00%
30.00%
20.00%
10.00%
0.00%
125U 250U 500U 750U 1000U
Activity Units
IMCS Temperature
Sample Name Temperature Actual Codeine Concentration Calculated Concentration
50% N/A 25 27.06
CO N/A 50 45.26
3x N/A 150 153.54
10x N/A 500 473.93
15x N/A 750 736.03
25x N/A 1250 1359.56
50x N/A 2500 2429.62
LCU N/A 30 35.05
MCU N/A 50 46.41
HCU N/A 150 165.57
2500ng/mL Codeine 35C B-Gluc 35°C 2500 262.24
IMCSzyme
30.00%
25.00%
Percent Recovery
20.00%
15.00%
10.00%
5.00%
0.00%
35°C 40°C
0.00%
35°C 40°C
Accuracy
108.25
90.52
102.36
94.79
98.14
108.76
97.18
116.83
92.82
10.490%
10.288% 10.53% 0.26% 2.44%
10.798%
17.834%
18.791% 17.97% 0.76% 4.23%
17.291%
24.596%
22.379% 23.35% 1.13% 4.86%
23.067%
24.519%
25.177% 25.45% 1.10% 4.31%
26.661%
16.872%
19.165% 17.59% 1.36% 7.75%
16.742%
8.092%
7.231% 7.70% 0.44% 5.67%
7.789%
Kura Time
50% N/A 25 24.39
CO N/A 50 53.41
3x N/A 150 146.3
10x N/A 500 469.76
15x N/A 750 781.15
25x N/A 1250 1181.23
50x N/A 2500 2496.27
LCU N/A 30 168.77
MCU N/A 150 169.87
HCU N/A 2500 170.97
Accuracy
97.54
106.82
97.53
93.95
104.15
94.5
99.85
562.57
113.25
6.827%
6.835% 6.823% 0.014% 0.207%
6.808%
16.838%
16.313% 16.315% 0.522% 3.197%
15.795%
27.003%
26.700% 26.622% 0.426% 1.600%
26.162%
29.664%
30.684% 29.500% 1.273% 4.315%
28.154%
40.860%
41.253% 41.104% 0.213% 0.517%
41.198%
47.007%
47.168% 47.741% 1.134% 2.376%
49.047%
Figure 3. Kura Biotec Incubation Time Trials

60.000%
50.000%
Percent Recovery
40.000%
30.000%
20.000%
10.000%
0.000%
15 min 30 min 45 min 60 min 90 min 120 min
Incubation Time
Kura Activity (Volume)
50% 0U 25
CO 0U 50
3x 0U 150
10x 0U 500
15x 0U 750
25x 0U 1250
50x 0U 2500
LCU 0U 30
MCU 0U 50
HCU 0U 150
24.42 97.7
48.2 96.4
160.93 107.29
494.18 98.84
1079.39 143.92
1247.26 99.78
1872.01 74.88
34.68 115.62
57.95 115.89
64.89 2.596%
71.25 2.850% 2.525% 0.366% 14.497%
53.21 2.128%
157.02 6.281%
145.32 5.813% 6.320% 0.528% 8.356%
171.67 6.867%
404.81 16.192%
416.15 16.646% 16.034% 0.704% 4.394%
381.6 15.264%
599.97 23.999%
568.16 22.726% 23.582% 0.741% 3.143%
600.54 24.022%
594.69 23.788%
426.14 17.046% 20.417% 4.767% 23.350%
N/A N/A Inject did not work
Figure 1. Kura Biotec Activity Trials

25.000%
20.000%
Percent Recovery
15.000%
10.000%
5.000%
0.000%
125U 250U 500U 750U 1000U
Activity Units
Kura Temperature
50% N/A 25 24.36
CO N/A 50 53.55
3x N/A 150 163.03
10x N/A 500 435.11
15x N/A 750 1255.61
25x N/A 1250 1195.43
50x N/A 2500 2603.53
LCU N/A 30 126.1
MCU N/A 50 55.72
HCU N/A 150 171.51
2500ng/mL Codeine 40C B-Gluc 40°C 2500 366
Figure 2. Ku
35.000%
30.000%
25.000%
Percent Recovery
20.000%
15.000%
10.000%
5.000%
0.000%
35°C 40°C
5.000%
0.000%
35°C 40°C
Accuracy
97.42
107.1
108.68
87.02
167.41
95.63
104.14
420.33
111.44
8.557%
9.032% 8.773% 0.241% 2.742%
8.731%
14.640%
14.184% 13.998% 0.753% 5.382%
13.168%
16.946%
17.254% 17.310% 0.396% 2.288%
17.732%
23.661%
26.230% 25.014% 1.290% 5.157%
25.152%
29.971%
27.580% 28.821% 1.198% 4.158%
28.911%
0.376%
0.298% 0.359% 0.055% 15.434%
0.405%
Figure 2. Kura Biotec Temperature Trials

.000%
.000%
.000%
.000%
.000%
.000%
.000%
.000%
35°C 40°C 45°C 50°C 55°C 60°C
Temperature
.000%
.000%
35°C 40°C 45°C 50°C 55°C 60°C
Temperature
IMCS Activity (Volume)
50% 0U 25
CO 0U 50
3x 0U 150
10x 0U 500
15x 0U 750
25x 0U 1250
50x 0U 2500
Blank 0U 30
LCU 0U 30
MCU 0U 50
HCU 0U 150
IMCSzyme™ Kura Biotec

125U 17.57% 2.53%
250U 21.30% 6.32%
500U 33.24% 16.03%
750U 46.92% 23.58%
1000U 49.52% 20.42%
24.58 98.30
50.15 100.31
153.35 102.23
484.82 96.96
776.72 103.56
1230.44 98.43
2505.15 100.21
<0 N/A
32 106.68
35.22 70.44
436.03 17.441%
459.4 18.376% 17.57% 0.75% 4.26%
422.44 16.898%
551.18 22.047%
531.47 21.259% 21.30% 0.73% 3.44%
514.56 20.582%
789.26 31.570%
837.81 33.512% 33.24% 1.55% 4.65%
865.65 34.626%
1182.91 47.316%
1161.73 46.469% 46.92% 0.43% 0.91%
1174.08 46.963%
1242.1 49.684%
1257.11 50.284% 49.52% 0.86% 1.74%
1214.55 48.582%
Figure 1. Activity Units Trials

60.00%
50.00%
Percent Recovery
40.00%
30.00%
20.00%
10.00%
0.00%
125U 250U 500U 750U 1000U
Activity Units

Kura Activity (Volume)
50% 0U 25
CO 0U 50
3x 0U 150
10x 0U 500
15x 0U 750
25x 0U 1250
50x 0U 2500
LCU 0U 30
MCU 0U 50
HCU 0U 150
24.42 97.7
48.2 96.4
160.93 107.29
494.18 98.84
1079.39 143.92
1247.26 99.78
1872.01 74.88
34.68 115.62
57.95 115.89
64.89 2.596%
71.25 2.850% 2.525% 0.366% 14.497%
53.21 2.128%
157.02 6.281%
145.32 5.813% 6.320% 0.528% 8.356%
171.67 6.867%
404.81 16.192%
416.15 16.646% 16.034% 0.704% 4.394%
381.6 15.264%
599.97 23.999%
568.16 22.726% 23.582% 0.741% 3.143%
600.54 24.022%
594.69 23.788%
426.14 17.046% 20.417% 4.767% 23.350%
N/A N/A Inject did not work
1472.92 58.917%
1252.3 50.092% 55.089% 4.527% 8.218%
1406.49 56.260%
2010.08 80.403%
1751.22 70.049% 75.940% 5.323% 7.009%
1934.18 77.367%
2278.2 91.128%
2076.21 83.048% 86.404% 4.210% 4.873%
2125.86 85.034%
2594.12 103.765%
2399.07 95.963% 96.829% 6.546% 6.761%
2268.95 90.758%
2382.75 95.310%
2426.28 97.051% 95.075% 2.104% 2.213%
2321.57 92.863%
2365.93 94.637%
2243.28 89.731% 95.843% 6.796% 7.091%
2579.05 103.162%
IMCS Temperature
50% N/A 25 27.06
CO N/A 50 45.26
3x N/A 150 153.54
10x N/A 500 473.93
15x N/A 750 736.03
25x N/A 1250 1359.56
50x N/A 2500 2429.62
LCU N/A 30 35.05
MCU N/A 50 46.41
HCU N/A 150 165.57
2500ng/mL Codeine Control 2500 22.27
Accuracy Sample Name
108.25 50%
90.52 CO
102.36 3x
94.79 10x
98.14 15x
108.76 25x
97.18 50x
116.83 LCU
92.82 MCU
110.38 Average % Recovery Standard Deviation %CV HCU
0.891% 2500ng/mL Codeine 35C B-Gluc
0.796% 0.86% 0.06% 6.47% 2500ng/mL Codeine 35C B-Gluc
8.092%
7.231% 7.70% 0.44% 5.67%
7.789%
35°C
40°C
45°C
50°C
55°C
60°C
Kura Temperature
Temperature Actual Codeine Concentration Calculated Concentration Accuracy
N/A 25 24.36 97.42
N/A 50 53.55 107.1
N/A 150 163.03 108.68
N/A 500 435.11 87.02
N/A 750 1255.61 167.41
N/A 1250 1195.43 95.63
N/A 2500 2603.53 104.14
N/A 30 126.1 420.33
N/A 50 55.72 111.44
N/A 150 171.51 114.34 Average % Recovery
35°C 2500 213.92 8.557%
35°C 2500 225.81 9.032% 8.773%
35°C 2500 218.28 8.731%
40°C 2500 366 14.640%
40°C 2500 354.61 14.184% 13.998%
40°C 2500 329.21 13.168%
45°C 2500 423.64 16.946%
45°C 2500 431.35 17.254% 17.310%
45°C 2500 443.29 17.732%
50°C 2500 591.52 23.661%
50°C 2500 655.74 26.230% 25.014%
50°C 2500 628.8 25.152%
55°C 2500 749.28 29.971%
55°C 2500 689.49 27.580% 28.821%
55°C 2500 722.77 28.911%
60°C 2500 9.4 0.376%
60°C 2500 7.44 0.298% 0.359%
60°C 2500 10.12 0.405%
Figure 2. Temperature Trials

35.00%
10.53% 8.77% 30.00%
Percent Recovery
17.97% 14.00% 25.00%

20.00%
23.35% 17.31% 15.00%
25.45% 25.01% 10.00%
5.00%
17.59% 28.82%
0.00%
7.70% 0.36% 35°C 40°C 45°C 50°C 55°C
Temperature

Standard Deviation %CV
0.241% 2.742%
0.753% 5.382%
0.396% 2.288%
1.290% 5.157%
1.198% 4.158%
0.055% 15.434%
perature Trials
°C 50°C 55°C 60°C

mperature
Kura Biotec
IMCS Time
50% N/A 25 20.28
CO N/A 50 48.66
3x N/A 150 159.21
10x N/A 500 541.64
15x N/A 750 796.39
25x N/A 1250 1363.26
50x N/A 2500 2295.54
LCU N/A 30 27.11
MCU N/A 50 47.61
HCU N/A 150 172.04
Accuracy Sample Name
81.14 50%
97.32 CO
106.14 3x
108.33 10x
106.19 15x
109.06 25x
91.82 50x
90.36 LCU
95.22 MCU
114.69 Average % Recovery Standard Deviation %CV HCU
8.021% 2500ng/mL Codeine 15 min
8.901% 8.45% 0.44% 5.22% 2500ng/mL Codeine 15 min
16.849% 16.80% 0.69% 4.13% 2500ng/mL Codeine 30 min
24.027% 23.60% 0.68% 2.88% 2500ng/mL Codeine 45 min
24.534% 24.71% 0.37% 1.51% 2500ng/mL Codeine 60 min
29.693% 29.53% 1.79% 6.07% 2500ng/mL Codeine 90 min
38.118% 37.41% 0.90% 2.41% 2500ng/mL Codeine 120 min

15 min 8.45% 6.82%
30 min 16.80% 16.32%
45 min 23.60% 26.62%
60 min 24.71% 29.50%
90 min 29.53% 41.10%
120 min 37.41% 47.74%
Kura Time
Time Actual Codeine Concentration Calculated Concentration Accuracy
N/A 25 24.39 97.54
N/A 50 53.41 106.82
N/A 150 146.3 97.53
N/A 500 469.76 93.95
N/A 750 781.15 104.15
N/A 1250 1181.23 94.5
N/A 2500 2496.27 99.85
N/A 30 168.77 562.57
N/A 150 169.87 113.25
N/A 2500 170.97 6.84 Average % Recovery
15 min 2500 170.67 6.827%
15 min 2500 170.88 6.835% 6.823%
15 min 2500 170.19 6.808%
30 min 2500 420.95 16.838%
30 min 2500 407.83 16.313% 16.315%
30 min 2500 394.87 15.795%
45 min 2500 675.08 27.003%
45 min 2500 667.51 26.700% 26.622%
45 min 2500 654.05 26.162%
60 min 2500 741.59 29.664%
60 min 2500 767.09 30.684% 29.500%
60 min 2500 703.84 28.154%
90 min 2500 1021.51 40.860%
90 min 2500 1031.33 41.253% 41.104%
90 min 2500 1029.95 41.198%
120 min 2500 1175.17 47.007%
120 min 2500 1179.19 47.168% 47.741%
120 min 2500 1226.18 49.047%
Figure 3. Incubation Time Trials

60.00%
50.00%
Percent Recovery
40.00%
30.00%
20.00%
10.00%
0.00%
Incubation Time

0.00%
Incubation Time

Standard Deviation %CV
0.014% 0.207%
0.522% 3.197%
0.426% 1.600%
1.273% 4.315%
0.213% 0.517%
1.134% 2.376%

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IMCS Time

Sample Name Time Actual Codeine Concentration Calculated Concentration

IMCSzyme™ Incubation Time Trials

IMCSzyme™ Activity Units Trials

Figure 3. Kura Biotec Incubation Time Trials

Figure 1. Kura Biotec Activity Trials

Figure 2. Kura Biotec Temperature Trials

IMCSzyme™ Kura Biotec

Figure 1. Activity Units Trials

IMCSzyme™ Kura Biotec

Figure 2. Temperature Trials

17.97% 14.00% 25.00%

IMCSzyme™ Kura Biotec

°C 50°C 55°C 60°C

IMCSzyme™ Kura Biotec

Figure 3. Incubation Time Trials

IMCSzyme™ Kura Biotec

IMCSzyme™ Kura Biotec

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