Table 3 Matrix of the chromatographic and extraction conditions suggested by software and responses obtained experimentally for each run (mean ± SEM, n = 3)
Experiment number | Chromatographic conditions | |||||||
|---|---|---|---|---|---|---|---|---|
Factors | Responses | |||||||
A (%v/v) | B (%v/v) | C | D (℃) | E (mL/min) | PCZ RT (min) | IS RT (min) | Resolution | |
1 | 52 | 0 | 3 | 25 | 1.00 | 11.3 ± 0.1 | 11.3 ± 0.1 | 0.0 ± 0.0 |
2 | 60 | 0 | 3 | 25 | 0.70 | 13.3 ± 0.0 | 14.5 ± 0.0 | 1.3 ± 0.0 |
3 | 52 | 6 | 3 | 25 | 0.70 | 24.8 ± 0.1 | 20.8 ± 0.0 | 4.6 ± 0.1 |
4 | 60 | 6 | 3 | 25 | 1.00 | 8.6 ± 0.0 | 9.0 ± 0.0 | 0.9 ± 0.0 |
5 | 52 | 0 | 7 | 25 | 0.70 | 23.3 ± 0.7 | 18.3 ± 0.2 | 4.7 ± 0.4 |
6 | 60 | 0 | 7 | 25 | 1.00 | 8.3 ± 0.0 | 8.3 ± 0.0 | 0.0 ± 0.0 |
7 | 52 | 6 | 7 | 25 | 1.00 | 20.7 ± 0.0 | 13.4 ± 0.0 | 10.1 ± 0.1 |
8 | 60 | 6 | 7 | 25 | 0.70 | 16.3 ± 0.3 | 14.5 ± 0.3 | 2.3 ± 0.3 |
9 | 52 | 0 | 3 | 40 | 0.70 | 20.6 ± 0.0 | 16.9 ± 0.9 | 1.9 ± 0.1 |
10 | 60 | 0 | 3 | 40 | 1.00 | 7.9 ± 0.1 | 8.3 ± 0.0 | 1.0 ± 0.1 |
11 | 52 | 6 | 3 | 40 | 1.00 | 15.6 ± 0.0 | 11.9 ± 0.0 | 5.8 ± 0.1 |
12 | 60 | 6 | 3 | 40 | 0.70 | 15.0 ± 0.0 | 15.0 ± 0.0 | 0.0 ± 0.0 |
13 | 52 | 0 | 7 | 40 | 1.00 | 14.3 ± 0.3 | 10.7 ± 0.0 | 5.4 ± 0.1 |
14 | 60 | 0 | 7 | 40 | 0.70 | 13.5 ± 0.1 | 12.8 ± 0.1 | 0.7 ± 0.1 |
15 | 52 | 6 | 7 | 40 | 0.70 | 31.8 ± 0.0 | 20.2 ± 0.1 | 9.9 ± 0.0 |
16 | 60 | 6 | 7 | 40 | 1.00 | 9.7 ± 0.2 | 8.4 ± 0.2 | 2.2 ± 0.4 |
17 | 56 | 3 | 5 | 33 | 0.85 | 14.5 ± 0.0 | 12.3 ± 0.0 | 3.5 ± 0.2 |
18 | 56 | 3 | 5 | 33 | 0.85 | 14.6 ± 0.1 | 12.3 ± 0.1 | 3.5 ± 0.4 |
19 | 56 | 3 | 5 | 33 | 0.85 | 15.4 ± 0.1 | 12.8 ± 0.0 | 3.8 ± 0.2 |
20 | 56 | 3 | 5 | 33 | 0.85 | 14.2 ± 0.2 | 11.8 ± 0.3 | 3.8 ± 0.2 |
Experiment number | Extraction conditions | ||||||
|---|---|---|---|---|---|---|---|
Factors | Responses | ||||||
A´ (μL) | B´ | C´ (%w/v) | D´ (min) | E´ (min) | PCZ recovery (%) | IS recovery (%) | |
1´ | 500 | 3 | 0 | 1.0 | 10.0 | 55.7 ± 3.6 | 53.7 ± 1.2 |
2´ | 1500 | 3 | 0 | 1.0 | 1.0 | 67.7 ± 1.5 | 60.3 ± 6.6 |
3´ | 500 | 11 | 0 | 1.0 | 1.0 | 93.8 ± 3.0 | 58.2 ± 0.0 |
4´ | 1500 | 11 | 0 | 1.0 | 10.0 | 98.1 ± 2.6 | 72.7 ± 4.2 |
5´ | 500 | 3 | 10 | 1.0 | 1.0 | 36.4 ± 0.3 | 56.7 ± 1.6 |
6´ | 1500 | 3 | 10 | 1.0 | 10.0 | 46.6 ± 0.6 | 56.1 ± 1.3 |
7´ | 500 | 11 | 10 | 1.0 | 10.0 | 101.1 ± 2.1 | 57.0 ± 3.3 |
8´ | 1500 | 11 | 10 | 1.0 | 1.0 | 62.0 ± 0.1 | 57.3 ± 0.2 |
9´ | 500 | 3 | 0 | 10.0 | 1.0 | 56.6 ± 2.7 | 51.0 ± 2.0 |
10´ | 1500 | 3 | 0 | 10.0 | 10.0 | 81.7 ± 0.7 | 63.0 ± 6.9 |
11´ | 500 | 11 | 0 | 10.0 | 10.0 | 73.9 ± 0.8 | 44.9 ± 0.9 |
12´ | 1500 | 11 | 0 | 10.0 | 1.0 | 85.5 ± 4.6 | 63.6 ± 0.7 |
13´ | 500 | 3 | 10 | 10.0 | 10.0 | 30.3 ± 3.2 | 64.1 ± 4.0 |
14´ | 1500 | 3 | 10 | 10.0 | 1.0 | 51.4 ± 0.9 | 65.0 ± 1.4 |
15´ | 500 | 11 | 10 | 10.0 | 1.0 | 97.7 ± 1.5 | 80.9 ± 1.0 |
16´ | 1500 | 11 | 10 | 10.0 | 10.0 | 87.8 ± 0.1 | 69.8 ± 2.1 |
17´ | 1000 | 7 | 5 | 5.5 | 5.5 | 96.1 ± 0.4 | 77.6 ± 8.2 |
18´ | 1000 | 7 | 5 | 5.5 | 5.5 | 99.1 ± 0.9 | 70.8 ± 6.9 |
19´ | 1000 | 7 | 5 | 5.5 | 5.5 | 95.8 ± 1.9 | 76.8 ± 2.1 |
20´ | 1000 | 7 | 5 | 5.5 | 5.5 | 97.0 ± 0.8 | 75.0 ± 3.4 |