Table 3.
Method validation
|
CKs |
Recovery (%)a |
Determined spiked CK content (pmol)b |
Method precision (% RSD)b |
Method accuracy (% bias)b |
||
|---|---|---|---|---|---|---|
| 1 mg FW | 2 mg FW | 5 mg FW | ||||
|
tZ |
80 ± 10 |
63 ± 8 |
21 ± 4 |
0.99 ± 0.17 |
17.2 |
0.1 |
|
tZR |
72 ± 12 |
46 ± 7 |
8 ± 1 |
0.84 ± 0.09 |
10.4 |
16.1 |
|
tZ7G |
88 ± 6 |
57 ± 4 |
6 ± 1 |
0.91 ± 0.13 |
14.6 |
8.6 |
|
tZ9G |
59 ± 7 |
31 ± 3 |
8 ± 1 |
0.97 ± 0.09 |
9.6 |
2.5 |
|
tZOG |
85 ± 5 |
68 ± 6 |
11 ± 2 |
1.07 ± 0.19 |
18.1 |
−7.1 |
|
tZROG |
55 ± 4 |
30 ± 3 |
4 ± 1 |
0.82 ± 0.09 |
11.1 |
17.6 |
|
tZMP |
35 ± 6 |
11 ± 1 |
5 ± 1 |
0.85 ± 0.11 |
13.3 |
14.7 |
|
cZ |
75 ± 9 |
65 ± 5 |
24 ± 4 |
0.83 ± 0.02 |
2.5 |
17.0 |
|
cZR |
81 ± 13 |
44 ± 9 |
8 ± 1 |
0.96 ± 0.12 |
12.8 |
4.1 |
|
cZ9G |
74 ± 12 |
37 ± 5 |
5 ± 1 |
1.18 ± 0.13 |
11.3 |
−17.6 |
|
cZOG |
89 ± 6 |
66 ± 7 |
9 ± 2 |
1.09 ± 0.14 |
13.2 |
−9.1 |
|
cZROG |
52 ± 6 |
24 ± 2 |
3 ± 1 |
0.89 ± 0.11 |
12.9 |
11.2 |
|
cZMP |
32 ± 3 |
17 ± 1 |
2 ± 1 |
0.86 ± 0.15 |
17.9 |
13.9 |
| DHZ |
77 ± 13 |
61 ± 7 |
20 ± 3 |
0.90 ± 0.10 |
10.8 |
9.7 |
| DHZR |
88 ± 13 |
48 ± 8 |
12 ± 1 |
1.03 ± 0.06 |
5.6 |
−2.9 |
| DHZ7G |
89 ± 3 |
65 ± 3 |
8 ± 2 |
1.18 ± 0.05 |
4.3 |
−18.2 |
| DHZ9G |
78 ± 10 |
35 ± 6 |
6 ± 1 |
0.96 ± 0.07 |
7.2 |
3.7 |
| DHZOG |
77 ± 5 |
50 ± 7 |
9 ± 3 |
1.16 ± 0.06 |
4.9 |
−15.8 |
| DHZROG |
87 ± 8 |
42 ± 5 |
5 ± 1 |
0.90 ± 0.12 |
13.3 |
9.8 |
| DHZMP |
37 ± 1 |
12 ± 1 |
3 ± 1 |
0.96 ± 0.15 |
15.8 |
3.6 |
| iP |
76 ± 9 |
68 ± 3 |
26 ± 5 |
0.97 ± 0.06 |
5.9 |
3.3 |
| iPR |
84 ± 8 |
53 ± 4 |
17 ± 1 |
1.13 ± 0.06 |
5.6 |
−12.8 |
| iP7G |
83 ± 10 |
60 ± 5 |
7 ± 1 |
0.91 ± 0.15 |
16.5 |
9.4 |
| iP9G |
74 ± 8 |
49 ± 8 |
8 ± 2 |
0.97 ± 0.08 |
8.0 |
3.1 |
| iPMP | 78 ± 9 | 39 ± 9 | 9 ± 2 | 0.92 ± 0.16 | 17.0 | 8.4 |
a Values are means ± SD (n = 4); b Values are means ± SD (n = 12).
Recovery (%) is shown for different amounts of plant matrix together with determinations of the analytical precision and accuracy of whole procedure. Plant tissues (1–5 mg FW ofA. thalianaseedlings spiked with 1 pmol of authentic CK standards) were extracted in Bieleski buffer, purified by multi-StageTip microcolumn chromatography and directly analyzed by UHPLC-ESI(+)-MS/MS.