Table 5.
The intra-day precision for BoNTs C/D, D, A, B, E, and F after 4 and 24 h of incubation. BoNT C/D and D was spiked in chicken serum and the others in human serum.
| NTP/IS | CTP/IS | |||||
|---|---|---|---|---|---|---|
| Mean | RSDr (%) | x blank | Mean | RSDr (%) | x blank | |
| Blank 4h (24h) | 0.22 (0.24) | 9 (21) | - | 0.20 (0.32) | 17 (23) | - |
| 23ng BoNT-C/D 4h (24h) | 0.59 (1.02) | 18 (18) | 2.6 (4.2) | 1.08 (2.13) | 9 (12) | 5.5 (6.7) |
| 33ng BoNT-C/D 4h (24h) | 0.77 (1.37) | 14 (14) | 3.4 (5.6) | 1.41 (2.97) | 8 (17) | 7.2 (9.4) |
| 49ng BoNT-C/D 4h (24h) | 0.96 (1.91) | 15 (7) | 4.3 (7.8) | 2.16 (4.30) | 13 (11) | 11 (14) |
|
| ||||||
| Blank 4h (24h) | 0.02 (0.01) | 37 (23) | - | 0.03 (0.03) | 10 (17) | - |
| 0.1ng BoNT-D 4h (24h) | 0.12 (0.67) | 26 (47) | 7.4 (50) | 0.24 (0.90) | 36 (50) | 8.8 (35) |
| 0.9ng BoNT-D 4h (24h) | 1.04 (5.79) | 25 (48) | 66 (434) | 2.35 (4.77) | 18 (40) | 87 (185) |
| 1.8ng BoNT-D 4h (24h) | 2.34 (7.35) | 35 (55) | 149 (551) | 3.60 (6.03) | 18 (36) | 133 (234) |
|
| ||||||
| Blank 4h (24h) | 0.17 (0.17) | 12 (18) | - | 0.35 (0.31) | 15 (19) | - |
| BoNT-A 0.5MLD50 4h (24h) | 1.19 (1.96) | 5 (5) | 6.8 (11.2) | 2.07 (2.55) | 6 (6) | 6.0 (8.3) |
| BoNT-A 1MLD50 4h (24h) | 2.16 (3.28) | 15 (15) | 16 (24) | 3.40 (3.83) | 16 (20) | 14 (17) |
| BoNT-A 5MLD50 4h (24h) | 9.71 (15.34) | 13 (10) | 73 (114) | 15.36 (19.88) | 14 (12) | 65 (86) |
|
| ||||||
| Blank 4h (24h) | 0.01 (0.01) | 12 (14) | - | 0.11 (0.07) | 21 (15) | - |
| BoNT-B 0.5MLD50 4h (24h) | 0.05 (0.22) | 6 (7) | 5.0 (21) | 0.10 (0.10) | 9 (23) | 1.0 (1.4) |
| BoNT-B 1MLD50 4h (24h) | 0.07 (0.33) | 6 (5) | 7.1 (32) | 0.11 (0.10) | 19 (19) | 1.1 (1.4) |
| BoNT-B 5MLD50 4h (24h) | 0.24 (1.08) | 12 (8) | 25 (103) | 0.13 (0.17) | 21 (24) | 1.3 (2.3) |
|
| ||||||
| Blank 4h (24h) | 0.013 (0.011) | 28 (20) | - | 0.10 (0.13) | 24 (12) | - |
| BoNT-E 5MLD50 4h (24h) | 0.018 (0.026) | 15 (34) | 1.4 (2.3) | 0.16 (0.36) | 17 (16) | 1.7 (2.7) |
| BoNT-E 10MLD50 4h (24h) | 0.024 (0.034) | 12 (42) | 1.8 (3.1) | 0.23 (0.55) | 15 (39) | 2.4 (4.1) |
| BoNT-E 20MLD50 4h (24h) | 0.031 (0.068) | 20 (27) | 2.3 (6.1) | 0.34 (1.04) | 8 (13) | 3.4 (7.8) |
|
| ||||||
| Blank 4h (24h) | 0.002 (0.003) | 29 (124) | -- | 0.04 (0.04) | 11 (11) | - |
| BoNT-F 0.5MLD50 4h (24h) | 0.008 (0.006) | 24 (25) | 5.1 (2.2) | 0.24 (0.26) | 10 (12) | 6.2 (6.9) |
| BoNT-F 1MLD50 4h (24h) | 0.017 (0.012) | 45 (25) | 11 (4.4) | 0.37 (0.46) | 15 (15) | 9.8 (12) |
| BoNT-F 5MLD50 4h (24h) | 0.055 (0.048) | 39 (87) | 35 (17) | 1.33 (1.55) | 12 (10) | 35 (41) |
NTP/IS = The ratio between the intensities of the N terminal cleavage product and the internal standard peptide in the same spectrum.
CTP/IS = The ratio between the intensities of the C terminal cleavage product and the internal standard peptide in the same spectrum.
RSDr= the relative standard deviation in percent calculated from 6 replicates, each spotted and analyzed 3 times.
x blank = the mean NTP/IS (or CTP/IS) in a sample (6 replicates, 3 spots each) divided by the mean of the same ratio of the blank samples (6 replicates, 3 spots each).