Mouse serum IL-6 after acute sleep fragmentation (real data)
Nguyen, Fields & Ashley, Dryad doi:10.5061/dryad.tdz08kq3p (CC0). Groups = treatment × time; see notebooks/01 for the time-matched analysis. · generated 2026-09-29 13:14 · immunoassay v0.1.0
3/4
plates passed QC
IL-6
analytes
110
samples
103
flagged replicate groups
0
values measured in range
1 · Plate acceptance
A plate passes when R² ≥ 0.98, ≥ 75% of standard levels recover within 80–120% with CV ≤ 20%, a quantifiable range exists, and ≥ 67% of controls with expected ranges fall inside them. Limits are in-well concentrations.
| plate_id | analyte | verdict | model | R² | standards OK | lod | lloq | uloq | controls OK | flagged groups | reasons |
|---|---|---|---|---|---|---|---|---|---|---|---|
| plate1 | IL-6 | ✓ pass | 4PL | 0.9994 | 7/7 | 5.552 | 7.8 | 500.0 | n/a | 29 | |
| plate2 | IL-6 | ✗ fail | 4PL | 0.9934 | 4/7 | 3.019 | 62.5 | 500.0 | n/a | 30 | only 4/7 standard levels pass |
| plate3 | IL-6 | ✓ pass | 4PL | 0.9958 | 6/7 | 4.801 | 15.6 | 500.0 | n/a | 24 | |
| plate4 | IL-6 | ✓ pass | 4PL | 0.9957 | 6/7 | 4.071 | 15.6 | 500.0 | n/a | 20 |
2 · Standard curves
Weighted (1/y^2) least-squares fits. 5PL is chosen over 4PL only when its AICc is lower by more than 2. Weighted residuals should scatter evenly around zero; a curved pattern means the model is wrong for the data. The plate map shows raw OD. Excluded wells are marked ✗.
plate1 · IL-6
| model | a | a_se | b | b_se | c (EC50) | c_se | d | d_se | R² | aicc_4pl | aicc_5pl |
|---|---|---|---|---|---|---|---|---|---|---|---|
| 4pl | -0.00109 | 0.00162 | 1.1 | 0.03131 | 862.6 | 197.4 | 4.113 | 0.6841 | 0.9994 | -165.8 | -161.8 |
Standard recovery
| concentration | n_wells | n_quantified | mean_back_calc | cv_pct | recovery_pct |
|---|---|---|---|---|---|
| 7.8 | 2 | 2 | 7.32 | 18.0 | 93.8 |
| 15.6 | 2 | 2 | 15.6 | 9.281 | 99.7 |
| 31.3 | 2 | 2 | 31.9 | 2.477 | 101.9 |
| 62.5 | 2 | 2 | 62.5 | 0.3126 | 99.9 |
| 125.0 | 2 | 2 | 127.2 | 1.938 | 101.7 |
| 250.0 | 2 | 2 | 243.5 | 3.33 | 97.4 |
| 500.0 | 2 | 2 | 506.6 | 1.088 | 101.3 |
plate2 · IL-6
| model | a | a_se | b | b_se | c (EC50) | c_se | d | d_se | R² | aicc_4pl | aicc_5pl |
|---|---|---|---|---|---|---|---|---|---|---|---|
| 4pl | -0.00602 | 0.0135 | 0.8642 | 0.1063 | 453.5 | 357.6 | 3.726 | 1.442 | 0.9934 | -96.3 | -92.2 |
Standard recovery
| concentration | n_wells | n_quantified | mean_back_calc | cv_pct | recovery_pct |
|---|---|---|---|---|---|
| 7.8 | 2 | 2 | 5.23 | 30.2 | 67.1 |
| 15.6 | 2 | 2 | 20.5 | 1.818 | 131.5 |
| 31.3 | 2 | 2 | 40.5 | 10.5 | 129.4 |
| 62.5 | 2 | 2 | 57.5 | 19.6 | 92.0 |
| 125.0 | 2 | 2 | 115.5 | 12.0 | 92.4 |
| 250.0 | 2 | 2 | 261.1 | 4.823 | 104.5 |
| 500.0 | 2 | 2 | 514.2 | 6.241 | 102.8 |
plate3 · IL-6
| model | a | a_se | b | b_se | c (EC50) | c_se | d | d_se | R² | aicc_4pl | aicc_5pl |
|---|---|---|---|---|---|---|---|---|---|---|---|
| 4pl | 0.000273 | 0.00812 | 0.7803 | 0.08461 | 2,145.5 | 3,470.3 | 7.724 | 6.997 | 0.9958 | -102.8 | -99.4 |
Standard recovery
| concentration | n_wells | n_quantified | mean_back_calc | cv_pct | recovery_pct |
|---|---|---|---|---|---|
| 7.8 | 1 | 1 | 10.9 | – | 139.7 |
| 15.6 | 2 | 2 | 18.0 | 15.9 | 115.4 |
| 31.3 | 2 | 2 | 30.4 | 8.729 | 97.0 |
| 62.5 | 2 | 2 | 53.2 | 4.447 | 85.2 |
| 125.0 | 2 | 2 | 122.6 | 0.4025 | 98.1 |
| 250.0 | 2 | 2 | 277.3 | 2.512 | 110.9 |
| 500.0 | 2 | 2 | 509.9 | 2.658 | 102.0 |
plate4 · IL-6
| model | a | a_se | b | b_se | c (EC50) | c_se | d | d_se | R² | aicc_4pl | aicc_5pl |
|---|---|---|---|---|---|---|---|---|---|---|---|
| 4pl | 0.000484 | 0.0058 | 1.025 | 0.05893 | 436.2 | 143.2 | 4.319 | 0.8221 | 0.9957 | -122.3 | -118.4 |
Standard recovery
| concentration | n_wells | n_quantified | mean_back_calc | cv_pct | recovery_pct |
|---|---|---|---|---|---|
| 7.8 | 2 | 2 | 9.241 | 26.7 | 118.5 |
| 15.6 | 2 | 2 | 14.4 | 10.1 | 92.3 |
| 31.3 | 2 | 2 | 32.4 | 5.252 | 103.5 |
| 62.5 | 2 | 2 | 59.8 | 2.505 | 95.6 |
| 125.0 | 2 | 2 | 130.3 | 11.3 | 104.3 |
| 250.0 | 2 | 2 | 256.4 | 6.806 | 102.6 |
| 500.0 | 2 | 2 | 495.1 | 13.8 | 99.0 |
Excluded wells
| plate_id | well | type | sample_id | od | outlier_reason |
|---|---|---|---|---|---|
| plate3 | G2 | standard | STD7 | 0.185 | standard pair CV > 20% and deletion residual 12.2 SD > 3.5 |
3 · Controls and flagged samples
None.
Flagged replicate groups
below_lloq / above_uloq: outside the validated range (not reported as numbers). below_lod: not distinguishable from blank. cv_high: replicate CV above 20%.
| plate_id | analyte | sample_id | group | dilution | wells | od_mean | conc_well | cv_pct | flags |
|---|---|---|---|---|---|---|---|---|---|
| plate1 | IL-6 | M_NSF0h-1 | NSF 0h | 1 | A3,A4 | 0.0275 | 9.444 | 36.4 | cv_high |
| plate1 | IL-6 | M_NSF1h-7 | NSF 1h | 1 | A7,A8 | -0.003 | – | 70.7 | below_lloq;below_lod;cv_high |
| plate1 | IL-6 | M_ASF1h-5 | ASF 1h | 1 | A9,A10 | -0.0055 | – | 77.1 | below_lloq;below_lod;cv_high |
| plate1 | IL-6 | M_NSF0h-2 | NSF 0h | 3 | B3,B4 | 0.0025 | – | 396.0 | below_lloq;below_lod;cv_high |
| plate1 | IL-6 | M_NSF0h-10 | NSF 0h | 3 | B5,B6 | 0.002 | 1.206 | 109.1 | below_lloq;below_lod;cv_high |
| plate1 | IL-6 | M_NSF1h-8 | NSF 1h | 1 | B7,B8 | -0.0005 | 0.2752 | 0 | below_lloq;below_lod |
| plate1 | IL-6 | M_ASF1h-6 | ASF 1h | 1 | B9,B10 | -0.0045 | – | 0 | below_lloq;below_lod |
| plate1 | IL-6 | M_NSF0h-3 | NSF 0h | 3 | C3,C4 | -0.0105 | – | 107.7 | below_lloq;below_lod;cv_high |
| plate1 | IL-6 | M_NSF1h-1 | NSF 1h | 1 | C5,C6 | 0.0215 | 7.615 | 34.5 | below_lloq;cv_high |
| plate1 | IL-6 | M_NSF1h-9 | NSF 1h | 1 | C7,C8 | -0.0065 | – | 0 | below_lloq;below_lod |
| plate1 | IL-6 | M_ASF1h-7 | ASF 1h | 1 | C9,C10 | -0.0055 | – | 77.1 | below_lloq;below_lod;cv_high |
| plate1 | IL-6 | M_NSF0h-4 | NSF 0h | 3 | D3,D4 | 0.0055 | 2.481 | 0 | below_lloq;below_lod |
| plate1 | IL-6 | M_NSF1h-2 | NSF 1h | 1 | D5,D6 | -0.006 | – | 11.8 | below_lloq;below_lod |
| plate1 | IL-6 | M_NSF1h-10 | NSF 1h | 1 | D7,D8 | -0.004 | – | 265.2 | below_lloq;below_lod;cv_high |
| plate1 | IL-6 | M_ASF1h-8 | ASF 1h | 1 | D9,D10 | 0.03 | 10.2 | 23.0 | cv_high |
| plate1 | IL-6 | M_NSF0h-5 | NSF 0h | 3 | E3,E4 | 0.0055 | – | 231.4 | below_lloq;below_lod;cv_high |
| plate1 | IL-6 | M_NSF1h-3 | NSF 1h | 1 | E5,E6 | -0.0075 | – | 0 | below_lloq;below_lod |
| plate1 | IL-6 | M_ASF1h-1 | ASF 1h | 1 | E7,E8 | -0.01 | – | 77.8 | below_lloq;below_lod;cv_high |
| plate1 | IL-6 | M_ASF1h-9 | ASF 1h | 1 | E9,E10 | 0.015 | 5.595 | 20.1 | below_lloq;cv_high |
| plate1 | IL-6 | M_NSF0h-6 | NSF 0h | 3 | F3,F4 | -0.0055 | – | 25.7 | below_lloq;below_lod;cv_high |
| plate1 | IL-6 | M_NSF1h-4 | NSF 1h | 1 | F5,F6 | -0.007 | – | 90.9 | below_lloq;below_lod;cv_high |
| plate1 | IL-6 | M_ASF1h-2 | ASF 1h | 1 | F7,F8 | -0.0025 | – | 113.1 | below_lloq;below_lod;cv_high |
| plate1 | IL-6 | M_ASF1h-10 | ASF 1h | 1 | F9,F10 | -0.009 | – | 7.857 | below_lloq;below_lod |
| plate1 | IL-6 | M_NSF0h-7 | NSF 0h | 3 | G3,G4 | -0.007 | – | 30.3 | below_lloq;below_lod;cv_high |
| plate1 | IL-6 | M_NSF1h-5 | NSF 1h | 1 | G5,G6 | 0.0045 | 2.11 | 70.4 | below_lloq;below_lod;cv_high |
| plate1 | IL-6 | M_ASF1h-3 | ASF 1h | 1 | G7,G8 | -0.0065 | – | 108.8 | below_lloq;below_lod;cv_high |
| plate1 | IL-6 | M_NSF0h-8 | NSF 0h | 3 | H3,H4 | -0.007 | – | 10.1 | below_lloq;below_lod |
| plate1 | IL-6 | M_NSF1h-6 | NSF 1h | 1 | H5,H6 | 0.003 | – | 212.1 | below_lloq;below_lod;cv_high |
| plate1 | IL-6 | M_ASF1h-4 | ASF 1h | 1 | H7,H8 | -0.0045 | – | 31.4 | below_lloq;below_lod;cv_high |
| plate2 | IL-6 | M_NSF2h-1 | NSF 2h | 1 | A3,A4 | 0.029 | 2.131 | 86.3 | below_lloq;below_lod;cv_high |
| plate2 | IL-6 | M_NSF2h-9 | NSF 2h | 2 | A5,A6 | -0.014 | – | 242.4 | below_lloq;below_lod;cv_high |
| plate2 | IL-6 | M_ASF2h-7 | ASF 2h | 2 | A7,A8 | -0.0115 | – | 153.7 | below_lloq;below_lod;cv_high |
| plate2 | IL-6 | M_NSF6h-5 | NSF 6h | 1 | A9,A10 | -0.015 | – | 84.9 | below_lloq;below_lod;cv_high |
| plate2 | IL-6 | M_NSF2h-2 | NSF 2h | 1 | B3,B4 | 0.048 | 3.435 | 18.4 | below_lloq |
| plate2 | IL-6 | M_NSF2h-10 | NSF 2h | 1 | B5,B6 | -0.019 | – | 0 | below_lloq;below_lod |
| plate2 | IL-6 | M_ASF2h-8 | ASF 2h | 1 | B7,B8 | 0.3235 | 30.4 | 5.174 | below_lloq |
| plate2 | IL-6 | M_NSF6h-6 | NSF 6h | 1 | B9,B10 | -0.004 | – | 1,308.1 | below_lloq;below_lod;cv_high |
| plate2 | IL-6 | M_NSF2h-3 | NSF 2h | 1 | C3,C4 | 0.0085 | – | 357.7 | below_lloq;below_lod;cv_high |
| plate2 | IL-6 | M_ASF2h-1 | ASF 2h | 2 | C5,C6 | -0.013 | – | 424.3 | below_lloq;below_lod;cv_high |
| plate2 | IL-6 | M_ASF2h-9 | ASF 2h | 1 | C7,C8 | 0.3225 | 30.5 | 27.4 | below_lloq;cv_high |
| plate2 | IL-6 | M_NSF6h-7 | NSF 6h | 2 | C9,C10 | 0.011 | 0.9984 | 141.4 | below_lloq;below_lod;cv_high |
| plate2 | IL-6 | M_NSF2h-4 | NSF 2h | 2 | D3,D4 | 0.0055 | 0.5904 | 101.4 | below_lloq;below_lod;cv_high |
| plate2 | IL-6 | M_ASF2h-2 | ASF 2h | 2 | D5,D6 | -0.0155 | – | 205.3 | below_lloq;below_lod;cv_high |
| plate2 | IL-6 | M_ASF2h-10 | ASF 2h | 1 | D7,D8 | 0.129 | 10.2 | 35.0 | below_lloq;cv_high |
| plate2 | IL-6 | M_NSF6h-8 | NSF 6h | 1 | D9,D10 | 0.027 | 2.075 | 123.2 | below_lloq;below_lod;cv_high |
| plate2 | IL-6 | M_NSF2h-5 | NSF 2h | 2 | E3,E4 | 0.003 | – | 565.7 | below_lloq;below_lod;cv_high |
| plate2 | IL-6 | M_ASF2h-3 | ASF 2h | 1 | E5,E6 | -0.001 | – | 848.5 | below_lloq;below_lod;cv_high |
| plate2 | IL-6 | M_NSF6h-1 | NSF 6h | 2 | E7,E8 | -0.024 | – | 29.5 | below_lloq;below_lod;cv_high |
| plate2 | IL-6 | M_NSF6h-9 | NSF 6h | 1 | E9,E10 | -0.0335 | – | 6.332 | below_lloq;below_lod |
| plate2 | IL-6 | M_NSF2h-6 | NSF 2h | 1 | F3,F4 | -0.031 | – | 22.8 | below_lloq;below_lod;cv_high |
| plate2 | IL-6 | M_ASF2h-4 | ASF 2h | 1 | F5,F6 | 0.0985 | 7.541 | 40.7 | below_lloq;cv_high |
| plate2 | IL-6 | M_NSF6h-2 | NSF 6h | 1 | F7,F8 | 0.014 | 1.174 | 132.4 | below_lloq;below_lod;cv_high |
| plate2 | IL-6 | M_NSF6h-10 | NSF 6h | 1 | F9,F10 | 0.0375 | 2.666 | 17.1 | below_lloq;below_lod |
| plate2 | IL-6 | M_NSF2h-7 | NSF 2h | 1 | G3,G4 | -0.0105 | – | 370.4 | below_lloq;below_lod;cv_high |
| plate2 | IL-6 | M_ASF2h-5 | ASF 2h | 1 | G5,G6 | 0.0235 | 1.78 | 107.6 | below_lloq;below_lod;cv_high |
| plate2 | IL-6 | M_NSF6h-3 | NSF 6h | 1 | G7,G8 | -0.009 | – | 110.0 | below_lloq;below_lod;cv_high |
| plate2 | IL-6 | M_NSF2h-8 | NSF 2h | 1 | H3,H4 | 0.019 | 1.413 | 58.3 | below_lloq;below_lod;cv_high |
| plate2 | IL-6 | M_ASF2h-6 | ASF 2h | 1 | H5,H6 | 0.019 | 1.418 | 64.6 | below_lloq;below_lod;cv_high |
| plate2 | IL-6 | M_NSF6h-4 | NSF 6h | 2 | H7,H8 | -0.0035 | – | 707.1 | below_lloq;below_lod;cv_high |
| plate3 | IL-6 | M_ASF6h-9 | ASF 6h | 1 | A5,A6 | 0.114 | 9.819 | 2.426 | below_lloq |
| plate3 | IL-6 | M_NSF12h-7 | NSF 12h | 2 | A7,A8 | -0.0075 | – | 37.7 | below_lloq;below_lod;cv_high |
| plate3 | IL-6 | M_ASF12h-5 | ASF 12h | 1 | A9,A10 | 0.0305 | 1.79 | 41.6 | below_lloq;below_lod;cv_high |
| plate3 | IL-6 | M_ASF6h-10 | ASF 6h | 1 | B5,B6 | 0.222 | 23.6 | 27.2 | cv_high |
| plate3 | IL-6 | M_NSF12h-8 | NSF 12h | 1 | B7,B8 | 0.037 | 2.389 | 86.7 | below_lloq;below_lod;cv_high |
| plate3 | IL-6 | M_ASF12h-6 | ASF 12h | 2 | B9,B10 | 0.0145 | 0.7694 | 130.0 | below_lloq;below_lod;cv_high |
| plate3 | IL-6 | M_NSF12h-1 | NSF 12h | 2 | C5,C6 | -0.0015 | – | 2,262.7 | below_lloq;below_lod;cv_high |
| plate3 | IL-6 | M_NSF12h-9 | NSF 12h | 1 | C7,C8 | -0.0265 | – | 21.3 | below_lloq;below_lod;cv_high |
| plate3 | IL-6 | M_ASF6h-4 | ASF 6h | 1 | D3,D4 | 0.138 | 12.6 | 0.6699 | below_lloq |
| plate3 | IL-6 | M_NSF12h-2 | NSF 12h | 1 | D5,D6 | 0.003 | 0.09477 | 136.4 | below_lloq;below_lod;cv_high |
| plate3 | IL-6 | M_NSF12h-10 | NSF 12h | 1 | D7,D8 | -0.005 | – | 410.1 | below_lloq;below_lod;cv_high |
| plate3 | IL-6 | M_ASF12h-8 | ASF 12h | 2 | D9,D10 | 0.0835 | 6.63 | 45.6 | below_lloq;cv_high |
| plate3 | IL-6 | M_ASF6h-5 | ASF 6h | 1 | E3,E4 | 0.088 | 7.023 | 17.7 | below_lloq |
| plate3 | IL-6 | M_NSF12h-3 | NSF 12h | 2 | E5,E6 | 0.002 | – | 813.2 | below_lloq;below_lod;cv_high |
| plate3 | IL-6 | M_ASF12h-1 | ASF 12h | 1 | E7,E8 | 0.02 | 1.19 | 134.0 | below_lloq;below_lod;cv_high |
| plate3 | IL-6 | M_ASF12h-9 | ASF 12h | 2 | E9,E10 | 0.0665 | 4.908 | 35.6 | below_lloq;cv_high |
| plate3 | IL-6 | M_ASF6h-6 | ASF 6h | 2 | F3,F4 | 0.138 | 12.9 | 55.7 | below_lloq;cv_high |
| plate3 | IL-6 | M_NSF12h-4 | NSF 12h | 1 | F5,F6 | 0.008 | 0.3509 | 129.7 | below_lloq;below_lod;cv_high |
| plate3 | IL-6 | M_ASF12h-2 | ASF 12h | 1 | F7,F8 | 0.1575 | 15.0 | 23.4 | below_lloq;cv_high |
| plate3 | IL-6 | M_ASF12h-10 | ASF 12h | 2 | F9,F10 | 0.0515 | 3.513 | 28.3 | below_lloq;below_lod;cv_high |
| plate3 | IL-6 | M_NSF12h-5 | NSF 12h | 1 | G5,G6 | 0.0245 | 1.337 | 22.4 | below_lloq;below_lod;cv_high |
| plate3 | IL-6 | M_ASF12h-3 | ASF 12h | 2 | G7,G8 | 0.0915 | 7.4 | 24.0 | below_lloq;cv_high |
| plate3 | IL-6 | M_NSF12h-6 | NSF 12h | 1 | H5,H6 | 0.0165 | 0.8245 | 75.2 | below_lloq;below_lod;cv_high |
| plate3 | IL-6 | M_ASF12h-4 | ASF 12h | 1 | H7,H8 | 0.0315 | 1.996 | 106.3 | below_lloq;below_lod;cv_high |
| plate4 | IL-6 | M_NSF24h-1 | NSF 24h | 1 | A3,A4 | 0 | – | – | below_lloq;below_lod |
| plate4 | IL-6 | M_NSF24h-9 | NSF 24h | 2 | A5,A6 | -0.002 | – | 318.2 | below_lloq;below_lod;cv_high |
| plate4 | IL-6 | M_ASF24h-7 | ASF 24h | 1 | A7,A8 | 0.0565 | 6.37 | 30.0 | below_lloq;cv_high |
| plate4 | IL-6 | M_NSF24h-2 | NSF 24h | 1 | B3,B4 | 0.0115 | 1.289 | 25.1 | below_lloq;below_lod;cv_high |
| plate4 | IL-6 | M_NSF24h-10 | NSF 24h | 1 | B5,B6 | -0.0005 | – | 1,414.2 | below_lloq;below_lod;cv_high |
| plate4 | IL-6 | M_ASF24h-8 | ASF 24h | 1 | B7,B8 | 0.089 | 10.0 | 15.1 | below_lloq |
| plate4 | IL-6 | M_NSF24h-3 | NSF 24h | 1 | C3,C4 | -0.0145 | – | 39.0 | below_lloq;below_lod;cv_high |
| plate4 | IL-6 | M_ASF24h-1 | ASF 24h | 2 | C5,C6 | 0 | – | – | below_lloq;below_lod |
| plate4 | IL-6 | M_ASF24h-9 | ASF 24h | 1 | C7,C8 | 0.1275 | 14.4 | 13.4 | below_lloq |
| plate4 | IL-6 | M_NSF24h-4 | NSF 24h | 2 | D3,D4 | -0.0115 | – | 73.8 | below_lloq;below_lod;cv_high |
| plate4 | IL-6 | M_ASF24h-2 | ASF 24h | 3 | D5,D6 | -0.0105 | – | 175.1 | below_lloq;below_lod;cv_high |
| plate4 | IL-6 | M_ASF24h-10 | ASF 24h | 1 | D7,D8 | 0.0875 | 9.859 | 6.473 | below_lloq |
| plate4 | IL-6 | M_NSF24h-5 | NSF 24h | 2 | E3,E4 | -0.0145 | – | 9.753 | below_lloq;below_lod |
| plate4 | IL-6 | M_ASF24h-3 | ASF 24h | 2 | E5,E6 | 0.0145 | – | 156.1 | below_lloq;below_lod;cv_high |
| plate4 | IL-6 | M_NSF24h-6 | NSF 24h | 1 | F3,F4 | 0.0225 | – | 150.8 | below_lloq;below_lod;cv_high |
| plate4 | IL-6 | M_ASF24h-4 | ASF 24h | 2 | F5,F6 | 0.0445 | 5.02 | 19.0 | below_lloq |
| plate4 | IL-6 | M_NSF24h-7 | NSF 24h | 2 | G3,G4 | -0.025 | – | 25.5 | below_lloq;below_lod;cv_high |
| plate4 | IL-6 | M_ASF24h-5 | ASF 24h | 1 | G5,G6 | 0.04 | 4.514 | 15.9 | below_lloq |
| plate4 | IL-6 | M_NSF24h-8 | NSF 24h | 1 | H3,H4 | -0.019 | – | 11.2 | below_lloq;below_lod |
| plate4 | IL-6 | M_ASF24h-6 | ASF 24h | 3 | H5,H6 | -0.0035 | – | 606.1 | below_lloq;below_lod;cv_high |
4 · Dilution linearity
Samples run at more than one dilution should agree after correction (±20%). A concentration that rises with dilution points to matrix interference or the high-dose hook effect.
None.
5 · Inter-plate normalization
Bridge controls shared by every plate estimate a multiplicative plate factor, on the log scale with the average plate as reference. Corrected concentration = raw / factor.
| plate_id | analyte | factor | log_factor_se | n_bridges | bridge_ids |
|---|---|---|---|---|---|
| plate1 | IL-6 | 1 | – | 0 | |
| plate2 | IL-6 | 1 | – | 0 | |
| plate3 | IL-6 | 1 | – | 0 | |
| plate4 | IL-6 | 1 | – | 0 |
None.
6 · Group comparisons
Tests are run on log10-transformed concentrations. Shapiro-Wilk on every group decides between parametric tests (Welch t / Welch ANOVA) and rank tests (Mann-Whitney / Kruskal-Wallis). Omnibus p-values are Benjamini-Hochberg adjusted across analytes; pairwise p-values are Holm-adjusted within each analyte. Hollow points are censored values that were substituted.
| analyte | handling | n |
|---|---|---|
| IL-6 | excluded: cv_fail | 3 |
| IL-6 | excluded: plate failed QC | 30 |
| IL-6 | substituted: below common LLOQ 46.8 → /1.41 | 77 |
Omnibus tests
| analyte | groups | n | test | statistic | p | effect | value | shapiro |
|---|---|---|---|---|---|---|---|---|
| IL-6 | NSF 0h vs NSF 1h vs ASF 1h vs ASF 6h vs NSF 12h vs ASF 12h vs NSF 24h vs ASF 24h | NSF 0h=9, NSF 1h=10, ASF 1h=9, ASF 6h=9, NSF 12h=10, ASF 12h=10, NSF 24h=10, ASF 24h=10 | Kruskal-Wallis | – | – | epsilon² | – | NSF 0h: p=nan, NSF 1h: p=nan, ASF 1h: p=nan, ASF 6h: p=nan, NSF 12h: p=nan, ASF 12h: p=nan, NSF 24h: p=nan, ASF 24h: p=nan |
Pairwise comparisons
| analyte | reference | group | test | effect_name | estimate | 95% CI low | 95% CI high | std. effect | value | p (Holm) | significant |
|---|---|---|---|---|---|---|---|---|---|---|---|
| IL-6 | NSF 0h | NSF 1h | Mann-Whitney U | fold change (HL) | 1 | 1 | 1 | rank-biserial r | 0 | 1 | ✗ fail |
| IL-6 | NSF 0h | ASF 1h | Mann-Whitney U | fold change (HL) | 1 | 1 | 1 | rank-biserial r | 0 | 1 | ✗ fail |
| IL-6 | NSF 0h | ASF 6h | Mann-Whitney U | fold change (HL) | 1 | 1 | 1 | rank-biserial r | 0 | 1 | ✗ fail |
| IL-6 | NSF 0h | NSF 12h | Mann-Whitney U | fold change (HL) | 1 | 1 | 1 | rank-biserial r | 0 | 1 | ✗ fail |
| IL-6 | NSF 0h | ASF 12h | Mann-Whitney U | fold change (HL) | 1 | 1 | 1 | rank-biserial r | 0 | 1 | ✗ fail |
| IL-6 | NSF 0h | NSF 24h | Mann-Whitney U | fold change (HL) | 1 | 1 | 1 | rank-biserial r | 0 | 1 | ✗ fail |
| IL-6 | NSF 0h | ASF 24h | Mann-Whitney U | fold change (HL) | 1 | 1 | 1 | rank-biserial r | 0 | 1 | ✗ fail |
| IL-6 | NSF 1h | ASF 1h | Mann-Whitney U | fold change (HL) | 1 | 1 | 1 | rank-biserial r | 0 | 1 | ✗ fail |
| IL-6 | NSF 1h | ASF 6h | Mann-Whitney U | fold change (HL) | 1 | 1 | 1 | rank-biserial r | 0 | 1 | ✗ fail |
| IL-6 | NSF 1h | NSF 12h | Mann-Whitney U | fold change (HL) | 1 | 1 | 1 | rank-biserial r | 0 | 1 | ✗ fail |
| IL-6 | NSF 1h | ASF 12h | Mann-Whitney U | fold change (HL) | 1 | 1 | 1 | rank-biserial r | 0 | 1 | ✗ fail |
| IL-6 | NSF 1h | NSF 24h | Mann-Whitney U | fold change (HL) | 1 | 1 | 1 | rank-biserial r | 0 | 1 | ✗ fail |
| IL-6 | NSF 1h | ASF 24h | Mann-Whitney U | fold change (HL) | 1 | 1 | 1 | rank-biserial r | 0 | 1 | ✗ fail |
| IL-6 | ASF 1h | ASF 6h | Mann-Whitney U | fold change (HL) | 1 | 1 | 1 | rank-biserial r | 0 | 1 | ✗ fail |
| IL-6 | ASF 1h | NSF 12h | Mann-Whitney U | fold change (HL) | 1 | 1 | 1 | rank-biserial r | 0 | 1 | ✗ fail |
| IL-6 | ASF 1h | ASF 12h | Mann-Whitney U | fold change (HL) | 1 | 1 | 1 | rank-biserial r | 0 | 1 | ✗ fail |
| IL-6 | ASF 1h | NSF 24h | Mann-Whitney U | fold change (HL) | 1 | 1 | 1 | rank-biserial r | 0 | 1 | ✗ fail |
| IL-6 | ASF 1h | ASF 24h | Mann-Whitney U | fold change (HL) | 1 | 1 | 1 | rank-biserial r | 0 | 1 | ✗ fail |
| IL-6 | ASF 6h | NSF 12h | Mann-Whitney U | fold change (HL) | 1 | 1 | 1 | rank-biserial r | 0 | 1 | ✗ fail |
| IL-6 | ASF 6h | ASF 12h | Mann-Whitney U | fold change (HL) | 1 | 1 | 1 | rank-biserial r | 0 | 1 | ✗ fail |
| IL-6 | ASF 6h | NSF 24h | Mann-Whitney U | fold change (HL) | 1 | 1 | 1 | rank-biserial r | 0 | 1 | ✗ fail |
| IL-6 | ASF 6h | ASF 24h | Mann-Whitney U | fold change (HL) | 1 | 1 | 1 | rank-biserial r | 0 | 1 | ✗ fail |
| IL-6 | NSF 12h | ASF 12h | Mann-Whitney U | fold change (HL) | 1 | 1 | 1 | rank-biserial r | 0 | 1 | ✗ fail |
| IL-6 | NSF 12h | NSF 24h | Mann-Whitney U | fold change (HL) | 1 | 1 | 1 | rank-biserial r | 0 | 1 | ✗ fail |
| IL-6 | NSF 12h | ASF 24h | Mann-Whitney U | fold change (HL) | 1 | 1 | 1 | rank-biserial r | 0 | 1 | ✗ fail |
| IL-6 | ASF 12h | NSF 24h | Mann-Whitney U | fold change (HL) | 1 | 1 | 1 | rank-biserial r | 0 | 1 | ✗ fail |
| IL-6 | ASF 12h | ASF 24h | Mann-Whitney U | fold change (HL) | 1 | 1 | 1 | rank-biserial r | 0 | 1 | ✗ fail |
| IL-6 | NSF 24h | ASF 24h | Mann-Whitney U | fold change (HL) | 1 | 1 | 1 | rank-biserial r | 0 | 1 | ✗ fail |
7 · Settings and warnings
{
"model": "auto",
"weighting": "1/y^2",
"include_zero_standard": true,
"qc": {
"max_cv_pct": 20.0,
"recovery_low_pct": 80.0,
"recovery_high_pct": 120.0,
"edge_tolerance_pct": 5.0,
"lod_sd_multiplier": 3.0,
"outlier_z": 3.5,
"max_std_exclusions": 2,
"min_r_squared": 0.98,
"min_std_pass_frac": 0.75,
"min_control_pass_frac": 0.6666666666666666
},
"stats": {
"transform": "log10",
"censored": "substitute",
"harmonize_lloq": true,
"lloq_divisor": 1.4142135623730951,
"alpha": 0.05,
"normality_alpha": 0.05,
"exclude_failed_plates": true,
"value_col": "conc"
},
"normalize": false,
"linearity_max_deviation_pct": 20.0,
"group_order": null,
"control_ranges": null
}
- invalid value encountered in scalar divide