Table 4.
Calibration curve parameters, accuracies, and precisions of the described method
| Range (pmol) | Calibration curve equation (y = ax2 + bx + c) | Within-run accuracy (% bias) | Within-run precision (% CV) | Between-run accuracy (% bias) | Between-run precision (% CV) | |||||||||||||||||
|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|
| Name | LLOQ | ULOQ | a | b | c | R2 | LLOQ | Low | Mid | High | LLOQ | Low | Mid | High | LLOQ | Low | Mid | High | LLOQ | Low | Mid | High |
| 7-keto-25-OHC | 0.39 | 50 | 1.92E-04 | 0.0680 | 0.0092 | 0.975 | −6.6 | 8.6 | −12.2 | −9.5 | 5.5 | 14.8 | 3.7 | 2.5 | 7.7 | 6.1 | 5.1 | 3.5 | 17.8 | 18.0 | 10.9 | 10.1 |
| 7-keto-27-OHC | 0.39 | 50 | 3.09E-04 | 0.1509 | −0.0052 | 0.990 | 17.4 | 12.7 | 1.8 | 11.9 | 8.1 | 4.5 | 8.4 | 4.5 | 14.9 | 8.7 | 1.3 | −8.6 | 12.8 | 15.7 | 8.2 | 9.2 |
| 7α,24 (S)-diOHC | 1.95 | 250 | −3.51E-06 | 0.0038 | 0.0122 | 0.971 | 6.7 | 13.4 | 13.2 | 13.8 | 9.2 | 3.3 | 6.3 | 12.0 | 5.7 | 11.3 | −1.5 | −10.4 | 17.3 | 13.2 | 11.8 | 16.2 |
| 7α,25-diOHC | 1.95 | 250 | 2.83E-05 | 0.0110 | −0.0042 | 0.973 | 7.4 | 3.8 | −2.8 | −7.0 | 10.3 | 5.7 | 9.4 | 7.8 | 13.3 | 1.1 | 7.1 | 7.8 | 18.9 | 11.7 | 12.5 | 4.8 |
| 7α,27-diOHC | 1.95 | 250 | 1.69E-05 | 0.0050 | 0.0011 | 0.986 | 2.3 | 0.6 | −3.0 | 1.7 | 13.5 | 2.9 | 8.6 | 6.6 | 16.3 | 11.6 | 7.4 | 12.5 | 13.6 | 12.5 | 10.7 | 5.7 |
| 5α,6β-diOHC | 1.95 | 250 | −2.82E-06 | 0.0021 | 0.0035 | 0.980 | 5.4 | −1.3 | −28.2 | −14.0 | 17.1 | 16.5 | 10.1 | 12.6 | 24.6 | 12.0 | −7.9 | −17.6 | 21.9 | 13.8 | 11.9 | 5.8 |
| 7α,12α-diOHCone | 0.39 | 50 | −1.33E-05 | 0.0613 | 0.0101 | 0.969 | −12.8 | −12.6 | −11.7 | −6.7 | 24.2 | 4.5 | 15.3 | 15.2 | −15.8 | 18.1 | −5.3 | −14.6 | 13.7 | 15.7 | 24.9 | 17.5 |
| 7-ketochol | 1.95 | 250 | −5.97E-07 | 0.0032 | 0.0271 | 0.947 | - | - | - | - | - | - | - | - | - | - | - | - | - | - | - | - |
| 4β-OHC | 3.91 | 500 | −1.82E-06 | 0.0066 | 0.0241 | 0.978 | 12.8 | 21.2 | 1.3 | 13.9 | 4.1 | 7.7 | 7.2 | 1.1 | −18.4 | 10.3 | 0.8 | 8.7 | 16.3 | 17.3 | 15.4 | 11.8 |
| 7α-OHC | 1.95 | 250 | 1.56E-06 | 0.0042 | 0.0081 | 0.944 | −13.7 | −14.0 | 7.2 | 19.0 | 17.4 | 6.3 | 2.1 | 2.1 | −13.0 | −10.2 | 2.7 | 11.5 | 19.6 | 19.5 | 15.7 | 10.5 |
| 22 (R)-OHC | 1.95 | 250 | 1.05E-06 | 0.0028 | 0.0016 | 0.991 | 2.9 | 3.0 | −4.0 | −8.9 | 12.3 | 10.3 | 2.3 | 4.5 | 20.2 | 9.7 | 3.4 | 1.5 | 12.0 | 18.1 | 8.1 | 4.7 |
| 24 (S)-OHC | 1.95 | 250 | −5.63E-06 | 0.0066 | 0.0033 | 0.992 | −9.8 | 10.0 | 2.5 | 2.7 | 5.0 | 5.5 | 5.0 | 2.4 | −13.8 | −1.3 | 5.4 | 6.4 | 10.1 | 9.6 | 5.1 | 6.9 |
| 25-OHC | 1.95 | 250 | 2.31E-06 | 0.0035 | −0.0002 | 0.997 | 12.1 | 3.7 | 4.3 | 18.2 | 13.0 | 5.5 | 7.5 | 5.0 | 10.4 | −0.8 | 2.4 | 0.1 | 7.9 | 7.3 | 3.6 | 2.5 |
| 27-OHC | 1.95 | 250 | −5.84E-07 | 0.0078 | −0.0036 | 0.997 | 18.8 | 15.3 | 2.4 | 10.5 | 10.8 | 7.2 | 7.7 | 6.6 | 17.6 | 2.2 | 2.6 | −1.5 | 3.2 | 4.1 | 4.9 | 4.0 |
| 7α-OHCone | 1.95 | 250 | 1.23E-05 | 0.0062 | −0.0039 | 0.953 | - | - | - | - | - | - | - | - | - | - | - | - | - | - | - | - |
| 5α,6α-epoxychol | 3.91 | 500 | −8.45E-06 | 0.0227 | 0.0332 | 0.990 | −4.1 | 14.4 | −8.2 | 12.8 | 17.5 | 10.2 | 5.4 | 4.1 | −2.0 | 2.9 | −7.9 | 0.1 | 7.8 | 9.3 | 8.4 | 9.5 |
| 5β,6β-epoxychol | 3.91 | 500 | −2.13E-05 | 0.0451 | 0.1436 | 0.982 | 8.1 | 30.1 | 10.6 | 9.5 | 9.3 | 7.2 | 7.2 | 1.2 | −17.9 | 2.0 | −0.2 | −0.8 | 27.2 | 16.9 | 13.7 | 13.5 |
| 24 (S),25-epoxychol | 3.91 | 500 | 4.57E-07 | 0.0007 | 0.0003 | 0.984 | 14.2 | −1.8 | 13.8 | −4.6 | 17.0 | 22.7 | 8.0 | 12.8 | 10.3 | 16.1 | 6.2 | −4.8 | 18.3 | 11.2 | 10.8 | 15.2 |
| α-MCA | 1.95 | 250 | 1.85E-06 | 0.0048 | −0.0039 | 0.994 | 28.4 | 10.2 | 4.9 | −2.1 | 2.1 | 2.0 | 3.2 | 3.0 | 19.8 | 6.5 | 8.3 | 1.4 | 6.2 | 7.7 | 5.5 | 4.2 |
| β-MCA | 1.95 | 250 | 1.41E-05 | 0.0023 | 0.0027 | 0.967 | 32.5 | 26.5 | 7.1 | 8.0 | 2.6 | 4.5 | 6.7 | 2.1 | 15.6 | 16.5 | 4.9 | 12.4 | 19.8 | 7.4 | 5.6 | 2.1 |
| ω-MCA | 1.95 | 250 | 1.33E-05 | 0.0030 | 0.0029 | 0.985 | 15.4 | 19.5 | 8.9 | 3.9 | 6.9 | 4.0 | 4.9 | 1.9 | 3.0 | 12.1 | 4.1 | 8.2 | 19.9 | 11.5 | 5.1 | 3.6 |
| HCA | 1.95 | 250 | −1.29E-06 | 0.0073 | −0.0058 | 0.997 | 29.4 | 12.8 | 6.8 | −7.5 | 5.9 | 3.0 | 3.7 | 4.2 | 22.3 | 5.3 | 9.9 | 1.4 | 7.6 | 4.6 | 4.7 | 4.5 |
| CA | 1.95 | 250 | 1.49E-05 | 0.0078 | 0.0083 | 0.993 | −17.2 | 2.2 | 6.7 | 2.8 | 2.7 | 1.5 | 3.4 | 1.9 | −17.0 | −3.6 | 4.5 | 2.9 | 7.2 | 10.9 | 5.1 | 4.1 |
| CDCA | 1.95 | 250 | 3.09E-05 | 0.0125 | −0.0002 | 0.997 | 17.6 | 14.3 | 6.3 | 17.9 | 4.9 | 6.0 | 3.7 | 2.6 | 12.4 | 5.2 | 7.2 | 2.1 | 12.1 | 5.4 | 2.7 | 1.2 |
| HDCA | 1.95 | 250 | 2.77E-05 | 0.0035 | 0.0051 | 0.989 | 25.9 | 17.4 | 3.7 | 0.5 | 4.2 | 4.2 | 7.5 | 1.7 | 18.2 | 12.0 | 4.2 | 4.6 | 10.9 | 8.1 | 7.7 | 3.0 |
| MDCA | 1.95 | 250 | 2.85E-05 | 0.0079 | −0.0011 | 0.994 | −1.6 | 8.5 | 8.0 | 19.9 | 17.5 | 4.7 | 5.0 | 3.1 | 8.8 | 2.2 | 8.1 | 2.6 | 22.8 | 6.4 | 4.8 | 3.3 |
| UDCA | 1.95 | 250 | 6.53E-05 | 0.0091 | 0.0096 | 0.989 | 2.4 | 6.2 | 1.4 | −1.6 | 4.0 | 6.2 | 5.3 | 2.7 | 18.6 | 10.0 | 3.2 | 3.7 | 21.8 | 10.9 | 4.7 | 3.2 |
| DCA | 1.95 | 250 | 3.56E-05 | 0.0046 | 0.0052 | 0.986 | 19.4 | 16.3 | 4.9 | 2.1 | 8.6 | 4.3 | 4.7 | 1.9 | 22.6 | 12.8 | 3.2 | 5.3 | 9.9 | 15.7 | 3.3 | 4.5 |
| LCA | 1.95 | 250 | 2.37E-05 | 0.0151 | 0.0010 | 0.996 | −3.3 | 5.1 | 5.6 | 18.6 | 8.2 | 5.9 | 6.2 | 1.8 | 15.0 | 5.7 | 8.0 | 3.6 | 9.0 | 7.8 | 4.4 | 2.9 |
| Tα-MCA | 3.91 | 500 | 7.72E-06 | 0.0080 | −0.0103 | 0.983 | 9.9 | 8.1 | 1.1 | 4.3 | 3.3 | 4.3 | 4.1 | 1.3 | −9.0 | −9.6 | −0.1 | −1.5 | 12.5 | 13.1 | 8.9 | 6.0 |
| Tβ-MCA | 3.91 | 500 | 4.53E-06 | 0.0039 | 0.0017 | 0.990 | 2.6 | 10.9 | 8.5 | 1.9 | 5.4 | 2.7 | 1.3 | 8.4 | −0.3 | −1.5 | 3.9 | 4.0 | 16.7 | 18.5 | 6.6 | 5.5 |
| Tω-MCA | 3.91 | 500 | 2.19E-06 | 0.0017 | 0.0018 | 0.989 | 12.8 | 17.0 | 4.9 | 1.9 | 12.7 | 3.0 | 6.7 | 2.8 | 7.2 | 13.8 | 5.3 | 5.9 | 14.7 | 12.8 | 7.3 | 4.1 |
| TCA | 3.91 | 500 | 2.67E-06 | 0.0063 | −0.0028 | 0.998 | 6.2 | 2.0 | 3.8 | 18.1 | 6.6 | 4.8 | 3.8 | 1.6 | 9.9 | 3.3 | 4.8 | 3.2 | 12.8 | 3.5 | 3.5 | 1.3 |
| TCDCA | 3.91 | 500 | 5.32E-06 | 0.0053 | 0.0038 | 0.987 | −7.4 | 2.9 | 2.5 | −2.3 | 15.2 | 2.7 | 2.9 | 9.8 | −6.8 | −2.6 | 2.5 | 0.4 | 8.5 | 13.1 | 6.6 | 7.1 |
| THDCA | 3.91 | 500 | 6.14E-06 | 0.0035 | −0.0002 | 0.989 | −2.6 | 5.7 | 4.2 | −1.4 | 15.7 | 4.7 | 1.8 | 9.5 | −5.3 | −2.4 | 3.3 | 0.3 | 16.5 | 19.0 | 3.9 | 5.9 |
| TUDCA | 3.91 | 500 | 5.89E-06 | 0.0029 | −0.0036 | 0.971 | −17.2 | 18.6 | 6.4 | 11.0 | 8.3 | 3.7 | 5.0 | 1.9 | −4.0 | −10.8 | −0.1 | 0.2 | 11.7 | 17.4 | 11.8 | 6.5 |
| TDCA | 3.91 | 500 | 5.56E-06 | 0.0045 | −0.0054 | 0.979 | −7.8 | 23.5 | 6.5 | 10.1 | 17.9 | 5.3 | 6.5 | 1.5 | −8.0 | −11.2 | −0.9 | 0.0 | 12.0 | 13.9 | 7.3 | 5.2 |
| TLCA | 3.91 | 500 | 7.02E-06 | 0.0117 | 0.0071 | 0.982 | −1.8 | 2.3 | 4.2 | −3.7 | 18.1 | 4.9 | 4.2 | 11.3 | 1.6 | 9.1 | 0.5 | −0.2 | 9.9 | 16.2 | 12.3 | 10.8 |
| GCA | 3.91 | 500 | 1.90E-06 | 0.0061 | 0.0012 | 0.997 | 16.5 | 3.4 | 1.1 | 15.0 | 8.5 | 4.0 | 3.3 | 1.9 | 18.5 | 7.4 | 4.3 | 2.2 | 6.8 | 5.0 | 3.5 | 4.6 |
| GCDCA | 3.91 | 500 | −1.08E-06 | 0.0023 | 0.0021 | 0.986 | 19.9 | 18.2 | 8.8 | 1.3 | 15.5 | 6.2 | 7.3 | 12.2 | 8.0 | 12.4 | 3.9 | 6.1 | 16.8 | 6.9 | 17.7 | 15.9 |
| GHDCA | 3.91 | 500 | −1.72E-08 | 0.0013 | 0.0002 | 0.987 | 9.1 | 14.1 | 15.3 | 1.2 | 4.5 | 8.2 | 4.8 | 7.9 | 4.3 | 2.6 | 12.6 | 5.3 | 20.5 | 9.6 | 11.1 | 8.0 |
| GUDCA | 3.91 | 500 | 9.41E-07 | 0.0011 | −0.0011 | 0.989 | −11.8 | 5.7 | 13.1 | 15.1 | 16.0 | 13.2 | 4.6 | 1.8 | −7.8 | −3.5 | 6.6 | 2.0 | 16.9 | 11.9 | 6.4 | 5.4 |
| GDCA | 3.91 | 500 | 2.99E-06 | 0.0068 | −0.0045 | 0.983 | −6.3 | 2.0 | 2.3 | 6.8 | 9.4 | 4.0 | 5.9 | 2.4 | 4.8 | −2.9 | 6.9 | 6.8 | 12.3 | 11.4 | 8.5 | 9.3 |
| GLCA | 3.91 | 500 | 2.90E-06 | 0.0013 | 0.0003 | 0.975 | −16.5 | 13.7 | 0.8 | 14.6 | 20.2 | 3.4 | 10.6 | 1.8 | 3.4 | 9.9 | 7.3 | 8.1 | 17.3 | 7.0 | 6.5 | 6.0 |
| THCA | 3.91 | 500 | −1.91E-06 | 0.0099 | −0.0041 | 0.997 | 15.7 | −2.5 | −6.6 | −5.4 | 3.5 | 4.0 | 2.8 | 1.8 | 17.6 | 3.9 | 2.6 | 0.1 | 3.6 | 2.6 | 4.0 | 5.8 |
This table shows the dynamic range (pmol per sample) and parameters of each calibration curve, as well as the within-run and between-run accuracies (expressed as % bias), and precisions (expressed as % CV).