Table I.
Recovery tests of additive steroids and glucuronides into the blood and the testis in the determination method developed in this study.
| Blood (0.50 ml)a |
Testis (1.00 g)a |
Liver (1.00 g)a |
||||||
|---|---|---|---|---|---|---|---|---|
| Steroids | Added (ng/g) | Recovery (%) | RSD (%) | Recovery (%) | RSD (%) | Recovery (%) | RSD (%) | LOD (fmol/g) |
| PGN | 10.0 | 93.3 | 18.5 | 98.3 | 7.9 | 86.7 | 3.5 | |
| 5.0 | 91.0 | 18.4 | 104.3 | 14.8 | 85.0 | 15.3 | 14.2 | |
| 1.0 | 85.9 | 15.7 | 91.6 | 19.4 | 99.3 | 9.8 | ||
| PGT | 10.0 | 90.7 | 10.9 | |||||
| 5.0 | 94.6 | 8.2 | 92.0 | 10.5 | 86.6 | 16.0 | ||
| 1.0 | 90.1 | 6.4 | 97.7 | 16.5 | 107.1 | 15.3 | 3.2 | |
| 0.1 | 82.8 | 4.5 | 101.6 | 9.3 | ||||
| HPGT | 10.0 | 72.4 | 4.3 | |||||
| 5.0 | 110.3 | 2.3 | 105.2 | 7.0 | 75.8 | 19.8 | ||
| 1.0 | 86.0 | 9.4 | 89.4 | 10.4 | 89.7 | 6.7 | 2.8 | |
| 0.1 | 97.2 | 13.6 | 92.6 | 16.4 | ||||
| ADS | 10 | 105.2 | 7.0 | |||||
| 5.0 | 113.6 | 6.4 | 86.0 | 4.5 | 89.4 | 10.4 | ||
| 1.0 | 115.4 | 2.5 | 86.2 | 5.6 | 92.6 | 16.4 | 2.6 | |
| 0.1 | 100.7 | 7.6 | 102.0 | 5.3 | ||||
| TS | 10.0 | 98.6 | 4.5 | |||||
| 5.0 | 106.8 | 5.2 | 80.5 | 4.7 | 88.0 | 9.1 | ||
| 1.0 | 104.8 | 5.7 | 77.6 | 5.7 | 74.3 | 4.4 | 0.6 | |
| 0.1 | 110.6 | 7.8 | 96.4 | 15.1 | ||||
| 17α-Estradiol | 10.0 | 108.7 | 6.0 | |||||
| 5.0 | 91.9 | 14.1 | 108.0 | 1.0 | 89.7 | 20.5 | ||
| 1.0 | 73.3 | 2.1 | 109.5 | 4.0 | 89.0 | 7.9 | 25.3 | |
| 0.5 | 89.4 | 15.6 | 107.0 | 6.1 | ||||
| 17β-Estradiol | 10.0 | 99.3 | 3.1 | |||||
| 5.0 | 99.3 | 3.1 | 105.7 | 0.7 | 73.3 | 2.1 | ||
| 1.0 | 73.3 | 2.1 | 81.9 | 1.5 | 112.3 | 6.7 | 24.2 | |
| 0.5 | 112.3 | 6.7 | 96.9 | 18.4 | ||||
| CCS | 10.0 | 101.3 | 4.0 | |||||
| 5.0 | 82.7 | 9.4 | 84.8 | 16.4 | 88.9 | 9.6 | ||
| 1.0 | 80.7 | 4.3 | 100.6 | 13.9 | 104.3 | 2.9 | 0.4 | |
| 0.1 | 107.8 | 5.1 | 85.7 | 9.1 | ||||
| 11-DCC | 10.0 | 95.9 | 5.2 | |||||
| 5.0 | 78.6 | 7.4 | 116.9 | 1.7 | 97.6 | 2.0 | ||
| 1.0 | 90.0 | 11.3 | 78.1 | 4.3 | 100.7 | 21.5 | 9.2 | |
| 0.1 | 81.5 | 8.8 | 75.3 | 1.2 | ||||
| TS-17-glucuronide | 10.0 | 85.0 | 8.6 | |||||
| 5.0 | 91.8 | 18.8 | 85.0 | 2.6 | 95.4 | 16.4 | 34.6 | |
| 1.0 | 98.5 | 13.8 | 95.4 | 6.4 | 110.0 | 10.1 | ||
| 0.5 | 71.3 | 14.9 | 117.0 | 1.1 | ||||
| 17β-Estradiol-3-glucuronide | 10.0 | 103.5 | 4.3 | |||||
| 5.0 | 90.7 | 10.9 | 102.7 | 2.3 | 97.8 | 4.0 | 73.3 | |
| 1.0 | 86.6 | 16.0 | 109.2 | 7.7 | 119.1 | 0.5 | ||
| 0.5 | 107.1 | 15.3 | 79.1 | 12.3 | ||||
| 17β-Estradiol-3-sulfate | 10.0 | 90.3 | 17.1 | 106.4 | 1.4 | 70.2 | 6.5 | |
| 5.0 | 101.3 | 4.6 | 101.5 | 13.5 | 72.3 | 12.3 | 146.6 | |
| 1.0 | 82.3 | 13.2 | 91.3 | 12.5 | 71.2 | 15.3 | ||
Standard substrates ∼10-fold amount of the assay limits were added into the blood, liver and the testis. Standards substrates were extracted and determined by the assay method developed in this study. Recovery data obtained were shown with RSD. The accurate data having 70–120% recovery and RSD under 25% and LOD (signal/noise [S/N] > 3) were obtained, respectively, as previously indicated by FDA as a guideline (FDA 2001).
aData presented mean values of n = 3–5.