Validation of the HPLC/MS assay for cyclosporine and SDZ-RAD in monkey organ tissues
| Tissue | Linear Range | Cyclosporine | SDZ-RAD | |||||
|---|---|---|---|---|---|---|---|---|
| Regression Analysis | CV | Acc | Linear Range | Regression Analysis | CV | Acc | ||
| μg/l | % | μg/l | % | |||||
| Brainstem | 50–1000 | y = 1.02x − 7.5, r = 0.996 | 3.0 | −0.8 | 0.5–100 | y = 0.91x + 0.22, r = 0.999 | 8.3 | −5.5 |
| Cerebellum | 50–1000 | y = 1.04x − 9.5, r = 0.997 | 5.7 | −1.3 | 0.5–100 | y = 0.91x − 0.31, r = 0.999 | 4.7 | −2.4 |
| Cerebrum | 50–1000 | y = 0.98x − 4.6, r = 0.998 | 5.9 | −3.9 | 0.5–100 | y = 1.05x − 0.25, r = 0.995 | 7.1 | +1.4 |
| Kidney | 50–1000 | y = 1.01x − 4.1, r = 0.995 | 9.8 | +0.2 | 0.5–100 | y = 1.05x − 0.03, r = 0.997 | 12.3 | +6.0 |
| Liver | 50–1000 | y = 0.95x − 4.6, r = 0.992 | 13.9 | −10.3 | 0.5–100 | y = 1.04x − 0.37, r = 0.999 | 9.8 | +10.8 |
| Stomach | 50–1000 | y = 0.99x − 1.5, r = 0.993 | 4.1 | +1.1 | 0.5–100 | y = 1.05x − 1.38, r = 0.997 | 7.9 | −1.1 |
| Colon | 50–1000 | y = 1.01x + 3.9, r = 0.993 | 6.9 | +6.6 | 0.5–100 | y = 0.93x + 1.02, r = 0.997 | 11.5 | −2.8 |
| Ileum | 50–1000 | y = 1.01x − 5.7, r = 0.988 | 6.5 | −0.9 | 0.5–100 | y = 0.96x + 0.28, r = 0.997 | 7.9 | +2.8 |
| Duodenum | 50–1000 | y = 1.02x + 0.8, r = 0.990 | 11.8 | +10.2 | 0.5–100 | y = 1.01x + 0.42, r = 0.999 | 11.5 | −2.8 |
| Jejunum | 50–1000 | y = 0.91x + 0.8, r = 0.995 | 6.0 | −6.6 | 0.5–100 | y = 0.95x − 0.91, r = 0.997 | 7.7 | −2.7 |
| Pancreas | 50–1000 | y = 0.96x + 5.1, r = 0.998 | 9.2 | −6.6 | 0.5–100 | y = 1.06x − 1.67, r = 0.996 | 15.4 | −6.1 |
| Heart | 50–1000 | y = 1.07x − 5.9, r = 0.996 | 6.8 | +5.5 | 0.5–100 | y = 1.00x − 0.69, r = 0.999 | 9.0 | −5.6 |
| Lung (right) | 50–1000 | y = 1.02x − 2.6, r = 0.999 | 7.4 | +0.4 | 0.5–100 | y = 1.03x − 0.02, r = 0.997 | 7.6 | −2.7 |
| Lung (left) | 50–1000 | y = 1.06x + 2.2, r = 0.998 | 6.0 | +8.6 | 0.5–100 | y = 0.90x + 1.75, r = 0.997 | 4.5 | −3.7 |
| Spleen | 50–1000 | y = 1.07x − 2.1, r = 0.997 | 8.0 | +7.2 | 0.5–100 | y = 1.03x − 0.46, r = 0.999 | 4.3 | −0.8 |
| Testes | 50–1000 | y = 1.04x − 2.7, r = 0.995 | 4.4 | +4.7 | 0.5–100 | y = 0.99x − 1.25, r = 0.999 | 3.6 | −2.3 |
| Thymus | 50–1000 | y = 1.06x − 5.3, r = 0.996 | 6.3 | +3.5 | 0.5–100 | y = 0.96x − 0.5, r = 0.998 | 8.0 | +3.0 |
| Fat | 50–1000 | y = 1.02x − 1.4, r = 0.996 | 3.6 | +3.0 | 0.5–100 | y = 0.95x − 0.43, r = 0.996 | 5.1 | −3.4 |
| Lymph node | 50–1000 | y = 1.04x − 1.5, r = 0.995 | 5.5 | +2.7 | 0.5–100 | y = 0.98x± 0, r = 0.999 | 3.8 | −4.3 |
| Gall bladder | 50–1000 | y = 1.08x − 7.8, r = 0.999 | 10.9 | +2.1 | 0.5–100 | y = 1.11x − 0.47, r = 0.999 | 9.3 | −8.1 |
Tissue samples were collected from untreated monkeys. Tissues were homogenized and cyclosporine or SDZ-RAD was added and incubated at 37°C for 30 min to allow for distribution and protein binding. The samples were extracted as described in Experimental Procedures. A concentration was accepted as the lower or upper limit of quantitation when four of five samples (80%) were within ±20% of the nominal concentration and the coefficient of variation of all samples was ≤20%. The calibration curve consisted of four different concentrations (n = 3 for each concentration). Interassay coefficient of variation (CV) and accuracy (Acc) were measured at three different concentrations (n = 3 per concentration). Means (n = 9) are presented.