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PH46A, a new chemical entity developed by our group, has shown potent anti-inflammatory activities through various pre-preclinical studies. The aim of this work was to develop and validate a sensitive and robust LC-MS/MS analytical method to determine the levels of PH46 in human plasma, urine and faeces. The linearity (0.5–500 ng/mL for plasma/urine, and 10–2000 ng/g for human faeces), accuracy (within 100 ± 15% for plasma/urine or 100 ± 20% for faeces), precision (≤ 15% CV for plasma/urine or ≤ 20% CV for faeces) and the method’s specificity were demonstrated to be acceptable. No significant matrix effects or carry-over was observed for PH46 and IStd, and the recovery was consistent. About 10- and 100-fold dilutions in control matrix were found not to affect the assays’ performance. PH46 was proven to be stable: at room temperature for >24 hrs in plasma through 3 freeze-thaw cycles, at –20°C for 83 days in plasma/32 days in urine/33 days in faeces, and at –80°C for 154 days in plasma/33 days in faeces. The re-injection reproducibility of PH46 in matrix extracts was at least 239 hrs at 4°C in plasma/25 days in urine/6.5 days in faeces. This method was successfully applied to the pharmacokinetic evaluation of the Phase I clinical studies.

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Bioactive Indanes: Development, Validation of a Bioanalytical Method of LC-MS/MS for the Determination of PH46A, a New Potential Anti-Inflammatory Agent, in Human Plasma, Urine, Faeces