Development and validation of a high-performance liquid chromatography-tandem mass spectrometry assay quantifying
C M Nijenhuis1, L Lucas, H Rosing
1Department of Pharmacy & Pharmacology, Slotervaart Hospital/The Netherlands Cancer Institute, Louwesweg 6, 1066 EC, Amsterdam, The Netherlands.
Abstract:
Olaparib is an inhibitor of poly ADP ribose polymerase 1 (PARP-1). Phase I and II trials showed promising results of olaparib against tumours in BRCA mutation carriers. Currently an increasing number of clinical trials with olaparib in combination with other compounds or radiotherapy are conducted. To support these clinical trials an LC-MS/MS method was developed and validated for the quantification of olaparib in human plasma. Human plasma samples were collected in the clinic and stored at nominally -20°C. Olaparib was isolated from plasma by liquid-liquid extraction, separated on a C18 column with gradient elution and analyzed with triple quadrupole mass spectrometry in positive ion mode. A deuterated isotope was used as internal standard for the quantification. The assay, ranging from 10 to 5000ng/mL, was linear with correlation coefficients (r(2)) of 0.9994 or better. The assay was accurate and precise, with inter-assay and intra-assay accuracies within ±7.6% of nominal and inter-assay and intra-assay precision ≤9.3% at the lower limit of quantification and ≤5.7% at the other concentration levels tested. All results were within the acceptance criteria of the US FDA and the latest EMA guidelines for method validation. A quantitative method was developed and validated for the quantification of olaparib in human plasma. The method could successfully be applied for the pharmacokinetic quantification of olaparib in cancer patients treated with olaparib.
Insights
A new method accurately quantifies olaparib in human plasma, crucial for clinical trials of this PARP-1 inhibitor in cancer patients, especially BRCA mutation carriers.
Area of Science:
- Pharmacology
- Analytical Chemistry
- Oncology
Background:
- Olaparib, a PARP-1 inhibitor, shows promise in BRCA-mutated cancers.
- Clinical trials combining olaparib with other treatments are expanding.
- Accurate quantification of olaparib in plasma is essential for supporting these trials.
Purpose of the Study:
- To develop and validate an LC-MS/MS method for quantifying olaparib in human plasma.
- To ensure the method meets regulatory standards for clinical trial support.
Main Methods:
- Liquid-liquid extraction of olaparib from plasma.
- Separation using a C18 column with gradient elution.
- Analysis via triple quadrupole mass spectrometry (LC-MS/MS) in positive ion mode with a deuterated internal standard.
Main Results:
- The validated assay covers a range of 10-5000 ng/mL with excellent linearity (r² ≥ 0.9994).
- Assay accuracy was within ±7.6% and precision was ≤9.3% (LLOQ) and ≤5.7% (other levels).
- Method validation met US FDA and EMA guidelines.
Conclusions:
- A robust and validated LC-MS/MS method for olaparib quantification in human plasma was successfully developed.
- This method is suitable for pharmacokinetic analysis in cancer patients undergoing olaparib treatment.
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