Development and validation of an ultrafiltration-UPLC-MS/MS method for rapid quantification of unbound docetaxel in human plasma

• Docetaxel lipid microsphere is a novel formulation of docetaxel without Tween-80.
• Reliable ultrafiltration coupled with UPLC-MS/MS method is firstly developed.
• Low LLOQ (0.2 ng/mL) and short chromatographic run time (2.5 min) were achieved.
• Much smaller volume of human plasma (only 450 μL) is utilized.
• This validated method could be applied to clinical study of docetaxel lipid microsphere.

Docetaxel lipid microsphere (DT-LM) is a novel formulation of docetaxel without Tween-80. A sensitive, robust and reproducible ultrafiltration (UF) followed by UPLC-MS/MS method was developed and validated for the quantification of unbound docetaxel in human plasma using paclitaxel as IS. Ultrafiltrate samples were chromatographed on Acquity UPLC BEH C18 column (50 mm × 2.1 mm, 1.7 μm). The mobile phase was a mixture of 10 mM ammonium formate in water containing 0.2% formic acid (A) and acetonitrile containing 0.2% formic acid (B). The volume of plasma utilized was only 450 μL. The calibration curve was linear over the range of 0.2–200 ng/mL, with LLOQ of 0.2 ng/mL. The method was shown to be reliable and reproducible with intra- and inter-day precision and accuracy <±15%, and extraction recovery of 98.1–104.8%. Docetaxel was stable during stability studies, e.g., short term, post-preparation and freeze–thaw cycles. The validated method was utilized to support the pharmacological study of DT-LM in patients with advanced cancer.