HPLC–MS/MS method for quantification of paclitaxel from keratin containing samples

Abstract

Local drug delivery of paclitaxel is becoming ever more prevalent. As complex drug/excipient combinations are being developed and tested, new high performance liquid chromatography-mass spectrometry (HPLC-MS) techniques capable of quantifying paclitaxel from such formulations are needed. Here a method for quantifying paclitaxel from aqueous, protein and oil containing samples was developed and validated. Keratin, derived from human hair, is the protein component/paclitaxel excipient in the development and validation of said method. The novelty of this method is described by its ability to overcome water solubility issues and address clean-up of residual solvents in clinical grade paclitaxel injection composition. The method evaluates tert-butyl methyl ether and ethanol as extraction solvents with an extraction efficiency of 31.9 ± 2.3% and 86.4 ± 4.5% respectively. Upon evaporation and rehydration, samples were evaluated by HPLC-MS and a method was developed for paclitaxel quantification. The method developed had an inter-day precision of 9.1% relative standard deviation and an intra-day precision of 4.3% relative standard deviation normalized to a docetaxel internal standard. The described method is applicable to any aqueous paclitaxel sample containing protein and/or oils.

Arginine, glycine, aspartic acid peptide-modified paclitaxel and curcumin co-loaded liposome for the treatment of lung cancer: in vitro/vivo evaluation

“…The sample handling and determination process was referenced to previous literature by the LC-MS/MS method. [25][26][27][28] cell uptake…”

Conformation changes in human hair keratin observed using confocal Raman spectroscopy after active ingredient application

“…Several physical and biochemical techniques have already been used to test human hair and characterise keratin fibres, such as x‐ray diffraction, solid state nuclear magnetic resonance (NMR), circular dichroism, scanning electron microscopy and high‐pressure liquid chromatography–mass spectrometry (HPLC–MS) . Although biochemical and microscopic techniques are innovative, they present some technical limitations owing to protocol complexities, prior sample preparation, the need for extraction, and reagent costs.…”

In vitro and in vivo Assessment of Keratose as a Novel Excipient of Paclitaxel Coated Balloons

“…Paclitaxel-containing keratose hydrogels were formed at 12, 15, and 20% weight-to-volume ratios of keratose in phosphate buffered saline (PBS) (GE Healthcare Hyclone, Logan, UT, United States) as previously described ( Turner et al, 2017 ). Hydrogels were injected into microcentrifuge tubes.…”

“…Paclitaxel was quantified from PBS samples containing keratose by high-performance liquid chromatography tandem mass spectrometry (HPLC-MS). Prior to HPLC-MS, keratose was removed from the samples by ethanol extraction ( Turner et al, 2017 ). A Symmetry C18, 100 Å, 3.5 μm, 100 × 2.1 mm column (Waters Corporation, Milford, MA, United States) was used for separation.…”

“…A Symmetry C18, 100 Å, 3.5 μm, 100 × 2.1 mm column (Waters Corporation, Milford, MA, United States) was used for separation. Using a previously developed HPLC-MS method, paclitaxel was quantified with an LTQ Velos Orbitrap Mass Spectrometer with Electrospray Ionization (Thermo Fisher Scientific, Waltham, MA, United States) run in positive ion mode ( Turner et al, 2017 ). Paclitaxel was detected at 854 m/z.…”