Orthogonal method development and validation of reverse phase ultra-performance liquid chromatographic-mass spectrometry (using PDA and QDa mass detector) for quantification of temsirolimus in temsiro

In this study a sensitive, specific and stability-indicating Ultra performance liquid chromatographic (Waters ACQUITY UPLC H-class PDA with integrated QDa mass detector) assay method is developed and validated for the quantification of Temsirolimus in Temsirolimus injection. UPLC separation is achieved with a YMC Pack Pro C18 RS (100 mm x 4.6mm id x 3µm) as stationary phase and 0.05% Trifluoro acetic acid (pH adjusted to 3.0 with ammonia solution):Methanol : Acetonitrile (25:60:15, v/v) as eluent, with a flow rate 0.7 ml/min. UV detection was performed at 280 nm. The retention time of Temsirolimus peak is about 20 minutes. The method has been fully validated and is linear. Results of analysis are validated statistically and by recovery studies. The standard and sample solutions are stable up to 24 hours at 5˚C. Temsirolimus was found to degrade in all stress conditions. Purity of Temsirolimus was found to be less than purity threshold and no additional mass interference was found at Temsirolimus peak in all controlled and stressed samples. No interference was found from corresponding stressed blank with Temsirolimus peak. This method offers advantages over using photodiode-array UV detection (LC-PDA) for the determination of UPLC peak purity, namely components with similar UV spectra can be distinguished, the molecular mass of the impurity can be determined and structural data can be obtained by using QDa mass detector. The result of studies showed that the proposed RP-UPLC method is found to be precise, linear, accurate, rugged, selective, specific, and robust and stability indicating. Hence this method can be used for the routine analysis in bulk drug and in its pharmaceutical dosage forms.