Trace determination of lenalidomide in plasma by non-extractive HPLC procedures with fluorescence detection after pre-column derivatization with fluorescamine

Abstract

BackgroundLenalidomide (LND) is a new potent drug used for treatment of multiple myeloma. For its pharmacokinetic studies and therapeutic monitoring, a proper analytical method was required.ResultsIn this study, a non extractive and simple pre-column derivatization procedures have been proposed, for the for trace determination of lenalidomide (LND) in human plasma by HPLC with fluorescence detection. Plasma samples were treated with acetonitrile for protein precipitation then treated with copper acetate to form stable complexes with the biogenic amines and mask their interference with the derivatization reaction of LND. Treated plasma samples containing LND was derivatized with fluorescamine (FLC) in aqueous media at ambient temperature. Separation of the derivatized LND was performed on Hypersil BDS C18 column (250 × 4.6 mm, 5 μm particle size) using a mobile phase consisting of phosphate buffer (pH 4):methanol: tetrahydrofuran (70:10:20, v/v) at a flow rate of 1.0 mL/min. The derivatized samples were monitored at an emission wavelength of 495 nm after excitation at a wavelength of 382 nm. Under the optimum chromatographic conditions, a linear relationship with good correlation coefficient (r = 0.9997, n = 9) was found between the peak area and LND concentrations in the range of 2–100 ng/mL. The limits of detection and quantitation were 0.8 and 2.30 ng/mL, respectively. The intra- and inter-assay precisions were satisfactory and the accuracy of the method was proved. The recovery of LND from the spiked human plasma was 99.30 ± 2.88.ConclusionsThe proposed method had high throughput as the analysis involved simple sample pre-treatment procedure and a relatively short run-time (< 15 min). The results demonstrated that the method would have a great value when it is applied in the therapeutic monitoring and pharmacokinetic studies for LND.

Development and validation of a liquid chromatography-tandem mass spectrometric method for the determination of lenalidomide in human plasma and its application on bioequivalence studies

“…It is eliminated primarily in renal, and the mean half-life of lenalidomide is 3 h in healthy subjects and 3 to 5 h in patients (MYOPATHY n.d.;Richardson et al 2002) . Few analytical methods have been reported for the estimation of lenalidomide in bulk form and capsule dosage forms (Darwish et al 2012;Reddy et al 2012;Saravanan et al 2007;Sastry et al 2009); besides few analytical methods, determination of lenalidomide in human plasma and other biological matrices have been reported (Hasnain et al 2013;Khalil et al 2013;Ranganathan et al 2018;Saha 2019;Veeraraghavan et al 2014;Veeraraghavan et al 2015). Some of these reported methods have their individual advantages of short runtime and improved method sensitivity and sample treatment.…”

Development and Validation of an Environmentally Benign and Robust Stability Indicating Assay Method for Lenalidomide: Comprehensive Degradation Kinetics Study and Application of Synergistic Approach Involving Green Analytical Chemistry and Quality by Des

“…Though, there have been various methods available for LEN estimation, so far, to the best of our knowledge, there has not been any green method which involves combined application of GAC and QbD. [12][13][14][15] Most of these methods used ACN for method development which is not considered to be an environmentally favourable solvent or having complex mobile phase with high EAT scores. The present manuscript describes the involvement of two concepts GAC and QbD in method development and validation taking LEN as a model drug.…”

A Wide Linearity Range Method for the Determination of Lenalidomide in Plasma by High-Performance Liquid Chromatography: Application to Pharmacokinetic Studies

“…In the past years, fluorescence detection has been proposed as an alternative to mass detectors, but LND is not a fluorescent compound so precolumn derivatization with fluorescamine is necessary, which increases time of procedure and variability. 6,7 On the other hand, none of the published methods has a linearity range that allows therapeutic drug monitoring of LND for pharmacokinetic studies. Accordingly, the development of a new alternative analytical method for the determination of LND in plasma with adequate sensitivity and selectivity, low cost, and wider linearity range is needed.…”