ABSTRACT
A sensitive and accurate LC/MS method for the determination of elbasvir (ELB) and grazoprevir (GZP) in human plasma was established using daclatasvir (DCT) as an internal standard. The analytes were separated on a Waters Spherisorb phenyl column (150mm×4.6mm ID, 5µm particle size) maintained at 40°C±2°C. Gradient elution, at a flow rate of 0.8mLmin-1, was used. The mobile phase consists of 90% of acetonitrile mixed to 10% of a 5mM ammonium formate buffer (+0.1% v/v of trimethylamine, pH was adjusted to 3.2 by formic acid) as phase A and 10% of acetonitrile mixed to 90% of the same buffer as phase B. Liquid-liquid extraction with ethyl acetate solvent was used to recuperate compounds from plasma. The method was validated over a concentration range of 2 and 100ng/mL for GZP and between 1 and 50ng/mL for ELB. The intra- and inter-day precision and accuracy of the quality control samples at low, medium, and high concentration levels exhibited relative standard deviations (RSD)<15%, and the accuracy values ranged from 94.2 to 107.8%. The robustness of the method was established using a two-level full factorial design.
ABSTRACT
Many antiviral drugs were developed to counteract coronavirus disease, 2019 (COVID-19) with severe acute respiratory syndrome. Therefore, the scientific community's efforts have focused on the detection and quantification of antiviral compounds currently being tested for COVID-19 treatment. Cuttlefish bone powder (CFBP) has been used for the first time as solid-phase extraction (SPE) sorbent for the extraction of SARS CoV-2 antiviral drugs (chloroquine, ritonavir and indomethacin) from water samples. An effective and sensitive method was developed by combining SPE and liquid chromatography- UV detection (LC-UV). An experimental design was applied for the optimization of extraction process. Experimental variables were optimized using Doehlert matrix. The developed method included 50 mg of CFBP sorbent, 20 mL of water sample at pH = 9 and 5 mL of ACN/KH2PO4 buffer solution (80:20, v/v) in the elution step. For validation of the method, selectivity, linearity precision, and sensitivity were evaluated. Extraction recovery percentage of all Sars cov-2 antivirals were above 98.2%. The detection and quantification limits were between 0.1 and 0.5 µg L-1 and 0.6 and 2 µg L-1, respectively. The current study suggested that CFBP has the application potential for the enhanced SPE of SARS CoV-2 antiviral drugs from water samples.
