Dried blood spot analysis of (+) and (−) darunavir enantiomers on immobilized amylose tris‐(3, 5‐dimethylphenylcarbamate) LC and its application to pharmacokinetics

Abstract

Dried blood spot analysis is an innovative novel blood sampling technique gaining interest in drug discovery and development processes owing to its inherent advantages over the conventional whole blood, plasma or serum sample collection. The present manuscript describes the development and validation of a highly sensitive and precise method of evaluation of pharmacokinetics of (+) and (-) darunavir enantiomers on rat dried blood spots. The enantiomers on rat dried blood spots were extracted into methanol and separated by LC on a Chiralpak IA column using hexane and ethanol containing 0.1% DEA (75:25, v/v) as a mobile phase at 20°C; both the enantiomers were detected at 266 nm using a photodiode array detector. The method was validated in terms of selectivity, linearity, accuracy, precision and stability as per the US Food and Drug and Administration guidelines. The hematocrit effect on extraction recovery was evaluated and the mean recoveries of (-) and (+) enantiomers of darunavir from dried blood spots were found to be 85.76 and 88.91% respectively. The intra- and inter-day precision and accuracy were 3.1-8.4 and 0.8-4.8% respectively. The developed method was successfully applied to a pharmacokinetic study of (+) and (-) enantiomers of darunavir on rat dried blood spots.

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“…Furthermore, many studies have been devoted to the dried samples (i.e. blood, plasma, breast milk) spots [ [34] , [35] , [36] , [37] , [38] , [39] , [40] , [41] ]. However, a lack of standard validation guidelines for drug quantification in dried samples spots is ascertained [ 34 ].…”

“…A separate discussion is needed for the preparation of dried sample spots [ [34] , [35] , [36] , [37] , [38] , [39] , [40] , [41] ]. This low-cost technology has recently received a great deal of attention because of their distinct advantages of sample collection, less invasive sampling, simpler transfer, storage and shipping [ 104 , 105 ].…”

“…Recently, LC-UV methods have been also developed for drug enantiomers detection in dried blood spots, by using chiral columns, namely (±) darunavir and (±) emtricitabine [ 37 , 38 ]. In this case, a Chiralpak IA column (250 mm x 4.6 mm, 5 μm) has been employed and the chromatographic separation has been carried out isocratically with n-hexane-ethanol-diethyl amine (75:25:0.1) or with n-hexane-ethanol (65:35) as mobile phases.…”