Validation of an UPLC-MS/MS Method for Quantitative Analysis of Raltegravir in Human Plasma Samples

Abstract

This rapid and sensitive method was validated and could be applied to pharmacokinetic studies for the determination of RTG concentrations in human plasma samples.

Variability of Raltegravir Plasma Levels in the Clinical Setting

“…The method has been previously described in detail [21]. Briefly, plasma samples were extracted by liquid-liquid extraction followed by evaporation to dryness and reconstitution in mobile phase solution.…”

Selective and rapid determination of raltegravir in human plasma by liquid chromatography–tandem mass spectrometry in the negative ionization mode

“…All the reported methods employed positive ionization mode for LC–MS/MS analysis of RAL in different biological matrices [8] , [9] , [12] , [14] , [15] , [16] , [19] , [20] , [21] , [24] , [25] , [26] . Contrary to this approach, negative ionization mode was selected in the present work as it showed better selectivity without compromising the sensitivity.…”

“…The proposed method is more sensitive compared to several methods developed for determination of raltegravir in human plasma [8] , [9] , [10] , [11] , [13] , [14] , [15] , [18] , [20] , [24] , [25] , [26] . Further, it is more rapid than all other methods except one report [20] . The present method employs small plasma volume (100 µL) for processing, which is much less compared to many reported procedures.…”

“… Extraction technique Sample volume (µL) Linear range (ng/mL) Retention time (min); run time (min) Application Refs. LLE with hexane–DCM 200 2.0–1000 1.56; 3.5 To support 18 clinical studies during Phase I through Phase III [8] LLE with hexane–DCM 200 1.0–3000 3.80; 7.0 Determination of raltegravir in a single HIV-infected patient [12] LLE with hexane–DCM 500 10.0–7680 8.23; 16.0 – [13] PP with ACN–methanol 25 10–3000 1.65; 4.0 Analysis of human plasma samples [14] PP with ACN–methanol 50 50–10000 4.20; 10.0 Pharmacokinetic study in one HIV-infected patient [15] LLE 100 5.0–2560 –; 1.0 Pharmacokinetic study in human plasma samples [20] PP with ACN 50 2.0–2000 nmol/L 4.90; 9.0 Pharmacokinetic study with 400 mg raltegravir in 6 healthy subjects [21] LLE with hexane–DCM 100 2.0–6000 1.35; 2.0 Bioequivalence study with 400 mg raltegravir in 18 healthy subjects; Reanalysis of 87 incurred samples (% change within±17%) PW LLE: liquid–liquid extraction; PP: protein precipitation; SPE: solid phase extraction; DCM: dichloromethane; ACN: acetonitrile; PW: present work. …”

“…Selective and sensitive determination of anti-HIVs in plasma is essential for studying drug–drug interaction, pharmacokinetics, pharmacodynamics and therapeutic drug monitoring. Several methods are reported for the determination of RAL as a single analyte [8] , [9] , [10] , [11] , [12] , [13] , [14] , [15] , [16] , [17] , [18] , [19] , [20] or in combination with its glucuronide (Glu) metabolite [21] or other anti-HIV drugs [22] , [23] , [24] , [25] , [26] in different biological samples such as human cell extracts [15] , [19] , cerebrospinal fluid [16] , cervicovaginal fluid [17] , dried blood spots [15] , bile [9] , feces [9] and human plasma [8] , [10] , [11] , [12] , [13] , [14] , [18] , [20] , [21] , [22] , [23] , [24] , [25] , [26] . Mainly, liquid chromatography with UV [10] , [17] , [23] , photodiode array [18] , [22] , fluorescence [11] or mass spectrometry [8] , [9] , [12] , [13] , [14] , [15] , [16] , [19] , [20] , [21] , [24] , [25] , [26] detection has been used for the quantification of RAL in these matrices.…”