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

Abstract

A selective and rapid high-performance liquid chromatography–tandem mass spectrometry method was developed and validated for the quantification of raltegravir using raltegravir-d3 as an internal standard (IS). The analyte and IS were extracted with methylene chloride and n-hexane solvent mixture from 100 µL human plasma. The chromatographic separation was achieved on a Chromolith RP-18e endcapped C18 (100 mm×4.6 mm) column in a run time of 2.0 min. Quantitation was performed in the negative ionization mode using the transitions of m/z 443.1→316.1 for raltegravir and m/z 446.1→319.0 for IS. The linearity of the method was established in the concentration range of 2.0–6000 ng/mL. The mean extraction recovery for raltegravir and IS was 92.6% and 91.8%, respectively, and the IS-normalized matrix factors for raltegravir ranged from 0.992 to 0.999. The application of this method was demonstrated by a bioequivalence study on 18 healthy subjects.

Tandem mass spectrometry of small-molecule antiviral drugs: 1. HIV-related antivirals

“…The analysis of raltegravir ([M−H] – with m/z 443) in negative-ion mode has also been reported, using the product-ions with m/z 316 ([C 16 H 15 FN 3 O 3 ] – with m/z 316.110), which is due to the loss of 5-methyl-1,3,4-oxadiazole-2-carboxamide (C 4 H 5 N 3 O 2 ). In addition, in negative-ion mode, product ions are observed with m/z 276 ([C 13 H 11 FN 3 O 3 ] – with m/z 276.079) due to the loss of 5-methyl- N -(prop-1-en-2-yl)-1,3,4-oxadiazole-2-carboxamide (C 7 H 9 N 3 O 2 ) and with m/z 165, possibly an ion with m/z 165.067 ([C 8 H 9 N 2 O 2 ] – ) [ 96 , 97 ]. No accurate- m/z data were found.…”

Silica-Based Monolithic Columns as a Tool in HPLC—An Overview of Application in Analysis of Active Compounds in Biological Samples

“…Some reports presented the usefulness of monoliths in the analysis of urine, saliva [ 74 , 75 ], whole blood [ 76 ], plasma [ 75 , 77 ], serum, and human breast milk samples [ 78 ]. The papers described the chromatographic analysis of compounds from different therapeutic groups, such as antibiotics [ 25 , 77 , 79 , 80 ], diuretics [ 75 , 77 , 81 ], antidepressants [ 82 , 83 ], analgesics [ 74 ], antidiabetics [ 84 , 85 ], benzodiazepine derivatives [ 76 , 86 , 87 ], and anti-allergic [ 88 ] and antiviral drugs [ 26 , 89 ]. Table 2 presents examples of applications of monolithic columns in medical analysis.…”

Quantitative Reverse-phase High-performance Liquid Chromatographic Method for the Quantification of Raltegravir Potassium in Bulk and Dosage Forms

“…The mobile phase was ammonium formate buffer (pH 4.0): acetonitrile (30:70) in isocratic mode. 5 Pardhi SS and co-workers, in their review article, stated that Highly Active Antiretroviral Therapy (HAART), a combination drug therapy is a topic of current interest in the treatment of HIV and AIDS. Techniques for the analysis and the quality control of antiretroviral drugs, particularly in the drug combinations are vital in achieving the quality of these drugs and the treatments involved.…”