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

• Published in: Journal of pharmaceutical analysis 2015-04, Vol.5 (2), p.101-109
• Main Authors: Gupta, Ajay, Guttikar, Swati, Shah, Priyanka A., Solanki, Gajendra, Shrivastav, Pranav S., Sanyal, Mallika
• Format: Article
• Language: English
• Subjects: Bioequivalence study; C6000; Human plasma; LC–ESI–MS/MS; Negative ionization mode; Original; Raltegravir; 人血浆; 子模式; 快速测定; 生物等效性; 色谱选择性; 质谱法; 高效液相色谱
• ISSN: 2095-1779; 2214-0883
• DOI: 10.1016/j.jpha.2014.10.002

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 mL human plasma. The chromatographic separation was achieved on a Chromolith RP-18 e 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/m L.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.