A Robust LC-MS/MS Method for Quantitative Bioanalysis of siRNA Antisense Strand in Mouse Serum and Liver

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June 12,2026

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In this work, a robust and selective LC–MS/MS method was developed for the quantification of the antisense strand of a siRNA in mouse serum and liver homogenate. The assay was performed on a Triple Quad 6500+ mass spectrometer using negative electrospray ionization and multiple reaction monitoring. Chromatographic separation was achieved using ion-pair reversed-phase UPLC with hexafluoroisopropanol (HFIP) and diisopropylethylamine (DIPEA) on a peptide BEH C18 column. Nusinersen was employed as a structurally relevant internal standard to control extraction efficiency and instrumental variability.

A-Robust-LC-MMS-Method-for-Quantitative-Bioanalysis-of-siRNA-Antisense-Strand-in-Mouse-Serum-and-Liver.jpg

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