A quantitative LC-MS/MS method for the bioanalysis of oligonucleotides in tissues

CBF 2.0 2025 -- Oligonucleotides are currently being investigated as potential therapeutic agents at preclinical and clinical stages for various diseases. There is a growing need for efficient bioanalysis of oligonucleotide therapeutics. Most recently, LC-MS/MS methodologies have emerged as a powerful option for the bioanalysis of a variety of oligonucleotide therapeutics due to the high specificity and sensitivity of these assays. Compound A, a siRNA-GalNAc conjugated oligonucleotide, is designed to reduce serum complement factor B (CFB) protein levels and efficiently inhibit complement alternative pathway (CAP) activity and is currently under preclinical development. A sensitive and specific LC-MS/MS method was developed to effectively separate and quantify the antisense strand of Compound A and successfully applied to the bioanalysis of various tissue homogenates, including liver, heart, fat, muscle, adrenal gland, thymus, kidney, lung, stomach, pancreas, duodenum, submandibular lymph nodes, uterus, brain, spleen, testis, skin, ovary and mesenteric lymph nodes.