This study demonstrates the significant advantages of an automated SPE workstation based on the Bravo platform combined with a new generation triple quadrupole liquid chromatography-mass spectrometry 6495D LC-MS/MS in oligonucleotide bioanalysis. We provide a complete solution from sample pretreatment to analysis. Mipomersen [1] (trade name: Kynamro) in pig plasma ®） Taking quantitative analysis as an example, the excellent performance of the 6495D LC-MS/MS system in high sensitivity, low detection limit, and resistance to matrix interference was demonstrated, as well as the advantages of automated SPE workstations in reducing manual intervention time, improving operational consistency, and reliability. The results indicate that this integrated automated solution can replace manual operations in high-throughput oligonucleotide bioanalysis and provide excellent sensitivity, accuracy, and reproducibility.