[Case Study] Method Transfer for HPLC Purification and Analysis of Oligonucleotides
Streamlining the process from high-purity fractionation of double-labeled oligos to quality confirmation using HRAM-MS.
This application note presents a case study of transferring the purification and analysis workflow of synthetic oligonucleotides from a competitor's HPLC system to the Thermo Scientific Vanquish HPLC system. Focusing on dual-labeled oligonucleotide samples, the workflow was validated through purification via semi-preparative RP-HPLC, purity assessment using HPLC-UV, and identity confirmation with HRAM-MS. A comparison using the Hypersil GOLD column demonstrated the ability to isolate the target oligonucleotides with high purity, achieving efficient quality control. The note provides practical comparative data on retention behavior, peak shape, and collection capacity differences during method transfer. **Features** - Case study on the transfer from a competitor's LC purification system to the Vanquish analytical purification LC system. - Validation of purification, purity assessment, and identity confirmation of dual-labeled oligonucleotides as a complete workflow. - Comparison of separation performance between the Hypersil GOLD column and competitor columns. - Explanation of differences in peak shape and collection capacity with specific chromatograms. - Introduction of an efficient quality control method combining HPLC-UV and HRAM-MS.
basic information
Vanquish Analysis Refined LC System Configuration Pump: Thermo Scientific Vanquish Binary Pump F Autosampler: Thermo Scientific Vanquish Split Sampler FT Column Oven: Thermo Scientific Vanquish Column Compartment H Detector: Thermo Scientific Vanquish Diode Array Detector FG (Standard Biocompatible Flow Cell) Fraction Collector: Thermo Scientific Vanquish Fraction Collector
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Thermo Fisher Scientific K.K., the Japanese subsidiary of Thermo Fisher Scientific Incorporated, is a sales company for comprehensive system analysis instruments, bio-related equipment, and research reagents. In Japan, it has bases in Yokohama, Tokyo, Osaka, Fukuoka, and Nagoya, handling various analytical instruments including mass spectrometers, various bio-related equipment, measuring instruments, medical devices, lab software, and research reagents.