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[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.

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HPLC & UHPLC Comprehensive Catalog 2023-2024 Edition

Worldwide bestseller LC

Since Agilent launched the HP1010 LC system in 1973, it has consistently incorporated customer feedback and focused on improving system performance. During this time, there was a period when we had development and manufacturing bases in Germany and Japan (Yokogawa Electric Corporation, Yokogawa Analytical Systems), but now all LC systems are "Made in Germany." By integrating the latest technologies, we have been recognized as a worldwide bestseller in LC systems, addressing various needs such as pharmaceutical development, food safety assurance, chemical and material analysis, and fractionation, while also complying with global regulations. Currently, Agilent has formed a teamwork approach among LC instruments, columns, consumables, mass spectrometers, software, and service/support under the InfinityLab series, continuously evolving to maximize daily analysis, equipment, and lab efficiency.

  • Analytical Equipment and Devices
  • Contract measurement
  • HPLC

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