Mejora de la forma del pico de oligonucleótidos utilizando la función de pretratamiento automática (coinyección)

Beneficios para el usuario

- Sample dilution can be automated using the automatic pretreatment function “co-injection” for purified fractions of oligonucleotides containing high concentrations of salts. - The automatic pretreatment function “co-injection” enables the simultaneous injection of a sample and a selected co-injection solvent. The co-injection solvent and its injection volume can be easily set via LabSolutions MD interface.

Introducción

In HPLC analysis, the composition of the sample solvent is critical for achieving proper peak shape. Si el solvente de la muestra es un solvente de elución más fuerte que la fase móvil, la condensación de la banda de la muestra en la entrada de la columna puede ser insuficiente, lo que provocaría el ensanchamiento de la banda de la muestra. Por ejemplo, el aumento de la proporción de solvente orgánico en el solvente de muestra para disolver compuestos de baja polaridad puede deteriorar las formas pico de los compuestos de elución temprana en la cromatografía de fase inversa. Oligonucleotides are synthesized using the phosphoramidite method and subsequently purified by anion exchange chromatography or reversed-phase ion-pair chromatography (RP-IP). Purified fractions from anion exchange chromatography often contain high concentrations of salts, such as sodium chloride (NaCl) and sodium bromide (NaBr). When these purified fractions are analyzed using RP-IP, the high salt concentration in the sample solvent may interfere with the ion-pair formation capacity of the oligonucleotides. This interference can result in improper peak retention and adversely affect the chromatogram. This article introduces a case study demonstrating an improvement in oligonucleotide peak shape by suppressing the sample solvent effect through simultaneous injection with a desired solvent. This automatic pretreatment function “co-injection” is a standard feature included in the autosamplers of Nexera series. Furthermore, by utilizing LabSolutions MD, a dedicated software for supporting method development, comprehensive evaluation of co-injection solvent types and co-injection volumes was efficiently performed.

18 de noviembre de 2025 GMT