Hydrophilic additives enhancing electrospray ionization of oligonucleotides in hexafluoro-2-propanol-free reversed-phase ion-pair liquid chromatography/mass spectrometry

Miyazaki, Takashi; Asano, Natsuyo; Yamaki, Satoshi et al.

Journal of Chromatography A, 2025, 1765, 466485

アクセス数:67件（2026-02-16 04:11 集計）

固定URL: https://hdl.handle.net/11094/103521

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タイトル
Hydrophilic additives enhancing electrospray ionization of oligonucleotides in hexafluoro-2-propanol-free reversed-phase ion-pair liquid chromatography/mass spectrometry
著者
著者名
Miyazaki, Takashi
著者名
Asano, Natsuyo
著者名
Yamaki, Satoshi
NRID
1000080596601 1000080596601
ORCID
0000-0003-3180-0257 0000-0003-3180-0257
researchmap permalink
takao-yamaguchi takao-yamaguchi
Scopus Author ID
56322895500 56322895500
著者名
Yamaguchi, Takao
NRID
1000080243252 1000080243252
ORCID
0000-0002-6842-6812 0000-0002-6842-6812
researchmap permalink
read0014368 read0014368
Scopus Author ID
7003800550 7003800550
著者名
Obika, Satoshi
キーワード等
1,1,1,3,3,3-Hexafluoro-2-propanol
Electrospray ionization
Hydrophilic additive
Liquid chromatography/mass spectrometry
Oligonucleotide
Reversed-phase ion-pair chromatography
内容
Miyazaki T., Asano N., Yamaki S., et al. Hydrophilic additives enhancing electrospray ionization of oligonucleotides in hexafluoro-2-propanol-free reversed-phase ion-pair liquid chromatography/mass spectrometry. Journal of Chromatography A 1765, 466485 (2026); https://doi.org/10.1016/j.chroma.2025.466485.
抄録
The characterization of oligonucleotide therapeutics is commonly performed by liquid chromatography/mass spectrometry (LC/MS) using reversed-phase ion-pair (RP-IP) chromatography with 1,1,1,3,3,3-hexafluoro-2-propanol (HFIP) and triethylamine (TEA) due to its efficient electrospray ionization (ESI) performance. However, HFIP poses several challenges, including environmental concerns as a per- and polyfluoroalkyl substance (PFAS), limited mobile phase stability, and high reagent costs. Traditional HFIP-free alternatives, such as acetic acid (AA) and TEA, generally result in lower MS sensitivity. This study aimed to develop an HFIP-free analytical method for oligonucleotides and explored the use of AA/TEA-based mobile phase additives to improve MS sensitivity. An AA/glycine/TEA-based mobile phase prepared by adding glycine to an AA/TEA-based mobile phase, exhibited significantly enhanced MS sensitivity compared to the original AA/TEA system, reaching levels comparable to those achieved with HFIP/TEA. Comprehensive screening of amino acids revealed that aliphatic amino acids, particularly those with high hydrophilicity, contributed to enhanced ionization. These findings suggest that hydrophilic additives may promote the localization of oligonucleotides at the droplet surface and facilitate ion release, thereby enhancing ionization efficiency. Moreover, the AA/glycine/TEA system maintained MS sensitivity for a longer duration than the HFIP/TEA system. Although glycine is typically avoided in LC/MS due to its non-volatile nature, its use at sufficiently low concentrations did not lead to contamination or performance degradation. These findings support the potential of hydrophilic additives like glycine to enhance ionization efficiency in HFIP-free RP-IP-LC/MS for oligonucleotide analysis.
出版者
Elsevier B.V.
収録物名
Journal of Chromatography A
巻(号)
1765
ページ
466485
刊行年月
2025-10-29
言語
英語
ハンドルURL
https://hdl.handle.net/11094/103521
PISSN
00219673
eISSN
18733778
関連情報 (isIdenticalTo)
DOI (出版社版)
https://doi.org/10.1016/j.chroma.2025.466485
アクセス権
open access
権利情報
This article is licensed under a Creative Commons Attribution 4.0 International License.
出版タイプ
Version of Record
カテゴリ
学術雑誌論文 Journal Article