Monolithic alkylsilane column: A promising separation medium for oligonucleotides by ion-pair reversed-phase liquid chromatography

Qiao, Jun-qin; Liang, Chao; Zhu, Zhenyu; Cao, Zhao-ming; Zheng, Wei-juan; Lian, Hong-zhen

doi:10.1016/j.chroma.2018.07.062

Cited by 12 publications

(12 citation statements)

References 51 publications

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“…In the present derivatization method, 24 nucleotides were quantified with a total analysis cycle of 18 min, possessing more quantified analytes and shorter run time than previous ion-pairing and HILIC methods (Table ). − Moreover, no significant decrease in column performance was observed in this method which is superior to ion-pairing methods. Peak shapes of all analytes in this method were as good as that obtained in our previous method (Figure ) while peak tailing is hardly avoidable in HILIC methods. − In addition, positive and negative ion mode switch was required in the ion-pairing method in order to overcome the interference of ATP on dGTP quantification because they shared the same ion transition in negative ion mode.…”

Section: Discussionmentioning

confidence: 84%

“…Currently, the predominant method for analysis of cellular nucleotide is ion-pairing LC-MS method, − which is also used in our previous studies. − The difference between reported ion-pairing LC-MS methods mainly shows in the kind and amount of ion-pairing regents. Over the past several years, HILIC-MS method has been investigated and applied in nucleotide analysis as an alternative method. − Therefore, here we made a comparison of this derivatization method with ion-pairing and HILIC methods.…”

Section: Discussionmentioning

confidence: 99%

“…In addition, several derivatization methods have been introduced for the quantification of nucleotides using RPLC-MS/MS. , However, those derivatization reactions are complex and time-consuming and present limited applicability. Another widely used approach for nucleotide analysis is RPLC-MS/MS with ion-pairing agents. − We have also developed an ion-pairing LC-MS/MS method to determine RNs and dRNs and investigated their intracellular levels perturbed by different nucleoside analogues. − Although ion-pairing method demonstrates great reproducibility and good peak shapes for the nucleotides, ion-pairing agents are unfriendly to the LC-MS system due to diminished column life, inevitable system contamination, and notable ion suppression in the electrospray ionization source (ESI). Recently, a few hydrophilic interaction chromatography (HILIC) studies on the separation of nucleotides have been reported using different types of stationary phases. − However, to date HILIC still encounters some inevitable defects in analysis of nucleotides.…”

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confidence: 99%

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Method for Quantification of Ribonucleotides and Deoxyribonucleotides in Human Cells Using (Trimethylsilyl)diazomethane Derivatization Followed by Liquid Chromatography–Tandem Mass Spectrometry

Zhang

et al. 2018

Anal. Chem.

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Investigation into intracellular ribonucleotides (RNs) and deoxyribonucleotides (dRNs) is important for studies of the mechanism of many biological processes, such as RNA and DNA synthesis and DNA repair, as well as metabolic and therapeutic efficacy of nucleoside analogues. However, current methods are still unsatisfactory for determination of nucleotides in complex matrixes. Here we describe a novel method for the determination of RN and dRN pools in cells based on fast derivatization with (trimethylsilyl)diazomethane (TMSD) followed by quantification using liquid chromatography–tandem mass spectrometry (LC-MS/MS). Derivatization was accomplished in 3 min, and each derivatized nucleotide not only had a sufficient retention on reversed-phase column by introduction of methyl groups but also exhibited a unique ion transition which consequently eliminated mutual interference in LC-MS/MS. Chromatographic separation was performed on a C18 column with a simple acetonitrile–water gradient elution system, which avoided contamination and ion suppression caused by ion-pairing reagents. The developed method was fully validated and applied to the analysis of RNs and dRNs in cell samples. Moreover, results demonstrated that the applicability of this method could be extended to nucleoside analogues and their metabolites and could facilitate many applications in future studies.

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Section: Discussionmentioning

confidence: 84%

Section: Discussionmentioning

confidence: 99%

mentioning

confidence: 99%

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Method for Quantification of Ribonucleotides and Deoxyribonucleotides in Human Cells Using (Trimethylsilyl)diazomethane Derivatization Followed by Liquid Chromatography–Tandem Mass Spectrometry

Zhang

et al. 2018

Anal. Chem.

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“…These include C18 with polar end-capped groups [92], pentafluorophenyl-based stationary phases [86], as well as styrenedivinylbenzene [93] and phenyl-based resins [94]. are always less retained than random coils on monoliths, while no retention was observable with particle-packed columns [96]. This has been attributed to the fact that hairpins are more easily deformed into particle-packed columns, leading to more complex retention behaviours.…”

Section: Analytical Characterization Of Oligonucleotidesmentioning

confidence: 99%

Oligonucleotides: Current Trends and Innovative Applications in the Synthesis, Characterization, and Purification

Catani

Luca

Alcântara

et al. 2020

Biotechnology Journal

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Oligonucleotides (ONs) are gaining increasing importance as a promising novel class of biopharmaceuticals. Thanks to their fundamental role in gene regulation, they can be used to develop custom-made drugs (also called N-to-1) able to act on the gene expression at pretranslational level. With recent approvals of ON-based therapeutics by FDA, a growing demand for high-quality chemically-modified ONs is emerging and their market is expected to impressively prosper in the very next future. To satisfy this growing market demand, a scalable and economically sustainable ON production is needed. In this paper, the state of the art of the whole ON production process is illustrated with the aim of highlighting the most promising routes towards the auspicated market-size production. In particular, the most recent advancements in both the upstream stage, mainly based on solid-phase synthesis and recombinant technology, and the downstream one, focusing on chromatographic techniques, are reviewed. Since ON production is projected to expand to the large scale, automatized multi-column technologies will reasonably be required soon to replace the current ones based on batch single-column operations. Cutting edge purification solutions, based on continuous chromatography, will be thus presented in the last part of this review.

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“…Although the octadecyl silane (C18)‐modified monolithic silica column has been widely applied in protein/peptide separation by RPLC, it was difficult to separate polar and ionized solutes in RPLC . In order to better meet the requirements of numerous applications for pharmaceutical, biomedical and clinical analysis, monolithic columns enable to independent control their porous structure and surface property to offer diverse selectivities .…”

Section: Basic Strategy For Designing Monolithic Materialsmentioning

confidence: 99%

Challenges and Advances in the Fabrication of Monolithic Bioseparation Materials and their Applications in Proteomics Research

Wang

et al. 2019

Advanced Materials

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High‐performance liquid chromatography integrated with tandem mass spectrometry (HPLC–MS/MS) has become a powerful technique for proteomics research. Its performance heavily depends on the separation efficiency of HPLC, which in turn depends on the chromatographic material. As the “heart” of the HPLC system, the chromatographic material is required to achieve excellent column efficiency and fast analysis. Monolithic materials, fabricated as continuous supports with interconnected skeletal structure and flow‐through pores, are regarded as an alternative to particle‐packed columns. Such materials are featured with easy preparation, fast mass transfer, high porosity, low back pressure, and miniaturization, and are next‐generation separation materials for high‐throughput proteins and peptides analysis. Herein, the recent progress regarding the fabrication of various monolithic materials is reviewed. Special emphasis is placed on studies of the fabrication of monolithic capillary columns and their applications in separation of biomolecules by capillary liquid chromatography (cLC). The applications of monolithic materials in the digestion, enrichment, and separation of phosphopeptides and glycopeptides from biological samples are also considered. Finally, advances in comprehensive 2D HPLC separations using monolithic columns are also shown.

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Monolithic alkylsilane column: A promising separation medium for oligonucleotides by ion-pair reversed-phase liquid chromatography

Cited by 12 publications

References 51 publications

Method for Quantification of Ribonucleotides and Deoxyribonucleotides in Human Cells Using (Trimethylsilyl)diazomethane Derivatization Followed by Liquid Chromatography–Tandem Mass Spectrometry

Method for Quantification of Ribonucleotides and Deoxyribonucleotides in Human Cells Using (Trimethylsilyl)diazomethane Derivatization Followed by Liquid Chromatography–Tandem Mass Spectrometry

Oligonucleotides: Current Trends and Innovative Applications in the Synthesis, Characterization, and Purification

Challenges and Advances in the Fabrication of Monolithic Bioseparation Materials and their Applications in Proteomics Research

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