scholarly journals Attomol-level quantification of chemically modified ribonucleosides enabled by capillary porous graphitic carbon columns in nano LC-MS

2017 ◽  
Author(s):  
L. Peter Sarin ◽  
Sandra D. Kienast ◽  
Johannes Leufken ◽  
Robert L. Ross ◽  
Patrick A. Limbach ◽  
...  
Keyword(s):  
Biological Samples ◽  
Stationary Phases ◽  
Reversed Phase ◽  
Porous Graphitic Carbon ◽  
Graphitic Carbon ◽  
Detection Range ◽  
Rna Molecules ◽  
Chemically Modified ◽  
High Resolution Analysis ◽  
Resolution Analysis

ABSTRACTPost-transcriptional chemical modifications of (t)RNA molecules are crucial in fundamental biological processes, such as translation. Despite their biological importance and accumulating evidence linking them to various human diseases, technical challenges have limited the development of methods for reliable detection and accurate quantification of these modifications. Here, we present a sensitive capillary nanoflow liquid chromatography mass spectrometry (nLC-MS) pipeline for quantitative high-resolution analysis of ribonucleoside modifications from complex biological samples. We evaluated two porous graphitic carbon (PGC) materials as stationary phases for reversed-phase separation of ribonucleosides and found that both PGC matrices have excellent retention and separation characteristics, as well as the capability to separate structural isomers. Using PGC matrices in nLC-MS yielded excellent signal-to-noise ratios in a detection range spanning up to six orders of magnitude, allowing for the analysis of individual ribonucleosides down to attomol concentrations. Furthermore, normalizing the obtained signal intensities to a stable isotope labeled spike-in enabled direct comparison of ribonucleoside levels between different samples. In conclusion, capillary PGC columns coupled to nLC-MS constitute a powerful and sensitive tool for quantitative analysis of chemically modified ribonucleosides in complex biological samples. This setup will be invaluable for further unraveling the intriguing and multifaceted biological roles of RNA modifications.

Comparison of hydrophilic-interaction, reversed-phase and porous graphitic carbon chromatography for glycan analysis

2011 ◽  
Vol 1218 (1) ◽  
pp. 118-123 ◽  
Author(s):  
Michael Melmer ◽  
Thomas Stangler ◽  
Andreas Premstaller ◽  
Wolfgang Lindner
Keyword(s):  
Reversed Phase ◽  
Porous Graphitic Carbon ◽  
Graphitic Carbon ◽  
Hydrophilic Interaction ◽  
Glycan Analysis

Evaluation of reversed phase columns designed for polar compounds and porous graphitic carbon in “trapping” and separating neurotransmitters

2006 ◽  
Vol 1122 (1-2) ◽  
pp. 97-104 ◽  
Author(s):  
Didier Thiébaut ◽  
Jérôme Vial ◽  
Monika Michel ◽  
Marie-Claire Hennion ◽  
Tyge Greibrokk
Keyword(s):  
Polar Compounds ◽  
Reversed Phase ◽  
Porous Graphitic Carbon ◽  
Graphitic Carbon

scholarly journals Comparison of Various Chromatographic Systems for Analysis of Cytisine in Human Serum, Saliva and Pharmaceutical Formulation by HPLC with Diode Array, Fluorescence or Mass Spectrometry Detection

Molecules ◽  
2019 ◽  
Vol 24 (14) ◽  
pp. 2580 ◽  
Author(s):  
Karol Wróblewski ◽  
Anna Petruczynik ◽  
Tomasz Tuzimski ◽  
Dominika Przygodzka ◽  
Grzegorz Buszewicz ◽  
...  
Keyword(s):  
Mass Spectrometry ◽  
Liquid Chromatography ◽  
Cation Exchange ◽  
Biological Samples ◽  
Stationary Phases ◽  
Reversed Phase ◽  
Pharmaceutical Formulation ◽  
Diode Array ◽  
Strong Cation Exchange ◽  
Pre Treatment

Background: Identification and quantitative determination of cytisine, especially in biological samples and pharmaceutical formulations, is still a difficult analytical task. Cytisine is an alkaloid with a small and very polar molecule. For this reason, it is very weakly retained on reversed phase (RP) stationary phases, such as commonly used alkyl-bonded phases. The very weak retention of cytisine causes it to be eluted together with the components of biological matrices. Objective: Comparison and evaluation of various chromatographic systems for analysis of cytisine in different matrices—serum, saliva and pharmaceutical formulation—by high performance liquid chromatography (HPLC) with diode array (DAD), fluorescence (FLD) and mass spectrometry (MS) detection. Methods: The analyses were performed using HPLC in reversed phase (RP), hydrophilic interaction liquid chromatography (HILIC) and ion exchange chromatography (IEC) modes. Different sample pre-treatment methods were tested: Protein precipitation (with acetone, methanol (MeOH) or acetonitrile (ACN), and solid phase extraction (SPE) using cartridges with octadecyl (C18), hydrophilic-lipophilic balanced copolymer (HLB) or strong cation exchange sorbents (Strata X-C). Conclusion: Significant differences were observed in retention parameters with a change of the used chromatographic system. The various properties of stationary phases resulted in differences in analyte retention, peaks’ shape and systems’ efficiency. The weakest retention was observed using RP systems; however, the use of the Polar RP phase can be an alternative for application in green chromatography. In the strongest retention was observed using a strong cation exchange (SCX) phase. The most optimal systems were chosen for the analysis of cytisine in the pharmaceutical preparation, serum and saliva after sample pre-treatment with the new SPE procedure. Due to the sensitivity, the use of HPLC-DAD or HPLC-FLD is the most optimal for drug analysis in pharmaceutical preparations, whereas HPLC-MS is suitable for analysis of cytisine in biological samples.

Online Two-Dimensional Porous Graphitic Carbon/Reversed Phase Liquid Chromatography Platform Applied to Shotgun Proteomics and Glycoproteomics

2014 ◽  
Vol 86 (24) ◽  
pp. 12172-12179 ◽  
Author(s):  
Yun Zhao ◽  
Samuel S. W. Szeto ◽  
Ricky P. W. Kong ◽  
Chun Hin Law ◽  
Guohui Li ◽  
...  
Keyword(s):  
Liquid Chromatography ◽  
Shotgun Proteomics ◽  
Reversed Phase ◽  
Porous Graphitic Carbon ◽  
Graphitic Carbon ◽  
Two Dimensional ◽  
Reversed Phase Liquid Chromatography ◽  
Phase Liquid

scholarly journals Abnormal retention of s-triazine herbicides on porous graphitic carbon

2021 ◽  
Vol 41 (1) ◽  
pp. 1-9
Author(s):  
Oksana Grinevich ◽  
Zoya Khesina ◽  
Alexey Buryak
Keyword(s):  
Stationary Phase ◽  
High Performance ◽  
Reversed Phase ◽  
Retention Mechanism ◽  
Porous Graphitic Carbon ◽  
Graphitic Carbon ◽  
Chromatographic Behavior ◽  
Triazine Herbicides ◽  
Pesticide Monitoring ◽  
Modeling Of Adsorption

Abstract Porous graphitic carbon (PGC) is a widely used stationary phase for reversed-phase high-performance liquid chromatography (HPLC) that allows separation of structurally similar compounds retained in mixed form on a flat graphite surface. Such a stationary phase can be used in analytical chemistry to provide good separation and selectivity in pesticide monitoring. In this article, we studied the chromatographic behavior of five common triazine herbicides (simazine, atrazine, desmetryn, propazine, prometryn) on PGC vis-à-vis octadecyl-functionalized silica gel (ODS). It was found that the herbicides studied have an abnormal elution order on PGC compared to ODS. PGC was also characterized by higher selectivity of analyte separation. This behavior of triazine herbicides on PGC cannot be explained either with the help of existing theory or by mathematical modeling of adsorption processes on graphite. Therefore, we have proposed a possible retention mechanism, explaining the effects observed, due to the shielding of the amino group in the triazine ring by alkyl substituents, which decreases the “polar retention effect” of PGC. Satisfactory separation efficacy was obtained with the proposed analytical method, using convenient UV-detection and without resort to laborious techniques such as HPLC coupled with mass spectrometry.

Online combination of reversed-phase/reversed-phase and porous graphitic carbon liquid chromatography for multicomponent separation of proteomics and glycoproteomics samples

2011 ◽  
Vol 32 (21) ◽  
pp. 2930-2940 ◽  
Author(s):  
Maggie P. Y. Lam ◽  
Edward Lau ◽  
S. O. Siu ◽  
Dominic C. M. Ng ◽  
Ricky P. W. Kong ◽  
...  
Keyword(s):  
Liquid Chromatography ◽  
Reversed Phase ◽  
Porous Graphitic Carbon ◽  
Graphitic Carbon ◽  
Multicomponent Separation
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