Separation of DNA restriction fragments by ion-pair chromatography

Eriksson, Stig; Glad, Gunnar; Pernemalm, Per-Åke; Westman, Elisabeth

doi:10.1016/0021-9673(86)80080-2

Cited by 27 publications

(12 citation statements)

References 11 publications

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“…The introduction of ion-pair reagent-containing buffers for oligonucleotide separation follows two basic goals: (i) to improve separation selectivity and (ii) to introduce charge-charge interactions to the separation mechanism so that a regular retention of oligonucleotides can be realised according to their chain length. TEAA, an inexpensive and volatile organic salt, is the most commonly chosen ion-pair reagent and allows for a good separation of oligonucleotides without desalting the collected fraction [36]. In addition, as a weak ion-pairing system, TEAA can not completely eliminate the impact of oligonucleotide sequence on its retention [7].…”

Section: Hydrophobic Order Of a And Tmentioning

confidence: 99%

Study on Retention Behaviour of Homo-Oligonucleotides in IP-RPLC Using Dual-Point Retention Time Correction on “Standard Column” with Internal Standards

Yuan¹,

Han²,

Yang³

et al. 2012

CAC

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The retention behavior of homo-oligonucleotides was investigated in ion-pair reverse-phase high-performance liquid chromatography (IP-RPLC). Rectification of retention time was performed by introducing the concept of "standard column" with double-point retention time correction (DP-RTC) using two anchor oligonucleotides as internal standards to ensure the accuracy of the retention measurement. Through polynomial fitting, a quadric relationship was adopted between normalized retention time and chain length of (dA) 10-21 and (dT) 10-21 . The retention times of homo-oligonucleotides depended on the number of bases and increased with increasing hydrophobicity, namely, A

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Section: Hydrophobic Order Of a And Tmentioning

confidence: 99%

Study on Retention Behaviour of Homo-Oligonucleotides in IP-RPLC Using Dual-Point Retention Time Correction on “Standard Column” with Internal Standards

Yuan¹,

Han²,

Yang³

et al. 2012

CAC

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“…To obtain anal. pure starting material, 1 was purified by reversedphase ion-pair HPLC [26] [27]. (8 2 )); 2.35 (m, H''ÀC(7 1 )); 2.19 (m, H'ÀC(17 1 )); 2.19 (m, H'ÀC(8 1 )); 2.16 (m, H''ÀC(3 2 )); 1.85 (m, H''ÀC(8 1 )); 1.75 (m, H'ÀC(13 1 )); 1.71 (m, H'ÀC(3 1 )); 1.46 (m, H''ÀC(3 1 )); 1.36 (m, H''ÀC(13 1 )); 1.71 (m, H'ÀC(13 2 )); 1.70 (m, H''ÀC(17 1 )); 1.34 (m, H''ÀC(13 2 )); 1.33 (m, CH 2 (5)); 1.12 (s, MeÀC (7)); 1.00 (s, MeÀC (2)).…”

Section: Experimental Partmentioning

confidence: 99%

Derivatives of Coenzyme F430 with a Covalently Attached α‐Axial Ligand. Part II

Bauer

Jaun

2003

Helvetica Chimica Acta

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Dedicated to Professor Duilio Arigoni on the occasion of his 75th birthday Coenzyme F 430 pentamethyl ester 2 was partially hydrolyzed to a mixture of the five F 430 tetramethyl esters 7 ± 11, which were separated by HPLC and identified by means of a full NMR characterization. The tetramethyl ester with a free COOH group at the side chain at C(3) of F 430 was coupled to the N-terminus of the peptidic spacerÀligand construct 12 selected and studied as described before. The UV/VIS and NMR spectra in CH 2 Cl 2 /3,3,3-trifluoroethanol 6 : 1 show that the new derivative, the Ni II (3 3 -dehydroxy-8 3 ,12 2 ,13 3 ,18 2tetra-O-methyl-F 430-3 3 -yl)-l-prolyl-l-prolyl-N p -methyl-l-histidine methyl ester (13), is an intramolecular, pentacoordinate, paramagnetic complex. In the same solvent system, the parent 3 3 ,8 3 ,12 2 ,13 3 ,18 2 -penta-Omethyl-F 430 (2) is four coordinate and diamagnetic even in the presence of equimolar 1H-imidazole. Protonation of the axially coordinating histidine residue of 13 gave the diamagnetic tetracoordinate base-off form, which allowed us to establish the constitution of 13 by NMR.

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“…The feasibility of separating nucleic acids on commercial packing materials by means of IP‐RP‐HPLC was first tried by Eriksson et al. in 1986 . Since 1992, the separation of nucleic acids by IP‐RP‐HPLC has become more and more popular .…”

Section: Introductionmentioning

confidence: 99%

Retention of nucleic acids in ion‐pair reversed‐phase high‐performance liquid chromatography depends not only on base composition but also on base sequence

Qiao

Liang

Wei

et al. 2016

J of Separation Science

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The study on nucleic acid retention in ion-pair reversed-phase high-performance liquid chromatography mainly focuses on size-dependence, however, other factors influencing retention behaviors have not been comprehensively clarified up to date. In this present work, the retention behaviors of oligonucleotides and double-stranded DNAs were investigated on silica-based C stationary phase by ion-pair reversed-phase high-performance liquid chromatography. It is found that the retention of oligonucleotides was influenced by base composition and base sequence as well as size, and oligonucleotides prone to self-dimerization have weaker retention than those not prone to self-dimerization but with the same base composition. However, homo-oligonucleotides are suitable for the size-dependent separation as a special case of oligonucleotides. For double-stranded DNAs, the retention is also influenced by base composition and base sequence, as well as size. This may be attributed to the interaction of exposed bases in major or minor grooves with the hydrophobic alky chains of stationary phase. In addition, no specific influence of guanine and cytosine content was confirmed on retention of double-stranded DNAs. Notably, the space effect resulted from the stereostructure of nucleic acids also influences the retention behavior in ion-pair reversed-phase high-performance liquid chromatography.

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Separation of DNA restriction fragments by ion-pair chromatography

Cited by 27 publications

References 11 publications

Study on Retention Behaviour of Homo-Oligonucleotides in IP-RPLC Using Dual-Point Retention Time Correction on “Standard Column” with Internal Standards

Study on Retention Behaviour of Homo-Oligonucleotides in IP-RPLC Using Dual-Point Retention Time Correction on “Standard Column” with Internal Standards

Derivatives of Coenzyme F430 with a Covalently Attached α‐Axial Ligand. Part II

Retention of nucleic acids in ion‐pair reversed‐phase high‐performance liquid chromatography depends not only on base composition but also on base sequence

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