Synthesis and Characterisation of a New Hydrophilic Interaction/Reversed Phase Mixed-Mode Chromatographic Stationary Phase
A novel multifunctional stationary phase based the amide-silica was synthesised starting from L-isoleucine
and 2-aminoacetanilide. Hereby, compounds 1, 2, 3 and 4 were synthesised for the first time and 1, 2 and 4
were identified. The stationary phase was synthesised by a reliable and repeatable method and characterized
by elemental analysis, solid state 13C NMR, scanning electron microscope (SEM), and Brunauer, Emmett and
Teller (BET). This stationary phase possess four amide groups as polar cites, phenyl ring as aromatic non-polar
cite and isopropyl as an aliphatic non-polar cite. Therefore, it can act as both hydrophilic interaction (HILIC)
and reversed phase (RPLC) chromatographic stationary phase. The stationary phase has also chirality and can
be used in enantioseparation of racemic compounds in normal phase chromatography (NPLC).
___
- L. Zhang, Q. Dai, X. Qiao, C. Yu, X. Qin, H. Yan,
Mixed-mode chromatographic stationary phases:
Recent advancement and its aplications for highperformance
liquid chromatography, Trends Anal.
Chem., 82 (2016) 143-163.
- P. Janas, S. Bocian, P. Jandera, T. Kowalkowski, B.
Buszewski, Separation of flavonoids on different
phenyl-bonded stationary phases-the influence
of polar groups in stationary phase structure, J.
Chromatogr. A, 1429 (2016) 198-206.
- Y. Liu, Z. Guo, Y. Jin, X. Xue, Q. Xu, F. Zhang, et
al., Click oligo(ethylene glycol)”: an excellent
orthogonal stationary phase to C18 for twodimensional
reversedphase/ reversed-phase liquid
chromatography, J. Chromatogr. A, 1206 (2008) 153-
159.
- P. Jandera, Z. Kucˇerova, J. Urban, Retention
times and bandwidths in reversedphase gradient
liquid chromatography of peptides and proteins, J.
Chromatogr. A, 1218 (2011) 8874–8889.
- X. Guo, X. Zhang, Z. Guo, Y. Liu, A. Shen, G. Jin, et al.,
Hydrophilic interaction chromatography for selective
separation of isomeric saponins, J. Chromatogr. A,
1325 (2014) 121-128.
- P. Jiang, D.Wu, C.A. Lucy, Determination of void
volume in normal phase liquid chromatography, J.
Chromatogr. A, 1324 (2014) 63-70.
- A.J. Alpert, M. Shukla, A.K. Shukla, L.R. Zieske,
S.W. Yuen, M.A.J. Ferguson, A. Mehlert, M. Pauly, R.
Orlando, Hydrophilic-interaction chromatography of
complex carbohydrates, J. Chromatogr. A, 676 (1994)
191-202
- A.J. Alpert, Advances in Chromatography, 44 (2006)
317-329.
- M. R. Gama, R. G. C. Silva, C. H. Collins, C. B. G. Bottoli,
Hydrophylic interaction chromatography, Trends in
Anal. Chem. 37 (2012) 48-60
- M.A. Strege, S. Stevensen, S. M.Lawrence, Mixed-mode
anion−cation exchange/hydrophilic interaction liquid
chromatography−electrospray mass spectrometry as
an alternative to reversed phase for small molecule
drug discovery, Anal. Chem., 72 (2000) 4629-4633
- M.A. Strege, Hydrophilic interaction chromatography−
electrospray mass spectrometry analysis of polar
compounds for natural product drug discovery, Anal.
Chem., 70 (1998) 2439-2445
- B.Y. Zhu, C.T. Colin, R.S. Hodges, Hydrophilicinteraction
chromatography of peptides on
hydrophilic and strong cation-exchange columns, J.
Chromatogr., 548 (1991) 13-24.
- T. Saga, Y. Inoue, K. Yamaguchi, Determination
of carbohydrates by hydrophilic interaction
chromatography with pulsed amperometric detection
using postcolumn pH adjustment, J. Chromatogr. A,
625 (1992) 151-155.
- Q. W. Yu, B. Lin, Y. Q. Feng, F. P. Zou, Application of
humic acid bonded‐silica as a hydrophilic‐interaction
chromatographic stationary Phase in separation
of polar compounds, J. Liq. Chromatogr. Related.
Technol., 31 (2008) 64-78.
- X. Liu, C. Pohl, New hydrophilic interaction/
reversed-phase mixed-mode stationary phase and
its application for analysis of nonionic ethoxylated
surfactants, J. Chromatogr. A, 1191 (2008) 83–89.
- H. Hinterwirth, M. Lammerhofer, B. Preinerstorfer,
A. Gargano, R. Reischl, W. Bicker, et al., Selectivity
issues in targeted metabolomics: separation of
phosphorylated carbohydrate isomers by mixedmode
hydrophilic interaction/ weak anion exchange
chromatography, J. Sep. Sci., 33 (2010) 3273–3282.
- X. Liu, C.A. Pohl, HILIC behavior of a reversed-phase/
cation-exchange/anionexchange trimode column, J.
Sep. Sci., 33 (2010) 779-786.
- X. Liu, C.A. Pohl, Comparison of reversed-phase/
cation-exchange/anion-exchange trimodal stationary
phases and their use in active pharmaceutical
ingredient and counterion determinations, J.
Chromatogr. A, 1232 (2012) 190-195.
- X. Liu, C. Pohl, A. Woodruff, J. Chen, Chromatographic
evaluation of reversedphase/anion-exchange/cationexchange
trimodal stationary phases prepared by
electrostatically driven self-assembly process, J.
Chromatogr. A, 1218 (2011) 3407-3412.
- Y. Zhao, H.C. Law, Z. Zhang, H.C. Lam, Q. Quan, G.
Li, et al., Online coupling of hydrophilic interaction/
strong cation exchange/reversed-phase liquid
chromatography with porous graphitic carbon liquid
chromatography for simultaneous proteomics and
N-glycomics analysis, J. Chromatogr. A, 1415 (2015)
57-66.
- A.S. Feste, I. Khan, Separation of glucooligosaccharides
and polysaccharide hydrolysates by gradient elution
hydrophilic interaction chromatography with pulsed
amperometric detection, J. Chromatogr., 630 (1992)
129-139.
S.C. Churms, Recent progress in carbohydrate
separation by high-performance liquid
chromatography based on hydrophilic interaction, J.
Chromatogr. A, 720 (1996) 75-91.
- S.C. Lin, W.C. Lee, Separation of a fructooligosaccharide
mixture by hydrophilic interaction
chromatography using silica-based micropellicular
sorbents, J. Chromatogr. A, 803 (1998) 302-306.
- A.R. Oyler, B.L. Armstrong, J.Y. Cha, M.X. Zhou,
Q. Yang, R.I. Robinson, R. Dunphy, D.J. Burinsky,
Hydrophilic interaction chromatography on aminosilica
phases complements reversed-phase high
performance liquid chromatography and capillary
electrophoresis for peptide analysis, J. Chromatogr.
A, 724 (1996) 378-383.
- T. Yoshida, Peptide separation in normal phase liquid
chromatography, Anal. Chem., 69 (1997) 3038-3043.
- C.T. Mant, L.H. Kondejewski, R.S. Hodges, Hydrophilic
interaction/cation-exchange chromatography for
separation of cyclic peptides, J. Chromatogr. A, 816
(1998) 79-88.
- T. Yoshida, T. Okada, T. Hobo, R. Chiba, Calculation
of amino acid hydrophilicity indices for retention of
peptides on amide, diol and silica columns in normalphase
liquid chromatography, Chromatographia, 52
(2000) 418-424.
- Y. Guo, S. Gaiki, Retention behavior of small polar
compounds on polar stationary phases in hydrophilic
interaction chromatography, J. Chromatogr. A, 1074
(2005) 71-80
- Y. Kawachi, T. Ikeami, H. Takubo, Y. Ikeami, M.
Miyamoto, N. Tanaka, J. Chromatogr. A, 1218 (2011)
5903-5919.
- X. Qiao, L. Zhang, N. Zhang, X. Wang, X. Qin, H. Yan,
et al., Imidazolium embedded C8 based stationary
phase for simultaneous reversed-phase/hydrophilic
interaction mixed-mode chromatography, J.
Chromatogr. A, 1400 (2015) 107-116.
- M. Sun, J. Feng, C. Luo, X. Liu, S. Jiang, Benzimidazole
modified silica as a novel reversed-phase and anionexchange
mixed-mode stationary phase for HPLC,
Talanta, 105 (2013) 135-141.
- K. HU, W. Zhang, H. Yang, Y. Cui, J. Zhang, W. Zhao,
A. Yu, S. Zhang, Calixarene ionic luqiuid modified
silica gel: A novel stationary phase for mixed-mode
chromatography, Talanta, 152 (2016) 392-400.
- H. Aral, T. Aral, K.S. Çelik, G. Topal, Preparation of a
novel ionic hybdid stationary phase by non-covalent
functionalization of single-walled carbon nanotubes
with amino-derivatized silica gel for fast HPLC
separation of aromatic compounds, Talanta, 149
(2016) 21-29.
- T. Aral, H. Aral, B. Ziyadanoğulları, R. Ziyadanoğulları,
Synthesis of a mixed-model stationary phase derived
from glutamine for HPLC separation of structurally
different biologically active compounds: HILIC and
reversed-phase applications, Talanta, 131 (2015) 64-
73.
- H. Aral, T.Aral, B. Ziyadanoğulları, R. Ziyadanoğulları,
Development of a novel amide-silica stationary phase
for the reversed-phase HPLC separation of different
classes of phytohormones, Talanta, 116 (2013) 155-163.
- Q. Wang, Y. Long, L. Yao, L. Xu, Z.G. Shi, L. Xu,
Preparation, characterization and application of a
reversed phase liquid chromatography/hydrophilic
interaction chromatography mixed-mode C18-DDT
stationary phase, Talanta, 146 (2016) 442-451.
- D. Kotani, I. D’Acquarica, A. Ciogli, C. Villani, D.
Capitani, J. Chromatogr. A, 1232 (2012) 196-211.
- Y. Li, Y. Feng, T. Chen, H. Zhang, Imidazoline
type stationary phase for hydrophilic interaction
chromatography and reversed-phase liquid
chromatography, J. Chromatogr. A, 1218 (2011) 5987-
5994.
- S. Liu, H. Xu, J. Yu, D. Li, M. Li, X. Qiao, et al., Novel
imidazolium-embedded N,N-dimethylaminopropylfunctionalized
silica-based stationary phase for
hydrophilic interaction/reversed-phase mixed-mode
chromatography, Anal. Bioanal. Chem., 407 (2015)
8989-8997.
- X.D. Cheng, X.T. Peng, Q.W. Yu, B.F. Yuan, Y.Q. Feng,
Preparation and chromatographic evaluation of a
novel phosphate ester-bonded stationary phase with
complexation and hydrophobic interactions retention
mechanism, J. Chromatogr. A, 1302 (2013) 81-87.
- Y. Zhang, Y. Zhang, G. Wang, W. Chen, P. He, Q.
Wang, Simultaneous separation of polar and
non-polar mixtures by capillary HPLC based on
an ostadecylsilane and taurine derivatized silica
continuously packed column, Talanta, 161 (2016) 762-
768.
- Q. Wang, M. Ye, L. Xu, Z.G. Shi, A reversed-phase/
hydrophilic interaction mixed-mode C18-Diol
stationary phase for multiple applications, Anal. Chim.
Acta, 888 (2015) 182-190.
- T. Liang, Q. Fu, A. Shen, H. Wang, Y. Jin, H. Xin, Y. Ke,
Z. Guo, X. Liang, Preparation and chromatographic
evaluation of newly designed steviol glycoside
modified-silica stationary phase in hydrophilic
interaction liquid chromatography and reversed
phase liquid chromatography, J. Chromatogr. A, 1388
(2015) 110-118.
- H. Qui, M. Zhang, T. Gu, M. Takafuji, H. Ihara, A
Sulphonic-azobenzene–grafted silica amphiphilic
material: a versatile stationary phase for mixed-mode
chromatography, Chem. Eur. J., 19 (2013) 18004-
18010.
- S. Bocian, A. Nowaczyk, B. Buszewski, New-alkylphospate
bonded stationary phase for liquid
chromatographic separation of biologically active
compounds, Anal. Bioanal. Chem., 404 (2012) 731-740.
- Z. Liu, B.D. Cai, Y.Q. Feng, Rapid determination
of endogenous cytokinins in plant samples by
combination of magnetic solid phase extraction with
hydrophilic interaction chromatography-tandem
mass spectrometry, J. Chromatogr. B, 891-892 (2012)
27-35.
- T. Tang, W.B. Zhang, J.W. Xu, M. Z. Xia, X.D. Gong,
F.Y. Wang, T. Li, Synthesis, characterization, and
application of a novel multifunctional stationary
phase for hydrophilic interaction/reversed phase
mixed-mode chromatography, Chinese J. Anal. Chem.,
45 (2017) 56-60.
- L. Qiao, A. Duou, X. Shi, H. Li, Y. Shan, X. Lu, G. Xu,
Development and evaluation of new imidazoliumbased
zwitterionic stationary phases for hydrophilic
interaction chromatography, J. Chromatogr. A, 1286
(2013) 137-145.
- L. Qiao, X. Zhou, Y. Zhang, A. Yu, K. Hu, S. Zhang,
4-Chloro-6-pyrimidinylferrocene modified silica gel:
A novel multiple-function stationary phase for mixedmode
chromatography, Talanta, 153 (2016) 8-16.
- A. Shen, X. Li, X. Dong, J. wei, Z. Guo, X. Liang,
Glutathione-based zwitterionic stationary phase for
hydrophilic interaction/cation-exchange mixed-mode
chromatography, J. Chromatogr. A, 1314 (2013) 63-69.