Fusion 1.8µm UPLC & 3.5µm, 100Å columns & guards (slightly negative surface)
Fusion (+) 3.5µm, 100Å columns & guards (slightly positive surface)
Fusion (P) 5µm, 200Å columns (for high pH metabolomics, inorganic ions and nTP's)
iHILIC®-(P) Classic 5µm, 200Å columns (for metabolomics, inorganic ions and nTP methods developed on ZIC® pHILIC columns). Operating Instructions/Chemistry

Addenda:
pH Range of Buffers Suitable for LC-MS Operation

"Addressing a Common Misconception: Ammonium Acetate as Neutral pH "Buffer" for Native Electrospray Mass Spectrometry." Konermann, L., J Am Soc Mass Spectrom. 2017 Sep;28(9):1827-1835. doi: 10.1007/s13361-017-1739-3. Epub 2017 Jul 14.

Effect of Buffer Strength on iHILIC® columns

Publications:

iHILIC® Column Publications
iSPE® Publications
LCGC Application Notes
Posters
Application Library by Chemical Type

Applications:

• Amino Acids
  

  

  Direct Analysis of Amino Acids by HILIC–ESI-MS. Alexander Schriewer ¹, Katharina Johanna Heilen¹, Heiko Hayen¹, and Wen Jiang², LCGC, THE APPLICATION NOTEBOOK – July 2017. ¹Institute of Inorganic and Analytical Chemistry, University of Münster, Münster, Germany, ²HILICON AB

  
  
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• Sweetners
  Sucralose Acesulfame K Saccharin Neotame
  Neohesperidin Na Cyclamate Aspartame

  
  iHILIC® Fusion(+), Column: 2.1 x 150mm, 100Å, 3.5µm, 0.3 mL/min.
  Gradient elution: A) acetonitrile; B) 10mM ammonium formate, pH = 3.5; gradient elution from (95/5) A/B to (84/16) A/B in 8.5 min.
  Column temperature: 40 °C
  Injection volume: 5 µL
  Data Courtesy of University of Münster
  

• Mono- and di-saccharides

  
  iHILIC® Fusion(+), Column: 4.6 x 150mm, 100Å, 3.5µm, 1.0 mL/min.
  Isocratic 80:20 ACN 25mM ammonium acetate (v/v)
  Column temperature: 35 °C
  Injection volume: 10 µL
  Data Courtesy of Exact Scientific Services
  

• Glucuronides and Glucosides
  
  iHILIC® Fusion(+), 2.1 x 100mm, 100Å, 3.5µm, 0.4mL/min.
  Gradient 90:10 to 30:70; ACN:10mM, pH 3.4, ammonium acetate w/0.1% FA (v/v)), 10µL injection.
  Data Courtesy of Mike Karb, P&G.
  
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• UDP-Glucose
  
  iHILIC® Fusion, 2.1 x 100mm, 100Å, 3.5µm, 0.2mL/min.
  Gradient 80:20 to 60:40; ACN:100mM, pH 5.8, ammonium acetate (v/v))
  Data Courtesy of Narek Darabedian in Dr. Matt Pratt's lab, USC.
  Publication: "Metabolic chemical reporters of glycans exhibit cell-type selective metabolism and glycoprotein labeling." Anna R. Bhatt, et al., ChemBioChem 10.1002/cbic.201700020
  
  
  
• cGAMP
  
  iHILIC® Fusion, 2.1 x 100mm, 100Å, 3.5µm, 0.2mL/min.
  Gradient 90:10 to 50:50; ACN:50mM, pH 5, ammonium acetate (v/v))
  Data Courtesy of Dr. Michelle Dubuke in Dr. Scott Shaffer's lab, UMass Medical Center.
  
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• Mono-, Di- & Tri-phosphate Nucleotides
  
  iHILIC® Fusion, 2.1 x 100mm, 100Å, 3.5µm, 0.2mL/min.
  Nucleotides Separate at a lower buffer concentration on iHILIC® Fusion at pH 5.8, 70:30; ACN:100mM, pH 5.8, ammonium acetate (v/v) than at pH 4.5, 70:30; ACN:100mM, on ZIC® cHILIC due to less titrating to lower the pH.
  
  
  
• Enhanced sensitivity of phosphates by MS at high pH:
  
  iHILIC® Fusion(P), 4.6 x 150mm, 200Å, 5µm.
  Separation of nTP's at pH 9.3 with a volatile buffer: Gradient. Buffer A: 80:20 (v/v) Acetonitrile, 100mM ammonium bicarbonate and Buffer B: 30mM ammonium bicarbonate pH 9.3.
  
  Metabolomics (ATP/ADP ratios, acetyl-CoA). "Breast Cancer-Derived Lung Metastases Show Increased Pyruvate Carboxylase-Dependent Anaplerosis," Christen et al., 2016, Cell Reports 17, 837–848 October 11, 2016 and supplemental information. MS in negative mode. Column iHilic Fusion(P). Solvent: ACN and pH=9.3, 10mM ammonium acetate.
  
  
• Separation of 2'NADP from 3'NADP at high pH:
  
  iHILIC® Fusion(P), 4.6 x 150mm, 200Å, 5µm.
  Separation of NADP's at pH 9.3 with a volatile buffer: Gradient. Buffer A: 80:20 (v/v) Acetonitrile, 20mM ammonium bicarbonate and Buffer B: 20mM ammonium bicarbonate pH 9.3.
  Unpublished data courtesy of Corey D. Broeckling, Colorado State University.
  
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• Organic Acids:
  
  iHILIC® Fusion(P) PEEK, 2.1 x 50mm, 5µm, 200Å
  Isocratic separation of γ-hydroxybutyric acid (GHB), γ-aminobutyric acid (GABA) & succinic acid with a volatile buffer: Buffer A: 90:10 (v/v) Acetonitrile, 20mM ammonium acetate.
  Unpublished data courtesy of Patrick Belanger, INSPQ - Centre de Toxicologie (CTQ). See also: Effects of pH and Buffer Strength, an Organic Acids Study.
  
  
• Phospholipids HPLC:
  
  iHILIC® Fusion, 2.1 x 150mm, 3.5µm, 100Å.
  * Double peak of PA is caused by in-source fragmentation of the PS head group resulting in corresponding PA species.
  Journal of Chromatography A, 1565 (2018) 105–113 Eluent: A) Ammonium acetate solution (35 mM, pH 5.75) and acetonitrile (95:5, v/v); B) Acetonitrile. Gradient elution: 0-0.5 min, 97% B; 0.5-26.5 min, from 97% to 75% B; 27-33 min, 60% B; 35-45 min 97% B.
  Flow rate: 0.3 mL/min
  Column temperature: 40 °C, Injection volume: 10 μL
  Phospholipid standards: Bis(monoacylglycero)phosphate (BMP 36:2), phosphatedylglycerol (PG 36:2), phosphatidylcholine (PC 32:0), phosphatidylethanolamine (PE 32:0), phosphatidylserine (PS 32:0)

  
  " Separation and identification of phospholipids by hydrophilicinteraction liquid chromatography coupled to tandem high resolutionmass spectrometry with focus on isomeric phosphatidylglycerol andbis(monoacylglycero)phosphate" Christian Vosse¹, Carina Wienken¹, Cristina Cadenas², Heiko Hayen¹, ¹Institute of Inorganic and Analytical Chemistry, University of Münster, Corrensstr. 30, 48149 Münster, Germany ²Leibniz Research Centre for Working Environment and Human Factors, Ardeystr. 67, 44139 Dortmund, Germany
  

• Lipids SPE:
  Cardiolipin iSPE® HILIC Isolation
  
  Publication outlines a HILIC-based clean‐up method enabling complete separation of polar lipids containing four fatty acyl residues from non-polar lipid classes using iSPE® HILIC columns.
  

  
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• Biometals
  
  Enterobactin, a catecholate siderophore
  Siderophores are small, high-affinity iron-chelating compounds secreted by microorganisms such as bacteria and fungi and serving to transport iron across cell membranes. Siderophores are amongst the strongest soluble Fe3+ binding agents known.
  iHILIC® Fusion, 2.1 x 150mm, 3.5µm, 100Å.
  Six siderophores of Pseudomonas taiwanensis VLB120 and their iron(III)- and aluminum(III) complexes were separated by means of an iHILIC Fusion column. This technique enabled the separation of the siderophores according to their different acyl side chain, and additionally, according to their central ion. " Mass spectrometric characterization of siderophores produced by Pseudomonas taiwanensis VLB120 assisted by stable isotope labeling of nitrogen source" Karen Scholz¹ Till Tiso² , Lars M. Blank² , Heiko Hayen¹
  ¹ Institute of Inorganic and Analytical Chemistry, University of Münster, Münster, Germany, ² Institute of Applied Microbiology, RWTH Aachen, Germany
  
  
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• Enhanced sensitivity of inorganic ion detection with ELSD when using the polymeric Fusion(P):
  Elimination of silica leaching increases sensitivity.
  
  iHILIC® Fusion(P), 4.6 x 150mm, 200Å, 5µm.
  Separation of ammonia and sulfate:
  Mobile phase: A) Acetonitrile; B) 25 mM ammonium acetate, pH 6.8 (salt solution without pH adjustment)
  Gradient program: 0 min: 85% A, 6 min: 65% A, 6-9 min: 65% A, 9-16 min: 85% A.
  Flow rate: 0.2 mL/min. Detector: NQAD
  Peaks: #2: Ammonium, #3: Sulfate.

  iHILIC® Fusion(P), out performs ZIC-pHILIC for analysis of inorganic ions
  
  iHILIC® Fusion(P), 4.6 x 150mm, 200Å, 5µm.
  Gradient: 85% ACN/15% 25 mM ammonium acetate to 60% ACN/40% 25 mM ammonium acetate.
  When using iHILIC® Fusion(P), thiosulfate is a sharp symmetrical peak, sulfate elutes after thiosulfate as a sharp symmetrical peak and sodium is more retained than on the ZIC®pHILIC column (shown above).
  Data courtesy of Paul Kostel, Eurofins Advantar Labs., Inc.

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