1. Where Tradition Meets Tomorrow
Using Cogent TYPE-C Silica™
Where Tradition Meets Tomorrow
Presented by: Bill Ciccone
MicroSolv Technology Corporation
Thank you for inviting me to Watson, the title of my talk is:
My name is:
I am from MicroSolv Technology Corp:
Eatontown, NJ USA

2. MicroSolv’s Mission Statement
Our mission is to encourage, develop, promote, assist, and aid in the improvement and development of chemistry and biological technologies. To participate in the discovery of new technologies that will improve the living condition of people and animals all over the world.
It is within the scope of our mission that one day one of our customers using our technology will make a major improvement to the human condition.
Also, our mission is to provide an easy, effective and efficient way to purvey information and to become an exchange of ideas that promotes science in both the professional and amateur fields.
Just a little bit about MicroSolv and what we do:
Formed in 1992 for tech transfer
We take technology ideas and develop them, manufacture them, bring them to market and develop and suppor them

3. Formed to work with technology transfer departments at universities
About MicroSolv
MicroSolv started in 1992
Formed to work with technology transfer departments at universities
Role MicroSolv plays in product development
Tech Transfer
Product Development and Marketing R&D
Production Design QC
Marketing and Sales Distribution of Wide Array of Products
Education and Technical Support
Product Line Extension
Just a little bit about MicroSolv and what we do:
Formed in 1992 for tech transfer
We take technology ideas and develop them, manufacture them, bring them to market and develop and suppor them

4. First Introduced at Pittcon 2003
Cogent TYPE-C™ Silica HPLC Columns
First Introduced at Pittcon 2003
Today I am here to introduce you to Cogent TYPE-C Silica Based HPLC Columns. To avoid any confusion, TYPE-C is a trademark of Microsolv. I am not here to tell you to throw out all of the columns you are working with now and how our columns can replace everything you do. No, that is unrealistic and there will be times when our columns are not the best choice. But I think after you see what they do, why they are a great addition to your favorites to develop methods with.

5. Agenda:
1.What are TYPE-C™ Silica HPLC columns? 2. What do they do? 3. Why are they recommended for all chromatographers? Even for Reverse Phase? 4. Exciting Real life examples where problems were solved. To give you an idea why these columns are useful for your lab.
Today, I hope to accomplish answering some question that you might have… If you have any questions during the presentation, please feel free to ask. I would like to keep this talk somewhat informal.

6. 1. What are TYPE-C™ Silica based HPLC columns
1. What are TYPE-C™ Silica based HPLC columns? Unique Selectivity These are different from all Others. Efficiency Stability-Superior Versatility Reproducible
So to get started, the first question on the agenda is WHAT IS TYPE-C SILICA?

7. Type-C Silica Columns are “Silica-Hydride” Based
HPLC Stationary Phases:
First in Class HPLC Columns
For Solving Today’s Separation Problems
So to begin to answer the first question This technology was invented by Professor Joseph Pesek at San Jose State University in an attempt to bond cholesterol to the surface of silica particles and CE capillaries. Grants from NASA and NIH and 10 years of research and he came up with the current technology.
These columns are “first in class” and solve a lot of problems in routine daily use.

8. Ordinary Silica Standard; Well Known by all

9. Silica Hydride
Ordinary Silica
TYPE-C™ Silica M

10. Silica Hydride Ordinary Silica
How Does this structural difference impact the performance of the columns?
Silica Hydride
Ordinary Silica
TYPE-C™ Silica

11. How Does this structural difference impact the performance of the columns?
People often ask us, what is the biggest difference between the Cogent TYPE-C silica products and all other silica HPLC columns. The answer is how they both adsorb and desorb the mobile phase. Dessicant.

12. Stationary Phases Solvated by aqueous Mobile Phase
“Hydride” Silica
Ordinary Silica
Water Shell
“Water Layer”

13. In High Organic The “Shells” Can Differ
“Hydride” Silica
Ordinary Silica
Water Shell
Organic Layer

14. Some Examples of TYPE-C Silica Phases
Bidentate C18 and C8 Moieties Bonded To Hydride Silica M ( C H 2 ) 6 3 O S i
Bidentate C8
Cogent BDC8 16
Cogent BDC18
Another unique feature of TYPE-C columns is the way the bonded phase is attached to silica.

16. Bidentate C18 Direct Silicon-Carbon Bonds
TYPE-C Silica M

17. Bidentate C18 Direct Silicon-Carbon Bonds
TYPE-C Silica M
Two Points of Attachment

18. Bidentate C18 Direct Silicon-Carbon Bonds
TYPE-C Silica
Two Points of Attachment
Not Susceptible to Hydrolysis.

19. The Cholesterol Column

20. Adsorbed Mobile Phase on
Bidentate C18 and C8 Moieties Bonded To
TYPE-C Silica ( C H 2 ) 6 3 O S i
Bidentate C8
Cogent BDC8 16
Cogent BDC18

21. Stationary Phases
Cogent Silica-C™ –unmodified Silica-Hydride-Most likely to retain polar compounds in ANP.
Cogent Diamond Hydride ™ -small amounts of carbon in the silica. Excellent for amino acids Patent Applied for.
Cogent Bidentate C8 ™ - less hydrophobic than C18. Often best choice for complex mixtures.
Cogent UDC-Cholesterol ™ - liquid crystal phase. Shape selectivity as well as 3 modes of HPLC
Cogent Bidentate C18 ™ -Excellent for RP and ANP.

22. Are these Columns Stable?

23. Are these Columns Stable? What about in the presence of Water?
YES, These columns are VERY Stable…even in Water.

24. TYPE-C BASED COLUMN AFTER MORE THAN 2 YEARS OF
USE IN 0.05% Acidic MOBILE PHASE
uracil +
pyridine
phenol
uracil +
pyridine
phenol
A: initial conditions, B: after 2 years

25. Ordinary Silica
TYPE-C Silica
Nonendcapped Normal
C18 Bonded Phase

26. Ordinary Silica
TYPE-C Silica
Nonendcapped Normal
C18 Bonded Phase

27. 2. What do TYPE-C™ Silica HPLC Columns do
2. What do TYPE-C™ Silica HPLC Columns do?: Unique Selectivity Efficiency Stability-Superior Versatility Reproducible
So far we have seen what the structure and some of the chemical properties of TYPE-C silica are but…
Unique selectivity

28. Ordinary Silica
TYPE-C Silica
Non-endcapped Ordinary
C18 Bonded Phase

29. Comparison of the Retention of Basic Compounds on Ordinary Silica-Based and TYPE-C Silica-Based Columns
Reversed Phase Only
Reversed Phase
Aqueous Normal Phase
Flow Rate: 3ml/min
Wavelength: 254nm
Solvents A: Buffer
B: 100%ACN

30. Isn’t this HILIC? Developed for Bioanalytical Applications
Hydrophilic Interaction Chromatography

31. Two Different Mechanisms for Different Applications
NO!
Two Different Mechanisms for Different Applications

32. Silica Hydride-Based Column
DIFFERENCES BETWEEN AQUEOUS NORMAL PHASE AND HILIC
Aqueous Normal Phase
Silica Hydride-Based Column
Hydrophilic Interaction Chromatography (HILIC) uses ordinary Silica-Based Column
Does not require water to work
Retains nonpolar compounds by reversed phase mechanism
Retains polar compounds by normal phase mechanism
Both reversed phase and normal phase mechanisms can operate simultaneously
Can separate samples with both polar and nonpolar compounds
HILIC Requires a “water rich” environment to work
Retains polar compounds by a normal phase mechanism
Does not retain nonpolar compounds
Cannot usually separate samples having both polar and nonpolar compounds
Main differences are non polar and polar in isocratic run, reproducibility which we will see later and stability.

33. Comparison of the Retention of Acidic Compounds Ordinary Silica-Based (HILIC) and TYPE-C Silica-Based Columns
Aqueous Normal Phase
Under these conditions there is no RP Retention but there is ANP.
Solvents A: Buffer
B: 100%ACN
Flow Rate: 3ml/min
Wavelength: 254nm

34. Tech Note: ANP Retention is a Function of the Compound
The TYPE-C columns will all give you ANP but ANP is a function of the compound’s nature. Acids and bases will retain much better than neutrals. However since the TYPE-C columns can have bonded phases you retain polar and non polar compounds.

35. ANP Retention is a Function of the Compound & Some Compounds can be Retained in both ANP and RP

36. Nucleosides with Alkyl Chains
Some Compounds can Display Both Reversed Phase and ANP Behavior on the Same Column M
Nucleosides with Alkyl Chains
No RP Retention

37. Nucleosides with Alkyl Chains
Some Compounds can Display Both Reversed Phase and ANP Behavior on the Same Column
Nucleosides with Alkyl Chains
If you are looking to purify the nucleoside R3 from R1 you would use RP and if you were looking to quantitate both compounds you would use ANP.
Both RP and ANP Retention

38. What Other Capabilities to TYPE-C Columns Have?
In Addition to ANP
What Other Capabilities to TYPE-C Columns Have?

39. Only Cogent TYPE-C Silica™ based columns operate in 3 chromatographic modes. Without hysteresis or damage to columns Reverse Phase Aqueous Normal Phase Normal Phase with Non Polar Solvents

40. TYPE-C Based HPLC Columns are Excellent Reversed Phase Columns!

41. Steroid Mixture on Cogent UDC-Cholesterol Column
MOBILE PHASE: 50:50 MeOH/water
Column Dimensions:
4.6 x 150mm
Solutes: 1 = adrenosterone; 2 = corticosterone; 3 = 4-androstene-3,17-dione;
4= 11-acetoxyprogesterone; 5 = estrone; 6 = estradiol.

42. Use Bidentate C18 Columns with 100% Water No Loss of Rt over Time

43. Using 100% Aqueous Mobile Phase On Cogent Bidentate C18 HPLC Column
1- OXALIC ACID
2- FORMIC ACID
3- ACETIC ACID
4- SODIUM AZIDE
5- URACIL
6- FUMARIC ACID
7- PROPIONIC ACID
After 3 hours of 100% aqueous exposure, results are the same as with the first injection
100% 0.05% v/v H3PO4
Flow Rate:1mL/min
Column: 75 x 4.6mm id
UV: 215 nm

44. A Very Useful and Unique Feature Of Cogent TYPE-C™ HPLC Columns
Organic Normal Phase Chromatography without Hysteresis or Damage to The Column.

45. Normal Phase Separation of Substituted Phenolic Compounds on a C18 HPLC Column
- APCI+
Column: Bidentate C18, mobile phase: 95:5 Hexane/Ethyl Acetate
Flow rate: 1.0 mL/min.
Samples: 1 – phenol with aldehyde, 2 – parent phenol,
3 – phenol with ketone, 4 – phenol with acid

46. Cogent TYPE-C Columns for Analytical or Prep Normal Phase Gradients
A: Bidentate C18
B: Silica-C
A: min 100% hexane; min to 50:50 hexane/ dicholoromethane; 50:50 hexane/ dichoromethane to 10 min.
B: 0.0 to 1.0 min 100% dichloromethane; 1.0 to 3.0 min to 100% ethyl acetate
Analytical
Preparative

47. Stationary phase is un-modified silica hydride
Cogent Silica-C™ Column in Normal Phase Chromatography- Selectivity Advantage
Stationary phase is un-modified silica hydride

48. Generic Strategy is Very Easy and Fast
Where do I start?
Generic Strategy is Very Easy and Fast
1. Mobile Phase to Start with.
A. Water w/0.1% Formic Acid/.05% TFA
B. MeCN w/0.1% Formic Acid
2. Gradient starting at 95% A to 40% A over 20minutes for a 75mm column.
3. Equilibrate for one column volume.
4.Inverse Gradient starting at 95% B to 40% B over 20 minutes for a 75mm column.
5. Optimize depending on your results.

49. 3. Why are TYPE-C™ Silica HPLC columns recommended for all chromatography labs? Unique Selectivity Efficiency Stability-Superior Versatility (3 Modes) Reproducibility & much more….

50. Fast LC? Equilibrate Between Gradients in very little Time

51. Rapid Equilibration of Bidentate C18 HPLC Column
Reversed Phase Mode - Gradient Elution
Gradient program:
0-3 min ACN/water (50:50)
3-18 min to 100% ACN
18-23 min 100% ACN
Equilibration to 50:50 ACN/water
Retention Time (min)
Solute/Equilibration Time 25 min min min
Benzene
Naphthalene
Phenanthrene
Anthracene
Pyrene

52. Reproducibility

53. Peptides: Simple isocratic RP – HPLC analysis
An HPLC peptide standard mixture was resolved in under 5 minutes using a short (4.6 x 75 mm) Cogent BD C18 column and a simple isocratic RP-HPLC method. The separation was very reproducible. To achieve the separation presented on a conventional HPLC column, 4.6 x 250 mm, a gradient method is required.
If higher resolution is desired a longer column should be used. Columns from leading brands of manufacturers were evaluated using the same conditions and compounds 3 and 4 were never separated under RP-HPLC isocratic conditions.
Chromatogram is a composite of 8 consecutive injections
Mobile Phase: 25% acetonitrile/75% DI water + 0.1% formic acid
Flow rate: mL/minute
Detection UV 214 nm
1. Gly –Tyr; 2. Val – Tyr – Val; 3. Met-enkephalin
4. Leu-enkephalin; 5. Angiotensin II

54. Reproducibility of Cogent Silica-C Column in a real life example
Six Injections-Overlaid
Excellent Run to Run Precision
%RSD-0.3%

55. Retain and Separate Sugars in Reverse Phase
Because of the unique solvent shell around the base silica, you can get compounds to retain in RP (C18) that normally do not retain at all on ordinary RP columns.

56. SEPARATION OF CARBOHYDRATE STRUCTUAL ISOMERS
maltotriose a 1,4
panose a 1,4 + a 1,6
isomaltotriose a 1,6
Mobile Phase: 100% water
All compounds have MW = 504
Each compound detected by MS in APCI+ mode with single ion monitoring (SIM) using a specific fragment ion
cellotriose b 1,4

57. You Can and Should Avoid high pH when Possible Why Damage Your instrument when you do not have to?
Also, this is for orthogonality. Retain basic compounds at low pH and High pH

58. Every Chromatographer should have a Suite of the Cogent TYPE-C Columns for: 1.Problem Selectivity...Polar Compounds 2.Rapid Equilibration Ballistic Gradients 3. LCMS Increase Sensitivity 4. Non Robust Methods 100% Water 5. To Challenge Every Method with RP and Quickly convert to ANP without purging your system 6. Impurity Profiling. Orthogonality 7. Complex Mixtures/Metabolites Natural Products 8. When ordinary columns are not stabile enough 9. When samples adsorb to ordinary silica columns 10. When samples hyrdolize on ordinary silica columns
Point

59. 1 of each Phase in the Same Column Configuration
Suite of Columns for Scouting & Method Development:
1 of each Phase in the Same Column Configuration

60. 4. Real Life Examples of Problems Solved with: Unique Selectivity Efficiency Stability-Superior Versatility Reproducibility

61. GLUCOSAMINE on A Bidentate C18
Retain Sugars with High Organic Strength Mobile Phases

62. Separation based on size
Retention of Smaller Carbohydrates on a Cogent Bidentate C18™ Column
Separation based on size
Detection by MS in the APCI+ mode

63. Clinical Application 1 Samples:
Method Conditions
Column: Cogent Bidentate C18, 4m, 100A.
Catalog No.: P
Dimensions: x 75 mm
Mobile phase: A. 90:10 Acetonitrile/DI Water +0.5% formic acid
B. 85:15 Acetonitrile/DI Water % formic acid
Flow rate: mL/min.
Injection Volume: 10 mL
Samples:
1. Triacetylnormetanephrine (m/z 166.2)
2. Triacetylmetanephrine (m/z 180.2)
200 ng of each sample was dissolved in 1 mL of reverse osmosis water
Detection: APCI+
Single Ion Monitoring
In detection single ion monitoring (SIM) was used. Mass transition of m/z to m/z (triacetylnormetanephrine) and m/z to m/z (triacetylmetanephrine) that correspond to the fragmentation of the (M+H+) ions were monitored.
Separate Closely Related Compounds in clinical applications Metabolomics/Drug Metabolism

64. Clinical Application 2 Inverse Gradient
The powerful anticancer drug, methotrexate (4-amino-N10-methylpteroyl glutamic acid) acts as an antimetabolite and is used for the treatment of many neoplastic diseases including acute leukemia, osteosarcoma, non-Hodgkins lymphoma, and breast cancer. There is a great interest in pharmacological studies and clinical monitoring of methotrexate.
Inverse Gradient
A: DI Water + 0.5% Formic Acid
B: Acetonitrile
A quadrupole mass spectrometer operating in the positive – ion mode and an atmospheric pressure ionization (API) source was used for selective detection and assured that no interfering peaks affect the quantitative results. A bidentate C18 column was the column of choice for the ANP gradient analysis of the drug. The retention of the methotrexate is more than sufficient. The LC-MS method developed assures both high specificity and sensitivity.

65. Exploiting the Separation Power of TYPE-C Based Stationary Phases
METFORMIN
Aqueous Normal Phase
GLYBURIDE
Reversed Phase

66. Comparison of Bidentate C18 and Ordinary C18 Columns
RP and Aqueous Normal Phase Mode

67. FAST SEPARATION Metformin/Glyburide On Cogent Bidentate C18 WITH
UV DETECTION
Column dimensions: 2.1 x 20 mm
Mobile Phase:
A: 50:50 acetonitrile, DI water + 0.5% formic acid
B: 80:20 acetonitrile, DI water + 0.5% formic acid
C: 85:15 acetonitrile, DI water + 0.5% formic acid
Flow rate: 0.3 mL/minute
Injection Volume: 1 µL
Samples:
1. Metformin
2. Glyburide
100 µg/mL of each in the mobile phase
Detection: UV 254nm

68. FAST SEPARATION Metformin/Glyburide On Cogent Bidentate C18 WITH
UV DETECTION
Column dimensions: 2.1 x 20 mm
Mobile Phase:
A: 50:50 acetonitrile, DI water + 0.5% formic acid
B: 80:20 acetonitrile, DI water + 0.5% formic acid
C: 85:15 acetonitrile, DI water + 0.5% formic acid
Flow rate: 0.3 mL/minute
Injection Volume: 1 µL
Samples:
1. Metformin
2. Glyburide
100 µg/mL of each in the mobile phase
Detection: UV 254nm

69. Mobile Phase: 60:40 acetonitrile/water
Separation of a Mixture of Polar and Non Polar Compounds
Compound No. Compound Type Mol Wt ApKa BpKa Log P
Cytidine-R
Cytidine-R
Cytidine-R
Quinolinedione-R ,
Tetramic acid ,
Quinolinedione-R
Benzopyran
Mobile Phase: 60:40 acetonitrile/water

70. SEPARATION OF ACETYLCHOLINE/CHOLINE ON STANDARD
And Short Cholesterol Columns
Challenging quaternary
amine compounds
4.6x75 mm column (standard ID)
2.1 x 20 mm Column
90% ACN + 10% DI water 0.5% Formic Acid
92% ACN+ 8% DI water with 0.5% Formic Acid

71. Comparison of Choline and Acetylcholine Retention
Cogent UDC-Cholesterol &
BD C18 Columns
ANP Conditions

72. Retention of Nucleosides on a Cogent UDC-Cholesterol HPLC Column
Column: Cogent UDC Cholesterol 4.6 x 150 mm
Nucleoside Analog MW BpKa LogP
Compound
Compound
Column: Cogent UDC Cholesterol 4.6 x 150 mm
Nucleoside Analog MW BpKa LogP
Cytidine Derivative
Cytidine Derivative
Retain Nucleosides with Reverse Phase Solvents
Same pKa and LogP
Shape Recognition
Same pKa and Different LogP
Increased ANP

73. Retention of the Basic Drug “Tobramycin” in Acid Conditions

74. Comparison of Peak Shapes on TYPE-C Columns And Ordinary HPLC Columns
Tobramycin - High Efficiency on TC Column
Cholesterol Column
End-capped Commercial C18 Column
4.6 x 75 mm
4.6 x 150 mm

75. Separating Tetramic Acid derivative and a Hydrophilic Peptide
with similar Log P and acid dissociation constants
on the cholesterol column
Tetramic Acid Derivative -R

76. Retention can be Observed for Other Hydrophilic Peptides
General Peptide Structure: Ac-AXEXAHKAY-NH2

77. Detection: TOF-MS m/z = 166
Phenylalanine Retention on a Cogent Silica-C™ Column:
70:30 ACN/water + 0.1% FA
tR = 2.96 min t0 = 0.95 min
80:20 ACN/water + 0.1% FA
tR = 5.22 min
90:10 ACN/water + 0.1% FA
tR = min
Detection: TOF-MS m/z = 166

78. Succinic Acid Retention on a Cogent Silica-C™ Column
Mobile Phase: 95:5 ACN/water + 0.1% FA
Detection: m/z = 117

79. Complex Amino Acid Mixture on a Cogent Diamond Hydride™
Column: 2.1 x 150mm DH Gradient: 95%B – 60% B
Flow Rate: 0.4ml/min Column Temp: 20°C
Detection: Agilent TOF-MS Mobile Phase: A:0.1% Formic Acid B:MeCN 0.1% FA

80. Polar-Alkaloids on Diamond Hydride™
Tryptamine HCL
Column: Cogent Diamond Hydride™ HPLC Column, 4 mm, 100 Å
Dimensions: mm i.d. x 75mm packed
Mobile phase: :15 acetonitrile/DI water + 0.1% fa % TFA
Flow rate: mL/min.

81. Metabolite of Cyromazine on Diamond Hydride™
Melamine
Column: Cogent Diamond Hydride™ HPLC Column, 4 mm, 100 Å
Dimensions: mm i.d. x 75mm packed
Mobile phase: :40 acetonitrile/DI Water + 0.1% acetic Acid
70:30 acetonitrile/DI Water + 0.1% acetic Acid
Flow rate: mL/min.

82. Biogenic Amine on Diamond Hydride™
Serotonin & Analogs
5-hydroxy-3-indole acetic acid (5-HIAA), metabolite of serotonin 192 m/z
3,4-dihydroxyphenylacetic acid (DOPAC) 169 m/z
Serotonin 177 m/z
Epinephrine 184 m/
Column: Cogent Diamond Hydride™ HPLC Column, 4 mm, 100 Å
Dimensions: mm i.d. x 75mm packed
Mobile phase: A: DI water + 0.1% formic acid
B: acetonitrile + 0.1% formic acid
Flow rate: mL/min. Inverse Gradient

83. Carbohydrates on Diamond Hydride™
Glucose
Column: Cogent Diamond Hydride™ HPLC Column, 4 mm, 100 Å
Dimensions: mm i.d. x 150mm
Mobile phase: A: DI water + 0.1% acetic acid
B: acetonitrile + 0.1% acetic acid
Flow rate: mL/min. Inverse Gradient

84. Citric, Maleic & Trans-Aconitic Acid
Organic Acids on Diamond Hydride™
Citric, Maleic & Trans-Aconitic Acid
Column: Cogent Diamond Hydride™ HPLC Column, 4 mm, 100 Å
Dimensions: mm i.d. x 150mm
Mobile phase: A: DI water + 0.1% amm acetate
B: acetonitrile + 0.1% amm acetate
Flow rate: mL/min. Inverse Gradient

85. Sulfonamide on C18 Sulfonamide
Column: Cogent Bidentate C18™ HPLC Column, 4 mm, 100 Å
Dimensions: mm i.d. x 75mm
Mobile phase: A: DI water + 0.1% formic acid
B: acetonitrile
Flow rate: mL/min. RP Gradient

86. Sulfonamide Retention
Carbohydrates on C18
Sulfonamide Retention
Column: Cogent Bidentate C18™ HPLC Column, 4 mm, 100 Å
Dimensions: mm i.d. x 75mm
Mobile phase: A: DI water + 0.1% formic acid
B: acetonitrile
Flow rate: mL/min. Fast RP Gradient

87. Cleaning Validation for Guanidine
Strong Bases on C18™
Cleaning Validation for Guanidine
Column: Cogent Bidentate C18™ HPLC Column, 4 mm, 100 Å
Dimensions: mm i.d. x 150mm
Mobile phase: A: 100% DI water + 0.1% Phos acid + 1.5g/L
pentane sulfonic acid
Flow rate: mL/min. Isocratic, no loss of RT over time.

88. Guanfacine HCl & Degradants
USP Method on C8
Guanfacine HCl & Degradants
Column: Cogent Bidentate C8™ HPLC Column, 4 mm, 100 Å
Dimensions: mm i.d. x 150mm
Mobile phase: % Acetonitrile, 70% DI Water w/Phos Acid, SDS
Flow rate: mL/min. Isocratic 7 minute separation

89. Phosphorylated Sugars on Diamond Hydride™
Nucleotides by LCMS
ADP Glucose
Proprietary Sugar Nucleotide
CDP Glucose
UDP Hexanolamine
Column: Cogent Diamond Hydride™ HPLC Column, 4 mm, 100 Å
Dimensions: mm i.d. x 150mm
Mobile phase: A: DI water + 0.1% amm formate, pH 7.2
B: 90% acetonitrile + 10% DI Water +
0.1% amm formate, pH 6
Flow rate: mL/min. Inverse Gradient

90. Furazolidone: API & Related Compounds
Antibiotics on C18
Furazolidone: API & Related Compounds
Column: Cogent Diamond Hydride™ HPLC Column, 4 mm, 100 Å
Dimensions: mm i.d. x 150mm
Mobile phase: A: DI water + 0.1% acetic acid
B: acetonitrile + 0.1% acetic acid
Flow rate: mL/min. Inverse Gradient

91. Furazolidone: API & Related Compounds
Antibiotics on C18
Furazolidone: API & Related Compounds
Column: Cogent Bidentate C18™ HPLC Column, 4 mm, 100 Å
Dimensions: mm i.d. x 150mm
Mobile phase: % Acetonitrile, 88% DI Water w/ 10ml/L acetic acid
Flow rate: mL/min. Isocratic

92. Benefits of Silica Hydride Based Stationary Phases
Operate in 3 different modes of chromatography: normal phase, aqueous normal phase and reversed phases. Versatile and very convenient and useful.
Use in 100% aqueous mobile phases in reversed phase with no loss of Rt over time. More robust methods.
Bases are well retained at low pH so high pH mobile phases may not be necessary or useful as orthogonal.
Surface absorbs very little water so there is reproducible retention in the normal phase without drying mobile phases and rapid equilibration for gradient separations.
Use over a broad range of pH and can be used at high temperatures.
Increase MS Sensitivity with high organic MP and use TFA in low concentrations for MS Compatibility.
Expand your Reach without exotic solvents or additives.

93. Where can I get More Data?
Look under “Useful Information” in Top Menu Bar
Application Notes
HPLC
Cogent TYPE-C

94. You Can See there are MANY uses for the Cogent TYPE-C™ Silica Based HPLC Columns. These shown are only the beginning.

95. You Can See there are MANY uses for the Cogent TYPE-C™ Silica Based HPLC Columns. These shown are only the beginning.

96. Thank You.