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Flare_Diamond_Analytics

RP, IEC and HILIC Chromatography at Extremes of pH and Temp,

 

New: Flare Wide Pore C18 for Protein Analytics (up to 500kDa). See Applications

 

Introducing “FLARE”,  a novel nano-diamond core-shell enabling technology

In routine analytics Chromatographers  search for columns that cover a broad spectrum of analytes and process parameters.  This enables Chromatographers to improve their productivity substantially and to please their superiors. 80% of Chromatographers prefers to use reverse phase columns. Most column manufacturers want to be part of the large reverse phase market by launching new types of reverse phase columns. Market analysis reveal that many newly launched LC columns have attractive new names but very similar selectivity properties.

 Two brilliant  pioneering Chemists (see below) at Diamond Analytics applied nanotechnology with polycrystalline nano diamond  technology to evolve  a new to the world  reverse phase column  with  spectacular physical properties!  They named it “FLARE”.

Diamond Michael Vail

Diamond Matt Linford

Dr. Michael A. Vail

Dr Matthew R. Linford

 

What is FLARE?

Diamond Analytics is a division of  the large US Synthetic company  founded as synthetic diamond  manufacturers in 1978.  2006 Diamond Analytics was the first company to  combine nanotechnology  with reverse phase materials technology and so commenced the start of diamond based HPLC materials and columns. Diamond based  columns are unique, display amazing properties and are available world wide.  LCC  was engaged to create unique application and to build their HPLC column market in Europe.

How are Flare core shell particles manufactured?

Diamond Particle creation process

Carbon particles are being coated in a multi step process with Polyallylamine (PAAm)  and nano- diamonds,  then crosslinked and then functionalized  e.g. with a C18 ligand. The free amino groups on the cationic polymer surface can be protonated  (charged) /deprotonated at various pH values. Thus FLARE C18 is an exciting  multi-mode chromatography material for creative Chromatographers.  As multi mode materials Flare has two retention mechanisms

    • By Hydrophobic interaction
    • Ionic interaction
  • Ionic interaction  strength and peak shape can be controlled by :
    • pH (depends on analyte type and pKa)
    • Buffer type and concentration
    • Temperature
    • Solvents and additives used

 

Particle dimensions and shape are as follows:

Diamond Core Material

Diamond Particle construct

Diamond Finished Particles

Starting Carbon Core

 

Finished Particle 3.6 um

FLARE C18MM, C18+, HILIC

  • Total particle diameter: 3.6 um
  • Solid carbon core diameter:  3.4 um.
  • Porous shell thickness:  0.1 um
  • Surface area is 25 sqm/g which is the silica  equivalent of 40 sqm
  • Pore diameter : 120 Angstom
  • Loading Capacity (Amitryptiline): 5 ug(4.6 mm x 33 mm column)
  • Ion exchange capacity:  10 - 20 ueq/column (2.1x50mm,UTAS)
  • pH Stability:  1 -13
  • Temperature stability: 100 C

FLARE Wide Pore C18

  • Total particle diameter: 3.6 μm
  • Core diameter: 3.4 μm
  • Shell radius: 0.1 μm
  • Surface area: 10 m2/g
  • Pore diameter: 250
  • Loading capacity: 150 μg (4.6 x 33mm, BSA)
  • pH stability: 1 – 13
  • Temperature stability: 100C
  • Permeability: 9.8·10-11 m2
  • Porosity: 40%

Physico chemical properties for materials used in chromatography (25C)

Diamond Flare Physico Chemical Properties

 

When packed into columns they show excellent van Deempter properties as follow:

Diamond Van Deemter

FLARE Column also show excellent pH Stability over many injections

Diamond Column life at pH2

 

How does it compare?

The Hydrophobic Subtraction Model is a scientific tool to characterize and compare the physico-chemical properties  of a column. The triangle below encompasses the “Reverse Phase world”.

Diamond Unique column selectivity

The Reverse Phase World is crowded with phases of similar properties. The red dot in the triangle above represents the  FLARE C18 column. It show that this column has unique selectivity that can perform separations that other C18 columns in the market can’t do.

Enlarge the chromatographic spectrum

FLARE enables you to use LC System as GC method! With FLARE you can use temperature as additional gradient. Go to MatrixEx for the choice of oven click here

How to order:

Product Range& Article No.

.

Product

ID (mm)

Length 33 mm

Length 50 mm

Length 100 mm

Length 150 mm

Flare C18 MM

2.1

 

FL36-21050

FL36-21100

FL36-21150

Flare C18 MM

4.6

FL36-46033

FL36-46050

FL36-46100

FL36-46150

Flare C18 +

2.1

 

FP36-21050

FP36-21100

FP36-21150

Flare C18+

4.6

FP36-46033

FP36-46050

FP36-46100

FP36-46150

Flare HILIC

2.1

 

FH36-21050

FH36-21100

FH36-21150

Flare HILIC

4.6

FH36-46033

FH36-46050

FH36-46100

FH36-46150

Flare Kit

2.1

 

FK36-21050

FK36-21100

FK36-21150

Flare Kit

4.6

FK36-46033

FK36-46050

FK36-46100

FK36-46150

Flare WP C18

2.1

 

FW36-21050

FW36-21100

FW36-20150

Flare WP C18

4.6

 

FW36-46050

FW36-46050

FW36-46150

 

 

 

 

 

 

 

 Select Applications

FLARE C18 MM (Mixed Mode)

Retentions mechanism

 Diamond Flare MM Retentionmechanisms

 

Agrichemical Application

Natural Compounds

Pharmaceuticals

BioPharmaceuticals

FlareMixedModeColumn-TriazineHerbicides

Cannabinoids

Beta2-Agonists and Amphetamines

BSA-Apo LC-MS

Retention of Acidic Herbicides vs pKa Values

THC

Tricyclic Antidepressants (TCAs)

Separation of Apo-Transferrin and Bovine Serum Albumin (BSA) Proteins

Acidic Herbicides

Alkaloids

Estrogens

 

 

Eucalyptus Essential Oil

 

 

 

Melaleuca Essential Oil

 

 

 

Lavender Essential Oil

 

 

 

Peppermint Essential Oil

 

 

Our customers like to see differences in elution order, flexibility in using pH and  high temperatures.  Cannabinoids are for example stable at high pH where silica cannot be used.Drugs such as amphetamins or Beta2 antagonist possess primary and secondary amines that give these compounds pKa values of 9.3 to 9.8 .  At these pH the drugs would be ionized causing poor retention on a C18 column. At elevated pH the molecule would be neutral making good separation with a reversed phase material possible.

FLARE C18+

Food Additives

 

 

 

Methoxybenzoic Acid

 

 

 

 FLARE HILIC

Biopharmaceuticals

Cosmeticals

 

 

 

Nucleotides

Hydroxybenzoic Acids

 

 

 

Nucleobases

 

 

 

 

 

 

 

 

 

FLARE Wide Pore (WP) C18

Works primarily in reversed phase mode due to interaction of proteins with C18 ligands. Optimal pore size and permeability allows for retention of small to large-sized proteins (up to 500 kDa). Has been effectively used in analyzing mAbs, ADCs, insulin, haemoglobin,

Intact & Digested mAb Separations

Sample Preparation:

- Intact mAb aqueous solutions were prepared to a concentration of 2 mg/ml.
- Reduction of mAbs were performed by addition of 0.05 mg of dithiotreitol (DTT) to a 100 μl solution of mAb stock solution. The solution was incubated at 40oC for 60 minutes.
- Digestion was done by addition of 50 μl of papain (1.0 mg/ml) to the reduced sample. The solution was incubated at 40oC for 5 hours.

Sample Characterization:

- Cetuximab: The reduction process yielded two main fragments, the light chain (LC) of ca. 25 kDa and the heavy chain (HC) of ca. 50 kDa.
- Rituximab: The reduction and digestion processes yielded three main fragments of ca. 25 KDa, namely the native LC, the single-chain Fc (sFc) and the Fab portion of the HC (Fd)Diamond Flare WP C18 Rhituximap

Column Name: FLARE WP C18               

Column Dimensions: 2.1 x 100 mm

HPLC System: Acquity I-Class UPLC system

Injection Volume: 0.5 μl

Detection: FLD (280 - 360 nm)

Flow Rate: 0.15 ml/min

Mobile Phase: A: 0.5% TFA in H2O, B: 0.5% TFA in ACNDiamond Flare WP C18 Cetuximap

Gradient:

Time (mins)

%A

 %B

0.00

 75

25

10.00

 67

33

10.20

75

25

12.00

75

25 end

Temperature: 90C
Analyte: Intact Rituximab (MW= 144 kDa), Intact Cetuximab (MW= 146 kDa)

 

Trypsin inhibitor from Glycine(soybean) max  MW: 20.1 kDa

Column Name: FLARE WP C18
Column Dimensions: 4.6 x 100 mm
Column ID: 16636.1-3
HPLC System: Agilent 1200
Injection Volume: 5.0 μl
Detection: UV at 230 nm
Flow Rate: 1.0 ml/min
Mobile Phase: A: 5 ml ACN, 95 ml H2O, 0.1 ml TFA; B: 80 ml ACN, 20 ml H2O, 0.1 ml TFA
Gradient:

Time (mins) %A %B
0.00 100 0
12.00 60 40
12.01 100 0
20.00 100 0 end
Temperature: 55 C

Diamond Flare WP C18 Trypsin Inhibitor

Histone from calf thymus II-A (MW: 11-21 kDa)

Column Name: FLARE WP C18
Column Dimensions: 4.6 x 100 mm
Column ID: 16636.1-3
HPLC System: Agilent 1200
Injection Volume: 5.0 μl
Detection: UV at 230 nm
Flow Rate: 1.0 ml/min
Mobile Phase: A: 5 ml ACN, 95 ml H2O, 0.1 ml TFA; B: 80 ml ACN, 20 ml H2O, 0.1 ml TFA
Gradient:
Time (mins) %A %B
0.00 100 0
12.00 60 40
12.01 100 0
20.00 100 0 end
Temperature: 35 C
mAU230 nm
Time (min) 12

Diamond Flare WP C18Histone

Deoxyribonuclease I from bovine pancreas (MW: 39 kDa

Column Name: FLARE WP C18
Column Dimensions: 4.6 x 33 mm
Column ID: 16636.2-3-1
HPLC System: Agilent 1200
Injection Volume: 5.0 μl
Detection: UV at 230 nm
Flow Rate: 1.2 ml/min
Mobile Phase: A: 5 ml ACN, 95 ml H2O, 0.1 ml TFA; B: 80 ml ACN, 20 ml H2O, 0.1 ml TFA
Gradient:
Time (mins) %A %B
0.00 100 0
3.00 30 70
3.01 100 0
6.00 100 0 end
Temperature: 30, 55, 60 C

Diamond Flare WP C18 Desoxyribonuclease

Ribonuclease A from bovine pancreas (MW: 13.7 kDa)

Column Name: FLARE WP C18
Column Dimensions: 4.6 x 33 mm
Column ID: 16636.2-3-1
HPLC System: Agilent 1200
Injection Volume: 5.0 μl
Detection: UV at 230 nm
Flow Rate: 1.2 ml/min
Mobile Phase: A: 5 ml ACN, 95 ml H2O, 0.1 ml TFA; B: 80 ml ACN, 20 ml H2O, 0.1 ml TFA
Gradient:
Time (mins) %A %B
0.00 100 0
3.00 30 70
3.01 100 0
6.00 100 0 end
Temperature: 30, 55, 60 C

Diamond Flare WP C18 Ribonuclease

Hemoglobin human (MW: 60.4 kDa)

Column Name: FLARE WP C18
Column Dimensions: 4.6 x 100 mm
Column ID: 16636.1-3
HPLC System: Agilent 1200
Injection Volume: 5.0 μl
Detection: UV at 230 nm
Flow Rate: 1.0 ml/min
Mobile Phase: A: 5 ml ACN, 95 ml H2O, 0.1 ml TFA; B: 80 ml ACN, 20 ml H2O, 0.1 ml TFA
Gradient: Time (mins) %A %B
0.00 100 0
14.00 60 40
14.01 100 0
22.00 100 0 end
Temperature: 35 C

Diamond Flare WP C18 Haemoglobin human

Hemoglobin Bovine (MW: 67 kDa)

Column Name: FLARE WP C18
Column Dimensions: 4.6 x 100 mm
Column ID: 16636.1-3
HPLC System: Agilent 1200
Injection Volume: 5.0 μl
Detection: UV at 230 nm
Flow Rate: 1.0 ml/min
Mobile Phase: A: 5 ml ACN, 95 ml H2O, 0.1 ml TFA; B: 80 ml ACN, 20 ml H2O, 0.1 ml TFA
Gradient: Time (mins) %A %B
0.00 100 0
14.00 60 40
14.01 100 0
22.00 100 0 end
Temperature: 35 C

Diamond Flare WP C18 Haemoglobin bovine

 

Please contact us to discuss your particular application Click here !

 

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