FlexFire UHPLC and HPLC Columns - Develosil USA

UHPLC and HPLC Columns

The HPLC and UHPLC Columns That Help you Focus on Discovery

  • Detect a 1-base difference in miRNA
  • Metal free columns
  • pH forgiveness

FlexFire's Application

FlexFire’s Application in Biopharma

Reliable analysis of oligonucleotides is becoming increasingly more important as the results of analysis is more directly tied to the final product.

FlexFire columns provide flexibility and reliability you deserve with our variety of particle sizes, all offering the same high resolution.

The ability to reliably detect monoclonal antibodies (mAb) has grown in popularity as the demand for analysis of miRNA has increased. The most recent notable use is the SARS-CoV-2 vaccine. FlexFire’s Column mAb-RP, 2.6 µm (2.0×50 mm) provides the ability to detect up to a 1-base difference in miRNA.

Detect up to a 1-base difference in miRNA


The FlexFire mAb-RP, 2.6 µm (2.0×50 mm) offers excellent resolution and allows for the detection of a 1-base difference in RNA, as depicted in the chromatogram above. For the best resolution and retention, we suggest using Diisopropanolamine (DIPA) as your ion-pair. For comparison, we have also displayed our results while using triethylamine (TEA).

Column: FlexFire Column mAb-RP, 2.6 µm (2.0×50 mm)

Temperature: 60°C
Detection: UV260 nm
Mobile phase:


A) 100 mM HFIP + 10 mM TEA

B) (A) / Methanol=50/50


A) 100 mM HFIP + 10 mM DIPA

B) (A) / Methanol=50/50


Pore Size: 1000 Å

Injection volume: 2.0 µL

Mixer: 100 µL


     20nt: 5′-GU ACG CGG AAU ACU UCG AdTdT-3′

     21nt: 5′-CGU ACG CGG AAU ACU UCG AdTdT-3′

FlexFire’s Metal-Free Columns

Metal-free columns have received a great deal of attention not only in the analysis of peptides and proteins, but also in all fields.

Metal-free columns prevent absorption by a lining of packing material between the stainless steel column and the compound. This has an additional beneficial effect on the resolution and detection.

How it Works

FlexFire’s metal-free column has a PEEK tube inside of a stainless steel tube. This makes it highly durable, even under the high pressure of UHPLC columns with a 1.6µm particle size.


The PEEK interior prevents the compounds from coming into contact with the stainless steel exterior of the tube.

UHPLC Metal-free PEEK Lining

In the chromatogram below, we demonstrate a comparison of results between a traditional column and a metal-free column. The metal-free column detects ATP and ADP that the traditional column did not. Additionally, the AMP readings resulted in a sharper peak shape when using the metal-free column.

Metal vs Metal-Free
Metal vs Metal-Free Conditions

FlexFire Specifications

In the world of research, you need to count on your UHPLC column to deliver precise, high-resolution, reproducible separations—especially if you work with large molecules, such as monocolonal antibodies, or use high-pH mobile phases.

In the world of production, you need your HPLC column to work just as hard. But why must your protocols differ from those upstream in R&D?

They don’t have to. Not anymore. Not with FlexFire.

FlexFire UHPLC and HPLC chromatography columns not only provide industry-leading resistance to highly-basic applications, but they’re the only brand which allows for similar protocols for UHPLC and HPLC, allowing for seamless transferability from R&D to QC, regardless of functional group or particle size.

You won’t find this combination of robustness and flexibility anywhere else on the market.


Who makes the strongest HPLC column?

pH Forgiveness

We understand the restrictions pH level can put on the separation process. In order to help you in your journey to discover, the  FlexFire series of columns offers a higher variance in pH, ranging from 1 pH to 10 pH.


With this higher range of viable pH levels, you have more personalization during the separation process.


See how FlexFire out-performs well-known competitors with a pH of 10.5!

  • 20% Stronger Columns
  • Smaller Particles, Greater Longevity
Column Functional Group C18
Size 2.0 x 50 mm
Mobile Phase Acetonitrile / 10 mM NH4OH, pH10.5=60/40
Flow Rate 0.5 mL/min
Temperature 40°C
Detection UV 254 nm
Sample Naphthalene (0.1 mg/mL)
Injection Volume 0.16 µL
Column Types Particle Size (µm)
Develosil FlexFire 1.6
Brand A 1.7
Brand B 1.7
Brand C 1.8
Brand D 1.9


Unprecedented transferability between UHPLC and HPLC

The FlexFire series provides 1.6, 2.6, and 5 µm particle sizes which work for your analytical systems in both UHPLC and HPLC. This gives you unprecedented power to employ the same line of chromatography columns in different departments, from R&D to QC.

Think of the advantages to your operation: FlexFire dramatically reduces the required time for tweaking the optimal condition from one particle size to another, thanks to the quality and homogeneity of its silica gel packing, end-capping, and bonding of functional groups. That’s the key to FlexFire’s predictable consistency across all columns.

Don’t believe it? Just look at the figures below, showing HETP and column pressure at different flow rates. FlexFire’s smaller particle size provides higher resolution across a wider range of flow rates.

Pressure and HETP vs Flow Rate for FlexFire C18

Column Develosil FlexFire C18 (2.0 x 50 mm)
Mobile Phase Acetonitrile/Water = 60/40
Temperature 40°C
Detection UV 254 nm
Sample Naphthalene
Injection Volume 0.16 µL

Different particle sizes, same resolution

Witness the flexibility to transfer between UHPLC and HPLC

FlexFire provides the same surface area ratio of 340 m2/g, along with the same physical properties, and the same bonding of functional group and end-capping—on three different particle-size silica substrates.

As a result, by choosing the correct column dimensions, you can maintain equivalent resolution, even when shifting between particle sizes. That’s the “Flex” in “FlexFire”: You can now easily determine the flow rate and gradient for different particle sizes with your chromatography software: No trial-and-error. No adjustments.

If you’re using UHPLC during R&D and HPLC during QC, FlexFire will save time previously spent re-adjusting the analytical conditions to find the best resolution. And if you’re planning to adopt UHPLC in the future, you can simply start with FlexFire 2.6 µm or 5 µm columns, knowing you’ll be “future-proofed” for UHPLC.

FlexFire C18, 5 µm (2.0 x 150 mm)

Gradient min mL/min %A %B Curve
0.00 0.189 90 10 6
8.44 0.189 75 25 6
16.88 0.189 75 25 6
17.06 0.189 90 10 6

Injection Volume: 1.2 µL

FlexFire C18, 2.6 µm (2.0 x 50 mm)

Gradient min mL/min %A %B Curve
0.00 0.364 90 10 6
1.46 0.364 75 25 6
2.93 0.364 75 25 6
2.96 0.364 90 10 6

Injection Volume: 0.4 µL

FlexFire C18, 1.6 µm (2.0 x 50 mm)

Gradient min mL/min %A %B Curve
0.00 0.592 90 10 6
0.90 0.592 75 25 6
1.80 0.592 75 25 6
1.82 0.592 90 10 6

Injection Volume: 0.4 µL
System:Waters ACQUITY UPLC H-Class Plus
Mixer: 100 µL

Mobile Phase A) 25mM HCOONH4, pH3.0
B) Acetonitrile
Temperature 40°C
Detection UV 260 nm
  1. Gly-Tyr
  2. Val-Tyr-Val
  3. Met-Enk
  4. Angiotensin II
  5. Leu-Enk

Superior silica at work

The FlexFire series of reverse-phase columns take advantage of Develosil’s premium silica: It’s the highest grade, uniform, high-density, low-metal silica chemistry on the market, as evidenced by its unparalleled performance and durability in high-pH applications.

  • Durable. Stable at pH 1 to pH 10
  • Flexible. Transfer from UHPLC to HPLC at 1.6, 2.6, and 5 µm, with functional groups from C1 to C30 and Hilic

We invite you to try out FlexFire, obligation-free, for 7 days. Click here to get started!

Specifications and Line Up

Function Particle Size Surface Area Pore Diameter Pore Volume Carbon Load  pH Range  Temp Tolerance
C18 1.6, 2.6, and 5 µm 340 m2/g 11 nm 1.0 mL/g 22.00% 1-10 <60°C
C30 11.00%
C8 12.00%
AQ C18 8.50%
C1 5.50%
C4 mAb 2.5 and 5 µm 100 nm 1.4 mL/g 0.60% <80°C
mAb-RP 24 m2/g 100 nm 0.8 mL/g 1.30%
120 SEC  5 µm 340 m2/g 12 nm 1.0 mL/g 9.00% 2-10
300 SEC 170 m2/g 30 nm 1.4 mL/g 6.00%
WP C1 5 µm 3.00% 1-10
WP C4 5.00%
WP C8 7.00%
WP C18 1.6, 2.6, and 5 µm 15.00%

Pressure Tolerance

Particle Size For all Pore Sizes
1.6 µm 800 bar
2.6 µm 600 bar
5 µm 300 bar



Experience FlexFire For Yourself

You can’t appreciate FlexFire’s durability, robustness in high-pH environments, or unique ability to transfer between R&D and QC, without experiencing it firsthand.