InertSustain AX-C18 Mixed-Mode Column

InertSustain AX-C18 is a reversed-phase mixed-mode column with an octadecyl group and an alkylamine group.

It is possible to retain polar acidic components that are generally not retained without the use of ion-pair reagents, and by setting the ion exchange capacity weaker, the buffer mobile phase used in LC / MS can be sufficiently eluted. Is designed for. It also has excellent inter-lot reproducibility, which tends to be a problem with columns.

Specifications:

Chemical bond group silica gel: High-purity spherical silica gel (ES silica)
Particle size: 3µm, 5µm
Surface area: 200m 2 / g
Pore size: 200Å
Pore volume: 1.00 mL / g
Chemical bond group: Octadecyl group + tertiary amino group
End-capping: Yes
Carbon Loading: 8%
USP Code: L1, L78
Recommended PH: 1 to 9

Recommended when

You want to slightly change the separation pattern from C18 columns
You want to analyze acidic compounds and anionic compounds
You are having trouble with reproducibility and lot-to-lot variation in anion exchange columns

Feature 1: Retention behavior that combines both reversed-phase and anion exchange

Compared to typical C18 columns ➤➤➤Increased retention of acidic compounds

Analysis of nucleobases and monophosphates

Conditions:

Column: 3 μm, 150 × 2.1 mm I.D.

Eluent: 25 mM NaH2PO4 in H2O

Flow Rate: 0.2 mL/min

Col. Temp.: 40 ℃

Detection： UV 254 nm

Feature 2: Excellent retention for acidic compounds

No ion paid reagent in eluent➤➤➤ Highly polar acidic compounds retained

Analysis of sugar‐nucleotides

Eluent ： 20 mM HCOONH4 in H2O

(Ion pair not used)

Conditions

Column： 3 μm, 150 × 2.1 mm I.D.

Flow Rate： 0.2 mL/min

Col. Temp.： 40 ℃

Detection： UV 254 nm

Sample： 1. UDP‐Glu 2. UDP‐Gal 3. GDP‐Man

Strongly retains highly polar acidic compounds which cannot normally be retained by C18 column for an analysis of highly polar compounds!

Even if the salt concentration of the eluent is low ➤➤➤Acidic compounds strongly retained & Compatible with MS analysis

Analysis of monophosphoric acid

Eluent ：

A) 10 mM HCOONH4 in CH3OH

B) 10 mM HCOONH4 in H2O

A/B = 5/95 ‐ 10 min ‐ 100/0 ‐ 5 min ‐ 100/0, v/v

Conditions

Column：3 μm, 150 × 2.1 mm I.D.

Flow Rate：0.2 mL/min

Col. Temp.：40 ℃

Inj. Volume：5 μL

Detection：MS/MS (ESI, Positive, MRM)

Sample: Q1 > Q3

1. CMP: 324.90 > 111.95 (+) 0.5 mg/L

2. GMP: 364.90 > 153.00 (+) 0.5 mg/L

3. UMP: 325.90 > 96.90 (+) 1.0 mg/L

4. AMP: 348.90 > 136.95 (+) 0.05 mg/L

Strongly retains highly polar acidic compounds even when the salt concentration is as low as 10 mM! Sharp elution peaks! Can be analyzed at low salt concentration. Compatibility with MS!

Feature 3: Excellent reproducibility and quality

Reproducibility issues with mixed-mode ➤➤➤ are solved by GL Sciences’ original packing and manufacturing technology

Shipping inspection for evaluation of lot‐to‐lot variation

Conditions

Column ： 3 μm, 150 × 4.6 mm I.D.
Eluent ： CH3CN/H2O = 65/35, v/v
Flow Rate ： 1.0 mL/min
Col. Temp.：40 ℃
Detection ： UV 254 nm

Sample ： 1. Uracil 2. Acetophenone 3. Benzene 4. Toluene 5. Naphthalene

Evaluation of lot‐to‐lot variation in acidic compounds

Conditions

Column ： 3 μm, 150 × 4.6 mm I.D.
Eluent ： CH3CN/0.1 % HCOOH = 10/90, v/v

Flow Rate ： 1.0 mL/min

Col. Temp. ： 40 ℃

Detection ： UV 254 nm

Sample ： 1. AMP

Excellent reproducibility of retention time in mixed‐mode columns where reproducibility is difficult to control!

InertSustain AX-C18 Mixed-Mode Column