HPLC, LC/MS Columns

InertSustain AQ-C18®

Maximizing retention for highly polar compounds inreversed phase methods with highly aqueous mobile phases

Physical Properties
Silica :Particle Size :Surface Area :Pore Size :Pore Volume :Bonded Phase :End-capping :Carbon Loading :USP Code :pH Range :ES (Evolved Surface) Silica Gel
1.9 μm, 3 μm, 5 μm
350 m2/g
100 Å (10 nm)
0.85 mL/g
Octadecyl Groups
Complete13.0 %
L1, L961～10InertSustain AQ-C18®

Benefits
1. Exceptional retention for highly polar compounds (hydrophilic)
2. Highly inert packing material results in less tailing of peaks for virtually any type of analytes
3. Extreme resistance to low and high pH mobile phases
4. Endlessly reproducible from column-to-column and batch-to-batch
InertSustain AQ-C18 columns are designed to achieve strong retention for highly polar compounds, which is themost challenging goals in developing reversed phase methods. The optimization of bonding of the C18 groups atequal distance to the silica gel enable InertSustain AQ-C18 to offer significant retention for highly polar compounds
even under water rich mobile phases.
As illustrated in the following plot, InertSustain AQ-C18 provided exceptional retention for highly polar compounds
even under water rich mobile phases without dewetting or phase collapse.

Retention Factor of acidic, basic and neutralcompounds using 100% water mobile phase

Retention Properties of InertSustain AQ-C18
Remarks:

InertSustain AQ-C18
Aquasil C18

1.

The horizontal and vertical retention factor
values were calculated with reference to theretention factor values obtained fromInertSustain AQ-C18.

2.

The horizontal values represents the retention
factor of amylbenzene using
methanol/water=80/20.

3.

The vertical values represents the retention
factor of acidic, basic and neutral compounds
using 100% water mobile phase. (For furtherdetails, please see the following chart)

Atlantis T3
Inertsil ODS-4

Inertsil ODS-SP

Inertsil ODS-3
InertSustain C18

CAPCELL PAK C18-AQ
Synergi Fusion RP
XSelect CSH C18
Retention Factor of amylbenzene using methanol/water=80/20

InertSustain AQ-C18 provided strong retention for all basic, neutral and acidic
compounds under 100% water mobile phase
Remarks:
Pyridoxamine
Basic
5-methyluridine

Dopamine

NeutralInertSustain AQ-C18

UracilAcetic AcidAcidic
Succinic Acid

1

Generally, the retention for acidic compounds become
stronger than basic compounds when the surface of thesilica gel is basic.
The retention for basic compounds become stronger
than acidic compounds when the surface of the silica gel
is acidic.

CAPCELL PAK C18-AQ
Atlantis T3
Synergi Fusion RPHighly Polar Compound Retention with 100% Aqueous Mobile Phases
It is indeed difficult to retain highly polar samples by reversed phase mode as the polar samples tend to elute near
the t0 (void volume) with water rich mobile phases. On the other hand, polar embedded C18 columns or C18
columns with hydrophilic endcapping are available in the market for retaining highly polar compounds. However,they often show tailing or poor peak shapes due to the secondary interaction from their embedded functionalgroups.
InertSustain AQ-C18 not only provide strong retention for highly polar compounds, but also delivery symmetricpeak shapes for virtually any type of analytes.
1

InertSustain AQ-C18t0Conditions
Column： 5 μm, 150 × 4.6 mm I.D.
Eluent： 0.1 % HCOOH in H2O
Flow Rate ： 1.0 mL/min
Col. Temp. ： 40 ℃
Detection ： UV 210 nm

425361Atlants T3

t0Synergi Fusion RPt03+42

146

13+4

t0 2

InertSustain C182: Not eluted

53

Sample:1. Pyridoxamine
2. Thiamin
3. Nicotinic acid
4. Pyridoxal
5. Nicotinamide
6. Pyridoxine

65(Vitamin B6)(Vitamin B1)(Vitamin B3)(Vitamin B6)(Vitamin B3)(Vitamin B6)

65(Conventional C18 column)0246

Time(min)8Retentivity Under 20% Organic Solvent
As shown below, InertSustain AQ-C18 delivered stronger retention of catechin sample even under 20% organic
solvent mobile phase with exceptional peak shapes, while competitive column brand failed. Furthermore,
InertSustain AQ-C18 can prevent the co-elution between the targeted polar analytes and solvent peaks or sample
matrices due to its enhanced retentivity.
Complete separation
between solvent peak
1

InertSustain AQ-C1803 4224Time (min)5

2.0 2.2 2.4 2.6Time (min)66810Elution with solvent peak

12Atlants T302Conditions
Column： 5 μm ,150 ×4.6 mm I.D.
Eluent： A) 0.1 %HCOOH in H2OB) CH3CN
A/B= 80/20, v/v
Flow Rate ：1.0 mL/min
Col. Temp. ：40℃
Detection ：UV 280 nm
Sample:1. Gallocatechin (-) (GC)
2. Catechin (+) (C)
3. Epicatechin (-) (EC)
4. Epigallocatechin gallate (-) (EGCg)
5. Epicatechin gallate (-) (ECg)
6. Catechin gallate (-) (Cg)

2.0 2.2 2.4 2.6Time (min)4534Time (min)66Poor peak shape8102InertSustain AQ-C18®

Applications
High-Throughput Analysis of Drugs
InertSustain AQ-C18 (1.9 μm)
5.0(x1,000,000)

Acquity BEH C18 (1.7 μm)
5.0

1

4.5

4.5

Increase in Sensitivity

4.0(x1,000,000)

4.0

3.5

3.5

3.0

3.0

2.5

12.5

32 4

2.02 4

2.0

1.5

1.5

1.0

1.0

0.5

0.5

0.0

30.0
0.0

0.5

Conditions
Column Size
EluentFlow Rate
Detection
Injection Vol.

1.0

min

0.0

0.5

: 50 × 2.1 mm I.D.
: A) 0.05 % HCOOH in H2O: B) CH3CN
A/B= 70/30 - (0.5 min) - 10/90 - (0.5 min hold): 0.6 mL/min: LC/MS/MS(ESI,SRM): 5 μL

1.0minSample:1. Propranolol(+)
2. Carbamazepine(+)
3. Salicylic acid(-)
4. Ketoprofen(-)（each 100 μg/L in 50 % ACN）(+): Positive mode(-): Negative mode

Analysis of Nucleotide in Fish Meat

1: InertSustain AQ-C18(5 μm, 150 × 4.6 mm I.D.)
Eluent: 50 mM K2HPO4 in H2O (pH 7.0, H3PO4)*
Flow Rate: 1.0 mL/min
Col.Temp.
: 40 ℃
Detection: UV 260 nm
Injection Vol. : 1 μL

0.83-0.2

40.0mAU420.26mAU
0.40.681Conditions
Column502468102Time (min)356402010Time (min)20

Sample:
1.IMP
2.ATP
3.ADP
4.AMP
5.Hyp
6.Ino（each 5 mg/L）* Equilibrate the column with CH3CN/H2O=1/1,v/v after the analysis.
When storing the column for a long period of time, store it with
100 % CH3CN 100 %.

3

Applications
Analysis of Water-Soluble Vitamins

1.9 μm, 50 ×2.1 mm I.D.
0.4 mL/min

Conditions：InertSustain AQ-C18Column：A)0.1% H3PO4 B) CH3CN*Eluent
Col. Temp. ：40℃：UV 210 nm
Detection

5 μm, 150 ×4.6 mm I.D
1.0 mL/min

4384156892

1.012

2.02

3.0010Time (min)Time (min)Sample：1. Pyridoxamine
5. Nicotinamide
9. Folic Acid

2. Thiamin
3. Nicotinic Acid
6. Pyridoxal
7. Pyridoxine
10. Cyanocobalamin 11. RiboflavinTime(min)02
3.3126＜5 μm＞＜1.9 μm＞

7111＊Gradient Time Program

9 10

3 51071120A%9980
99B%
1201Time(min)05202768A%

B%

9999801991120991

4. Ascorbic acid
8. Pantonenic acid
12. Biotin

Analysis of Organic Acids
Conditions
ColumnEluent

3Flow Rate
Col. Temp.
Detection
Inj. Vol.

7211485169100324
Time (min)6： 5 μm, 150 × 4.6 mm I.D.
： 50 mM NaH2PO4 in H2O(pH 2.1, H3PO4)：1.0 mL / min：40℃：UV 210 nm：10 μL

Sample：1. Oxalic acid
50 mg/L
2. Tartaric acid
500 mg/L
3. Formic acid1000 mg/L
4. Malic acid1000 mg/L
5. Lactic acid1000 mg/L
6. Acetic acid1000 mg/L
7. Maleic acid10 mg/L
8. Citric acid1000 mg/L
9. Pyroglutamic acid 100 mg/L
10. Succinic acid1000 mg/L
11. Fumaric acid10 mg/L

8

InertSustain AQ-C18®

Applications
Analysis of Catecolamines
Conditions
ColumnEluentFlow Rate
Col. Temp.
Detection
Inj. Vol.

1： 5 μm, 150 × 4.6 mm I.D.
： 0.1% H3PO4 in H2O：1.0 mL / min：40℃：UV 210 nm：1 μL

Sample：1. Norepinephrine
3. Dopamine
5. L-(-)-Tyrosine

23HO4

2. L-Adrenaline
4. L-DOPA

HONH2123OHOHOH

5NH2N
HOHOHOHOO
HO02468101214Time (min)OHOH
NH2HO4NH2HO

5Analysis of Fish Freshness, K-Value
Conditions
ColumnEluentFlow Rate
Col. Temp.
Detection
Inj. Vol.
4Sample1. ATP
2. ADP
3. IMP
4. Hypoxanthine
5. AMP
6. Inosine

1
2 3： 5 μm, 150 × 4.6 mm I.D.
： 20 mM NaH2PO4 in H2O：1.0 mL / min：40℃：UV 254 nm：1 μL：100 ppm

56010Time (min)204Applications
Analysis of β-blocker
1Conditions
ColumnEluentFlow RateCol. Temp.
Detection
Inj. Vol.

523Sample：
1. Atenolol
3. Oxprenolol
5. Propranolol(100 μg/mL each)4602468Time (min)1012： 5 μm, 150 × 4.6 mm I.D.
： A: CH3CN
B: 0.1% H3PO4 in H2O
A/B=25/75, v/v：1.0 mL / min：40℃：UV 220 nm：1 μL
2. Acebutolol
4. Labetalol
6. Alprenolol

14Analysis of Nucleoside and Nucleic Acid Base
Conditions
ColumnEluentFlow Rate
Col. Temp.
Detection
Inj. Vol.

60

2Sample：1. Cytosine
2. Uracil
3. Guanine
4. Adenine
5. Cytidine
6. Uridine
7. Thymine
8. Adenosine
9. Guanosine
10. Thymidine(100 mg/L each)

4mAU401320765： 5 μm, 250 × 4.6 mm I.D.
： 0.1 M KH2PO4 in H2O + 0.2 M NaClO4 in H2O(pH2.0, H3PO4)：1.0 mL / min：40℃：UV 260 nm：1 μL

8901005102030Time (min)405060InertSustain AQ-C18®

Applications
Analysis of Nucleotides via LC/MS/MSSamples such as nucleotides have several phosphate groups which is sensitive to stainless steel hardware. Asshown below, the combination of highly inert packing material of InertSustain AQ-C18 and usage of a new *SteelCoated-PEEK hardware (metal-free) deliver excellent peak shapes with higher sensitivity for phosphatecompounds WITHOUT the formation of phosphate-iron complexes found with stainless steel column hardwares.
*Ordering information on Steel-Coated-PEEK hardware is available at page 10.
Conditions
SystemColumn size
Eluent: Nexera LCMS-8030 plus system: 3 μm HP, 150 x 2.1 mm I.D: A) 5 mM Ammonium formate in H2OB) CH3CN
Time (min)

B%020.5

23.025

3.01

27.00

2Flow Rate
Col. Temp.
Detection
Injection Vol.

: 0.3 mL/min: 40 ℃: LC/MS/MS (ESI) , Positive: 2 μL

Sample: 1.ATP 500 μg/L
2.ADP 500 μg/L
3.AMP 500 μg/L

Steel-Coated-PEEK hardware
Packed with
InertSustain AQ-C18
PEEK

Packing Material

Stainless Steel Hardware
Packed with
InertSustain AQ-C18
Packing Material

Stainless Steel

4.0

Stainless Steel(x10,000)

4.0(x10,000)

RT= 1.41 min

RT=1.54 min

33.5

3.5

3.0

3.0

2.5

2.5

2.0

Adsorption to thestainless steel hardware

2.0

Higher Sensitivity
without adsorption

21.5
1.0

1.5
1.0

10.5
0.0
0.0

0.5

t0
2.5

min

0.0
0.0

t0
2.5

min
6

Ordering Information
Analytical Columns
Particle Size：1.9 μm
Max. Operating Pressure：
80 MPa (800 Bar)End-fitting Format

Length/ I.D. (mm)

2.1

3.0

50

5020-89938

5020-89941

1005020-89939

5020-89942

150

5020-89940

5020-89943

2.4 mm

UP Type End-fittings※ End-fittings are UP Type end-fittings.

HP Series
Particle Size：3 μm
Max. Operating Pressure：
50 MPa (500 Bar)

Particle Size： 3 μm
Max. Operating Pressure：
20 MPa (200 Bar)

Particle Size： 5 μm
Max. Operating Pressure：
20 MPa (200 Bar)

7

Length/ I.D. (mm)305075100150250

Length/I.D. (mm)305075100150250
Length/I.D. (mm)
30
5075
100
125150250
Length/I.D. (mm)305075100150250
Length/I.D. (mm)
30
5075
100
125150250

2.15020-89920
5020-89921
5020-89922
5020-89923
5020-89924
5020-89925

3.0
5020-89926
5020-89927
5020-89928
5020-89929
5020-89930
5020-89931

1.05020-89871
5020-89872
5020-898735020-89874
5020-89875
5020-89876
2.1
5020-89831
5020-89832
5020-89833
5020-89834
5020-89835
5020-89836
5020-89837
1.05020-89741
5020-89742
5020-89743
5020-89744
5020-89745
5020-89746
2.1
5020-89701
5020-89702
5020-89703
5020-89704
5020-89705
5020-89706
5020-89707

1.5
5020-89877
5020-89878
5020-89879
5020-89880
5020-89881
5020-89882
3.0
5020-89839
5020-89840
5020-89841
5020-89842
5020-89843
5020-89844
5020-89845
1.55020-89747
5020-897485020-89749
5020-89750
5020-89751
5020-89752
3.0
5020-89709
5020-89710
5020-89711
5020-89712
5020-89713
5020-89714
5020-89715

4.6
5020-89932
5020-89933
5020-89934
5020-89935
5020-89936
5020-89937

4.0
5020-89847
5020-89848
5020-89849
5020-89850
5020-89851
5020-89852
5020-89853

4.6
5020-89855
5020-89856
5020-89857
5020-89858
5020-89859
5020-89860
5020-89861

4.0
5020-89717
5020-89718
5020-89719
5020-89720
5020-89721
5020-89722
5020-89723

4.6
5020-89725
5020-89726
5020-89727
5020-89728
5020-89729
5020-89730
5020-89731

InertSustain AQ-C18®

Ordering Information
Guard Column for UHPLC (Max. Operating Pressure 80 MPa, 800 Bar)
Replacement Cartridge(2 pcs)

Cartridge (2 pcs) + Holder (1 pcs) Set

Replacement Cartridge(2 pcs)

Cartridge (2 pcs) + Holder (1 pcs) Set

Particle Size 1.9 μm

Particle Size 1.9 μm

Particle Size 3 μm

Particle Size 3 μm

1.55020-89944

5020-89947

5020-89824

5020-89827

2.15020-89845

5020-89948

5020-89825

5020-89828

3.0

5020-89946

5020-89949

5020-89826

5020-89829

I.D. of theAnalytical
Length I.D.
Column Applicable (mm) (mm)(mm)
1.01.5, 2.1

102.1, 3.0

Holder for Guard Column for UHPLC

5020-08630

Cartridge Guard Columns
I.D. of the Analytical
Column Applicable(mm)Length(mm)Replacement Cartridge E (2 pcs)

I.D.
(mm)

Cartridge E (2 pcs) + Holder (1 pcs) Set

Particle Size

Particle Size

3 μm

5 μm

3 μm

5 μm

1.0

5020-89910

5020-89808

5020-89911

5020-89809

1.55020-89912

5020-89810

5020-89913

5020-89811

3.0

5020-89908

5020-89806

5020-89909

5020-89807

4.0,4.6

4.05020-89906

5020-89804

5020-89907

5020-89805

2.1,3.0

3.05020-89916

5020-89814

5020-89917

5020-89815

4.0

5020-89914

5020-89812

5020-89915

5020-89813

1.0
1.5,2.110

2.1,3.020

4.0,4.6

Holder for Cartridge Guard Column EFor 10 mm Length5020-08500

For 20 mm Length5020-08550

Metal-Free PEEK Columns
Length/ I.D. (mm)2.130

Please inquire-33

Please inquire

Please inquire

50

Please inquire

Please inquire

75-

Please inquire

100

Please inquire

Please inquire

150

Please inquire

Please inquire

200-Please inquire

250

Please inquire

Please inquire

300

Please inquire

Please inquire

PEEK Columns
Particle Size：5 μm
Max. Operating Pressure：
20 MPa (200 Bar)

4.6

Length/ I.D. (mm)

2.1

4.6

50

Please inquire

Please inquire

100

Please inquire

Please inquire

150

Please inquire

Please inquire

250

Please inquire

Please inquire

Steel-Coated PEEK Columns
Particle Size：3 μm
Max. Operating Pressure：
50 MPa (500 Bar)

GL Sciences disclaims any and all responsibility for any injury or damage which may be caused by this data directly or indirectly. We reserve theright to amend this information or data at any time and without any prior announcement.

GL Sciences, Inc. JapanGL Sciences B.V.

GL Sciences (ShangHai) Ltd.

GL Sciences, Inc. USA

22‐1 Nishishinjuku 6‐ChomeShinjuku‐ku, Tokyo,
163‐1130, Japan
Phone: +81‐3‐5323‐6620
Fax: +81‐3‐5323‐6621
Email: world@gls.co.jp
Web: www.glsciences.com

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Phone: +31 (0)40 254 95 31
Email: info@glsciences.eu
Web: www.glsciences.eu

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