Evaluation of Five Core Shell Columns Based on BothSeparation Behavior and Physical Property
www.chromanik.co.jp

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Norikazu Nagae , Tomoyasu Tsukamoto , Vinay Gaitonde , Anders Grahn , Ralf Jutvik

1) ChromaNik Technologies Inc., 6-3-1 Namiyoke, Minato-ku, Osaka, Japan3) Biotech AB, Box 133, 439 23, Onsala, Sweden

www.biotech-sweden.com

Abstract
A column packed with core shell particles has been widely used on HPLC and UHPLC, because it showed not only excellent column efficiency butalso lower back pressure than sub-2 um column. More than 10 kinds of core shell column are available in the market. It is said that two types ofcore shell silica particle are used. One is a mono-layer structure as a porous silica layer like a core shell silica manufactured by Advanced Materials
Technology and another is a multi-layer structure like that manufactured by Phenomenex.
In this study, a separation behavior and physical property of Kinetex C18, Accucore C18, PoroShell C18 EC, Ascentis Express C18 and SunShell C18
were evaluated. Retention factor, hydrogen bonding capacity, hydrophobicity and steric selectivity were measured using Tanaka method. Not only
peak shape of neutral, acidic and basic compounds but also loading capacity of amitriptyline under neutral and acidic conditions were alsomeasured. Furthermore stability under acidic pH1 and basic pH10 conditions was evaluated. Regarding physical property, carbon loading of eachC18 packing material, and specific surface area, pore volume, pore diameter of each core shell silica which was deleted alkyl chains by sintering at
600 degree Celsius for 8 hours were measured. As a result, the big difference was recognized among 5 kinds of core shell C18 for separation
behavior, stability and physical property. This difference is considered to be due to each manufacturing method and bonding technique as well as
fully porous silica C18s. SunShell C18 showed the largest retention factor and the highest stability though its carbon loading is not the highest,
while Kinetex C18 showed the lowest retention factor, the lowest carbon loading and the lowest specific surface area.

Physical Property

123123102

Company S C1822.2 MPak6=9.7645SunShell C18
21.8 MPa 4468135 (130)b

Accucore C188.8 (9)b130 (130)b

Kinetex C1812(Caffeine/Phenol)

14k6=10.4

16182022(Amylbenzene/Butylbenzene) (Triphenylene/o-Terphenyl)

0.48Company P C18

1.54

1.20

Company T C18

0.35

1.56

1.50

Company A C18

0.42

1.57

1.25

Company S C18

0.44

1.60

1.31

10SunShell C18

0.39

1.60

1.46

924

Retention time/min

4.9(12 effective)b102(200 effective)b

Number/%Company P C183

Company T C18

232

Company A C18

132

Company S C1812323456879Retention time/min

Company A C18Sunniest C18 3um

8000

Company P C18

6000

654Sunniest
C18 3mm(fully porous)

TF=2.43

P C18(core shell)

Company T C18
Company S C18

4000

T C18(core shell)

TF=3.2100.001

SunSehll C18 (2.54 μm)c
Ascentic Express C18 (2.52 μm)c
Accucore C18 (2.53 μm) c
Kinetex C18 (2.34 μm) c
PoroShell C18 (2.34 μm) c

7

TF=1.25

2000

0.01

0.1

110

S C18(core shell)

TF=4.38

Theoretical plate was calculated by 5σ methodusing peak width at 4.4% of peak height.

1234567Retention time/min

8910Sample: 1=Uracil, 2=Propranolol,
3= Nortriptyline, 4=Amitriptyline

16000

TF=2.0914000
12000

30Column:
Company P C18, 2.6 mm 150 x 4.6 mm
Company T C18, 2.6 mm 150 x 4.6 mm
Company A C18, 2.7 mm 150 x 4.6 mm
Company S C18, 2.7 mm 150 x 4.6 mm
SunShell C18, 2.6 mm 150 x 4.6 mm
Mobile phase: CH3OH/H2O=30/70
Flow rate: 1.0 mL/min
Temperature: 40 ºC
Detection: UV@250nm
Sample: 1 = Uracil
2 = Pyridine3 = Phenol

10000

A C18(core shell)

TF=1.4201

2Particle diameter/μm

48100 times

SunShell C18

10000

Company A C18Sunniest C18 3um

8000

T C18(core shell)

Company P C18

6000

Company T C18
Company S C18

TF=3.21

4000
2000*Measured using Beckman Coulter Multisizer 3 after C18 materials were sintered at
600 degree Celsius for 8 hours. The measured value of each sintered core shell silica
is considered to be different from that of the original core shell silica.

00.001

0.01

0.1

1100

12345678910Retention time/minSample weight/μgc. Median particle size
Mobile phase: Acetonitrile/10mM ammonium acetate pH6.8=(40:60)
14000

101112Stability under pH1 and 10 Conditions

13Durable test condition
Column size: 50 x 2.1 mm
Mobile phase: CH3CN/1.0% TFA, pH1=10/90
Flow rate: 0.4 mL/min
Temperature: 80 ºC

Metal Chelating Compound12

TF=1.24Company P C18

2

Company T C18

1

TF=3.942

1Company A
C18

TF=2.07

2

1

Company S C18

TF=2.04
2

1
TF=1.21

1234Column:
Company P C18, 2.6 mm 150 x 4.6 mm
Company T C18, 2.6 mm 150 x 4.6 mm
Company A C18, 2.7 mm 150 x 4.6 mm
Company S C18, 2.7 mm 150 x 4.6 mm
SunShell C18, 2.6 mm 150 x 4.6 mm
Mobile phase: CH3CN/20mM H3PO4=10/90
Flow rate: 1.0 mL/min
Temperature: 40 ºC
Detection: UV@250nm
Sample: 1 = 8-Quinolinol (Oxine)
2 = Caffeine

SunShell C18

5678910Retention time/min

Comparison of Formic Acid12

TF=1.47

13Company P C18

32Company T C18

TF=1.8612

TF=1.45

1Company A C18

32

TF=3.21Company S C18

1
TF=1.46

13Column:Company P C18, 2.6 mm 150 x 4.6 mm
Company T C18, 2.6 mm 150 x 4.6 mm
Company A C18, 2.7 mm 150 x 4.6 mm
Company S C18, 2.7 mm 150 x 4.6 mm
SunShell C18, 2.6 mm 150 x 4.6 mm
Mobile phase: CH3CN/0.1% H3PO4=2/98
Flow rate: 1.0 mL/min
Temperature: 40 ºC
Detection: UV@210nm
Sample: 1 = Formic acid
2 = Acetic acid
3 = Propionic Acid8060

SunShell C18
Company S C18

40

TF=1.20

SunShell C18(core shell)

Company P C1820

Company A C180204080

100120Sunshell C18
Company A C18
Company P C18
Company T C18Company S C18

8000
6000

TF=2.61

4000

TF=2.7300.001

0.01

0.1

10

Durable test condition
Column size: 50 x 2.1 mm
Mobile phase:
CH3OH/20mM Sodium borate/10mM
NaOH=30/21/49 (pH10)
Flow rate: 0.4 mL/min
Temperature: 50 ºC

SunShell C18
Company S C18
Company P C18
Company T C18

Measurement condition
Column size: 50 x 2.1 mm
Mobile phase: CH3OH/H2O=70/30
Flow rate: 0.4 mL/min
Temperature: 40 ºC
Sample: 1 = Butylbenzene

Company A C18

20pH10, 50oC

00123Sample weight/μg45678910Retention time/min

Sample: 1=Uracil, 2=Propranolol,
3= Nortriptyline, 4=Amitriptyline

Mobile phase: Acetonitrile/0.1% formic acid=(30:70)
4.5

100

40

S C18(core shell)

10

USP tailing factor
Company S C1860

P C18(core shell)

T C18(core shell)

TF=3.24

480

A C18(core shell)

TF=1.89

30 times

2000

Time/h1,000 2,000 3,000 4,000 5,000 6,000

3.5
3Company T C18

3.6

Company P C18

2.9

Company A C18
2.5

SunShell C18

2.4
7 times

21.8

1.5

1.3

10.00140.01

0.15 min

61Sample weight/μgIn the case of using acetonitrile /0.1% formic acid as a mobile phase, amitriptyline peak shows
more tailing because a loading capacity decreases in an acidic, low-ionic-strength mobile phase.

Elution volume/mLAcidiccondition pH 1

Basic condition pH range written in each
pH 10
brochure

SunShell C18◎◎

1.5 - 10

Ascentis Express C18○○2- 9PoroShell C18 EC△△2-9

Accucore C18△△1 - 11

Kinetex C18△△

1.5 - 10

32pH1, 80oC6010000

Measurement condition
Column size: 50 x 2.1 mm
Mobile phase: CH3CN/H2O=60/40
Flow rate: 0.4 mL/min
Temperature: 40 ºC
Sample: 1 = Uracil, 2 = Butylbenzene

Company T C18

0

Relative plate of butylbenzene/%

Comparison of Oxine,

Relative retention/%

10009.25 (10)b100 times

SunShell C18

12000

100.237

12000

SunShell C18

100.273

8.39 (8)b

SunShell C18(core shell)

TF=1.18

14000

1210.414

12.3 (12)b16000

23

10.2788.20 (9)bSample weight/μg8Comparison of Pyridine

8.34 (9)ba. Measured after C18 materials were sintered at 600 degree Celsius for 8hours. The measured value of each sintered core shell silica is considered
to be smaller than that of the original core shell silica.
b. Value written in each brochure or literature

Hydrogen bonding Hydrophobicity Steric selectivity

76

10

PoroShell C18 EC

8.5 (8)bAscentis Express C18

752
3

8.0

133 (150)b0.261

Theoretical plate

3125 (150)bMobile phase: Acetonitrile/20mM phosphate buffer pH7.0=(60:40)
Column dimension: 150 x 4.6 mm, Flow rate: 1.0 mL/min, Temp.: 40oC

USP tailing factor

127.3 (7)bSunShell C18

Pore diameter a(nm)Theoretical plate

Column:7Company P C18, 2.6 mm 150 x 4.6 mm (26.1 MPa)
5Company P C18
6Company T C18, 2.6 mm 150 x 4.6 mm (22.7 MPa)426.1 MPa
Company A C18, 2.7 mm 150 x 4.6 mm (30.6 MPa)
Company S C18, 2.7 mm 150 x 4.6 mm (22.2 MPa)
7
5SunShell C18, 2.6 mm 150 x 4.6 mm (21.8 MPa)k6=7.4Company T C18
Mobile phase: CH3OH/H2O=75/25
4622.7 MPa
Flow rate: 1.0 mL/min
Temperature: 40 ºC
7Sample: 1 = Uracil, 2 = Caffeine, 3 = Phenol,
5Company A C18
k6=9.04 = Butylbenzene, 5 = o-Terphenyl,
4630.6 MPa
6 = Amylbenzene, 7 = Triphenylene
k6=5.4211 3

Specific surface Pore volume aarea a (m2/g)(mL)

Carbonloading (%)

Theoretical plate

Comparison of Standard Samples

Loading Capacity of Amitriptyline

Column name
1. Company P C18, 2.6 mm: Kinetex c18
2. Company T C18, 2.6 mm: Accucore C18
3. Company A C18, 2.7 mm: PoroShell C18 EC
4. Company S C18, 2.7 mm: Ascentis Express C18
5. SunShell C18, 2.6 mmSunShell C18
23456Retention time/minConclusion

SummaryPressurea Retentionb Pyridine Oxine Formic acid Point
SunShell C18◎21.810.4◎◎◎

12Ascentis Express C18◎22.29.7△△×5PoroShell C18 EC

×30.6

9.0

◎

△

◎7Accucore C18◎22.7

7.4××△4Kinetex C18

△26.1

5.4×◎

◎

7a. Mobile phase, methanol:water=75:25, 40 oC, 1mL/min 150 x 4.6mm
b. Retention factor of amylbenzene◎: 3 point, ○: 2 point, △: 1 point, ×: 0 point The big difference concerning retention, residual silanol groups, metal impurity, loading
capacity and stability among core shell C18s was confirmed as well as fully porous C18s.
 SunShell showed the longest retention, the lowest pressure, the most symmetrical peak andthe highest stability.
 The measured value of physical property is not as same as a value shown in a brochure and aliterature in some cases.