Usability of a core shell column using high performance liquid
chromatograph for a routine analysis
Tomoyasu Tsukamoto1), Norikazu Nagae1), Harald Dibowski2), Annette Dibowski2),Anders Grahn3), Ralf Jutvik3)
and Vinay D. Gaitonde3)
1) ChromaNik Technologies Inc., 6-3-1 Namiyoke, Minato-ku, Osaka, Japan2) di2chrom GmBh, Gräwenkolstraβe 66, D-45770 Marl, Germany3) Biotech AB, Box 133, 439 23, Onsala, Sweden

Abstract

About SunShell

752 3
10.5 mm

0.5 mm

1.6 mmPlate/pressureSunniest C18 –HT 2.0 mm
Brand B C18 1.9 mm
Brand C C18 1.8 mm
Brand D C18 1.7 mm

9,900
7,660
10,10011,140

16.7
16.3
19.6
32.0593470515348

SunShell C18 2.6 mm

9,600

9.7990

SunShell C18 used as core shell column
showed lower back pressure than sub-2μm fully porous column.

Brand B C18 1.9 mm

Plate/Pressure Valued of SunShell was
twice or three times as high as sub-2 μm
fully porous column

Brand C C18 1.8 mm
Brand D C18 1.7 mm
SunShell C18 2.6 mm
05,000 10,00001020300250 500 750 1000

Column: 50 x 2.1 mm C18, Mobile phase: Acetonitrile/water=(70/30), Temperature: 25 oC

Figure. 6 Comparison of performance by plate/pressure

Plate height (μm)

168060Brand K40SunShell C1800204060

80

100120Time (h)Brand E sub-2 μm 40 ℃
(Fully porous type)8

Comparing between SunShell and sub-2 μmcolumn • • •SunShell (core shell column) was••••

40
0.2

0.4

0.6

0.88 10 12 14 16 18 20 22 24
Retention time/min

0.48

1.54

1.20

Brand A

0.44

1.60

1.31

SunShell C180.40

1.59

1.47Measurement condition
Column Size: 50 x 2.1 mm
Mobile phase:
CH3OH/H2O=70/30
Flow rate: 0.4 mL/min
Temperature: 40 ºCSample: 1 = Butylbenzene

8060
Brand K

40

Brand ASunShell C182000

1,000

2,000 3,000 4,000
Elution volume(mL)

5,000

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
6,000

Figure 4. Stability under basic pH condition
Mobile phase: Acetonitrile/20mM phosphate buffer pH7.0=(60:40)
Column dimension: 150 x 4.6 mm, Flow rate: 1.0 mL/min, Temp.: 40oC
16000
14000

10000
8000
6000

Comparing between SunShell andother brand columns • • •SunShell has

SunShell C18(core shell)

12000
100 times

Brand K
Brand ASunShell C18
Sunniest C18 3um

Sunniest
C18 3mm(fully porous)

4000• Different selectivity• High stability under both acidic
and basic pH conditions• Good peak shape for
amitriptyline• High Loading capacity

Brand K C18(core shell)

00.001

0.01

0.1

1

Brand A C18(core shell)

10Sample weight/mg

0123456Retention time/min

78910Sample: 1=Uracil, 2=Propranolol,
3= Nortriptyline, 4=Amitriptyline

Theoretical plate was calculated by 5σ method using peakwidth at 4.4% of peak height.
4.4%

Sunshell 2.6 μm 40℃

0Different Selectivity
from other brandcolumns

Brand K

Figure 3. Stability under acidic pH condition

Sunshell 2.6 μm 25 ℃

1276100

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: Butylbenzene

Brand A2065Figure 2. Comparison of standard samples

2000

SunShell column kept the low plate heightunder the low flow rate conditions.
But fully porous column couldn‘t keepColumn: C18, 50 x 2.1 mm
Mobile phase:
Acetonitrile/water=(60/40)
Sample : Naphthalene

204

4 = Butylbenzene, 5 = o-Terphenyl,
6 = Amylbenzene, 7 = Triphenylene

Hydrogen bonding
Hydrophobicity
Steric selectivity(Caffeine/Phenol) (Amylbenzene/(Triphenylene/Butylbenzene)
o-Terphenyl)

Durable 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

And SunShell column showed similar
theoretical plate comparing with sub-2 μm
fully porous column.

Sunniest C18 –HT 2.0 mm

2silica particle

Relative retention (%)

Pressure(MPa)40Brand A Sample: 1 = Uracil, 2 = Caffeine, 3 = Phenol,

6SunShell C18

1Porous Silica

Pore volume: 0.30 mL/g, Specific surface area: 150m2/g,
Pore diameter: 9 nm
The ratio
porous silicadiagram
volume: 77%
Figure1.ofSchematicof a core shell

Theoretical PlatePlates

2 3Mobile phase: CH3OH/H2O=75/25
Flow rate: 1.0 mL/min
Temperature: 40 ºC

741CoreCore100

Core shell vs Sub-2 μm fully porous

52 32.6 mm

Column:Brand K C18, 2.6 μm 150 x 4.6 mm
Brand A C18, 2.7 μm 150 x 4.6 mm
SunShell C18, 2.6 μm 150 x 4.6 mm

Brand K
642.6 mm

Relative plate of butylbenzene(%)

Superficially porous particle (core shell particle) has been available as an alternative to using sub2 μm particle for HPLC or UHPLC. Core shell particles are composed of a 1.2 to 1.9 μm solidcore encircled a 0.25 μm to 0.5 μm porous layer. Especially a 2.6 μm core shell particle shows ahalf back pressure and the almost same efficiency to compare with sub-2 um particle because ofa large particle and reducing mass transfer due to a thin porous silica layer. In this study, a 2.6μm core shell silica with a non-porous core approximately 1.6 μm in diameter and a superficiallyporous layer of 0.5 μm was used as a based material. A core shell silica bonded with C18 andend-capped was evaluated for a routine analysis, which is done using conventional 5 μm particle
column sized 150 or 250 x 4.6 mm i.d. A core shell C18 column size 100 x 4.6 mm and aconventional C18 column sized 250 x 4.6 mm were compared for an analysis of analgesics using
Hitach LaChrom ELITE HPLC under a isocratic mode. Both columns showed the same efficiency
and an analysis time by a core shell C18 columns decreased to one third to compare with aconventional C18 column without changing of conditions except for a column, a same instrument,a same flow rate, a same mobile phase. In case of gradient separation of catechins, the almost
same result was obtained as well as under a isocratic mode.

1

Flow rate mL /minLower back pressure
Similar theoretical plate
High Plate/Pressure value
lower plate height under low flowrate conditions

Figure. 7 Comparison of plate height

Figure 5. Loading capacity of amitriptyline

HPLC

3121Brand F C18, 5 μm 250 x 4.6 mm

0241/3 or 1/2 of analysis time
SunShell C18, 2.6 μm 100 x 4.6 mm

54N(4)=18,444

543268101412N(4)=20,287

1618Column: SunShell C18, 2.6 μm 150 x 4.6 mm
Mobile phase: CH3CN/H2O=70/30
Flow rate: 1.0 mL/min
Pressure: 13.5 MPa
Temperature: 25 ºC

2101280% performance
to compare with UHPLCStill 1.5 times more separation thanthat of a 3 mm column

4UHPLC

N(4)=24,124

HPLC: Hitachi LaChrom ELITE(using 0.25 mm i.d. tubing)
UHPLC: Jasco X-LC
N(4)=22,424

Analysisi time decreases to 1/6 to comparewith a 5mm column sized 250x4.6mm.

Retention time/min

Figure. 9 Examples of transfer (isocratic separation)

Conclusions

4Brand H C18, 5 μm 150 x 4.6 mm

HPLC43046810Retention time/min

4121463234161889

5520

76879Retention time/min

4UHPLC

621738950

1

20 min 15 min 20 min
2%
25%
25%

for SunShell C18

11Time%B

1/2 of Analysis time

89

52089

56 7
32Column:
Brand H C18, 5 μm 150 x 4.6 mm
SunShell C18, 2.6 μm 75 x 4.6 mm
Mobile phase:A) 0.1% Phosphoric acidB) CH3CN
Gradient program for Brand G C18

762SunShell C18, 2.6 μm 75 x 4.6 mm

21In case of gradient separation of catechins, the almost same result was obtained
as well as under a isocratic mode.
SunShell was a good tool for high-throughput analysis and environmental
friendship.

Column:Brand F C18, 5 μm 250 x 4.6 mm
SunShell C18, 2.6 μm 100 x 4.6 mm
Mobile phase:
CH3CN/20mM Phosphoric acid = 45/55
Flow rate: 1.0 mL/min,1.8 mL/min at the lowest chromatogram
Temperature: 25 ºC
Pressure: 9.5 MPa for Brand F C18 5 mm
13.4 MPa for SunShell C18 2.6 mm
Detection: UV@230 nm
Sample: 1 = Benzydamine
2 = Ketoprofen
3 = Naproxen
4 = Indomethacin
5 = Ibuprofen

6

1.8 mL/min

Figure. 8 Comparison between HPLC and UHPLCAn analysis time by a core shell C18 columns decreased to one third or half
comparing with a conventional C18 columns without changing of analyticalconditions.

5

1.0 mL/min

1100 × 4.6 size SunShell showed the same efficiency for 250 × 4.6 size 5 μm fully
porous column under the isocratic mode.

2422543Retention time/min

Sample:1 = Uracil
2 = Toluene
3 = Acenaphthene
4 = BUtylbenzene

20N(4)=20,287
N(4)=20,287

3Appling for HPLC

54Brand G C18, 3 μm 150 x 4.6 mm

321

N(4)=19,313

3456Retention time/min

Figure. 10 Examples of transfer (gradient separation)

22Time%B

240 min 7.5 min 10 min
2%
25%
25%

Flow rate: 1.0 mL/min,
Temperature: 25 ºC
Detection: UV@250 nm
Sample: Oolong tea
1 = Gallocatechin, 2 = Epigallocatechin,
3 = Catechin, 4 = Caffeine, 5 =Epicatechin,
6 = Epigallocatechin gallate, 7 =Gallocatechin gallate, 8 = Epicatechingallate, 9 = Catechin gallate
HPLC: Hitachi LaChrom ELITE(using 0.25 mm i.d. tubing)
UHPLC: Jasco X-LC