RP C18 column with a feature of a silanol group

Silanol Activity Controlled C18 Column

Comparison of separation forethylbenzene and xylene
32C30

413,42C18

13,42PFP
Sunrise Triacontyl （C30)

11 = Ethylbenzene
2 = o-Xylene
3 = m-Xylene
4 = p-Xylene

Sunrise Octacosyl （C28)
Sunrise Octadecyl-SAC （C18-SAC) has an interaction of silanol groups

ChromaNik Technologies Inc. TEL: 06-6581-0885 http://chromanik.co.jp

HPLC column

Sunrise C30, C28
Sunrise C18-SAC
Name

Stationary phase

Carbon content

Ligand density

Particle sizeC30 Triacontyl

18%1.7 μmol/m2

3 μm, 5 μm

C28 Octacosyl

18%1.7 μmol/m2

3 μm, 5 μm

Silica support
Surface area：340 m2/g
Pore volume：1.0 mL/g
Pore diameter：12 nm

C18-SAC Octadecyl

14%

2.1 μmol/m2

3 μm, 5 μm

End-capping
Trimethylsilyl group (TMS)

pH range of C30 and C28: pH2～pH8, C18-SAC: pH2～pH7.5◆ Characteristics of end-capping type Sunrise series■ Separation of Basic compounds

C30, C28: (C30 and C28 shows the same separation.)■ A long alkyl chain improves both separation offat-soluble compounds to compare with C18 phaseand an excellent reproducibility in retention underhigh aqueous conditions.
■ Furthermore, a suitable ligand density of C28
allows to be obtained a shape peak shape even if
more than 50% aqueous mobile phase is used.
■ Different selectivity

5Column size: 150 x 4.6 mm
Mobile phase:80/20=CH3OH/25mM P.B. pH7.5
Flow rate: 1.0 mL/min
Temperature: 40 ºC
Sample: 1 = uracil
2 = propranolol

Sunrise C18
1423CH30246810 12retention time / min

1416

18ONHCH3OH

3 = toluene
4 = nortriptyline

5Suniest PhE

NHCH3Reverse
41

5 = amitriptyline

2CH3

3

C18： ( C18 has stopped production. Sunniest C18

01

is recommended as a replacement.)■ Conventional C18 phase with full end-capping

2NCH3

3
4
5
6
retention time / min78■ Separation of Vitamin EPhE：(Sunrise PhE has stopped production. Sunniest PhE
is recommended as a replacement)■ Interaction based with p-electron such as p-pinteraction■ p-electron also interacts with a polar site of acompound, so that phenyl phase improves
separation of polar compounds. Ethylene chain
between silica surface and phenyl group allows a
movable sphere of a phenyl group to be wide. Achain with more than three carbons shows morehydrophobic interaction, so that p-electroninteraction decreases relatively.
■ Phenethyl (PhE) group is a suitable phenyl phase.

Isomer can be separated by C28

Sunrise C30 or C28

Column size: 250 x 4.6 mm
Mobile phase: 97/3=CH3OH/H2O
Flow rate: 1.0 mL/min
Temperature: 25 ºC
Detection: UV at 295nm
Sample: 1=δ-tocopherol

1234HOO051015202530
Retention time / minConventional C18135

2=γ-tocopherol
HOO

2, 3

3=β-tocopherol
4HOO

4=α-tocopherol
HO05

10152025
Retention time / min

30OSunrise C30, C28■ Separation of xylene isomers
225 ºC
4

156

305

10

15
Retention time/min

202530215 ºC

54

163051015
Retention time/min

20

Column: Sunrise C30 or C28, 3 µm 4.6 x 250 mm
Mobile phase: CH3OH/H2O=70:30
Flow rate: 0.7 mL/min
Temperature: 25 ºC and 15 ºC
Detection: UV@250nm
Sample: BTEX
1 = Benzene
2 = Toluene
3 = Ethylbenzene
CH3
4 = o-Xylene
5 = m-Xylene
CH3
6 = p-Xylene

25Sunrise C30 or C28 can separate
xylene isomers although conventional
C18 can’t. The lower a temperature,
the better a separation of isomers.

30■ Separation of vitamin K1 isomers
1Column: Sunrise C30 or C28 3 µm, 150 x 4.6 mm
Company A C18 3 µm, 150 x 4.6mm
Mobile phase: Methanol
Flow rate: 1.0 mL/min
Temperature: 15 oC
Detection: UV@250 nm
Sample: Vitamin K1 isomers (trans and cis)

Sunrise C30, C28
205101520

Retention time/min

1,21 = transO
H 3C

C18 can not eparate
vitamin K1 isomers.

C18HH 3CHCH3HCH 3CH 3OCH 3

2 = cisOO051015202530

CH3Retention time/min

CH 3HCH3HCH 3

HCH 3CH 3■ Separation of ter-phenyl isomers
2

Conventional C18Column: Conventional C18 150 x 4.6 mm
Sunrise C30, or C28, 3 µm 150 x 4.6 mm
Mobile phase: Methanol/water=90:10
Flow rate: 1.0 mL / minTemperature: 25 oC
Detection: UV@250 nm
Sample: 1 = 0-Terphenyl, 2 = m-Terphenyl, 3 = p- Terphenyl

３1012345Retention time / min

Sunrise C30 ,C28

672310123456

Retention time / min78910Sunrise C28
Sunrise C18-SAC

HPLC column■ Comparison of stationary phases
Column size: 150 x 4.6 mm

Sunrise C18 70% CH3OH

6243

1Mobile phase: H3OH/H2O5Flow rate: 1.0 mL/min
Temperature: 40 °CSample:
1 = Methylparaben

Sunrise C30 or C28 69% CH3OH
12432 = Ethylparaben

63 = Benzene
4 = Toluene
5 = Naphthalene

56 = Ethylbenzene
6Sunnist PhE 56% CH3OH
41 2Retention increased due to ππ interaction.

53Sunrise C18-SAC 66% CH3OH
21Retention increased due πelectron interaction between
naphthalene and silanol groups.

6435Retention time / min■ Comparison of separation of α, β - carotene
21Conventional
C18

The mobile phase including
chloroform makes alkyl chains brash
up because chloroform can enter
among alkyl chains. Consequently
retention times of C30 or C28
became 2 times longer than C18.

Column size: 250 x 4.6 mmMobile phase: CH3CN/CHCl3 = 80:20
21Flow rate: 1.0 mL/min
Temperature: 30 ºCDetection: UV at 450nm
Sample: 1 = α-Carotene
CHH3H3C

Sunrise C30 or C28

CH3 H

H

H

H

H

CH3H

CH3H

H

CH3H

H H3C

CH3H
H3C

CH3H

10H

2 = β-Carotene

H3C0CH3H

H

20Retention time / min

3040CH3 H

H

CH3H

H

H

CH3CH3H

H

H

H

CH3H

H

H H3CH
H3CCH3Sunrise C18-SAC
Silanol Activity Controlled C18 HPLC Column◆ New generation reversed-phase utilized silanol groups■ Silanol group and peak tailing

5th generation
Hybrid type

It is generally said that residual silanol groups
on a stationary phase such as C18 (ODS)
causes absorption or peak tailing for a sample.
Especially silanol groups near a hydrophobic
site don’t solvate with water completely, sothat they show high absorption for basic
compounds. Its peak shows terribly tailing.
Several end-capping techniques have beendeveloped to solve these problems for manyyears.

Sunniest C18

4th generation
Highly endcapped typeSunrise C18

3rd generation
High carbon andend-capped type2nd generation

Silanol ActivityControl
Technology

Bonded typeSunriseC18-SAC

1st generation
Pellicular type

■ Silanol activity control technology
ChromaNik developed the technique that
decreased only silanol groups with highabsorption activity to a basic compound andremained effective silanol groups on thestationary phase. Silanol activity control andno end-capping led the existence of silanolgroups with high hydration which created anew and unique reversed-phase separation
mode including hydrogen bond and ionexchange interaction. Furthermore, silanol
activity controlling, then end-cappingtechnique improved a peak shape of a basic
compound exceedingly.

Changeover tosiloxane bond

Silanol groups with Silanolhigh absorption
Activityactivity
CH3HSiOHOO H OOSiSiSiSi

CH3Control
TechnologySiOOSiSiHOO
SiSi

Silanol activity control◆ Feature of Sunrise series
Sunrise C18•The “1st Choice” column as a fully end-capped C18column•Full end-capping after silanol activity control•Reducing adsorption of a basic compound extremely•A good peak shape for a metal chelating compound•Widely available for general reversed-phase separation■ The elution order of pyridineA) Sunrise C18

1Column size: 150 x 4.6 mm
Mobile phase:
CH3OH/H2O =30/70
Sample: 1 = uracil(2) = pyridine
3 = phenol3(end-capped)(2)B) Conventional no end-capped C18
3Sunrise C18-SAC•The "2nd Choice" column which takes advantage ofeffective silanol groups interaction•Reducing silanol groups with high adsorption activity•The new separation mechanism including hydrogenbond and ion-exchange interaction•Effective for separation of a basic compound and apolar compound•Different selectivity and improvement of separation
without changing a mobile phase1(2)C) Sunrise C18-SAC(non end-capped)
103(2)

510
retention time / min1520

Sunrise C18-SAC
Silanol Activity Controlled C18 HPLC Column◆ Sunrise series create an unique separation✻ Effectiveness of silanol activity control: Comparison between Sunrise C18 and C18-SAC
Sunrise C18 is the so-called fully end-capped C18 column.
It shows the same separation behavior as a conventional
C18 column.
On the other hand, Sunrise C18-SAC shows hydrogenbond and ion-exchange interactions based on a residual
silanol on the silica support in addition to reversed-phaseseparation. For example Sunrise C18 column separates abasic compound similarly as a conventional C18, while

Sunrise C18-SAC makes retention of a basic compound
be large because an ion-exchange interaction works
although a non-ionic compound shows the almost same
retention on both Sunrise C18 and C18-SAC. Furthermore,
Sunrise C18-SAC shows large retention for a polar
compound such as caffeine.

■ Comparison of selectivity for basic compounds■ Comparison of peak shape and retentionA) Sunrise C18A) Sunrise C18
pH3.04Column size: 150 x 4.6 mm
Mobile phase:
CH3CN/20mM H3PO4 = 10:90
Flow rate: 1.0 mL/min
Sample: 1 = 8-Quinolinol (Oxine)
2 = Caffeine

212315B) Sunrise C18-SAC4B) Sunrise C18-SAC

pH3.0123152A) Sunrise C18
4pH4.502462381012 14 16 18 20 22

Retention time / min

15B) Sunrise C18-SAC
1■ Comparison of caffeine

pH4.524A) Sunrise C18335240246810121416

1

Retention time / minColumn size: 150 x 4.6 mm
Mobile phase:
CH3CN/20mM Phosphate buffer pH3.0 or pH4.5 = 50:50
Flow rate: 1.0 mL/min
Temperature: 40 ºC
Sample: 1 = UracilB) Sunrise C182
SAC

Column size: 150 x 4.6 mm
Mobile phase:
CH3OH/20mM Phosphatebuffer pH4.5 = 30:70
Flow rate: 1.0 mL/min
Temperature: 40 °C
Sample:1 = Theobromine
2 = Theophylline
3 = Caffeine
4 = Phenol

43HNONH

2 = Propranolol

O

1CH3O

NHCH3OH

3 = Nortriptyline
NHCH34 = Amitriptyline
CH3N

5 = Toluene

CH3CH30 1234 5 6 7 8 9 10 11Retention time / minSunrise C18-SAC
Silanol Activity Controlled C18 HPLC Column◆ Multiple mode separation is achieved on Sunrise series✻ Silanol groups controlled its activity functions as ion-exchange groups
Separation on Sunrise C18-SAC is done includinghydrogen bond and ion-exchange interaction
based on silanol groups except for hydrophobicinteraction. Control of pH and salt concentration ofa mobile phase can regulate retention.

■ Separation of basic compounds with ammonium

acetate: Effect of salt concentration(Sunrise C18-SAC)
253

4200 mM21534100 mM
213retention(Sunrise C18-SAC)

Column size: 150 x 4.6 mm
Mobile phase:
CH3CN / acetate ammonium(pH4.1) = 70:30
Flow rate: 1.0 mL/min
Temperature: 40 °C
Sample:1 = Uracil
2 = Toluene
3 = Propranolol
5 4 = Nortriptyline
5 = Amitriptyline

4N (Amitriptyline)
1.2

12,000

1.0

10,000

Amitriptyline
0.8

8,000

0.6

Nortriptyline

0.4

50 mM
2114,000

1.4

log k1■ Relationship between buffer concentration and4,000

400.0
0.5

1.02321.5

2.02.53.0log [Buffer concentration]

54log k = −10 mM
02,000525 mM
1Propranolol

Toluene0.2

36,000468 101214161820222426zBz
V
1log[ A] + B log[ A] + log r +log K ABzAzAVm z A

A：ion in eluent，B：ion of sample， k：retention of B，K：selection coefficient

Retention time / min

ZA, ZB：charge，Vr, Vm：resin volume and mobile phase volume in column■ Chromatograms under different pH conditions (Sunrise C18-SAC)
1

2

pH2.7

4

3
1

pH3.5

2

345under acetic acid solution, pH was 2.7
and there no ammonium ion. A basic
compound shows tailing under pH 2.7 (noammonium ion).

5
N=13,000Column size: 150 x 4.6 mm
Mobile phase:
CH3CN / 100 mM acetate ammonium = 70:30
Flow rate: 1.0 mL/min
Temperature: 40 °C
Sample:1 = Uracil
HNO

1pH4.1

1pH5.02 3NHO

CH352 = Toluene

42

3 = Propranolol
354CH3ONH
OH

4 = Nortriptyline
12NHpH5.931pH6.854CH35 = Amitriptyline
N2CH3CH330N [Amitriptyline]
N(Amitriptyline)

Sunrise C18-SAC is bonded with octadecylsilane ona pure silica gel and controlled its silanol activity
without end-capping. Its carbon content is 14%.

5410152025
Retention time / min530

N=11,500
3540

CH3Sunrise C30, C28, C18-SAC✻ Sunrise series Analytical and Preparative Columns
Inner diameter

lengthSunrise C30, 3µm

Sunrise C30, 5µm

Sunrise C28, 3µm

Sunrise C28, 5µm[mm][mm]Cat. No.

Cat. No.

Cat. No.

Cat. No.

2.050

SM2241SM3241

ST2241

ST324175

SM2251－ST2251－100

SM2261SM3261ST2261

ST3261150

SM2271SM3271

ST2271

ST3271250

SM2281

SM32281

ST2281

ST3281

4.610

SM2411

SM3411

ST2411

ST341150

SM2441SM3441

ST2441

ST344175

SM2451－

ST2451－

100

SM2461SM3461

ST2461

ST3461150

SM2471

SM3471

ST2471

ST3471250

SM2481

SM3481

ST2481

ST3481

10.0250－

SM3781－

ST3781

20.0250－

SM3881－

ST3881

Inner diameter

lengthSunrise C18-SAC,3µm

Sunrise C18-SAC,5µm

[mm]

[mm]

Cat. No.

Cat. No.

2.050

SA2241

SA3241

4.675

SA2251－100

SA2261

SA3261

150

SA2271

SA327110SA2411

SA3411

50

SA2441

SA3441

75

SA2451－

100

SA2461

SA3461150

SA2471

SA3471

250－SA3481

10.0

250－SA3781

20.0

250－SA3881

2203

ChromaNik Technologies Inc.
6-3-1 Namiyoke, Minato-ku, Osaka 552-0001 Japan TEL: +81-6-6581-0885 FAX: +81-6-6581-0890 Email: info@chromanik.co.jp