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PEAK SHAPE PERFORMANCE UHPLC PERFORMANCE WITH ANY LC INSTRUMENT

SUNSHELL

HARDCORE SHELL TECHNOLOGY

GLOBAL DISTRIBUTOR BIOTECH AB www.biotech.se1

PEAK SHAPE PERFORMANCE

HARDCORE

SUNSHELL

SUPERFICIALLY POROUS SILICA

2

EFFICIENT

*1.6 μm and 3.4 μm of core and 0.5 μm and 0.6 μm of superficially p silica layer *Same efficiency and high throughput as a Sub 2 μm and 3 μm par *Same pressure as a 3 μm and 5 μm par *Same chemistry as Sunniest technology (reference figure 1) *Good peak shape for all compounds such as basic, acidic and chela ng compounds *High stability ( pH range for SunShell C18, 1.5 to 10) * Low breeding

WHAT IS SUNSHELL? THE NEXT GENERATION HARDCORE SHELL PARTICLE 0.5 µm

0.5 µm

C o re

2.6 µm

1.6 µm

2.6 µm C o re

Porous silica Secure your analysis with SunShell hardcore column technology Unique bonding technology combined with core shell particles gives you faster performance and more Schematic diagram of a core shell silica particle, 2.6 µm reliable results. The SunShell technique assures top efficiency with all kinds of LC and UHPLC systems.

FEATURES OF SUNSHELL 2.6 µm AND 5 µm Van Deemter Equation

. 1.6 μm and 3.4 μm of core and 0.5 μm and 0.6 μm of superficially porous silica layer. . Same efficiency and high throughput as a Sub-2 μm and 3 μm particle. . Same pressure as a 3 μm and 5 μm particles. . Same chemistry as Sunniest s. technology (reference figure below). . Good peak shape for all compounds such as basic, acidic and chelating compounds.

Superficially porous silica . High stability (pH range for SunShell C18, 1.5 to 10). orous . Low bleeding.

A term : Eddy diffusion(dp is particle diameter) B term diffusion 0.5: Longitudinal μm (Dm is diffusion coefficient) C term : Mass transfer

2.6 μm

Van Deemter plot

1.6 μm

CORE

H

ell particle

O

H

Si O Si

Si O

O

O

Si O

O O

O

O

O Si

Si O

Si

Si

Si O Si O

OO Si O

O

Si

Si O

SiO Si O

O

O

Si

O

Si

O O Si

O

Si O O OH O Si O O O O O Si Si Si O H OO OO O O O Si O Si O O Si Si Si O O O Si O O O O Si O O O O Si

O O

Si O

Si

Si

Si

Schematic diagram of bonding of SunShell C18

C term

POROUS SILICA

u MPa

45 SunShell C18 shows same efficienSunniest C18-HT 2.0 µm 40 cy as a Sub 2 μm C18. In comparBrand A C18 1.9 µm 35 ison between fully porous 2.6 μm Brand B C18 1.8 µm 30 and core shell 2.6 μm (SunShell), SunShell shows lower values for 25 Brand C C18 1.7 µm A term, B term and C term of Van 20 Brand D C18 2.6 µm Deemter equation. The core shell 15 SunShell C18 2.6 µm structure leads to higher perfor10 Column dimension: 50 x 2.1 mm mance compared with the fully 5 Mobile phase: Acetonitrile/water=(70/30) Temperature: 25 C porous structure.

0 Furthermore back pressure of 0 0,5 1 1,5 Flow rate, mL/min SunShell C18 is less than a half Comparison of back pressure for high throughput columns compared to Sub-2 μm C18s.

Back pressure, MPa

Final TMS

B term A term

o

3

1

me chemistry as *Same *Same Sunniest chemistry chemistry technology as asSunniest Sunniest (reference technology technology figure 1) (reference (reference figure figure 1) 1) ood peak shape*Good *Good for all peak compounds peak shape shapefor such forall allas compounds compounds basic, acidic such such andas asbasic, basic, acidic acidic and and la ng compounds chela chela ng ng compounds compounds 2.6 µm gh stability ( pH*High *High rangestability stability for SunShell ((pH pHrange range C18, 1.5 for forSunShell to SunShell 10) C18, C18, 1.5 1.5 to toSUN 10) 10) SHELL ow breeding **Low Low breeding breeding

C o re

2.6 µm

Final TMS

0.5 µm

0.5 0.5 µm µm

Porous silica

CCoore re 1.6 µm

2.6 µm

2.6 2.6 µm µm

C o re 1.6 1.6 µm µm

CCoore re

Si SiSi Si SiSi O OO SiSiOOO O OO SiSi Si O OOOO SiSi SiSi SiSiSi OO Si SiSi O OO O O Si Si Si O O O O O OO OO OO SiSi Si Si SiSi SiSi Si O Si Si OSi O O O OO OH OH OH Si SiSi Si SiSi SiSi O OO O OSi OO O OO SiSi OOSi OOO Si SiSi SiO Si Si O OO O Si O O Si Si Si Si Si O O O Si SiSi O H OO HH OOOO O O O OO Si SiSi Si O OO O OOOOOO OO O O O Si SiSi O OO O O SiSi O OO Si SiSi O OO O SiOO O Si SiSiSi SiSi Si SiSi SiSi OO Si O SiSi OO OO O Si O O O OO SiSi OOOO OO O OO O OOO Si SiSi O O O O O O O OO O O OO OO O OO O OO OO Si SiSi O OO O OO O OO

Porous Poroussilica silica

Van Deemter plot

Van Van Deemter Deemterplot plot

H 14 HETP, µm

12 B term

10

A term

O O

Back pressure, MPa

uu

40

40 40

Sunniest C18-HT 2.0 µm

35

35 35

14 14

Fully porous 3 µm

Fully Fullyporous porous33µm µm

12 12

Fully proous 1.8 µm

Fully Fullyproous proous1.8 1.8µm µm

10 10

SunShell C term 2.6 µm

SunShell SunShell2.6 2.6µm µm

88

Fully porous 2.6 µm Fully Fully porous porous 2.6 2.6 µm µm

Brand A C18 1.9 µm

20 15

S

Mobile phase: Acetonitrile/water=(60/40) Mobile Mobilephase: phase:Acetonitrile/water=(60/40) Acetonitrile/water=(60/40C 2Temperature: 2 25 oC Temperature: Temperature:25 25ooCC c Sunniest C18-HT 2.0 µm Sample : Naphthalene Sample Sample: :Naphthalene Naphthalene

00

Brand A C18 1.9 µm

Brand D C18 2.6 µm

v 15 15e p

F t

SunShell C18 2.6 µm

SunShell C18 shows SunShell SunShell same efficiency C18 C18 shows shows assame same a subefficiency efficiency 2 μm as as aa sub sub 2 10 Column dimension: 50 x 2.1 mm C18.5 In comparisonC18.

C18. between In InMobile comparison comparison fully porous between between 2.6 μm fully fully and porous porous 2.6 2.6 μm μm phase: Acetonitrile/water=(70/30) Temperature: 25 oC (SunShell), core shell 2.6 μm (SunShell), core core shell shell 2.6 2.6 SunShell μm μm (SunShell), shows lower SunShell SunShell shows shows lo lo Sunniest SunniestC18-HT C18-HT2.0 2.0µm µm 0 values0 for A term, B values values term and for for A A C term, term, term B B of term term Van and Deemter and C C term term of of Van Van De De 0,5 1 1,5 Brand BrandAAC18 C181.9 1.9µm µm equation. The coreequation. equation. shell structure The The core core leads shell shell higher structure structureleads leads higher higher Flow rate, mL/min Narrow particle distribution performance to compare performance performance with the to to fully compare compare porous with with structure. the the fully fully porous porous st s Brand BrandBBC18 C181.8 1.8µm µm (Core Shell silicaD /D =1.15)

Wide particle distribution Brand B C18 1.8 µm 30 30 (Conventional silica gel D90/D10=1.50) Back Back pressure, pressure, MPa MPa

Fully Fullyporous porous55µm µm

5 00 10 55 15 10 10 reducing Eddy Diffusion Brand B C18velocity, 1.8 µm mm/sec Mobile phase velocity, mm/sec Mobile Mobile phase phase velocity, mm/sec C C term term (multi-path diffusion) as the A term in between the particles in the column is 25 Comparison of plate Brand C C18 1.7 µm Comparison Comparison height of plate plate height height plots plotsof plots Van Deemter Equation. reduced and efficiency increases by

45 45

Comparison of back pressure for 90 high 10 throughput columns

25 25

Brand C C18 1.7 µm

Brand BrandCCC18 C181.7 1.7µm µm Furthermore

20 20

Brand D C18 2.6 µm

Brand BrandDD

SunShell C18 2.6 µm

SunShell SunShellC18 C182.6 2.6µm µm

10 10

Fully porous 5 µm

4Column: 4 C18, 50 x 4.6 mm C18Column: Column:C18, C18,50 50xx4.6 4.6mm mmC18 C18

35 0 porous particle, so that the space in 30

45

15 15

16 16

4

0

u

18 18

66u

40

B termThe size distribution BB term termof a core shell C term (SunShell) particle is much narrower A term AA term term than that of a conventional totally

4 10

Si

6

A TERM452

H H

15

Si

Van Deemter plot

HETP, HETP, µm µm

18

8

20

Si O

A term : Eddy diffusion(dp is particle diameter) B term : Longitudinal diffusion (Dm is diffusion coefficient) diagramSchematic Schematic of a core shell diagram diagram silica of ofparticle, aa core core shell shell 2.6 µm silica silica particle, particle,2.6 2.6 µm µm C term : Mass transfer

A term : Eddy diffusion(dp AAterm term is particle ::Eddy Eddy diameter) diffusion(dp diffusion(dp isisparticle particlediameter) diameter) B term : Longitudinal diffusion BBterm term ::Longitudinal Longitudinaldiffusion diffusion (Dm is diffusion coefficient) (Dm (Dmisisdiffusion diffusioncoefficient) coefficient) C term : Mass transfer CCterm term ::Mass Mass transfer transfer

25

O

Schematic diagramSchematic Schematic of bondingdiagram diagram of SunShell of of bonding bonding C18 of of SunShel SunShe VAN DEEMTER EQUATION

16

30

O

Van Deemter Equation

Van Deemter Equation Van Van Deemter DeemterEquation Equation

H

HH

SiSi

OO Schematic diagram of a core shellOsilica particle, 2.6 µm

0.5 0.5 µm µm

Porous silica

Schematic

OO

Si

O

C o re2.6 2.6 µm µm

H

HOW DOES SUNSHELL WORK? NARROW PARTICLE DISTRIBUTION

0.5 µm

Final Final TM TM

C o re

1.6 µm

O

6 µm

Back pressure, MPa

0.5 µm

back pressure Furthermore Furthermore of SunShell back back pressure pressure C18 isof of less SunShell SunShell C18 C18 is is l 1 than a half to compare than than with a a half half sub-2 to to compare compare μm C18s. with with sub-2 sub-2 μm μm C18s.

C18s.

C18 C182.6 2.6µm µm

Column dimension: 50 x 2.1 mm Column Columndimension: dimension:50 50xx2.1 2.1mm mm

/D10=1.59

50

Sunniest, 2 μm D : 2.63 µm

Wide particle distribution (Conventional silica gel D /D =1.50) reduces and efficiency D /D =1.15 among particles in the column Narrow particlein distribution (core shell silica D /D Equation =1.15) D :D1.75 µmµm All terms A term : 2.82 Van Deemter reduce.

The size distribution of a core D :D2.01 /D µm =1.15

D90: 2.65 µm D90/D10=1.59

90

10

50

10

90

50

90

90

10

90

10

reduces andby efficiency increases reducing Eddy (SunShell) particle is much increasesshell by reducing Eddy Diffusion (multi-path diffusion) narrower than that of a D : 1.75 µm Diffusion (multi-path diffusion) CompanyAF, term 2 μm as the The Aconventional term in Vantotally Deemter size distribution of aporous core D : 2.01 µm SunShell, 2.6 μm as the A term in Van Deemter D : 2.31 µm D : 1.67 µm shell (SunShell) particle is much .

Equation arison of Particle Size Distribution : 2.46 µm D D /D =1.32 Packing state of core shell and fully porous silica Flow of mobile phase : 2.09 µm Equation.particle, so that the space Comparison ofD Particle Size Distribution 10

D90: 2.31 µm 2 μm D90/D10Sunniest, =1.32

Wide particle distribution (Conventional silica gel D90/D10=1.50)

10

B TERM

50

10

90

50

90

D90: 2.65 Company µm F, 2 μm D90/D10=1.59 D10: 1.67 µm D50: 2.09 µm D90: 2.65 µm D90/D10=1.59

10

10

D50: 2.63 µm Packing state of core shell and fully porous silica D90: 2.82 µm SunShell, 2.6 μm Narrow particle distribution (core shell silica D90/D10=1.15) D90/D10=1.15 D10: 2.46 µm Flow of mobile phase D50: 2.63 µm

narrower than that of a among particles in the column conventional totally porous reduces and efficiency particle, so that the space increases by inreducing Eddy among particles the column Diffusion diffusion) Column: SunShell C18, 2.6(multi-path µm x 2.1 Column: SunShell C18, 2.650 µm 50mm x 2.1 mm reduces and efficiency Totally porous 2 µm250 x 2.1 Totally porous µm 50mm x 2.1 mm increases reducing Eddy as the Abyterm in Van Deemter Mobile phase: Acetonitrile/water=(60/40) Mobile phase: Acetonitrile/water=(60/40) Sample : Naphthalene Diffusion (multi-path diffusion) Sample : Naphthalene .

Equation 25 degree C Core shell 2.6 µm Deemter as the A term in Van 25 degree C Core shell 2.6 µm 40 degree.C Core shell 2.6 µm Equation 40 degree C Core shell 2.6 µm

D : 2.82 µm 90 10 ion of aDiffusion solute isof a solute 20 20 B term D /D =1.15 Bis term blocked by the ed by the Diffusion of a solute is A solute diffuses in a pore as well as outside of particles.

Totally porous silica Totally porous silica A solute diffuses in a pore as well as outside of particles.

16 16 ofby a the core, so nce of aexistence core, so blocked existence Comparison of Particle Size Distribution Packing state of core shell and fully porous silica that a solute diffuses solute diffuses of a core, so that a solute 12 12 less in a core shell silica Comparison of Particle Size Distribution Packing state of core shell and fully porous silica 40 degree C Totally porous 2 µm diffuses less in a core n a corecolumn shell silica than in a totally 40 degree C Totally porous 2 µm 8 shell silica column than mn than porous in a totally Diffusion of a solute is 8 20 silica column.

B term Column: SunShell C18, 2.6 µm 50 x 2.1 mm A core without pores blocks diffusion of a solute.

Core shell silica Totally porous 2 µm 50 x 2.1 mm blocked by the in a totally porous silica 4 s silica Consequently column.

A solute diffuses in a pore as well as outside of particles.

B term in of shell Totally porous silica Diffusion a solute is A core without pores blocks diffusion of a solute.

Mobile phase: Acetonitrile/water=(60/40) Core silica 20 B term Column: SunShell C18, 2.6 µm 50 x 2.1 mm 16 Sample 4 existence of a core, Totally: Naphthalene porous 2 µm 50 x 2.1 mm equentlyVan Bcolumn. term inConsequently blocked by theso Deemter Equation Totally porous silica A solute diffuses in a pore as well as outside of particles.

Mobile phase: Acetonitrile/water=(60/40) 0 16 that acore solute diffuses 25 degree C Core shell 2.6 µm : Naphthalene existence of a core, so B termin inthe Van Deemter 0 0,2 12 0,4Sample0,6 0,8 1 Deemterreduces Equation 40 degree C Core shell 2.6 µm 0 less in athat core shell ainsolute diffuses 25 degree C Core shell 2.6 µm shell silica column.

Equation reduces the silica Flow rate (mL/min) 12 40 degree C Totally porous 2 µm es in the core 0 0,2 0,4 40 0,6 0,8 1 degree C Core shell 2.6 µm shell silica column less thanininaacore totally 8 40 Plates degree C Totally porous 2 µm core shell silica column.

Difference in longitudinal diffusion Plot of Flow rate and height Difference of longitudinal diffusion silica column. porous silica column than in a totally 8 Flow rate (mL/min) column.

Narrow particle distribution (core shell silica D /D =1.15)

90

10

A core without pores blocks diffusion of a solute.

Plate height (μm)

Core shell silica

Plate height (μm)

Plate height (μm) Plate height (μm)

90

porous B silica column.

4 Consequently term in Difference of A core without pores blocks diffusion of a solute. Plot Core shell silica diffusion PlotofofFlow Plate height Flowheight rate rate and vs Plates longitudinal 4 Consequently B term Asinshown in the left figure, a core shell Van Deemter Equation C term 0 Van Deemter Equation particle has a core so that the diffusion path 0 reduces in the core 0 0,2 0,4 0,6 0,8 1 reduces in the core 0 0,2 0,4 Diffusion 0,6 0,8 1 Diffusion As shown the leftshortens figure, and a core shell of in samples mass transfer shell silica column.

Sample Flow rate (mL/min) Ca. 2.0 µm shell silica column.

2.0 µm Flow rate (mL/min) emanates in a becomes fast. means that the C term particle has a core soThis that the diffusion path Plot of Flow and Plates particlePlot after it Flow Difference of longitudinal diffusion of raterate andDiffusion Plates heightheight Difference of longitudinal diffusion Diffusion in Van Deemter Equation reduces.

In other enters. sample sample of samples shortens and mass transfer Sample Ca. 2.0 µm 2.0 µm HETPmeans (theoretical plate) kept emanates in a becomeswords, fast. This theleft Cisterm As rate shown inthat the left core As increases. shown in the a coreshell shell particle after it even if flow A figure, 2.6figure, µmacore C term C term in Van Deemter Equation In other enters. sample particle has athe core so that thediffusion pathsample particle hasreduces. a in core so that the path As shown left figure, adiffusion core shell shell particle shows as same column Diffusion Diffusion Diffusion Diffusion of samples shortens and masstransfer transfer Sample words, HETP (theoretical plate) is kept of samples shortens and mass Sample Ca. 2.0 µm efficiency asparticle a totallyhas porous sub-2 µm the diffusion 2.0 µm a core so that Ca. 2.0 µm emanates in a 2.0 µm becomes fast.

This means that the C term emanates in a becomes fast.

This means that the C term even if flow rate increases.

A 2.6 µm core particle after it C o r e S h e ll particle. particle afterT ito ta lly path of samples shortensreduces. and mass in Van Deemter Equation In other enters. sample sample in Van Deemter Equation reduces.

In other enters. sample sample p o r ou s shell particle shows as same column The right figure shows that a diffusion width words, HETP (theoretical plate) is keptthat transfer becomes fast.plate) This means words, HETP (theoretical is kept of a sample in a 2.6 µm core shell particle efficiency as a totally porous sub-2 µm even if flowin rate increases.

AEquation 2.6 µm core the term Van Deemter reeven ifCflow rate shows increases.

A 2.6 µm core porous particle.

Both shell particle as same column C o r e S h e ll particle. and a 2 µm totally ta lly T o µm 2.0 duces.

In other words, HETP (theoretical shellefficiency particle shows as same column as a totally porous sub-2 µm diffusion widths are almost same.

The 2.6 p o r ou s width The right figure shows that a diffusion 1.6 µm plate) is kept even if flow rate increasas aistotally porous sub-2 µm C o r e S h e ll T o ta lly µm core efficiency shellparticle. particle superficially of a sample in a 2.6 µm core shell particle p o r ou particle. es.

A 2.6 μm core shellthat particle shows The right figure shows a diffusion width 2.6 µmC o r e S h e ll T so ta lly porous, so that the diffusion width becomes p o r ou s and a 2 µm totally porous particle.

Both of a sample in a 2.6 µm core shell particle width The right figure shows that a diffusion the same column efficiency as a totally Diffusion of sample in core shell and totally porous silica narrower than particle size. Same diffusion 2.0 µm Comparison of diffusion path and a 2 µm totally porous particle.

Both of aporous sample in a 2.6 µm core shell particle diffusionmeans widthssame are almost same.

The 2.6 2.0 µm Sub-2 μm particle. efficiency. diffusion widths porous are almost same.Both The 2.6 1.6 µm 1.6 µm and a 2isµm totally particle. µm core shell particle superficially 2.0 µm µm core shell particle is superficially diffusion widths are almost same. The 2.6 2.6 µm 1.6 µm porous, so that the diffusion width becomes The rightso figure shows the diffusion porous, that the diffusion width becomes 2.6 µm µm core shell particle is superficially in coreand shell totally and totally poroussilica silica Diffusion sampleofinsample core shell porous Comparison of Performance by Plate/Pressure narrower particle Same diffusion narrower than particle size.

Same diffusion width of that a than sample in asize.

2.6width μm core shell of Diffusion Diffusion so diffusion becomes arison of diffusion path Comparison of diffusion path porous, 2.6 µm of a sample in core shell means sameathe efficiency. means same efficiency. particle and 2 μm totally porous parDiffusion of sample in core shellsilica and totally porous silica and totally porous narrower than particle size. press.

Same diffusion Plate Back press. (MPa) Plate/back Comparison of diffusion path ticle.

Both diffusion are almost Under a constant back pressure means efficiency.widths Sunniest C18 –HT 2.0 µm 9,900 16.7same 593 Brand A C18 1.9 µm 7,660 16.3 the same.

2.6 μm 470 core shell particle condition, SunShell C18 showed Comparison of Performance byThe Plate/Pressure Brand B C18 1.8 µm 10,100 19.6 515 is superficially porous, so that the dif- more than 2 times higher mparison ofC C18 Performance

by Plate/Pressure Brand 1.7 µm 11,140 32.0 348 Plate Back press. (MPa)990 Plate/back press. performance to compare with SunShellComparison C18 2.6 µm 9,600 9.7 fusion width becomes narrower than of Performance

by Plate/Pressure Under a constant back pressure Sunniest C18 –HT 2.0 µm 9,900 16.7 593 totally sub-2µm porous C18s.

Brand A C18 1.9 µm 7,660 16.3 470 particle size.

Same diffusion means Plate Back press. (MPa) Plate/back press.

erm

C TERM

Sunniest C18 –HT 2.0Brand µm B C18 1.8 µm nniest C18 –HT 2.0 Aµm 9,900 Brand C18 1.9 µmC18 Brand–HT C C18 Sunniest 2.01.7 µmµm and A C18 1.9Brand µm B C18 1.8 µm SunShell 7,660C18 2.6 µm Brand A C18 1.9 µm and B C18 1.8Brand µm C C18 10,100 Brand B C18 1.8 µm 1.7 µm Sunniest C18 –HT 2.0 µm and C C18 1.7SunShell µm 11,140 Brand C C18 1.7 µm C18 2.6 µm Brand A C18 nShell C18 2.6 µm SunShell C189,600 2.6 µm 1.9 µm

Plate10,100 Back press.19.6 (MPa) 16.7 593 same efficiency.

9,90011,140 16.732.0 16.3 9,600 470 9.7 7,660 16.3 19.6 515 10,100 19.6 32.0 348 11,140 32.0 9.7 990 9,600 9.7

0 µm 5 000 10 000 Brand B C18 1.8

0

10

20

30

condition, SunShell C18 showed

515press.Under a constant back pressure Plate/back more than 2 times higher Under a constant back pressure 348 593 performance to compare with condition, SunShellSunShell C18 showed 990 470 condition, C18 showed sub-2µm porous C18s.

515 more thantotally 2 times higher more than 2 times higher 348 performance to compare with with performance to compare 990

0 250 500 750 1000

5

SUNSHELL

C18 - 2.6 µm HIGHEST RETENTION / LARGEST STERIC SELECTIVITY / LOWEST BACKPRESSURE

SunShell C18, 2.6μm

Retention of standard samples and back pressure were compared for five kinds of core shell type C18s. Company A C18 showed only a half retention in comparison with SunShell C18. Steric selectivity becomes large when ligand density on the surface is high. SunShell C18 has the largest Characteristics of SunShell C18 steric selectivity as well as the highest ligand density leading toL1) the longest retention time.

Core shell silica C18 (USP SunShell C18

Particle size

Pore diameter

Specific surface area

Carbon content

Bonded phase

End-capping

Maximum operating pressure

Available pH range

2.6 µm

9 nm

150 m2/g

7%

C18

Sunniest End-capping

60 MPa or 8,570 psi

1.5 - 10

SUNSHELL C18 COMPARISON

Comparison of standard samples between core shell C18s k6=5.4 1

5

2 3

4

5

1 2

5

3

1

3

1

0

6

4

2

4

SunShell C18 21.8 MPa 4

6

8

7

k6=9.0 6

5

23 2

Company B C18 22.7 MPa

7

k6=7.4 6

4

Column: Company A C18, 2.6 μm 150 x 4.6 mm (26.1 MPa) Company B C18, 2.6 μm 150 x 4.6 mm (22.7 MPa) Mobile phase: CH3OH/H2O=75/25 Company C C18, 2.7 μm 150 x 4.6 mm (30.6 MPa) Flow rate: 1.0 mL/min, 40°ºC Company D C18, 2.7 μm Temperature: 150 x 4.6 mm (22.2 MPa) SunShell 2.6 μm x 4.6 mm3(21.8 MPa) Sample: 1 =C18, Uracil, 2 =150 Caffeine, = Phenol, O=75/25 phase: CH3OH/H 4 Mobile = Butylbenzene, 5 = 2o-Terphenyl, Flow rate: 1.0 mL/min, Temperature: 40 ºC 6 = Amylbenzene, 7 = Triphenylene Sample: 1 = Uracil, 2 = Caffeine, 3 = Phenol, 4 = Butylbenzene 5 = o-Terphenyl, 6 = Amylbenzene, 7 = Triphenylene

Company A C18 26.1 MPa

6

1 2 3

7

k6=9.7

Company C C18 30.6 MPa 7

6

Company D C18 22.2 MPa

5 4

6

7

k6=10.4

10 12 14 16 Retention time/min

18

20

22

Hydrogen bonding

Hydrogen bonding

Company A C18 (Caffeine/Phenol) 0.48 Company B C18 Company A C18 Company C C18 Company B C18 Company C C18 Company D C18 Company D C18 Sunshell C18 24 SunShell C18

0.480.35 0.350.42 0.420.44 0.440.39 0.39

Hydrophobicity

Hydrophobicity

Steric selectivity

Steric selectivity

1.54 1.20 (Amylbenzene/Butylbenzene) (Triphenylene/o-Terphenyl) 1.56 1.54 1.57 1.56 1.57 1.60 1.60 1.60 1.60

1.50 1.20 1.25 1.50

1.25 1.31 1.31 1.46

1.46

Retention of standard samples and back pressure were compared for five kinds of core shell type C18s. Company A C18 showed only a half retention to compare with SunShell C18. Steric selectivity becomes large when ligand density on the surface is high.

SunShell C18 has the largest steric selectivity so that it has the highest ligand density. This leads the longest retention time.

Mobile phase: Acetonitrile/10mM acetate ammonium pH6.8=(40:60)

Mobile phase: Acetonitrile/20mM phosphate buffer pH7.0=(60:40)

14000

16000

BEST PEAK SHAPE AVAILABLE 12000

12000 10000

100 倍

SunShell C18 Company C C18 Sunniest C18 3um Company A C18 Company B C18 overloads much Company D C18

8000

Theoretical plate

Theoretical plate

14000

6000 Amitriptyline more at acetonitrile/buffer mobile phase than 4000 methanol/buffer which causes tailing. 2000 Five kinds of core shell C18s were 0 0,001

0,01 3

1 2

1

TF=1.42

TF=2.43

6000

30倍

better sensitivity and accuracy of the method.

0 0,001

0,01

0,1

Sample: 1 = Uracil (0.07µg) 2 = Propranolol (1.53µg) 3 = Nortriptyline (0.32µg) 4 = Amitriptyline (0.32µg)

C C18 (core shell)

1

2

3

TF=1.20

Sunniest C18 3 μm (fully porous)

2

3

4 5 6 7 8 9 Retention time/min

SunShell C18 (core shell)

TF=2.61

A C18 (core shell)

TF=2.73 TF=3.24

D C18 (core shell)

TF=4.38 1

10

4

TF=1.89

TF=3.21 B C18 (core shell)

0

1

Sample weight/μg

SunShell C18 (core shell)

TF=1.25

8000

10

Sample weight/μg 4

Sunshell C18 Company A C18 Company P C18 Company T C18 Company S C18

compared as refers to loading capacity 4000 of amitriptyline. Thanks to the unique bonding technology Sunshell gives ex2000 traordinary peak shape, which means

0,1

TF=1.18

10000

10

0

Theoretical plate was calculated by 5σ method Mobile Phase: using peak width at 4.4% of peak height.

• Acetonitrile/20 mM phosphate buffer pH 7.0 (60/40)

4.4%

1

2

3

4

C C18 (core shell) A C18 (core shell) B C18 (core shell) D C18 (core shell)

5 6 7 8 9 Retention time/min

10

Mobile Phase: • Acetonitrile/10 mM ammonium acetate pH 6.8 (40/60)

Mobile phase: Acetonitrile/0.1% formic acid=(30:70) 4,5 4

USP tailing factor

Company D C18 Company B C18 Company A C18: Kinetex C18 3,5 Company A C18 C18 Company B C18: Accucore Company C C18 C18 EC 3 Company C C18: PoroShell SunShell C18 Company D C18: Ascentis Express C18 2,5 7倍 2

USP Tailing factor 3.6 2.9 2.4

1. Kinetex C18, 2.6 μm 2. Accucore C18, 2.6 μm 3. PoroShell C18 EC, 2.7 μm 4. Ascentis Express C18, 2.7 μm 5. SunShell C18, 2.6 μm

1.8 1.3

1,5 1

Comparison column

Amitriptyline overloads at low weight when acetonitrile/0.1% formic acid

7

EXPANDED pH RANGE DUE TO THE SUNSHELL BONDING TECHNOLOGY ◆Evaluation of Stability

SUNSHELL C18 STABILITY

Accelerated acidic test

100 ◆Evaluation of Stability

SunShell C18

80 40

Company D C18 Company B C18

60 20 40 0 20

1% TFA 80 ºC

Company A C18 SunShell C18 Compnay CompanyCDC18 C18 Company B C18 0

20 Company 40 A C1860 Compnay C C18

Durable test condition 0 0 20 40

Time/h 60

1% TFA 80 100 80 ºC 80

100

120

120

Column size: 50 x 2.1 mm Time/h Mobile phase: CH3CN/1.0% TFA, pH1=10/90 Durable test condition Flow rate: 0.4 mL/min Temperature: 80 ºC Column size: 50 x 2.1 mm Mobile phase: CH3CN/1.0% TFA, pH1=10/90 Measurement Flow rate: 0.4 condition mL/min Temperature: 80 ºC Column size: 50 x 2.1 mm Mobile phase: CH3CN/H2O=60/40 Measurement condition Flow rate: 0.4 mL/min Stability under acidic Temperature: 40 ºCpH condition was evaluated at 80°C Column size: 50 x 2.1 mm using acetonitrile/1% acid solution (10:90) as Sample: 1 = Uracil trifluoroacetic (t0) CN/H Mobile phase: CH 3 2O=60/40 mobile phase.

100% aqueous mobile phase expels from the 2 = Butylbenzene rate:packing 0.4 mL/min poresFlow of C18 materials by capillarity and packing Temperature: 40 ºC materials do not deteriorate.

Adding 10% acetonitrile to the Sample: 1 = Uracil (taccurate 0) mobile phase enables evaluation.

Stability under acidic pH condition was evaluated at 80 ºC using 2 = Butylbenzene

8

Alkaline test

Accelerated acidic test

100 60

acetonitrile/1% trifluoroacetic acid solution (10:90) as mobile phase. 100% aqueous mobile phase expels from the pore of packing materials by capillarity and packing materials doesn’t Stability under pH condition was evaluated at 80 deteriorate.

10%acidic acetonitrile in a mobile phase allows anºC using 1-3) acetonitrile/1% trifluoroacetic acid solution (10:90) as mobile accurate evaluation.

Relative plate of butylbenzene/% Relative plate of butylbenzene/%

Relative retention/% Relative retention/%

80

100 80

SunShell C18

100 60

Company D C18 Company A C18

80 40

Company B C18 SunShell C18 Company CompanyCDC18 C18

60 20 40 0 20

Alkaline test

Company A C18 Company B C18 0

1 000

2 000 Company 3 000 C4C18 000

Elution volume/mL

0 1 000 2 000 Durable0test condition

3 000

4 000

Elution volume/mL

pH10 50 ºC 5pH10 000 6 000

50 ºC

5 000

6 000

Column Size: 50 x 2.1 mm Mobile phase: Durable test condition CH3OH/20mM Sodium borate/10mM NaOH=30/21/49 (pH10) Flow rate: 0.4 mL/min Column Size: 50 x 2.1 mm Temperature: 50 ºC Mobile phase: CH3OH/20mM Sodium borate/10mM NaOH=30/21/49 (pH10) Measurement condition Stability under pH condition was evaluated at 50°C Flow rate: 0.4basic mL/min using methanol/Sodium borate buffer pH 10 (30:70) as Temperature: 50 ºC Column Size: 50 x 2.1 mm mobile phase.

Sodium borate is used as an alkaline stanMobile phase:for CHpH 3CN/H 2O=60/40 dard solution meters, which allows for a high buffer Measurement condition Flow rate: 0.4 mL/min capacity.

Elevated temperature of 10°C reduces column life 40 other ºC toTemperature: one third.

The company shows stability when tested Column Size: 50 x 2.1 mm 1 (room) = Butylbenzene atSample: ambient temperature.

Mobile phase: CH3CN/H2O=60/40If room temperature is 25°C, column life is Flow rate: 0.4sixteen mL/min times longer than at 50°C. Temperature: 40 ºC Stability under basic pH condition was evaluated at 50 ºC using Sample: 1 = Butylbenzene methanol/Sodium borate buffer pH 10 (30:70) as mobile phase.

BLEEDING TEST USING LC/MS The high stability of the SunShell columns also means low bleeding in LC/MS analysis as shown here.

TIC of Q1

Column size: 50 x 2.1 mm Mobile phase: A) 0.1% Acetic acid B) CH3CN Gradient: Time: 0min 1min 5min 7min %B: 5% 5% 100% 100% Flow rate: 0.4 mL/min Temperature: 40 째C MS: ABI API-4000 Ionization: Turboionspray (cation) Measurement mode: Q1 Scan m/z 100-1000

9

SUNSHELL C18 EFFICIENCY Efficiency of SunShell C18 Column: SunShell C18, 2.6 μm 50 x 2.1 mm 3 2

1

0

Column: SunShell C18, 2.6 μm 50 x 2.1 mm Mobile phase: CH3CN/H2O=60/40 Flow rate: 0.3 mL/min Pressure: 7 MPa Temperature: 23 ºC UHPLC: Jasco X-LC

Plates (3)=10,300 Plate(3)=10,300 4

1

2

3 4 Retention time/min

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

5

Column: SunShell C18, 2.6 μm 150 x 4.6 mm

Column: SunShell C18, 2.6 μm 150 x 4.6 mm Mobile phase: CH3CN/H2O=70/30 Flow rate: 1.0 mL/min Pressure: 15.5MPa Temperature: 25 ºC UHPLC: Jasco X-LC

1

Plates (3)=38,000 Plate(3)=38,000 2 4

3

0

2

4 6 Retention time/min

8

Efficiency=253,000 plate/m EFFICIENCY = 253,000 plates/m

10

Examples of transfer from a conventional 5 µm column to SunShell column Inner diameter (mm)

ORDERING INFO OF SUNSHELL Isocratic separation

HPLC

1

Length Brand F C18, 5 µm 250 x 4.6(mm) mm

3 2

Sunshell C18, 2.6 µm 3 1

10 0

SunShell C18, 2.6 µm 100 x 4.6 mm

2

5

4

2

4

6

30 50 75 100 150 250

1/3 of analysis time

8

10 12 14 16 Retention time/min 3

18

4

1.0

2.1

3.0

4.6

USP category

Catalog no

Catalog no

Catalog no

Catalog no

Catalog no

N(4)=19,313

Column: CB6931 CB6331 CB6431 Brand F C18,CB6341 5 μm 250 x 4.6CB6441 mm CB6941 SunShell C18, 2.6 μm 100 x 4.6 mm CB6951 CB6351 CB6451 CB6961 CB6361 CB6461 Mobile phase: CB6971 CB6371 CB6471 Phosphoric acid = 45/55 CH3CN/20mM --CB6481 Flow rate: 1.0CB6381 mL/min,

---5 CB6141 --CB6161 CB6171 --N(4)=20,287

20

22

N(4)=20,287

24

L1

1.8 mL/min at the lowest chromatogram Temperature: 25 ºC Pressure: 9.5 MPa for Brand F C18 5 μm 13.4 MPa for SunShell C18 2.6 μm Detection: UV@230 nm

SUNSHELL

SunShell C18, 5 μm SunShell C18, 5 μm Characteristics of SunShell C18, 5 μm Characteristics of SunShell C18, 5 μm C18 - 5 µm Core shell silica

Particle Pore Core shell silica

size diameter Particle Pore Specific 4.6 µmsurface 9 nm SunShell C18 size diameter area

SunShell C18

4.6 µm

90 m2/g

9 nm

C18 (USP L1)

Specific surface area Carbon 90 m2/g content

5.5 %

Carbon content Bonded 5.5 % phase

Bonded Maximum operating C18 (USP L1) End-capping phase pressure Maximum operating Available pH End-capping C18 Sunniest End-capping pressure 60 MPa or 8,570 range psi

C18

Sunniest End-capping

Available pH range

1.5 - 10

1.5 - 10

60 MPa or 8,570 psi

Comparison Can of retention using HPLC be used inand anyplate L1 method - but with improved performance.

Comparison of retention and plate using HPLC 1 1

1

2

SunShell C18 5 µm

Column size: 150 x 4.6 mm

N6=20,000

Column size: 150Mobile x 4.6phase: mmCH OH/H O=75/25 3 2 Column size: 150 x 4.6 mm N 5 6=20,000 Mobile phase: CH3OH/H2O=75/25 Flow rate: 1.0 mL/min 4 Mobile phase: CH3OH/H2O=75/25 6 7 Temperature: 40 ºC Flow Flow rate:rate: 1.01.0 mL/min 5 mL/min 3 4 Sample: 1 = Uracil 6 Temperature: 40 ºC Temperature: 40°ºC N6=31,000 2 = Caffeine SunShell C18 2.6 µm Sample: 1 = Uracil 7 Sample: 1 = Uracil 3 = Phenol 2 5 N =31,000 2 = Caffeine 6 21 4 SunShell 2.6MPa µm 1 C18 4 = Butylbenzene 76 3

2 = Caffeine 3 = Phenol 2 5 5 = o-Terphenyl 21 MPa 4 6 4 = Butylbenzene

3 = Phenol 3 6 = Amylbenzene 5 = o-Terphenyl

4 = Butylbenzene 7 = Triphenylene N6=14,000 6 = Amylbenzene 6 MPa HPLC: Hitachi LaChrom ELITE

5 = o-Terphenyl 7 = Triphenylene (Tubing, 0.25 mm i.d.) =14,000 N 6 6 MPa HPLC: LaChrom ELITE

6 =Hitachi Amylbenzene (Tubing, 0.25 mm i.d.)

7 = Triphenylene HPLC: Hitachi ELITE (Tubing, 0.25 mm i.d.)

Totally porous silica Core shell silica Core shellLaChrom silica 8 MPa 2 SunShell C18 5 µm 8 MPa 3

7

Sunniest C18, 5 μm SunShell C18, 2.6 μm SunShell C18, 5 μm Totally porous silica Core shell silica Core shell silica There is a little 2/g Specific surface area Sunniest Totally porous shell silica Core 150 m2/g 90shell m2/gsilica C18, 5silica μm 340 mSunShell C18,Core 2.6 μm SunShell C18, 5 μm difference of k between Sunniest C18,Retention 5 µm SunShell C18, 2.6 µm SunShell C18, 5 µm There is a little Retention Retention Retention Specific surface area 150 m2/gRetention 90 m2/gRetention 340 m2/g totally porous and core factor (k) time (tR) factor (k) time (t ) factor (k) difference of k between 2 time (tR) 2 2 R 90 m /g Specific surface area 340 m /g 150 m /g Retention Retention Retention Retention Retention Retention shell particles.

There is a small difference 1) Uracil 1.70 0 1.34 1.30(k) 0totally porous and core time (tR) factor (k) time (tR) factor (k) time0(tR) factor Retention Retention Retention Retention Retention Retention of k between totally porous shell particles.

6) Amylbenzene time1.70 19.96 10.74 time (tR) 16.56 11.36 13.43 9.33 (k) (tR) factor (k) factor (k) time factor 1) Uracil 0 1.34 0 1.30 0 (tR) 6) AmylbenzeneRelative value of Amylbenzene 19.96 1) Uracil 1.70

100% 10.74 0

100% 83% 16.56 1.34 11.36

106% 13.43 0

67% 9.33 1.30

value of Amylbenzene 6)Relative Amylbenzene

100% 10.74

83%

67% 11.36

87% 13.43

9.33

67%

87%

100% 19.96

16.56 106%

Comparison between normal and semi-micro HPLC Comparison between normal and semi-micro35,150 HPLC Relative value of Amylbenzene

100%

100%

ORDERING INFO OF SUNSHELL

Flow cell Flow cell Normal Semi-micro

42,053 Response Sampling Tubing ID

Normal Sampling 0.1 secTubing 0.4 ID sec Response Sunshell C18, µm0.4 0.1 sec50.25 Semi-micro 0.1 sec 0.4 sec mmsec

Semi-micro 0.1 sec 0.40.05 sec sec0.130.05 mmsec

Semi-micro 0.05 sec

0.05 sec

0.13 mm 0 2

34,462 2.1 4.6 USP category 40,3203.0 30,564 34,462 41,043 34,052 40,320 30,564 Catalog no40,139 Catalog no Catalog no Catalog no Catalog no 41,043 34,05237,854 40,139 41,610 37,854 39,255CB3471 ----CB3371 41,610 L1 39,255 ----CB3381 CB3481

150 250

0.13 mm

1

106%

Inner diameter 42,053 35,150 (mm) 13,993 Length (mm)

0.25 mm

0.13 mm 0

83%

1 3

and core shell particles.

87% 0

2 4

13,993 1.0

3 5

4 6

5 6 7 Retention time/min 7 8 9

8

9

10

11

10

11

11

SUNSHELL

SunShell C18-WP, RP-AQUA, C8, Phenyl, PFP, 2.6 μm SunShell C18-WP, RP-AQUA, C8, Phenyl, PFP, 2.6 μm (Pentafluoropheny)

(Pentafluoropheny)

◆ Characteristics of SunShell ◆ Characteristics ofCore SunShell shell silica

ULTIMATE SELECTIVITY FOR YOUR ANALYSIS

Specific Carbon surface Bonded phase USP L line size diameter content Specific area Carbon Particle Pore surface Bonded phase USP L line content 7% μm C18 L1 SunShell C18size 2.6diameter 9nm area 150 m2/g

Core shellPore silica Particle

Maximum Available operating pH range Maximum pressure Available End-capping operating pH range Sunniest endcapping 60 MPa 1.5 - 10 pressure End-capping

2.6 μm2.6 μm SunShell nm m2/g 90 m2/g 7% SunShell C18 C18-WP 9nm 16 150 2.6 μm2.6 μm SunShell RP-AQUA 90 m2/g 5% SunShell C18-WP 16 nm 16 nm 90 m2/g

5% L1 Sunniest Sunniest endcapping C18 C18 L1 60 MPa60 MPa 1.5 - 101.5 - 10 C18 -WP / RP-AQUA / C8 /endcapping PFP - 2.6 µm 4% C28 / PHENYL Equivalent toSunniest L62 Sunniest endcapping MPa C18 L1 endcapping 60 MPa60 1.5 - 10 2 - 8 a)

4.5% L7 Sunniest Sunniest endcapping SunShell C82.6 μm2.6 μm /g 150 m24% C28 C8 Equivalent to L62 endcapping 60 MPa60 MPa2 - 8a) 1.5 - 9 SunShell RP-AQUA 16 nm 9nm 90 m2/g 2/g 2.6 μm2.6 μm 5% L11 Sunniest Sunniest endcapping SunShell C8Phenylhexyl L7 endcapping 60 MPa60 MPa1.5 - 9 1.5 - 9 SunShell C8 Phenyl /g m4.5% 9nm 9nm 150 m2150 4.5% Phenylhexyl Pentafluorophenyl L11 L43 Sunniest TMS endcapping60 MPa60 MPa /g endcapping 1.5 - 9 2 - 8 SunShellSunShell Phenyl PFP2.6 μm2.6 μm /g m25% 9nm 9nm 150 m2150 2 2.6 μm 4.5% Pentafluorophenyl L43 TMS endcapping MPa 2 - 8 condition SunShell PFP 9nm 150 m /g a)60 Under 100% aqueous

◆ Comparison of standard samples ◆ Comparison of standard samples 1 2,3 5 7 1 2,3

SunShell PFP

4 6 7

5

SunShell PFP

4 6 2,3 1

5,7

2,3

1

4 4 1

1

2

1

1

1

5 6

2

1

2

SunShell RP-AQUA SunShell RP-AQUA

5

4 5

4 2

7

(Caffeine/Phenol)

SunShell C18

3

SunShell C18

SunShell C18-WP 5

4

7 7

6

5

4

6

0

12

2

2 4

4 6

6 8

8

10

12

14

16

18

10 12Retention 14 16time/min 18 20 Retention time/min

Separation of peptides Separation of peptides

20 22

22 24

(Amylbenzene/Butylbenzene)

Hydrophobicity

1.31 (Amylbenzene/Butylbenzene)

Hydrogen bonding

Hydrophobicity

(Triphenylene/o-Terphenyl)

Steric selectivity

2.38 (Triphenylene/o-Terphenyl) Steric selectivity

1.00 1.00

1.31 1.48

1.00 0.32 1.00

1.48 1.46 1.48

0.32 0.52 0.32

1.46 1.46 1.52

1.08 1.08

1.30

RP-AQUA RP-AQUA C18-WP

0.52 0.52 0.40

1.52 1.52 1.55

1.30 1.30

1.35

C18-WP C18-WP SunShell C18 24

0.40 0.40 0.39

1.55 1.55 1.60

1.35

0.39

1.60

PhenylC8 C8

3

0

1.00 (Caffeine/Phenol)

PFP Phenyl

SunShell C18-WP

7 6

6

Hydrogen bonding

PFP

7

2 3

1

SunShell Phenyl SunShell C8

6

4

SunShell Phenyl

SunShell C8

5 7 6

54

3

2 3

1

74

2 3

7 5 6

2

4

3

6

5,7

6

3

Column: SunShell C18, C18-WP,a) RP-AQUA, C8, Phenyl, PFP, 2.6 Under 100% aqueous condition μm Column: SunShell C18, C18-WP, RP-AQUA, C8, Phenyl, PFP, 2.6 μm 150 x 4.6 mm 150 x 4.6 mm RP-AQUA, C8, Phenyl, PFP, 2.6 μm Column: Mobile SunShell C18, C18-WP, phase: CH3OH/H2O=75/25 OH/H Mobile phase: CH 3 2O=75/25 150 x 4.6 mm Flow rate: 1.0 mL/min Flow rate: 1.0 mL/min Mobile phase: CH3OH/H2O=75/25 Temperature: Temperature: Flow rate: 1.0 mL/min40 ºC40°ºC Sample: 1 =ºCUracil Sample: 1 = Uracil Temperature: 40 2 = Caffeine Sample: 12==Uracil Caffeine 3 = Phenol 23==Caffeine Phenol 4 = Butylbenzene 3 = Phenol 4 = Butylbenzene 5 = o-Terphenyl 4 = Butylbenzene 6 = Amylbenzene o-Terphenyl 55==o-Terphenyl 7 = Triphenylene Amylbenzene 66==Amylbenzene 77==Triphenylene Triphenylene Hydrogen bonding Hydrophobicity Steric selectivity

PFP Phenyl

RP-AQUA C8

Sunshell C18

SunShell C18

1.00

0.39

Separation of amitriptyline using C8 Separation of amitriptyline using C8

1.31

1.60

2.38 1.01

2.38

1.01 1.08

1.01

1.35 1.46

1.46

1.46

Separation of basic compounds

C18 -WP / RP-AQUA / C8 / PHENYL / PFP - 2.6 µm

Inner diameter (mm)

1.0

2.1

3.0

4.6

USP category

Length (mm)

Catalog no

Catalog no

Catalog no

Catalog no

Catalog no

Sunshell C8, 2.6 µm

30 50 75 100 150

-----------

CC6931 CC6941 CC6951 CC6961 CC6971

CC6331 CC6341 CC6351 CC6361 CC6371

CC6431 CC6441 CC6451 CC6461 CC6471

L7

Sunshell PFP, 2.6 µm

30 50 75 100 150

-----------

CF6931 CF6941 CF6951 CF6961 CF6971

CF6331 CF6341 CF6351 CF6361 CF6371

CF6431 CF6441 CF6451 CF6461 CF6471

L43

Sunshell C18-WP, 2.6 µm

30 50 75 100 150

-----------

CW6931 CW6941 CW6951 CW6961 CW6971

CW6331 CW6341 CW6351 CW6361 CW6371

CW6431 CW6441 CW6451 CW6461 CW6471

L1

Sunshell RP-AQUA, 2.6 µm

30 50 75 100 150

--CR6141 --CR6161 CR6171

CR6931 CR6941 CR6951 CR6961 CR6971

CR6331 CR6341 CR6351 CR6361 CR6371

CR6431 CR6441 CR6451 CR6461 CR6471

Sunshell Phenyl, 2.6 µm

30 50 75 100 150

-----------

CP6931 CP6941 CP6951 CP6961 CP6971

CP6331 CP6341 CP6351 CP6361 CP6371

CP6431 CP6441 CP6451 CP6461 CP6471

ORDERING INFO OF SUNSHELL

Equivalent to L62

L11

13

SUNSHELL

SunShell HFC18-16, HFC18-30, 2.6 μm separation of peptides and proteins HFC18 - 16 / HFC18 -For30 - 2.6 µm

Characteristics of SunShell HFC18

C18 (USP L1)

Core shell silica

Particl e size

SunShell

Specific surface area

2.6 μm

Carbon content

Maximum operating pressure

End-capping

Ligand density

Available pH range

5% 2.5 μmol/m2 Sunniest endcapping 60 MPa or 8,570 psi 1.5 - 10 16 nm 90 m /g High speed separations of proteins and peptides.

2.5% 1.2 μmol/m2 Sunniest endcapping 60 MPa or 8,570 psi 1.5 – 9 90 m2/g HFC18-16 2.6 μm 16 nm What is HFC18? Hexa-Functional C18 has six functional groups.

1.3% 1.2 μmol/m2 Sunniest endcapping 60 MPaa or 8,570 psia 1.5 - 9 40 m2/g HFC18-30 2.6 μm 30 nm The HFC18 is much more stable under acidic conditions. a: 50MPa, 7141psi for 4.6 mm i.d. column

SunShell C18-WP SunShell

Pore diameter

2

What is HFC18?

Hexa-Functional C18 has six functional groups.

This HFC18 is much more stable under acidic condition.

Sunniest Bonding Technology Hexamethydichlorotrisiloxane

+

Trimethylchlorosilane (TMS)

Si O

O

(X: Cl, OCH3, OC2H5)

Si

Si O O O Si O O

Si O

O O Si O

O

Si

Si

O O

O

Si

Si

O

Si O

O

Si O

O

Si Si O O O Si O O O

HO O O

Si O

Si Si

Si O

O

O

Si Si O

O

0,8 0,7 0,6 0,5 0,4 0,3 0,2 0 1

10

100

Pore Diameter (nm)

1000

Pore distribution of core shell particle

Si Si O

O O Si O

O O

Schematic diagram of the state of bonding on silica surface

Proteins/peptides are often analysed 100 atCore acidic shell 30pH. nm The wide pore SunShell 80 Core shell are 16 nmoptimized for superior life phases 60 time at extreme conditions.

0,9

0,1

14

Si

O

O Si

Relative retention (%)

Desorption Dv (log d) (cc/g)

Schematic diagram of reagent

O

40

95% line

Durable test condition Column : SunShell HFC18-16 2.6 μm, 50 x 2.1 mm Mobile phase: CH3CN/0.1% formic acid, pH2.6=40/60 Flow rate: 0.4 mL/min Temperature: 70 ºC

0.1% formic acid, pH2.6 70 ºC

20 0 0

200

400

600

Time (h)

800

1000

Stability under LC/MS mobile phase condition

Measurement condition Mobile phase: CH3CN/H2O=60/40 Flow rate: 0.4 mL/min Temperature: 40 ºC Sample: 1 = Uracil 2 = Butylbenzene

Technology Hexamethydichlorotrisiloxane

+

Trimethylchlorosilane (TMS)

HFC18 - 16 / HFC18 - 30 - 2.6 µm Si O

O

(X: Cl, OCH3, OC2H5)

0,9

Si

O

O

Si O O O Si O O

Si O

O O Si O

O

Si

Si

Si

Si

O O

O

Si

Si

O

Si

O

O

Si O

O

Si Si O O O Si O O O

Si

HO O O

Si O

Si

Si O

O

O

Si Si O

O

Si

Si O

O O Si O

O O

Schematic diagram of the state of bonding on silica surface

100

0,8

Core shell 30 nm

0,7

Core shell 16 nm

0,6 0,5 0,4 0,3 0,2

Relative retention (%)

Desorption Dv (log d) (cc/g)

Schematic diagram of reagent

O

0,1 0 1

10

100

Pore Diameter (nm)

80 60 40

Durable test condition Column : SunShell HFC18-16 2.6 μm, 50 x 2.1 mm Mobile phase: CH3CN/0.1% formic acid, pH2.6=40/60 Flow rate: 0.4 mL/min Temperature: 70 ºC

0.1% formic acid, pH2.6 70 ºC

20 0

1000

Pore distribution of core shell particle

95% line

0

200

400

600

Time (h)

800

1000

Stability under LC/MS mobile phase condition

Measurement condition Mobile phase: CH3CN/H2O=60/40 Flow rate: 0.4 mL/min Temperature: 40 ºC Sample: 1 = Uracil 2 = Butylbenzene

Separation of peptides Column: SunShell HFC18-16, 2.6 μm (16 nm) 150 x 4.6 mm, SunShell C18-WP, 2.6 μm (16 nm) 150 x 4.6 mm Mobile phase: A) 0.1% TFA in Acetonitrile/water(10:90) B) 0.1 % TFA in Acetonitrile Gradient program:

SunShell HFC18-16

ORDERING INFO OF SUNSHELL

Inner diameter (mm) Length (mm)

SunShell C18-WP

Sunshell HFC18-16, 2.6 µm

Sunshell HFC18-30, 2.6 µm

2 50 peaks 100 150

1.0

2.1

3.0

Flow rate: 1.0 mL/min Temperature: 25 ºC Catalog no Catalog no Catalog no Detection: UV@210 nm --- Sample: Tryptic CG6941 CG6341 C digest of cytochrome

50 100 150

4.6

USP category

Catalog no

Catalog no L1

L1

-----

CG6961 CG6971

CB6361 CB6371

CG6441 CG6461 CB6471

-------

C46941 C46961 C46971

C46341 C46361 C46371

C46441 C46461 C46471

10

15

SUNSHELL

SunShell 2-EP, 2.6 μm

For Supercritical fluid Chromatography HARDCORE SFC SEPARATIONS 2.6 μm core shell column shows only one third of back pressure to compare with 1.7 μm fully porous column although both show almost same efficiency. By such low back pressure, a difference of density of supercritical fluid between an inlet and an outlet of the column is reduced. Consequently, .

2.6 μm core shell column performs a superior separation for SFC.

2 -EP (ETHYLPYRIDINE) - 2.6 µm

Characteristics of SunShell 2-EP Core shell silica Particle Pore Specific Carbon size column diameter same shell showssurface areathe content

Bonded phase

End-

Maximum operating

Available pH

capping rangeμm core shell column perefficiency. By such low back pressurequently, 2.6 The 2.6 μm core 2.6 μm 2.5% 2-Ethylpyridine no 2 – 7.5 SunShell 2-EP 9 nm 150 m2/g 60 MPa or 8,570 psi forms a superior separation for SFC. pressure, a difference of density of suonly one third of back pressure in compercritical fluid between an inlet and an parison with the 1.7 μm fully porous outlet of the column is reduced. Consecolumn. However, both show almost

Comparison between SunShell 2-EP and 1.7 μm fully porous 2-EP Figure 1

Figure 1: Chromatogram of the separation for he 17component mix using the Sun Shell 2-EP 150 x 3.0 mm column. A methanol gradient of < 2 minutes was used Figure 1: Chromatogram of the separation on the Agilent 1260 Infinity SFC system.

SFC conditions: flow rate: 4.0mL/min; outlet pressure 160 mix using the Sun for the 17-components bar; column temperatureShell 55⁰C. Gradient program: 2-EP 150 x 3.05.0mm column. A meth7.5% in 0.20 min, then 7.5-20% in 1.3 min and held at anol gradient of < 2 minutes was used on 20% for 0.2 min.

the Agilent 1260 Infinity SFC system. SFC

Figure 2: Chromatogram of the separation for the 17conditions: flow2-EP rate:100 4.0mL/min; outlet component mix using Acquity UPC2 Viridis x 3.0 mm column. 16 ofpressure the 17 components were 160 bar; column temperature resolved. A methanol gradient < 2 minutes was 55°C.ofGradient program: 5.0-7.5% in 0.20 used on the Agilent 1260 Infinity SFC system. SFC then 7.5-20% 1.3 min and held at conditions: flow rate 3.5min, mL/min; outlet pressurein 160 bar; and column temperature Gradient 20%70⁰C. for 0.2 min. program: 5.0-12.5% in 1.0 min, 12.5% for 0.25 min, then 12.5-20% in 0.75 min.

Figure 2

16

component mix using the Sun Shell 2-EP 150 x 3.0 mm column. A methanol gradient of < 2 minutes was used on the Agilent 1260 Infinity SFC system. SFC conditions: flow rate: 4.0mL/min; outlet pressure 160 bar; column temperature 55⁰C. Gradient program: 5.07.5% in 0.20 min, then 7.5-20% in 1.3 min and held at 20% for 0.2 min.

Figure 1

2 -EP - 2.6Figure µm 2: Chromatogram of the separation for the 17-

component mix using Acquity UPC2 Viridis 2-EP 100 x 3.0 mm column. 16 of the 17 components were resolved. A methanol gradient of < 2 minutes was used on the Agilent 1260 Infinity SFC system. SFC conditions: flow rate 3.5 mL/min; outlet pressure 160 bar; and column temperature 70⁰C. Gradient program: 5.0-12.5% in 1.0 min, 12.5% for 0.25 min, then 12.5-20% in 0.75 min.

Figure 2

Courtesy of Pfizer Inc.

ORDERING INFO OF SUNSHELL

Sunshell 2-EP, 2.6 µm

Figure 2: Chromatogram of the separation for the 17-components mix using Acquity UPC2 Viridis 2-EP 100 x 3.0 mm column. 16 of the 17 components were resolved. A methanol gradient of < 2 minutes was used on the Agilent 1260 Infinity SFC system. SFC conditions: flow rate 3.5 mL/min; outlet pressure 160 bar; and column temperature 70°C. Gradient program: 5.0-12.5% in 1.0 min, 12.5% for 0.25 min, then 12.5-20% in 0.75 min.

Courtesy of Pfizer Inc.

Inner diameter (mm) 11

1.0

2.1

3.0

4.6

USP category

Length (mm)

Catalog no

Catalog no

Catalog no

Catalog no

Catalog no

30 50 75 100 150

-----------

CE6931 CE6941 CE6951 CE6961 CE6971

CE6331 CE6341 CE6351 CE6361 CE6371

CE6431 CE6441 CE6451 CE6461 CE6471

17

SunShell HILIC-Amide, 2.6 μm For Hydrophilic Interaction Chromatography Characteristics of SunShell HILIC-Amide SUNSHELL

Amide (USP L68)

Core shell silica Particle size

Pore diameter

Specific surface area

Carbon content

Bonded phase

End-capping

Maximum operating pressure

Available pH range

2.6 μm

9 nm

150 m2/g

3%

Amide

no

60 MPa or 8,570 psi

2-8

SunShell HILIC-Amide, 2.6 μm

SunShell HILIC-Amide

HILIC -AMIDE - 2.6 µm

Stationary phase of HILIC-Amide

For Hydrophilic Interaction Chromatography

Characteristics of SunShell HILIC-Amide

R: Hydrophilic group Amide (USP L68) For Hydrophilic Interaction Chromatography.

Specific Carbon Bonded Maximum operating Available pH End-capping surface area content phase pressure range Highly efficient separation of very polar compounds.

Rapid equilibration.

2.6 μm 3% Amide no 60 MPa or 8,570 psi 2-8 9 nm 150 m /g Core shell silica

Particle size

SunShell HILIC-Amide

Pore diameter

2

stationary GROUP, phase ofso SunShell Stationary phase of HILIC-Amide Stationary phase of SunShell HILIC-Amide consists of AMIDE and The HYDROPHILIC that this HILIC-Amide stationary consists of AMIDE and HYDROPHILIC GROUP, phase is more polar than an individual group. High speed separation is leaded by core shell structure that derives high efficiency and fast equilibration. so that this stationary phase is more polar than R: Hydrophilic group

an individual group. High speed separation is a result of core shell structure that derives high efficiency fast equilibration.

Separation of Nucleic acid bases: Comparison of the other coreand shell hilic columns Stationary phase of SunShell HILIC-Amide consists of AMIDE and HYDROPHILIC GROUP, so that this stationary 2 an individual phase is more polar than group.

High speed separation is leaded by core shell structure that derives SunShell HILIC-Amide high efficiency and fast equilibration.

Column: 4

SunShell HILIC-Amide, 2.6 μm 100 x 4.6 mm, Coreshell polyol, 2.7 μm 100 x 4.6 mm, Core shell μm 100 x 4.6 mm Separation of Nucleic acid bases: Comparison of the other core shellSilica, hilic2.7columns Mobile phase: 2 2 SunShell HILIC-Amide Acetonitrile/20 mM ammonium acetate(pH4.7) = 8/2 S Company Column: Flow rate: 1.0 mL/min 4 3 4 2.6 μm 100 1 3 Core shell Polyol SunShell HILIC-Amide, 5 Temperature: 40 oxC4.6 mm, 1 5 Coreshell polyol, 2.7 μm 100 x 4.6 mm, Detection: UV@250 nm Core shell Silica, 2.7 μm 100 x 4.6 mm Mobile phase:Sample: 1 = Thymine, 2 = Uracil, 3 = Uridine, 4 = Cytosine, 5 = Cytidine 3

1

5

22

4

3

1

1

3

54

S Company Core shell Polyol

A Company Core shell Silica

5

2

Acetonitrile/20 mM ammonium acetate(pH4.7) = 8/2 Flow rate: 1.0 mL/min Temperature: 40 oC Detection: UV@250 nm Sample: 1 = Thymine, 2 = Uracil, 3 = Uridine, 4 = Cytosine, 5 = Cytidine

3 A Company 1 Retention5 time/min Core shell Silica 4

ORDERING INFO OF time/min SUNSHELL Retention Separation of Cyanuric acid and

Regarding retention of cytidine, SunShell HILIC-Amide showed 30% higher retention factor than S core shell polyol.

Inner diameter 1.0 2.1 3.0 4.6 Melamine Separation of water- soluble vitamins (mm)

2 Separation of Cyanuric acid and Melamine

Sunshell HILIC-Amide, 2.6 µm

2

1

18

1

Regarding retention of cytidine, SunShell HILIC-Amide showed 30% higher retention factor than S core shell polyol.

Length (mm) 30 50 75 100 150

Catalog no

Catalog no

Catalog no

Separation of water- soluble vitamins2 -----------

1

CH6931 CH6941 2 CH6951 CH6961 CH6971

3 1

CH6331 CH6341 CH6351 CH6361 CH6371

3

USP category

Catalog no

Catalog no

CH6431 CH6441 CH6451 CH6461 CH6471

L68

SunShell RP Guard Filter ＜Cartridge Type,

＜Cartridge Type,

Bonded with C18 and End-Capped with TMS

Available as a guard column for reversed phase

Bonded with C18 and End-Capped with TMS＞

Available as a guard column for reversed phase

Tubing 0.13mmID, 60mm length

Holder

Tubing 0.13mmID, 60mm length

Holder

Before

SUNSHELL

Before After Cartridge filter bonded with C18

PROTECT YOUR COLUMNS RP GUARD  The filter isFILTER made of porous glass sized 4 mm i.d. and 4 mm thickness.

After

Cartridge filter bonded with C18

 Pore diameter is 2 µm.  Low dead volume structure  The filter is made of porous glass sized 4 mm .i.d.

4ismm thickness. and Back pressure on glass filtersized is ca.4 mm 0.1 MPa at 41.0 mL/min of flow rate.

The filter made of porous glass i.d. and mm thickness  Pore diameter is 2 µm.Tubing  Upper pressure limit is more than 60 MPa . Pore size is 2 μm Holder  Low dead volume structure 0.13mmID,  Available for structure 2.1 mm i.d to 4.6 mm i.d. column . Low dead volume  Back pressure on 50mm glasslength filter is ca. Before 0.1 MPa at. 1.0 mL/min of flow rate.

Back pressure on glass filter is ca 0.1 MPa at 1.0 mL/min of flow rate Evaluation of SunShell RP Guard Filter  Upper pressure limit is more than 60 MPa . Upper pressure limit is more than 60 MPa  Available for 2.1 mm i.d to 4.6 mm i.d. column. Available for 2.1 mm i.d to 4.6 mm i.d. columns Cartridge filter bonded with C18

Hex 14 mm

Evaluation of SunShell RP Guard Filter

SunShell C18, 2.6 μm 50 x 2.1 mm

After

SunShell

tR(3)= 2.46 min N(3) = 9,239

tR(3) = 2.57 min N(3) = 8,786 5% decrease of plate

1

tR(3)= 2.46 min Mobile phase: N(3) 9,239150 x 4.6 mm C18, 2.6= μm CH CN/H O=60/40 for 2.1 mm i.d.

3

2

CH3CN/H2O=70/30 for 4.6 mm i.d.

Flow rate: tR(3)= 3.24 min 0.3 mL/min for 2.1 mm i.d.

Mobile phase: N(3) = 39,345 1.8 mL/min for 4.6 mm i.d.

CH3CN/H2O=60/40 for 2.1 mm i.d.

Temperature: 25 ºC tR(3) = 2.57 min CH3CN/H2O=70/30 for 4.6 mm Withi.d.

Guard Filter Detection: UV@250nm N(3) = 8,786 Flow rate: 5% decrease of plate Sample: 1 = Uracil 0.3 mL/min for 2.1 mm i.d.

2 = Toluene 1.8 mL/min for 4.6 mm i.d.

3 = Acenaphthene Temperature: 25 ºC tR(3) = 3.26 min With Guard Filter 4 = Butylbenzene N(3) = 38,940 Detection: UV@250nm Sample: 1 = Uracil Little change change of of plates plate 0 1 2 3 4 5 Small 2 = Toluene Retention time/min 3 = Acenaphthene 4 = Butylbenzene Without Guard Filter

Without Guard Filter

0

Without Guard Filter

Without Guard Filter

SunShell C18, 2.6 μm 50 x 2.1 mm

With Guard Filter

SunShell C18, 2.6 μm 150

2 3 Retention time/min

4

5

0

1

2

3

Price of SunShell RP Guard Filter Retention time/min

ORDERING INFO OF SUNSHELL RP GUARD FILTER

4

Name

(available as a guard column for reversed phase because of C18 bonding)

Price of SunShell RP Guard Filter

With Guard Filter

0

1

2 3 Retention time/mi

5

quantity

Part

Holder: 1 piece,No RP Guard filter : 1 piece Tubing: 1piece, Nut: 2 pieces, Ferrule: 2 CGGAAKN pieces (pressure Max: 9000 psi , 62MPa) Part number Photo CBGAAC

SunShell RP Guard Filter Starter Kit Sunshell RP Guard Filter Starter Kit (holder, cartridge, tubing)

Name

Sunshell RP Guard Filter for exchange (5 pcs) Sunshell RP Guard Filter holder

SunShell RP Guard Filter Starter Kit

quantity

RPfilter Guard Holder: 1 piece,SunShell RP Guard : 1 Filter piece For exchange Tubing: 1piece, Nut: 2 pieces, Ferrule: 2 CBGAAK pieces (pressure Max: 9000 psi , 62MPa)

CB

CBGAAH

5 pieces

CBG

19

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20