Sunniest C18-HT 2um

biotech AB presents Sunniest C18-HT 2Îźm HPLC column

Biotech in Partnership with

Biotech Sunniest C18-Ht 2 μm Reatures • Low back pressure and high efficiency by precisely classified particle • High pressure packing (10,000 psi) using hard silica gels with high pressure resistant • leads long column life without any void.

• Unique bonding technique for Sunniest (patent pending) • The most suitable inner surface of column by special grinding

Sunniest technique to 2 μm particle!

Narrow Particle Distribution and Low Back Pressure Measured by Coulter Counter method Classified particle Classified particle D10: 1.75 mm Classified particle D50: 2.01 D10:mm 1.75 mm D90: 2.31 D10: 1.75 D50:mm 2.01 mm D90/D10: D90:1.32 2.31 mm D50: 2.01

D90: 2.65 mm D90/D10: 1.59 Conventional Conventional particle particle Classified Classified particle D90: 2.31 particle mm D90/D10: 1.32 D90/D10: 1.59 D90/D10: 1.32 D10: 1.67 D10: mm 1.67 particle mm Conventional Conventional particle Classified Classified particle particle D10: 1.75 D10:Narrow mm 1.75 mm particle D50: 2.09 D50:mm 2.09 mm D50: 2.01 D50:distribution mm 2.01 mm D10: 1.67 D10: mm 1.67 mm D90: 2.65 D90:mm 2.65 mm Narrow particle D10: 1.75 D10: mm 1.75 D90: 2.31 D90: mm 2.31mm mm ( )*' D50: 2.01 D50: 2.09 D50: mm 2.09 Narrow particle D90/D10: D90/D10: 1.59 mm 1.59 distribution D50: mm 2.01 D90/D10: D90/D10: 1.32 mm 1.32 D90: 2.65 D90: mm 2.65 mm distribution D90: mm 2.31 mm (D90: )*' 2.31 D90/D10: D90/D10: 1.59 1.59 1.32 1.32 (D90/D10: )*' D90/D10:

Conventional 2 μm silica gel particle was classified again.

Narrow Narrow particle particle 20% volume was cut off from both sides respectively. distribution distribution Consequently column back pressure reduced more than NarrowNarrow particleparticle ( )*' ( )*' 15%. Our 2 μm silica gel particle showsdistribution a halfdistribution pressure to compare with the other ( )*'sub-2 ( )*'μm silica gel particle.

（MPa （MPa ） ） Pressure （MPaPressure ） Pressure Pressure （MPa）

Conventional particle D10: 1.67 mm Conventional particle D50: 2.09 mm D10: 1.67 mm D90: 2.65 mm D10: D50: 1.67 2.09 mm D90/D10: D90: 2.65 D50:1.59 2.09 mm

Pressure （MPa） Pressure （MPa） Pressure （MPa）

Conventional particle

Column pressure using methanol or acetonitrile and water

Methanol Acetonitrile Methanol Methanol Acetonitrile Acetonitrile Concentration of organic solvent (%)

Concentration of organic solvent (%) Methanol Methanol Concentration of organic solvent (%) Acetonitrile Acetonitrile Column: Sunniest C18-HT, 2 μm 50 x 2.1 mm Methanol Methanol Mobile phase: Acetonitrile CH3OH/H2O, CH3CN/H2O Acetonitrile Flow rate: mL/min Concentration Concentration of organic of organic solvent solvent (%) 0.5 (%) Temperature: 40 ºC Concentration Concentration of organic of organic solventsolvent (%) (%)

• An Unique Modification (Patent Pending as Sunniest series) A Methanol

5,061

N(4) =7,670 6,108 5,061 1 A Methanol N(4) =8,068 6,008 TF(4) =1.50 N(4) =7,670 6,108 5,061 TF(4) =1.13 31 A2 Methanol 5,785 N(4) =8,068 4 6,008 N(4) 6,108 TF(4)=7,670 =1.50 4 6,209 1 2 3 N(4) TF(4)=8,068 =1.13 5,785 4 =1.50 6,008 TF(4) 8,068 4 TF(4) =1.13 2 3 6,209 5,785 4 Other Comparison of plate number 4 6,209 8,068 5,061 5,061 A Methanol A Methanol C18 B Acetonitrile 8,068 N(4) =7,670 N(4) =7,670 6,108 Other 1.586,108 Comparison of plate number 1 1 1 N(4) =8,068 N(4) =8,068 C18=1.50 1.72 5,061 5,0616,008 6,008 2 TF(4) =1.50 TF(4) Other B Acetonitrile Comparison of plate number A Methanol A Methanol 1.58 N(4) TF(4) =1.13 TF(4) =1.13 1.49 3 =7,529 3 2 1 3 2 N(4) N(4)=3,068 N(4)4C18 =7,670 6,108 5,785 6,108 5,785 4=7,670 1.581.72 1 B Acetonitrile 12 N(4) =8,068 N(4) 1.63 TF(4) 4 =8,068 4TF(4)=4.26 6,209 6,209 N(4) =3,068 6,008 =1.50 TF(4) =1.50 1 3 TF(4) =1.19 1.496,008 4 1.72 N(4) =7,529 1.57 2 TF(4) 3 =1.13 4 =1.13 4 TF(4) 2 3 2 TF(4) 8,068 5,785 1.49 5,785 N(4)4 =3,068 1.63 8,068 =4.26 3 1.13 TF(4)=7,529 =1.19 4N(4) 4 4 6,209 number 6,209 1.57 Other=4.26 Other 4 1.63 Comparison Comparison of plate of number plate TF(4) Other TF(4) =1.19 2.0 1.0 1.13 8,068 8,068 1.57 C184 C18 4 B Acetonitrile B Acetonitrile C18 1.58 1.58 Comparison of tailing factor Other Comparison 2.0 Other Other 1 1 Comparison of1.0 plate1.13 ofnumber plate number 1.72 1.72 2 2 C18 C18 Other Comparison of tailing factor 2.0 N(4) =3,068 N(4)C18 =3,068 B Acetonitrile BRetention 1.0 1.49 1.49 3 Acetonitrile 3 time (min) 1.58 1.58 TF: USP tailing factor N(4) =7,529 N(4) =7,529 1 1 Comparison of tailing 1.63factor 1.63 TF(4) =4.26 TF(4)C18 =4.26 1.72 1.72 2 2 Retention TF(4) =1.19 TF(4) (min) =1.19 time 4N(4) =3,068 4 TF: USP tailing factor N(4)4=3,068 1.49 1.571.49 1.57 3 3 N(4) =7,529 4 N(4) =7,529 Retention time (min) 1.13 1.13 1.63 TF: USP tailing factor 1.63 TF(4) =4.26 TF(4) =4.26 TF(4) =1.19 TF(4) =1.19 4 4 Other 1.57 1.57 Other 2.0 2.0 4 4 1.0 1.0 1.13of tailing 1.13 C18 C18 Comparison Comparison of tailing factorfactor It is difficult to end-capOther on sub 2 μm or 21.0μm silica gel particle as2.0 Other 2.0 1.0 C18 time (min) TF: USPMost TF: tailing USP sub tailing factor Comparison Comparison of tailing of tailing factor factor wellRetention as 3Retention μm or(min) 5time μm silica gelC18 particle.

2 factor μm or 2 μm C18

columns show smaller and higher tailing factor than Retention Retention time (min) time (min) plate number TF: USP TF: tailing USPfactor tailing factor Sunniest C18-HT. Sunniest C18-HT 2 μm shows good peak shape for amitriptyline under not only metahnol/phosphate buffer mobile phase but also acetonitrile/phosphate buffer mobile.

Column: Sunniest C18-HT, 2 mm 50 x 2.0 mm Mobile phase: A) CH3OH/20mM Phosphate buffer pH7.5 = 80/20 B) CH3CN/20mM Phosphate buffer pH7.0 = 60/40 Flow rate: 0.4 mL/min Pressure: A) 19.5 MPa, B) 13.5 MPa Temperature: 40 ºC Sample: 1 = Uracil, 2 = Propranolol,

3 = Nortriptyline, 4 = Amitriptyline,

Uracil

Nortriptyline

Propranolol

Amitriptyline

Biotech Sunniest C18-Ht 2 μm 3 3

Sunniest C18 5 5m,C18 150 x 4.6 mm Sunniest C18 m, 150 xm,4.6 mm 4.6 mm Sunniest 5 Sunniest C18 5 m, 150 x150 4.6xmm 1 1 1 1 plate/m N(4)=80,000 plate/mplate/m 2 2 N(4)=80,000 N(4)=80,000 plate/m C18 5 m, x 4.6 mm 2C18 25 N(4)=80,000 SunniestSunniest m, 150 4.6150 mm 3 3 x4 3 4 3 1 1 4 4

1 12 2 1 2 1 30 30 x 2.1 mmmm x 2.1

N(3)= 5,375 N(3)= 5,375 N(3)= 5,375 3 N(3)= 5,375 1

3

2

2 1

N(4)=80,000 2 N(4)=80,000 plate/m plate/m 3 3 4 4

50 x 2.1 mm N(3)= 9,294 N(3)= 9,294 N(3)= 9,294 N(3)= 9,294 2

High throughput High throughput N(4)=175,000 plate/m N(4)=175,000 plate/m N(4)=175,000 plate/m N(4)=175,000 plate/m N(4)=175,000 N(4)=175,000 plate/m plate/m Sunniest C18-HT 2 C18-HT mm Sunniest C18-HT 2m, 50 m, x50 x m, 2.1 mm Sunniest 22.1 50 xmm 2.1 mm Sunniest C18-HT 2 m, 50 xm, 2.1 Sunniest C18-HT 2 50 Sunniest C18-HT 2 m, 50 x 2.1 mmx 2.1 mm

21 2 2

2

4

3 3

2 3

3

3

3 4 4

4

4

4

4

4 01 1 1 1 2 2 23 3 3 3 2 1 4 3 2 Retention time (min) 4 time (min) 0 3 32 Retention time 1 3 3 Retention Retention 3 4 Retention time (min) time (mi 21 3 2 Retention time (min) 4 2 1 4 75 75 x 2.1 mm x 2.1 mm 4 4 75 x mm 2.1 mm 4 x x2.1 mm 75 2.1 75 x 2.175 mm 0 0 0

0

1 12 2 1 231

N(3)= 14,021

14,021 N(3)= 14,021 N(3)= 14,021 N(3)=N(3)= 14,021 N(3)= 14,021

Retention time (min) Retention time (min) Retention time (min) Retention time (min) Retention time Retention time (min)(min)

3

CH3CN/0.1% Formic acid = 20/80 1.0 mL/min for 150 x 4.6 mm 0.6 mL/min for 50 x 2.1 mm 40 ºC UV@230 nm 1 = Acetaminophen, 2 = Antipyrine, 3 = Aspirin, 4 = Ethenzamide Aspirin

3 4

4

4

N(3)= 9,303 N(3)= 9,303

10 10 12 12 1010 12 12 10101212

Antipyrine

3 3 3 43 4 2

2

2 3

1 50 50 x 3.0 mm x 3.0 mm 50 x mm 3.0 mm 50 x 3.0 1 2 N(3)= 9,303 N(3)= 9,303 1 N(3)= N(3)= 9,303 x 3.0 mm 9,303 50 x 3.050 mm

High throughput High throughput throughput HighHigh throughput

Acetaminophen

4

4

x 2.1 mm 30 x 2.130 mm 2 2 1 1 50 50 x 5,375 2.1 mm N(3)= 5,375 x 2.1 mm N(3)= 50 x 2.1 mm1 1 1 x 2.1 1mm 50 x 2.150 mm N(3)= 9,294 1 1 N(3)= 9,294

Mobile phase: Flow rate:

Temperature: Detection: Sample:

2 4 4

4 4 no.

Comparison of plate 2.1 mm 30 x30 2.1x mm

Separation of Analgesics

2

3

3

33

1 mm 12 11 2 2 2 1 2.1 100 2.1 mm 100x100 x100 2.1 mm x x2.1 mm 2 42 1 100 x 2.1 mm N(3)= 100 x19,802 2.1 mm N(3)= 19,802

N(3)= 19,802 N(3)= 19,802 N(3)= 19,802 N(3)= 19,802

3

3 4

3 4 4

4

4

Retention time (min) Retention time (min)

Retention time (min) Retention time (min)

Retention Mobile phase: CH3CN/H2O = 60/40 Retention timetime (mi Flow rate: 0.6 mL/min for 2.1 x 30 mm and 2.1 x 50 mm

1.0 mL/min for 3.0 x 50 mm

0.4 mL/min for 2.1 x 75 mm and 2.1 x 100 mm Temperature: 40 ºC Detection: UV@250 nm Sample: 1=Uracil, 2=Toluene,

3=Acenaphthene, 4=Butylbenzene

Ethenzamide

2 μm particle allows to reduce retention time because high efficiency is kept under high flow rate conditions. As shown the above chromatograms, analytical time reduced 1/8 without sacrifices of separation by using 2 μm, 50 x 2.1 μm column instead of 5 μm 150 x 4.6 μm column.

Characteristics of Sunniest C18-Ht 2 μm Silica gel support

C18

Packings

Particle size

Pore diameter

Specific surface area

Sunniest C18-HT

2.0 μm (Coulter counter)

10 nm

340 m2/g

It is very important for 2 μm particle to have a capacity to resist pressure because of high column back pressure. The larger a pore volume of silica gel, the weaker a capacity to resist pressure. The silica gel with 0.85 ml/g of pore volume is used for Sunniest C18-HT, 2 μm, so that it have a high capacity to resist pressure and a high operating pressure.

Carbon content

Bonded phase

16 %

C18

Maximum operating pressure 70 MPa or 10,000 psi

Available pH range 1.5 - 10

Narrow Particle Distribution and Low Back Pressure Parameter of surface roughness Ra: Average roughness from center line Rz: Roughness calculated from 10 points average (5 points of maximum and 5 points of minimum )

Y company

R

Schematic diagram of surface roughness

It is considered that surface roughness affects column performance. Surface asperity of ChromaNik Technologies column is 1/30 to 1/20 to compare with that of GL Sciences, YMC, Shimadzu and Waters

G company

Center line

Ra

columns. ChromaNIk Technologies provides a column with very smooth surface which is the most suitable for 2 μm particle packing.

S1 company

S2 company

W company

ChromaNik Technologies

Ra

0.34 μm

0.32 μm

0.37 μm

0.03 μm

0.20 μm

0.01 μm

Rz

1.88 μm

1.62 μm

1.91 μm

0.19 μm

0.90 μm

0.10 μm 2 μm

2 μm

particle

particle

Inner surface of column of A, B, C companies

Inner surface of ChromaNik Technologies

Ordering Information of Sunniest C18-Ht 2 μm Length (mm)

30

50

75

100

Inner diameter (mm)

Cat. No.

Cat. No.

Cat. No.

Cat. No.

2.1

EB1931

EB1941

EB1951

EB1961

3.0

EB1331

EB1341

EB1351

EB1361

biotech AB P.O. Box 133 SE-439 23 onsala sweden tel +46 (0)300-56 91 80 fax +46 (0)300-56 91 81 www.biotech.se