Sunniest C18-HT 2um by Biotech Fluidics

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

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

3 3

2 3

3 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

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)

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

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

10 10 12 12 1010 12 12 10101212

Antipyrine

3 3 3 43 4 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

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

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 4

3 4 4

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

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

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

0.34 μm

0.32 μm

0.37 μm

0.03 μm

0.20 μm

0.01 μm

1.88 μm

1.62 μm

1.91 μm

0.19 μm

0.90 μm

0.10 μm 2 μm

2 μm

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)

100

Inner diameter (mm)

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