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SunShell - The 2nd generation Core Shell material

SunShell - The 2nd generation Core Shell material for UHPLC performance by using standard HPLC systems. History of Core Shell particle. Current trends in HPLC column usage. LCGC Europe Jan 1, 2012 By: Ronald E. Majors. Two versions of typical Core Shell particles.

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SunShell - The 2nd generation Core Shell material

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  1. SunShell - The 2nd generation Core Shell material for UHPLC performance by using standard HPLC systems

  2. History of Core Shell particle

  3. Currenttrends in HPLC column usage LCGC EuropeJan 1, 2012By: Ronald E. Majors

  4. Two versions of typical Core Shell particles One reaction monolayer porous silica structure Multilayer porous silica structure

  5. Schematic diagram of Core Shell silica (SunShell) 0.5mm Monolayer structure 2.6mm 0.5mm Core Core 2.6mm 1.6mm Porous silica Particle diameter: 2.6 mm, core diameter: 1.6 mm, porous silica thickness: 0.5 mm Pore volume: 0.30 mL/g, specific surface area: 150 m2/g, pore diameter: 9 nm; Ratio of porous silica volume: 77% Porous silica

  6. Comparison of plate height plots porous Column: 50 x 4.6 mm C18 Mobile phase: Acetonitrile/water=(60/40) Temperature: 25 °C Sample: Naphthalene

  7. Comparison of back pressure for high throughput columns Column dimension: 50 x 2.1 mm Mobile phase:Acetonitrile/water=(70/30) Temperature: 25 °C

  8. Kinetic plot analysis at 40 MPa 1000 Calculated values were plotted for 1.4, 2.0, 3.0, 5.0 μm fully porous particles. Experimental values were plotted for a 2.6 μm Core Shell particle. 5 mm 3 mm 2 mm 1.4 mm 100 t0/N2 (nS) 10 This figure shows, that the separation on a Core Shell material is faster than on a fully porous particle and that the Core Shell material has a higher plate count than the fully porous material at the same analysis time (t0). Core Shell 2.6 mm t0=10000 s t0=1000 s 40 MPa 1 t0=100 s t0=10 s 316,228 3,162,278 31,623 3,162 t0=1 s 100,000 10,000 1,000,000 N The curves for particulate columns were obtained by assuming η=0.00046 Pa s, φ=700, Dm=2.22x10-9 m2/s, Knox equation, h=0.65ν1/3+2/ν+0.08ν, Dp: Totally porous 1.4, 2.0, 3.0, 5.0 μm, Core Shell 2.6 μm.

  9. Why does a 2.6 mm Core Shell particle showthe same performance as a Sub-2 mm particle?Narrow particle distribution Sunniest,2 mm Wide particle distribution (Conventional silica gel D90/D10=1.50) D10: 1.75 mm D50: 2.01 mm D90: 2.31 mm D90/D10=1.32 Company F, 2 mm SunShell,2.6 mm D10: 1.67mm D50: 2.09 mm D90: 2.65 mm D90/D10=1.59 D10: 2.46 mm D50: 2.63 mm D90: 2.82 mm D90/D10=1.15 Flow of mobile phase Narrow particle distribution (Core Shell silicaD90/D10=1.15) 1 0.5 2 (mm) 4 Packing state of Core Shell and fully porous silica Comparison of Particle Size Distribution The size distribution of a Core Shell (SunShell) particle is much narrower than that of a conventional fully porous particle, so that the space among particles in the column reduces and efficiency increases by reducing Eddy Diffusion (multi-path diffusion) as being the A term in the Van Deemter equation.

  10. Difference of diffusion in column axis direction A solute diffuses outside a particle as well as into the pores. Fully porous silica Core Shell silica Cores block the path of diffusion of a solute. B term decreases.

  11. Short diffusion path based on thin porous silica layer porous

  12. Comparison between Core Shell and fully porous particles Diffusion area or Diffusion height 2.0 mm sample sample Core Shell Totally porous 2.0 mm 1.6 mm 2.6 mm Ca. 2.0 mm HETP is related to diffusion height, not particle diameter.

  13. Comparison of performance by plate/pressure Column: 50 x 2.1 mm C18, mobile phase: Acetonitrile/water=(70/30), temperature: 25 °C

  14. SunShell C18 2.6 mm 3 5 7 2 1 4 6 2 SunniestC18 5 mm 7 3 5 1 4 6 0 5 10 15 20 25 30 Comparison of retention between fully porous silica C18 and Core Shell silica C18s Mobile phase: Methanol/water(75:25) Temperature: 40 °C Column dimension: 150 x 4.6 mm Flow rate: 1.0 mL/min Retention time/min

  15. Chromatogram of standard samples on SunShell C18 3 Column:SunShell C18, 2.6 mm100x 2.1 mm Mobile phase: CH3CN/H2O=60/40 Flow rate: 0.3 mL/min Pressure: 12.5 MPa Temperature: 25 ºC Plates(3)=21,300 1 2 4 Sample: 1 = Uracil 2 = Toluene 3 = Acenaphthene 4 = Butylbenzene 1 Plates(3)=38,000 Column:SunShell C18, 2.6 mm 150 x 4.6 mm Mobile phase: CH3CN/H2O=70/30 Flow rate: 1.0 mL/min Pressure: 11.5 MPa Temperature: 25 °C 2 4 3 Retention time/min

  16. C18 silyl-reagent (HMODTS) Hexamethyloctadecyltetrasilane Step 2 Step 1 toluene toluene + H2O + 80°C, 4h 30°C, 2h

  17. Bonding state ofHMODTS on silica Final TMS An arm of HMODTS moves like a geometrid caterpillar, so that a functional group on the tip of that arm can bond to any silanol group.

  18. Comparison of standard samples 7 5 Kinetex C18 Column: Kinetex C18, 2.6 mm 150 x 4.6 mm Ascentis Express C18, 2.7 mm 150 x 4.6 mm SunShell C18, 2.6 mm 150 x 4.6 mm Mobile phase: CH3OH/H2O=75/25 Flow rate: 1.0 mL/min Temperature: 40 °C Sample: 1 = Uracil, 2 = Caffeine, 3 = Phenol, 4 = Butylbenzene 5 = o-Terphenyl, 6 = Amylbenzene, 7 = Triphenylene 6 2 3 4 1 5 7 2 3 Ascentis Express C18 4 6 1 2 3 SunShell C18 5 7 1 4 6 0 2 4 6 8 10 12 14 16 18 20 22 24 Retention time/min

  19. Comparison of pyridine Kinetex C18 3 Column: Kinetex C18, 2.6 mm 150 x 4.6 mm Ascentis Express C18, 2.7 mm 150 x 4.6 mm SunShell C18, 2.6 mm 150 x 4.6 mm Mobile phase: CH3OH/H2O=30/70 Flow rate: 1.0 mL/min Temperature: 40 °C Detection: UV@250nm Sample: 1 = Uracil 2 = Pyridine 3 = Phenol 2 1 3 Ascentis Express C18 2 1 3 2 SunShell C18 1 0 1 2 3 4 5 6 7 8 9 10 11 12 13 Retentiontime/min

  20. Comparison of oxine, a metal chelating compound 2 1 Kinetex C18 Column: Kinetex C18, 2.6 mm 150 x 4.6 mm Ascentis Express C18, 2.7 mm 150 x 4.6 mm SunShell C18, 2.6 mm 150 x 4.6 mm Mobile phase: CH3CN/20mM H3PO4=10/90 Flow rate: 1.0 mL/min Temperature: 40 °C Detection: UV@250nm Sample: 1 = 8-Quinolinol (Oxine) 2 = Caffeine 2 Ascentis Express C18 1 2 1 SunShell C18 0 1 2 3 4 5 6 7 8 9 10 Retention time/min

  21. Comparison of formic acid 3 1 2 Kinetex C18 Column: Kinetex C18, 2.6 mm 150 x 4.6 mm Ascentis Express C18, 2.7 mm 150 x 4.6 mm SunShell C18, 2.6 mm 150 x 4.6 mm Mobile phase: CH3CN/0.1% H3PO4=2/98 Flow rate: 1.0 mL/min Temperature: 40 ºC Detection: UV@210nm Sample: 1 = Formic acid 2 = Acetic acid 3 = Propionic Acid 2 3 Ascentis Express C18 1 1 2 3 SunShell C18 0 1 2 3 4 5 6 Retention time/min

  22. Comparison of basic compounds I Column: Sunniest C18, 5 mm 150 x 4.6 mm Re: Brand E C18, 5 mm 150 x 4.6 mm Mobile phase: A)CH3OH/20mM Phosphate buffer pH7.5 = 80/20 B) CH3OH/20mM Phosphate buffer pH6.0 = 80/20 C)CH3CN/20mM Phosphate buffer pH7.0 = 60/40 Flow rate: 1.0 mL/min Temperature: 40 ºC for A, C and D, 22 ºC for B Sample: 1 = Uracil 2 = Propranolol 3 = Nortriptyline 4 = Amitriptyline TF: USP tailing factor 4 A N(4) =11,000 TF(4) =1.03 N(4)=10,000 TF(4)=1.50 4 1 3 2 BrandE C18 3 4 1 B N(4) =9,000 TF(4) =1.06 N(4)=5,800 TF(4)=2.86 4 2 BrandE C18 2 1 3 C N(4) =12,500 TF(4) =1.04 4 N(4)=4,300 TF(4)=5.24 4 BrandE C18

  23. Comparison of basic compounds II Column size: 150 x 4.6 mm for A, D and E 100 x 2.1mm for B and C Mobile phase: CH3CN/20mM Phosphate buffer pH7.0=60/40 Flow rate: 1.0 mL/min for A, D, and E 0.4 mL/min for B and C Temperature: 40 ºC Sample: 1 = Uracil 2 = Propranolol 3 = Nortriptyline 4 = Amitriptyline A Sunniest C18 (HMODTS), 5 mm 1 4 3 TF(4)=1.04 2 B Sunniest C18-HT(HMODTS), 2 mm 2 3 1 4 TF(4)=1.21 SunShell C18(HMODTS), 2.6 mm Core Shell type C 3 4 1 2 TF(4)=1.20 D Brand D C18, 2.6 mm (Kinetex) Core Shell type 3 4 1 2 TF(4)=2.24 1 Brand E C18, 5 mm E ３ 2 TF: USP tailing factor 4 TF(4)=5.24

  24. Loading capacity of amitriptyline I Mobile phase: Acetonitrile/20mMphosphate buffer pH7.0=(60:40) Column dimension: 150 x 4.6 mm, Flow rate: 1.0 mL/min, Temp.: 40°C 100 times SunShell C18 (Core Shell) Sunniest C18 3mm (fully porous) 0 1 2 3 4 5 6 7 8 9 10 Kinetex C18 (Core Shell) Retention time/min Ascentis Express C18(Core Shell) Theoretical plate was calculated by 5σ method using peak width at 4.4% of peak height. Sample: 1=Uracil, 2=Propranolol, 3=Nortriptyline, 4=Amitriptyline 4.4%

  25. Loading capacity of amitriptyline II Mobile phase: Acetonitrile/10mMammonium acetate pH6.8=(40:60) Column dimension: 150 x 4.6 mm, flow rate: 1.0 mL/min, Temp.: 40 °C SunShell C18 30times Kinetex C18 0 1 3 4 5 6 7 8 9 10 2 Retention time/min Ascentis Express C18 Sample: 1=Uracil, 2=Propranolol, 3=Nortriptyline, 4=Amitriptyline

  26. Loading capacity of amitriptyline III Mobile phase: Acetonitrile/0.1% formic acid=(30:70) Column dimension: 150 x 4.6 mm, Flow rate: 1.0 mL/min, Temp.: 40oC 7 times 4 5 min 6 In the case of using acetonitrile /0.1% formic acid as mobile phase, the amitriptyline peak shows a larger tailing because the loading capacity decreases in an acidic, low-ionic-strength mobile phase.

  27. Stability under acidic pH conditions Durability test conditions Column size: 50 x 2.1 mm Mobile phase: CH3CN/1.0% TFA, pH1=10/90 Flow rate: 0.4 mL/min Temperature: 80 °C Measurement conditions Column size: 50 x 2.1 mm Mobile phase: CH3CN/H2O=60/40 Flow rate: 0.4 mL/min Temperature: 40 °C Sample: 1 = Uracil 2 = Butylbenzene

  28. Stability under basic pH conditions Durable test conditions 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 °C Measurement conditions Column Size: 50 x 2.1 mm Mobile phase: CH3OH/H2O=70/30 Flow rate: 0.4 mL/min Temperature: 40 °C Sample: 1 = Butylbenzene

  29. Bleeding test using LC/MS Q1: 5.997 min to 7.999 min of Sample 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 °C MS: ABI API-4000Ionization: Turboionspray (cation)Measurement mode: Q1 Scan m/z 100-1000 Brand L C18 SunShell C18

  30. Characteristics of SunShell a) This value was evaluated under 100% aqueous conditions, because SunShell RP-Aqua shows a reproducible retention under 100% aqueous conditions.

  31. Comparison of standard samples 1 2 7 5 SunShell C18-WP (16nm) 4 6 3 Column dimension: 150 x 4.6 mm Mobile phase: CH3OH/H2O=75/25 Flow rate: 1.0 mL/min Temperature: 40 °C Sample: 1 = Uracil, 2 = Caffeine, 3 = Phenol, 4 = Butylbenzene, 5 = o-Terphenyl, 6 = Amylbenzene, 7 = Triphenylene 1 7 2 5 SunShell RP-Aqua 4 6 3 1 2,3 7 5 SunShell PhE (phenylethyl) 4 6 2,3 7 5 1 SunShell PFP 6 4 1 7 2 SunShell C8 5 4 6 0 2 4 6 8 10 12 14 16 18 20 22 24 3 Retention time/min 7 1 5 2 SunShell C18 4 6 3

  32. Comparison of retention timefor 8-quinolinol and caffeine SunShell C18-WP (16nm) 2 1 SunShell RP-Aqua 1 2 Column dimension: 150 x 4.6 mm Mobile phase: CH3CN/20mM H3PO4=10/90 Flow rate: 1.0 mL/min Temperature: 40 °C Detection: UV@250nm Sample: 1 = 8-Quinolinol (Oxine) 2 = Caffeine SunShell PhE (phenylethyl) 2 1 1 SunShell PFP 2 SunShell C8 1 2 SunShell C18 1 2 6 0 1 2 3 4 5 7 8 9 10 Retention time/min

  33. Retention stability under 100%aqueous conditions at 40 °C Change of retention of thymine at 40 °C (measurement at every stop flow (1 hour step)) Separation of nucleic acid bases 2 5 1 40 °C 3 4 2 1 5 25 °C 3 4 Column: SunShell RP-Aqua, 2.6 mm 75 x 4.6 mm Mobile phase: 10mM Phosphate buffer pH7.0 Flow rate: 1.0 mL/min Temperature: 40 ºC and25 ºC Sample: 1 = Cytosine, 2 = Uracil, 3 = Thymidine, 4 = Uridine, 5 = Thymine SunShell RP-Aqua showed more than 97% reproducibility for the retention of thymine using 100% aqueous buffer as mobile phase. 0 1 2 3 4 Retention time/min

  34. Separation of peptides Column: SunShell C18-WP, 2.6 mm (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: %B 0% – 100% in 35 min Flow rate: 1.0 mL/min, temperature: 25 °C, detection: UV@210 nm Sample: Tryptic digest of myoglobin

  35. Separation of xanthines 3 SunShell C18 SunShell C18, PFP 2.6 mm 150 x 2.1 mm Mobile phase: CH3OH/water or buffer=30/70 Flow rate: 0.3 mL/min Temperature: 25 °C Detection: UV@250nm Sample: 1 = Theobromine 2 = Theophyline 3 = Caffeine 4 = Phenol Water 1 2 4 3 1 2 SunShell PFP Water 4 3 1 2 20mM Phosphate buffer pH7 SunShell PFP 4 3 2 50mM Phosphate buffer pH7 1 SunShell PFP 4 Xanthines experience a stronger retention on SunShell PFP than on SunShell C18. The higher the buffer concentration the longer the retention time. 4 0 1 2 3 5 Retention time/min

  36. Separation of xylene isomers SunShell C18, PFP 2.6 mm 150 x 2.1 mm Mobile phase: CH3OH/water=75:25 for SunShell C18 CH3OH/water=60:40 for SunShell PFP Flow rate: 0.3 mL/min Temperature: 25 °C Detection: UV@250nm Sample: 1 = o-Xylene 2 = m-Xylene 3 = p-Xylene 3 SunShell PFP shows a different selectivity in comparison to SunShell C18. SunShell C18 2 1 3 SunShell PFP 2 1 0 1 2 3 4 5 Retention time/min

  37. Separation of cresol isomers 1,2 Column: SunShell C18, 2.6 mm 150 x 4.6 mm SunShell PFP, 2.6 mm 150 x 4.6 mm Mobile phase: CH3OH/H2O=40/60 Flow rate: 1.0 mL/min Temperature: 25 ºC Sample: 1 = p-Cresol 2 = m-Cresol 3 = o-Cresol SunShell C18 3 2 3 1 SunShell PFP

  38. Separation of water-soluble vitamins SunShell RP-AQUA N(4)=30,800 1 N(3)=31,800 4 3 Column: SunShell RP-AQUA, 2.6 mm 150 x 4.6 mm Mobile phase: 40mM Phosphate buffer pH6.8 Flow rate: 1.0 mL/min Temperature: 40 °C Detection: UV@250nm Sample: 1 = Nicotinic acid 2 = Pyridoxal 3 = Pyridoxine 4 = Nicotinamide 2

  39. Separation of nucleic acid bases SunShell RP-AQUA 2 Column: SunShell RP-AQUA, 2.6 mm 150 x 4.6 mm Mobile phase: Water Flow rate: 1.0 mL/min Temperature: 24 °C Sample: 1 = Cytosine 2 = Uracil 3 = Thymidine 4 = Thymine 5 = Uridine N(3)=37,900 1 3 4 5 N(5)=30,000

  40. Comparison of separation of nucleic acid bases Sunniest RP-AQUA Column: Sunniest RP-AQUA, 5 mm 150 x 4.6 mm SunShell RP-AQUA, 2.6 mm 150 x 4.6 mm Mobile phase: 10mM Phosphate buffer pH7.0 Flow rate: 1.0 mL/min for Sunniest 1.5 ml/min for SunShell Temperature: 40 ºC Sample: 1 = Cytosine 2 = Uracil 3 = Thymidine 4 = Uridine 5 = Thymine 5 2 1 3 4 SunShell RP-AQUA 2 1 3 5 4

  41. Separation of nucleotides SunShell RP-AQUA 2 N(2)=37,900 Column: SunShell RP-AQUA, 2.6 mm 150 x 4.6 mm Mobile phase: 20mM Phosphate buffer pH6.0 Flow rate: 1.0 mL/min Temperature: 25 °C Detection: UV@250nm Sample: 1 = 5’-GDP 2 = 5’-ATP 3 = 5’-ADP 4 = 5’-AMP 1 N(4)=30,000 4 3

  42. Comparison between HPLC and UHPLC UHPLC Column:SunShell C18, 2.6 mm 150 x 4.6 mm Mobile phase: CH3CN/H2O=70/30 Flow rate: 1.0 mL/min Pressure: 13.5 MPa Temperature: 25 °C 1 Plates(3)=38,000 2 4 3 Sample: 1 = Uracil 2 = Toluene 3 = Acenaphthene 4 = Butylbenzene Retention time/min Plates(3)=31,000 3 HPLC 1 4 2 80%performance compared to UHPLC Still a 1.5 times better separation as with a 3 mm column Retention time/min

  43. Separation of standard samples using HPLC HPLC Column:SunShell C18, 2.6 mm 100 x 4.6 mm 150 x 4.6 mm Mobile phase: CH3CN/H2O=70/30 Flow rate: 1.0 mL/min Pressure: 9.5 MPa, 13.5 MPa Temperature: 25 °C Sample: 1 = Uracil 2 = Toluene 3 = Acenaphthene 4 = Butylbenzene 3 Plates(3)=20,000 3 HPLC Plates(3)=31,000 The same efficiency as with 5 mm, 250 x 4.6 mm 4 1 1 2 4 2 Saving 60% analysis time and solvent consumption Retention time/min Retention time/min

  44. Transfer examples (isocratic separation) ACEC18, 5 mm 250 x 4.6 mm Column: Brand F C18, 5 mm 250 x 4.6 mm SunShell C18, 2.6 mm 100 x 4.6 mm Mobile phase: CH3CN/20mM Phosphoric acid = 45/55 Flow rate: 1.0 mL/min, 1.8 mL/min for the lowest chromatogram Temperature: 25 °C 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 HPLC: Hitachi LaChrom ELITE (using 0.25 mm i.d. tubing) UHPLC: Jasco X-LC HPLC N(4)=19,313 3 1 2 5 4 3 SunShell C18, 2.6 mm 100 x 4.6 mm N(4)=20,287 5 1 4 2 1/3 of analysis time 3 N(4)=20,287 5 2 1 4 UHPLC 3 N(4)=24,124 1 1.0 mL/min 0 2 4 6 8 10 12 14 16 18 20 22 24 2 5 4 Retention time/min 3 1.8 mL/min N(4)=22,424 1 2 5 4 Analysis time decreases to 1/6 compared to a 5mm column, 250x4.6mm. 0 1 2 3 4 5 6 Retention time/min

  45. Transfer examples (gradient separation) Column: Sunniest C18, 5 mm 150 x 4.6 mm SunShell C18, 2.6 mm 75 x 4.6 mm Mobile phase: A) 0.1% Phosphoric acid B) CH3CN Gradient program for Brand G C18 for SunShell C18 Flow rate: 1.0 mL/min, Temperature: 25 °C Detection: UV@250 nm Sample: Oolong tea 1 = Gallocatechin, 2 = Epigallocatechin, 3 = Catechin, 4 = Caffeine, 5 = Epicatechin, 6 = Epigallocatechin gallate, 7 = Gallocatechin gallate, 8 = Epicatechin gallate, 9 = Catechin gallate HPLC: Hitachi LaChrom ELITE (using 0.25 mm i.d. tubing) UHPLC: Jasco X-LC HPLC 4 Sunniest C18, 5 mm 150 x 4.6 mm 7 6 2 1 3 8 9 5 SunShell C18, 2.6 mm 75 x 4.6 mm 4 7 6 2 1/2 of Analysis time 1 3 8 9 5 4 0 2 4 6 8 10 12 14 16 18 20 22 24 Retention time/min 7 6 2 1 3 8 9 5 7 8 9 UHPLC 4 7 6 2 1 3 8 9 5 0 1 2 3 4 5 6 0 1 2 3 4 5 6 Retention time/min Retention time/min

  46. High-throughput separation 2 7 Column: SunShell C18 2.6 mm, 30 x 3.0 mm. Mobile phase: A) Water, B) Acetonitrile; Gradient (Acetonitorile %), 0.00 min - 35%, 0.40 min - 100%, 0.80 min - 100%, 0.85 min - 35%, 1cycle; 1.8min, (High-pressure gradient). Flow rate: 1.0 mL/min. Temperature: 40 °C. Injection volume: 1 µL. Wavelength: 200 - 500nm, CH-9, 215 - 500nm (max abs.). Sample: Mixture of ultraviolet absorbers 1 = 2,2’,4,4’-Tetrahydroxybenzophenone 2 = Ethyl p-aminobenzoate 3 = 2, 4-Dihydroxybenzophenone 4 = 2,2’-Dihydroxy-4-methoxybenzophenone 5 = 2,2’-Dihydroxy-4,4’-dimethoxybenzophenone 6 = 2-Hydroxy-4-methoxybenzophenone 7 = 2-(2’-Hydroxy-5’-metylphenyl) benzotriazole 8 = 4-tert-Butylphenyl salicylate Courtesy of Jasco. 4 8 3 6 5 1

  47. SunShellRP Guard Filter Holder Tubing 0.13mmID, 60mm length Cartridge type, bonded with C18and end-capped with TMS Cartridge filter bonded with C18 Before After • The filter is made of porous glass with 4 mm i.d. and 4 mm thickness. • The pore diameter is 2 mm. • Low dead volume structure. • Back pressure of the glass filter is about 0.1 MPa at 1.0 mL/min. • Upper pressure limit is more than 60 MPa. • Available for 2.1 mm i.d. up to 4.6 mm i.d. columns.

  48. SunShell RP Guard Filter Evaluation of SunShell RPGuard Filter SunShell C18, 2.6 mm 50 x 2.1 mm SunShell C18, 2.6 mm 150 x 4.6 mm Without Guard Filter Without Guard Filter tR(3)= 2.46min N(3) = 9,239 tR(3)= 3.24min N(3) = 39,345 Mobile phase: CH3CN/H2O=60/40 for 2.1 mm CH3CN/H2O=70/30 for 4.6 mm Flow rate: 0.3 mL/min for 2.1 mm 1.8 mL/min for 4.6 mm Temperature: 25 °C Detection: UV@250nm Sample: 1 = Uracil 2 = Toluene 3 = Acenaphthene 4 = Butylbenzene With Guard Filter With Guard Filter tR(3) = 2.57min N(3) = 8,786 tR(3) = 3.26min N(3) = 38,940 1% decrease of N 5% decrease of N

  49. Conclusion • Core Shell C18 2.6 mm shows the same performance as Sub-2 mm C18. • A Core Shell particle bonded with polyfunctional C18 (SunShell C18) shows a 2 times higher retention factor than Kinetex C18. • SunShell C18 has a 7 to 100 times higher loading capacity of amitriptyline than other Core Shell C18s. • Furthermore, SunShell C18 is not only stable under acidic and basic pH conditions in comparison to other Core Shell C18s, but also shows less column bleeding under typical LC/MS conditions. • Numerous SunShell selectivities are available. • The SunShell RP guard filter can be used as guard for any reversed phase column.

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