HPLC column. Sunniest. C18, C18-HT, RP-AQUA, C8, PhE, PFP. ChromaNik Technologies Inc.

Similar documents
Sunniest C18 Sunniest RP-AQUA Sunniest C8 Patent pending

HPLC column C18 C8 PFP. SunShell. Core Shell Particle

RP C18 column with feature of a silanol group

C18, C18-WP, HFC18-16, HFC18-30,RP-AQUA, C8, PFP,

Inertsil ODS-EP Technical Information

An HPLC column offering moderate retentivity with superb peak shape Inertsil ODS Sprint

Silanol. Groups Liquid Chromatography

InertSustain AQ-C18. HPLC, LC/MS Columns. Maximizing retention for highly polar compounds in reversed phase methods with highly aqueous mobile phases

InertSustainSwift C8

InertSustainSwift C8

Fast Separation of Vastly Different Compounds by Isocratic HPLC

Method Development Kits

State-of-the-art C18 HPLC Columns

Packed columns for reversed-phase chromatography TSKgel ODS-100V and 100Z

ProntoSIL C18-EPS Reversed-Phase HPLC Columns

Inertsil Technical Library

C18, C18-WP, HFC18-16, HFC18-30,RP-AQUA, C8, PFP,

C18, C18-WP, HFC18-16, HFC18-30,RP-AQUA, C8, C30, PFP, Phenyl, C8-30, C8-30HT, C4-30, HILIC-Amide and 2-EP. Core Shell Particle

Alltech Alltima HP Introduction

A C18 Silica Column With Exceptional Temperature and ph Stability

penta-hilic UHPLC COLUMNS

Packed Column for Ultra-Fast Reversed-Phase Liquid Chromatography, TSKgel Super-ODS. Table of Contents

Reversed Phase Solvents

Shodex TM ODP2 HP series columns

Overview Applications of NUCLEODUR HPLC Phases

Packings for HPLC. Packings for HPLC

ISOSCELES PELLICULAR CORE

penta-hilic UHPLC COLUMNS

INERTSIL (GL Sciences)

Technical Guide Thermo Scientific Syncronis HPLC Columns

Comparison of different aqueous mobile phase HPLC techniques

Welcome to our E-Seminar: Choosing HPLC Columns for Faster Analysis Smaller and Faster

LC and LC/MS Column Selection Flow Chart

AN HPLC COLUMN InertSustain C1 AN HPLC COLUMN InertSustain C1 Inertsil continues to evolve to InertSustain Physical Properties

Setting the Standard for ph Method Development

Thermo Scientific Accucore XL HPLC Columns. Technical Manual

Orosil HPLC Columns. OroSil HPLC columns are designed for the separation of polar, semi-polar, and nonpolar compounds at low to medium ph.

Too Polar for Reversed Phase What Do You Do?

ColumnsAcclaim PolarAdvantage II (PA2) HPLC Columns

William E. Barber, Ph.D. Applications Chemist October

Acclaim Mixed-Mode WAX-1 Columns

STUDY OF LOADABILITY AND SELECTIVITY OF PHARMACEUTICAL COMPOUNDS ON RPLC COLUMNS

Analysis - HPLC A.136. Primesep 5 µm columns B.136

AQUASIL C18 Columns. AQUASIL 18 Columns TG Compatible with 100% aqueous mobile phase

The Secrets of Rapid HPLC Method Development. Choosing Columns for Rapid Method Development and Short Analysis Times

It s hot! ZIC -chilic

State - of - the -art C18 HPLC Columns

TCI Dual ~Mixed-Mode Mode column~

Advantages of polymerbased. an alternative for ODS

Put the. C18 aside. and take a look... Reasons. to try the. NEW Gemini C6-Phenyl

ACE Excel SuperC18. ACE UHPLC and HPLC Columns. Ultra-Inert UHPLC and HPLC Columns with Extended ph Stability

The Evolving HPLC Column Packings

Gemini. Second-Generation TWIN-NX Technology. TWIN (Two-In-One) Technology. U.S. Patent No. 7, 563, 367 Setting the Standard for ph Method Development

Ion Exchange Columns ---- Inertsil AX 078 Inertsil CX 080. HPLC Column Catalog

The Retention Behavior of Reversed-Phase HPLC Columns with 100% Aqueous Mobile Phase

ACE SuperC18. ACE UHPLC and HPLC Columns. Ultra-Inert UHPLC and HPLC Columns with Extended ph Stability

The Selection of an HPLC Column October

Tracer Excel TRACER EXCEL ODS-A. Total deactivation. Maximum Stability

HPLC Column Classification

COMPARISON GUIDE. to C18 Reversed Phase HPLC Columns. Comparison Data on 60 Commonly Used C18 Phases. Stationary Phase Specifications

Setting the Standard for ph Method Development

ACE SuperC18. ACE UHPLC and HPLC Columns. Ultra-Inert UHPLC and HPLC Columns with Extended ph Stability

CLIPEUS - A Small Shield for a Big Job

Choosing Columns and Conditions for the Best Peak Shape

H P L C C O L U M N S

P R O D U C T B U L L E T I N. Fused-Core particle technology for hyper-fast and super-rugged HPLC columns

HICHROM. Chromatography Columns and Supplies. LC COLUMNS SeQuant ZIC-HILIC. Catalogue 9. Hichrom Limited

Zirconia-Based Phases as a Powerful Complement to Silica-Based Phases for LC and LC-MS under Non-extreme Mobile Phase Conditions

Kromasil. for your analytical HPLC. Kromasil. The way to peak performance in liquid chromatography

columns Acclaim Mixed-Mode WCX-1 for Separating Basic Molecules

Capital HPLC Columns and HPLC Cartridges. Optimal HPLC Range

TCI Chiral HPLC Column

Product Bulletin. ACE LC/MS and Rapid Analysis HPLC Columns. HPLC Columns

Analyze Nucleotides, Nucleosides, Purine, and Pyrimidine Bases Simultaneously with the Ultra IBD Column

ODS-UG,HG,MG,SR columns

Method Development in Solid Phase Extraction using Non-Polar ISOLUTE SPE Columns for the Extraction of Aqueous Samples

NUCLEOSHELL core-shell silica for HPLC

LC Columns with Liquid Separation Cell Technology

Comparison Guide to C18 Reversed Phase HPLC Columns

Hypersil BDS Columns TG 01-05

Part 2. Overview of HPLC Media

Multi-mode Separations Using Zirconiabased Stationary Phases

Passion. Power. Productivity.

Acclaim Mixed-Mode WCX-1

Optimisil HPLC & UPLC Column catalogue

Barry E. Boyes, Ph.D. Consumables and Accessories Business Unit May 10, 2000

The Standard for ph Method Development

Ahead of the Curve for Large, Multiclass Lists by Mass Spec

Reversed Phase Chromatography: Mobile phase Considerations

( )( ) Selectivity Choices in Reversed-Phase Fast LC. Introduction. R s = 1 a 1 k 4 a 1 + k

HPLC UHPLC Columns. Dikma Technologies Inc.

for Acclaim Mixed-Mode WCX-1

High Performance Liquid Chromatography

Trends in Method Development for HPLC and UHPLC... the movement toward using solid-core particles

Gemini-NX Performance paired with Axia Technology

Novel LC/MS-Compatible Stationary Phase with Polar Selectivity

Optical Isomer Separation Columns and Packing Materials

ACE Ultra Inert Base Deactivated HPLC Columns

QUALITY HPLC COLUMNS PARTICLE PORE PAGE SIZE SIZE. C18 Bidentate C18 Extremely long column life and rugged 4um 100Å 3

Quantitative Analysis of Water-Soluble B-Vitamins in Cereal Using Rapid Resolution LC/MS/MS. Application. Authors. Abstract.

Transcription:

HPLC column Sunniest C8, C8-HT, RP-AQUA, C8, PhE, PFP Chromaik Technologies Inc.

A ovel Bonding Technique (patent pending) The State of Art trifunctional silyl-reagent was developed as shown in Fig.. This Unique silyl-bonded reagent (HMDTS)can bond with any silanol groups on lica Sorbent surface as shown in Fig..It can expand and contract by itself in Caterpillar manner. This technique can substantially minimise the contribution of residual silanol groups on Reverses phase stationary phase. Finally an end-capping was done with trimethylsilyl-reagent (TMS). X H C CH n X X n:,, or X: Cl, CH, CHCH etc. H H H TMS 化 Fig. Schematic diagram of bonding of novel silylreagent on silica surface Features Fig. ovel C8 reagent Little residual silanol groups by an unique bonding technique Excellent stability, especially under acidic ph conditions Sharp peak shape for acidic, basic and chelating compounds RP-AQUA with C8 bonding offers Performance in 00% aqueous conditions, and shows enhanced retention of polar compounds. Characteristics of Sunniest Particle size (mm) Pore diameter (nm) Specific surface area (m /g) Carbon content (%) Bonded phase ph range and 0 C8. - 0 -HT 0 0 C8. - 0 and 0 C8-8 and 0 0 C8. - 9 Sunniest PhE and 0 0 Phenylethyl. - 8 Sunniest PFP 0 0 Pentafluorophenyl - 8

Evaluation of End-capping Comparison of plates number () and USP tailing factor (TF) of amitriptyline メタノール CH H, ph7., 0 ºC () =,000 TF() =.0 () =0,00 TF() =.07 Column size:. x 0 mm Particle size: µm Mobile phase: CH H/0mM Phosphate buffer ph7. or.0 =80/0 CH C/0mM Phosphate buffer ph7.0 =0/0 Flow rate:.0 ml/min Temperature: 0 ºC or ºC Sample: = Uracil = Propranolol CH H CH H A 社 C8 B 社 C8 () =8,90 TF() =.7 () =0,000 TF() =.0 = ortriptyline = Amitriptyline アセトニトリル CH C, ph7.0, 0 ºC H CH CH CH 0 8 0 () =,00 TF() =.0 メタノール CH H, ph.0, ºC () =9,000 TF() =.09 () =,000 TF() =.09 () =,00 TF() =.0 () =8,000 TF() =.0 () =0,000 TF() =. Sunniest PhE A 社 C8 () =,900 TF() =.9 A 社 C8 () =,000 TF() =. B 社 C8 () =,800 TF() =.8 B 社 C8 () =,00 TF() =. 0 8 0 0 8 0 8 0 Amitriptyline is widely used to evaluate residual silanol groups on the C8 stationary phase. Peak shape of Amitriptyline was compared under kinds of conditions such as methanol/phosphate buffer/0 ºC, methanol/phosphate buffer/ ºC and acetonitrile/phosphate buffer/0 ºC. Majority of the HPLC columns offered good peak shapes under methanol/phosphate buffer/0 ºC conditions. However using Mobile phase of acetonitrile/phosphate buffer/0 ºC, most of the columns(refer column A and B) offered high extent of Tailing unlike Sunniest columns offering a symmetrical peak., RP-AQUA and C8 columns allow to use a wide range of mobile phase without peak tailing because of extremely low content of residual silanol groups on the stationary phase.

SunniestC8 Stability under acidic and basic ph conditions Relative retention of ethylbenzene (%) 00 90 80 70 0 0 0 0 0 Sunniest PhE B company C8 D compnay C8 0 0 0 0 80 00 0 Relative plate number of butylbenzene (%) 0 00 80 0 0 0 0, ph0 0ºC, ph9. 0ºC 0 00 00 00 800 Time (h)) Time (h) Durable test condition Column size: 0 x. mm Mobile phase: CH C/.0% TFA (ph) = 0/90 Flow rate:.0 ml/min Temperature: 80 ºC Measurement condition Column size: 0 x. mm Mobile phase: CH C/H =0/0 Flow rate:.0 ml/min Temperature: 0 ºC Sample: = Uracil = Ethylbenzene Durable test condition Column:, C8, mm 0 x. mm Mobile phase: C8: CH H/0mM Sodium borate/0mm ah=0//9 (ph0) C8: CH H/0mM Sodium borate (ph9.) =0/70 Flow rate:.0 ml/min Temperature: C8-0 ºC, C8-0 ºC Measurement condition Column:, C8, mm 0 x. mm Mobile phase: CH H/H =7/ Flow rate:.0 ml/min Temperature: 0 ºC Sample: = Butylbenzene Stability under acidic ph conditions was evaluated at 80 ºC using acetonitrile/% trifluoroacetic acid solution (0:90) as mobile phase. 00% aqueous mobile phase expels from the pore of packing materials by capillarity and packing materials doesn t deteriorate. 0% acetonitrile in a mobile phase allows an accurate evaluation. -) has kept 90% retention for 00 hours under severe conditions of acetonitrile /% trifluoroacetic acid solution (ph )at 80 deg C. ur Unique HMDTS bonding technique offers significant enhancement of column life, Considering the C8 ligand length the is less stable than. However, Sunniest RP- AQUA C8 column with HMDTS bonding along with end capping offers longer column life in comparison to other RP Aqua columns. ). agae, T. Enami and S. Doshi, LC/GC orth America ctober 00. ) T. Enami and. agae, American Laboratory ctober 00. ) T. Enami and. agae, BUSEKI KAGAKU, (00) 09. Stability under basic ph conditions was evaluated at 0 ºC using methanol/sodium borate buffer ph 0 (0:70) as mobile phase. Sodium borate is used as a alkaline standard solution for ph meter, so that its buffer capacity is high. Elevated temperature of 0 ºC makes column life be one third. When column is used at 0 ºC, column life becomes,000 hours. Most of the HPLC columns stability data is offered at ambient room temperature alternate ºC at ph -0 units..at temperature of ºC,the column life is sixteen times longer than that at 0 ºC. offers performance at elevated ph and temperature. Regarding stability under basic ph condition, there are very few C8 column like & Hybrid type C8 which can sustain and offer performance under such challenging conditions of high temperature and ph.it is considered that our double end-capping & base deactivation technique leads higher stability. has operational ph Range from. to 0.,Phenyl has operational ph Range. to 9 and Sunniest RP-Aqua and Pentaflorophenyl (PFP) at ph -8..

SunniestC8 Relationship between ph and retention of Acidic, Basic and eutral compounds Basic compound (B) B A ph Retention factor (k) eutral compound () A A B B ph7 ph0 Acidic compound (A) ph ph selectivity, ph. ph7 ph0 Column:, mm 0 x. mm Mobile phase: A) 0mM Phosphoric acid ph. A) 0mM Phosphate buffer ph7 A) 0mM Phosphate buffer ph0 B) Acetonitrile Time (min) 0 0 %B (%) Flow rate:.0 ml/min Temperature: 0 ºC Detection: UV@0 nm Sample: = Thiamine HCl Vitamin B = icotinamide = icotinic acid = Pyridoxine HCl Vitamin B = Folic acid = Riboflavin Vitamin B 0 0 0 0 It is interesting to note that the change in ph of mobile phase can offer different selectivity of ionic compounds. can be used at the ph range from. to 0, so that a suitable analytical method can be developed using Column..

Comparison data: Bleeding from column Comparison using Corona CAD Comparison using MS.E+08 F 社 C8 Area: 90,000.E+08.E+08.E+08 B 社 C8 B 社 C8 Area:,0,000 Intensity (cps).e+08.e+08.e+08.e+08 Area: 9,000.E+08.E+07 Column size: 0 x.0 mm Mobile phase: A) 0.% acetic acid B) CH C Gradient: Time: 0min min.min 8min 9min %B: % % 00% 00% % Flow rate: 0. ml/min Temperature: 0 ºC Detection: Corona CAD 0.E+00 0 0 0 Time (min) Column size: 0 x.0 mm Mobile phase: A) 0.% acetic acid B) CH C Gradient: Time: 0min min.min 8min 9min %B: % % 00% 00% % Flow rate: 0. ml/min Temperature: 0 ºC MS: ABI API-000 Ionization: Turboionspray (cation) Measurement mode: Q Scan m/z 00-000 Separation of antidepressants using Acetonitrile and Ammonium acetate for LC/MS Company E C8 Company F C8 Company G C8 Company H C8 =,00 TF=. =,00 TF=.0 =,000 TF=.8 =8,00 TF=. =,00 TF=. =,00 TF=. =8,00 TF=.0 Column size: 0 x. mm Particle size: mm Mobile phase: CH C/0mM Ammonium acetate ph.8=0/0 Flow rate:.0 ml/min Temperature: 0 ºC Sample: Amitriptyline CH CH 0 8 0

/C8 Stability of / C8 under 00% aqueous conditions Relative retetion of ethylbenzene (%) 00 90 80 70 0 0 0 0 0 Acidic ph test /C8 0 0 0 0 80 00 0 0 Time (h) Relative plate number of thtmine (%) 00 90 80 70 0 0 0 0 0 Basic ph test 0 00 00 00 00 Time (h) Durable test conditions Column:, mm 0 x. mm Mobile phase: 0.% TFA Flow rate:.0 ml/min Temperature: 0 ºC Durable test conditions Column:, mm 0 x. mm Mobile phase: 0mM Phosphate buffer ph8.0 Flow rate:.0 ml/min Temperature: 0 ºC Measurement conditions Column:, mm 0 x. mm Mobile phase: CH H/H =7/ Flow rate:.0 ml/min Temperature: 0 ºC Sample: = Uracil = Amylbenzene Measurement conditions Column:, mm 0 x. mm Mobile phase: 0mM Phosphate buffer ph7.0 Flow rate:.0 ml/min Temperature: 0 ºC Sample: = Thymine It is important that stability is evaluated for RP Aqua columns under 00% aqueous conditions because RP-Aqua column life becomes longer with incremental contents of organic solvent in a mobile phase. /C8 column can be used under 00% aqueous conditions from ph to ph 8. / C8 column can be used under 00% aqueous conditions from ph to ph 8. /C8 is one of the most stable aqua type column. /C8 column with HMDTS bonding along with end capping offers longer column life in comparison to other RP Aqua columns

/C8 Reproducibility of retention under 00% aqueous conditions C8 and C8 Rrversed phases exhibit decreased and poorly reproducible retention under more than 98% aqueous conditions as shown in Fig.. This problem traditionally has been explained as being the result of ligand collapse or a matting effect. agae - ascertained, however, that the mobile phase was being expelled from the pores of the packing material under 00% aqueous mobile phase conditions, as Fig. shows. When the surface of packing materials isn t wet by water, water used as a mobile phase expels from the pore of the packing material by capillarity. This is a reason why reproducibility in retention is low under 00% aqueous conditions. Reversely pressure around the packing material makes water permeate into the pore of the packing material to overcome a force worked by capillarity. In other words, the surface of a reversed phase like C8 isn t wet by water anytime even if water permeates into the pore of the packing material or not. So it is wrong that we say dewetting when water expel from the pore. Saying Depermeatiing is more appropriate. /C8 is a reversed stationary phase, so that it is not wet with water. However the contact angle of water on the surface of /C8 is less than that of a conventional C8. Expelling force (pressure) acted by capillarity on /C8 is less than atmospheric pressure. So, /C8 shows reproducible retention under 00% aqueous conditions. lica Pressure 0nm After stopping pump for hour. agae, T. Enami and S. Doshi, LC/GC orth America ctober 00. Fig. Retention behavior of a C8 column under 00% aqueous mobile phase conditions Water Water permeates into the pore. Permeating Stop flow Water expels from the pore by capillarity. Depermenating Fig. Schematic diagram of C8 particle ). agae, T. Enami and S. Doshi, LC/GC orth America ctober 00. ) T. Enami and. agae, American Laboratory ctober 00. ) T. Enami and. agae, BUSEKI KAGAKU, (00) 09. Separation of nucleic acid bases 7 0 ºC ºC 0 8 0 Column:,/C8 mm 0 x. mm Mobile phase: 0mM Phosphate buffer ph7.0 Flow rate:.0 ml/min Temperature: 0 ºC and ºC Sample: = Cytosine = Uracil = Thymidine = Uridine = Thymine Relative retention of thymine (%) Change of retention of thymine at 0 ºC (measurement every stop flow for hour) 00 90 80 70 0 0 0 0 0 0 0 Measurement number ev ery stop flow /C8 showed more than 97% of reproducibility in retention using 00% aqueous buffer as a mobile phase.

/C8 Separation of water-soluble vitamins Separation of nucleotides =9,800 0 7 8 9 0 Column:,/C8 mm 0 x. mm Mobile phase: 0mM Phosphate buffer ph.8 Flow rate:.0 ml/min Temperature: 0 ºC Detection: UV@0nm Sample: = icotinic acid, = Pyridoxal = Pyridoxine, = icotinamide 0 7 8 9 0 Column:, /C8 mm 0 x. mm Mobile phase: 0mM Phosphate buffer ph.0 Flow rate:.0 ml/min Temperature: 0 ºC Detection: UV@0nm Sample: = -GDP, = -GMP, = -ATP, = -ADP, = -AMP separation of olong tea Separation of water-soluble vitamins using mobile phase for LC/MS Caffeine xalic acid Theanine Guanylic acid 0 0 0 0 0 0 0 8 0 8 0 Column: /C8, mm 0 x. mm Mobile phase: A) H /H P (99.9:0.) B) CH C Time (min) 0 0 0 %B (%) 0 0 0 Flow rate:.0 ml/min Temperature: 0 ºC Detection: UV@0 nm Injection volume: 0 ml Sample: olong tea = (-)-Epigallocatechin = (+)-Catechin = (-)-Epigallocatechin gallate = (-)-Epicatechin = (-)-Epicatechin gallate Column: /C8i mm 0 x. mm Mobile phase: A) 0mM Ammonium acetate Flow rate:.0 ml/min Temperature: 0 ºC Detection: UV@0 nm Sample: = icotinic acid B) Acetonitrile/ A solution (0:80) Time (min) 0 0 %B (%) 0 0 00 00 = Pyridoxine HCl Vitamin B = icotinamide = Thiamine HCl Vitamin B = Folic acid = Riboflavin Vitamin B 8

Separation of standard samples 7 Separation of pyridine and phenol 7 7 a(pyridine/phenol) = 0.8 a(pyridine/phenol) = 0.0, 7 Sunniest PhE a(pyridine/phenol) = 0., 0 0 0 0 Column:, µm.x0 mm, µm.x0mm, µm.x0 mm Sunniest PhE, µm.x0 mm Sunniest PFP, µm.x0 mm Mobile phase: CH H/H =7/ Flow rate:.0 ml/min Temperature: 0 ºC Pressure:. MPa Sample: = Uracil, Hydrophobicity 7 = Caffeine, = Phenol, Sunniest PFP = Butylbenzene, (=,700) = o-terphenyl, (=,000) = Amylbenzene, (=,00) 7 = Triphenylene, (=,00) is plate number of the above. C8 RP-AQUA C8 PhE PFP a(amylbenzene/butylbenzene).....9 Hydrogen bonding capacity a(caffeine/phenol) 0. 0.9 0..00.00 Steric selectivity a(triphenylene/o-terphenyl).7.. 0.9. Residual silanol activity a(pyridine/phenol) 0.8 0.0 0. 0. ----- Sunniest PhE a(pyridine/phenol) = 0. 0 8 Column:, µm.x0 mm, µm.x0mm, µm.x0 mm Sunniest PhE, µm.x0 mm Mobile phase: CH H/H =0/70 Flow rate:.0 ml/min Temperature: 0 ºC Detection: UV@0nm Sample: = Uracil = Pyridine = Phenol Separation factor of pyridine and phenol is said to show the amount of residual silanol groups. The lower a value of separation factor, the less an effect of residual silanol groups. All Sunniest columns show one of the lowest value. shows not only high efficiency but also low column pressure. 9

Separation of a chelating compound Separation of acidic compounds Sunniest PhE Sunniest PFP Sunniest PhE 0 8 0 Column:, µm.x0 mm, µm.x0mm, µm.x0 mm Sunniest PhE, µm.x0 mm Sunniest PFP, µm.x0 mm Mobile phase: CH C/0mM H P =0/90 Flow rate:.0 ml/min Temperature: 0 ºC Detection: UV@0nm Sample: = 8-Quinolinol = Caffeine, RP-AQUA, C8,PhE and PFP are inert for a metal chelating compound and acidic and basic compounds, so that they show symmetrical peaks of each compound. 0 8 Column:, µm.x0 mm, µm.x0mm, µm.x0 mm Sunniest PhE, µm.x0 mm Mobile phase: CH C/0.% H P =/98 Flow rate:.0 ml/min Temperature: 0 ºC Detection: UV@0nm Sample: = Formic acid = Acetic acid = Propionic Acid Retention comparison between C8 and PFP Column size:. x 0 mm Mobile phase: CH C/0mM Phosphate buffer ph7.0 =0/0 Flow rate:.0 ml/min Sample: =Uracil, =Propranolol, =ortriptyline, =Amitriptyline Sunniest PFP PFP retains a cation such a nortriptyline much longer than a C8. 0

, C8-HT Features Low back pressure and high efficiency by precisely classified particle High pressure packing (0,000 psi) using hard silica gels with high pressure resistant leads long column life without any void. Unique bonding technique for Sunniest The most suitable inner surface of column by special grinding arrow Particle Distribution and Low Back Pressure Measured by Coulter Counter method Conventional particle D0:.7 mm D0:.09 mm D90:. mm D90/D0=.9 (mm) Classified particle D0:.7mm D0:.0mm D90:. mm D90/D0=. arrow particle distribution Pressure, MPa 0 0 0 0 0 Comparison of back pressure 0 0.. Flow rate, ml/min -HT.0 um Brand B C8.8 um Brand C C8.7 um Conventional mm silica gel particle was classified again. 0% volume was cut off from both sides respectively. Consequently column back pressure reduced more than %. ur mm silica gel particle shows a half pressure to compare with the other sub- mm silica gel particle. Column: Sunniest, Acquity and Zorbax Column dimension: 0 x. mm Mobile phase: Acetonitrile/water=(70/0) Temperature: o C An Unique Modification Leads Good Peak Shape eutral compounds Amitriptyline ( th peak) -HT mm () =,7 pl/m TF() =.9 Pressure =.8 MPa Acquity C8.7mm () = 8,77 pl/m TF() =.7 Pressure =. MPa Leading C8.mm A half of retention () = 7,9 pl/m TF() =.8 Pressure = 0.MPa Column dimension: 0 x. mm Mobile phase: Methanol/water=70/0 for neutral compounds Methanol/mM phosphate buffer (ph.0)=80/0 for antidepressants Flow rate: 0. ml/min Temperature: room temperature Sample: eutral compounds, =Uracil, =Toluene, =Biphenyl, =Penanthrene Antidepressants, =ortriptyline, =Toluene, = Imipramine, =Amitriptyline CH CH It is difficult to end-cap on sub mm or mm silica gel particle as well as mm or mm silica gel particle. Most sub mm or mm C8 columns show smaller plate number and larger tailing factor for a basic compound than -HT. -HT mm shows a good peak shape for amitriptyline under methanol/phosphate buffer mobile phase at room temperature. Furthermore -HT mm shows times longer retention time than the other brand columns.

, C8-HT Comparison of Plate umber 0 x. mm ()= 0,07 00 x. mm ()= 9,80 0 0 8 0 Retention time (min) Retention time (min) Mobile phase: CH C/H = 0/0 Flow rate: 0. ml/min for. x 0 mm and. x 0 mm, 0. ml/min for. x 7 mm and. x 00 mm Temperature: 0 ºC Detection: UV@0 nm Sample: =Uracil, =Toluene, =Acenaphthene, =Butylbenzene Separation of Analgesics mm, 0 x. mm ()=80,000 plate/m Mobile phase: CH C/0.% Formic acid = 0/80 Flow rate:.0 ml/min for 0 x. mm 0. ml/min for 0 x. mm Temperature: 0 ºC Detection: UV@0 nm Sample: =Acetaminophen, =Antipyrine, =Aspirin, =Ethenzamide CH H H CH CH H H C H CH Separation of Amino Acids derivatized with PA ()=7,000 plate/m 7 0 8 High throughput -HT mm, 0 x. mm 0 7 8 9 0 Retention time (min) 9 7 mm particle allows to reduce retention time because high efficiency is kept under high flow rate conditions. As shown the above chromatograms, analytical time reduced /8 without sacrifices of separation by using mm, 0 x. mm column instead of mm 0 x. mm column. Column: -HT mm, 00 x. mm Mobile phase: A) 0mM ap + 0mM ab7 + 0.mM a B) Acetonitrile/Methanol/Water (//0 %V) Time(min) 0 0. 7. 7.8 %B 0 00 Flow rate: 0.7 ml/min Temperature: 0 ºC Detection: UV@8 nm Sample: =Aspartic acid, =Glutaminc acid, =Serine, =Histidine, =Glycine, =Threonine, 7=Arginine, 8=Alanine, 9=Tyrosine, 0=Valine, =Methionine, =Tryptophan, =Pnehylalanine, =Isoleucine, =Leucine, =Lysine, 7=Proline 0 7 8 9 Retention time (min) Characteristics of -HT, mm lica gel support C8 Packings Particle size (mm) Pore diameter nm) Specific surface area (m /g) Carbon content (%) Bonded phase Maximum operating pressure Available ph range -HT.0 (Coulter counter) 0 0 C8 70 MPa or 0,000 psi. - 0 It is very important for mm 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 0.8 ml/g of pore volume is used for -HT, mm, so that it have a high capacity to resist pressure and a high operating pressure.

Sunniest Guard columns C8, RP-AQUA/C8, C8 PhE & PFP Guard Cartridge (0 x mm) Feature *mple structure *Low dead volume *Available for not only mm column but also mm column Filter Schematic Diagram of Cartridge 0xmm PEEK -ring Photo of Cartridge and Holder Analytical column only Both Guard and Analytical columns Comparison of chromatograms () =0,90 TF() =.0 () =0,770 TF() =.07 0 7 8 9 0 Column:, mm 0 x. mm Mobile phase: Guard cartridge 0 x mm CH H/0mM Phosphate buffer ph7. = 80/0 Flow rate:.0 ml/min Temperature: 0 ºC Pressure:.8 MPa,. MPa(+guard) Sample: = Uracil, = Propranolol, = ortriptyline, = Amitriptyline, TF: USP tailing factor Particle size Catalog o., mm Guard cartridge column (-pak + Holder) x 0 mm µm EBAH, mm Guard cartridge column (-pak + Holder) x 0 mm µm ERAH, mm Guard cartridge column (-pak + Holder ) x 0 mm µm ECAH, mm Guard cartridge (-pak) x 0 mm µm EBAC, mm Guard cartridge (-pak) x 0 mm µm ERAC, mm Guard cartridge (-pak) x 0 mm µm ECAC Sunniest Guard cartridge holder x 0 mm --- HLAC

, C8-HT Sunniest PhE Sunniest PFP Sunniest rdering information Inner diameter Length, µm, µm Sunniest RP- AQUA, µm Sunniest RP- AQUA, µm, µm, µm [mm] [mm] Catalog o. Catalog o. Catalog o. Catalog o. Catalog o. Catalog o. 0 EB EB ER ER EC EC 7 EB - ER - EC - 00 EB EB ER ER EC EC 0 EB7 EB7 ER7 ER7 EC7 EC7 0 EB8 EB8 ER8 ER8 EC8 EC8 0 EB EB ER ER EC EC 00 EB EB ER ER EC EC 0 EB7 EB7 ER7 ER7 EC7 EC7 0 EB8 EB8 ER8 ER8 EC8 EC8. 0 EB EB ER ER EC EC 0 EB EB ER ER EC EC 7 EB - ER - EC - 00 EB EB ER ER EC EC 0 EB7 EB7 ER7 ER7 EC7 EC7 0 EB8 EB8 ER8 ER8 EC8 EC8 0 0 - EB78 - ER78 - EC78 0 0 - EB8 - ER8 - EC8 0 - EB87 - ER87 - EC87 0 - EB88 - ER88 - EC88 Inner diameter Length Sunniest PhE, µm Sunniest PhE, µm Sunniest PFP, µm [mm] [mm] Catalog o. Catalog o. Catalog o..0 0 EP EP - 7 EP - - 00 EP EP - 0 EP7 EP7-0 EP8 EP8 -.0 0 EP EP - 00 EP EP - 0 EP7 EP7-0 EP8 EP8 -. 0 - EP - Inner diameter Length Sunniest C8-HT, µm [mm] [mm] Catalog o.. 0 EB9 0 EB9 0 EB9 7 EB9 00 EB9.0 0 EB 0 EB 0 EB 7 EB 00 EB 0 EP EP EF 7 EP - - 00 EP EP EF 0 EP7 EP7 EF7 0 EP8 EP8 EF8 0.0 0 - EP78-0.0 0 - EP8-0 - EP87-0 - EP88 -