(HILIC) Bill Champion Agilent Technologies, Inc opt 3/opt3/opt 2 HILIC - Agilent Restricted

Size: px
Start display at page:

Download "(HILIC) Bill Champion Agilent Technologies, Inc opt 3/opt3/opt 2 HILIC - Agilent Restricted"

Transcription

1 Hydrophilic Interaction Chromatography (HILIC) Bill Champion Agilent Technologies, Inc opt 3/opt3/opt 2 william_champion@agilent.com Page 1

2 HILIC A method of recent attention Bill Champion Agilent Technologies, Inc opt 3/opt3/opt 2 william_champion@agilent.com Page 2

3 HILIC but not the universal answer. Bill Champion Agilent Technologies, Inc opt 3/opt3/opt 2 william_champion@agilent.com Page 3

4 Objective - Outline Overview - Why HILIC is used Advantages/Disadvantages g How HILIC works Examples Method development Comments concerning use of HILIC Page 4

5 Why HILIC Is Used Retention of polar components Similar to NPLC but with aq mobile phase (General reversal of elution order from RPLC) Several different stationary phases available MS compatible May simplify sample prep Page 5

6 but there is always a price Slower equilibration than RPLC Peak distortion with mp-sample solvent mismatch (i.e., too much water in sample solvent) Poor retention of anions on silica Mechanism not well understood Page 6

7 Additional Benefits, Comments ACN has low viscosity allows high flow rates Van Deemter plot similar to RPLC Not truly orthogonal to RPLC, but dissimilar not just inverse separation complementary technique Type B Silica, high purity, less acidic less active but more reproducible Page 7

8 Type of stationary phase Parameters to Choose Organic solvent concentration ti Type of buffer Buffer (salt) concentration ph Temperature 8

9 Typical Stationary Phases - Polar Silica Amine Diol Amide Zwitterionic Page 9

10 Some Typical Polar Stationary Phases (-) Silica, Si-OH Si-O H (+) Amine, -(CH 2 ) 3 -NH 2 Diol, -(CH 2 ) 3 -O-CH 2 -(CHOH)CH 2 OH Amide, -(CH 2 ) n -(CO)NH 2 (+) (-) Zwitterionic, -(CH 2 ) n -N(Me) 2 -(CH 2 ) n -SO 3 Page 10

11 Typical Mobile Phase Water /ACN Buffer ph Page 11

12 Typical Mobile Phase Water (at least 2-3%, ~ 25%)/CAN Solvent strength: Water > MeOH > EtOH > IPA > ACN Buffer AmOAc, 5to20mM Buffer must be soluble in high CAN concentration ph 3 8 Page 12

13 Mobile Phase Need some water for hydration Water is strong solvent Increasing water, decreases retention 13

14 Buffer Controls Ionization of Silica Controls Ionization of Analyte Ammonium acetate, formate good solubility, MS friendly Phosphate - poor solubility at high % ACN 14

15 ph Controls Ionization of Silica Controls Ionization of Analyte Organic solvent affects the actual [H + ] 15

16 Temperature Higher Temperature Decreases Ret n Time Higher Temperature Increases Column Efficiency (increases N) Lower Temperature May Increase Selectivity 16

17 Separation Mechanism Page 17

18 Retention in Reversed-Phase Separations: Mechanism Similar to Liquid-Liquid Extraction In RPLC decrease retention by decreasing polarity of mobile phase Heptane Non-Polar Heptane Non-Polar H 2 0 Polar H 2 0/MeOH Less Polar Solvent Polarity: H2O > MeOH, ACN > EtOH > IPA >> Heptane Relatively non-polar analyte (e.g., toluene) favors non-polar phase. Decreasing polarity of aqueous phase increases affinity for analyte in H 2 O/MeOH phase. Page 18

19 Retention in Reversed-Phase Separations: Mechanism Similar to Liquid-Liquid Extraction In RPLC decrease retention by decreasing polarity of mobile phase Heptane Non-Polar Heptane Non-Polar H 2 0 Polar H 2 0/MeOH Less Polar Mobile Phase Strength: H2O < MeOH, ACN < EtOH < IPA << Heptane Relatively non-polar analyte (e.g., toluene) favors non-polar phase. Decreasing polarity of aqueous phase increases affinity for analyte in H 2 O/MeOH phase. Page 19

20 RPLC Comparison of HILIC and RPLC Non-polar stationary phase (e.g., C18) Polar mobile phase (i.e., H 2 0/MeOH, H 2 0/ACN, etc.) Decrease retention by decreasing polarity of mobile phase (e.g., increase ACN in mobile phase to decrease retention) HILIC Polar stationary ti phase (e.g., silica) Polar mobile phase (i.e., H 2 0/ACN) Decrease retention by increasing polarity of mobile phase (i.e., increase H 2 O in mobile phase to decrease retention) Page 20

21 Typical Elution Order for Test Compounds on HILIC Column RPLC Column Test Mixture mau chloronitrobenzene Column 4.6 x 150 mm Mobile phase 75% ACN: 25% H2O Flow Rate 0.5 ml/ml Phenol Col Temp 25 C 30 Injection Volume 5 µl Naphthalene Uracil 1. Notice Uracil normally unretained in RPLC retained ed in HILIC min 2. Notice completely different retention order than in RPLC 21

22 How Does HILIC Work on Silica Based Columns? A water layer must be adsorbed onto the stationary phase. The polar analyte partitions into and out of this adsorbed layer. A charged polar analyte can also undergo ion exchange with the charged silica molecules (i.e., cation exchange with amines) The combination of these mechanisms drives retention in HILIC. Retention/elution is from least to most polar the opposite of reversed-phase LC. HILIC offers more retention than reversed-phase for very polar bases. 22

23 How Does HILIC Work on Silica Based Columns? Potential Mechanisms H 2 O ACN ACN ACN ACN 1. ACN ACN ACN CH 2 CHCH 3 ACN NH + 3 H 2 O CH 2 CHCH 3 H 2 O H 2 O H 2 O - H 2 O H 2 O H 2 O -NH + O - O 3 2. O O - H 2 O ACN ACN 1. Partitioning in and out of adsorbed water layer 2. Ion exchange with silanols 23

24 Examples Page 24

25 Some Typical Analytes for HILIC Amino acids (when only a few are of interest) Nucleobases (purines and pyrimidines, adenine, guanine, thymine, thymine, cytosine, uracil) Nucleosides (Adenosine, cytosine etc.) Alkaloids Carbohydrates Polar compounds, small basic compounds Cytosine NH 2 CH 2 CHCH 3 N N Melamine Amphetamine NH 3 + H 2 N N NH 2 25

26 Comparison of Reversed-Phase and HILIC Structures of Drug Compounds Studied a) Paroxetine Antidepressant MW F O N O O b) Ranitidine Antiulcerative MW N O S N N NO 2 Basic portion of molecule, impacts HILIC retention 26

27 Chromatographic Conditions RPLC Instrument: Column: Mobile phase: Gradient: Column temp: 40 C Sample volume: Flow rate: Agilent Series 1100 HPLC ZORBAX Eclipse XDB-C18 (2.1 mm 150 mm, 5 μm) A: 8-mM HCO 2 NH 4 in water; B: 8-mM HCO 2 NH 4 in 95% ACN/5% water 5% B to 90% B in 10 min 5 μl 0.3 ml/min HILIC Everything same as for RPLC except for column and gradient conditions: Column: Gradient: ZORBAX Rx-SIL (2.1 mm 150 mm, 5 μm) 100% B to 50% B in 10 min 27

28 LC/MS/MS Separation of Paroxetine & Ranitidine on ZORBAX Eclipse XDB-C18 Column (RPLC Mode) RPLC separation ZORBAX Eclipse XDB-C18 (2.1mm 150mm) 100 ppb 5% B 90% B (10 min) gradient Paroxetine Abundanc ce RPLC has poor peak shape and limited sensitivity for Ranitidine! N Ranitidine O S N N m/z= F N O m/z= NO 2 O O min

29 LC/MS/MS Separation of Paroxetine and Ranitidine on ZORBAX Rx-SIL Column (HILIC Mode)- 100 ppb Level HILIC separation Zorbax RX-SIL Silica column (2.1mm 150mm) 100 ppb 100% B 50% B (10 min) gradient Paroxetine m/z= HILIC Separation has better peak shape and more sensitivity for ranitidine! F O O O N Ranitidine m/z= O N S N N NO min

30 HILIC Separations are Ideal for LC/MS Analysis of Melamine HILIC Separation Using ZORBAX Rx-Sil NH 2 Melamine 50ppt Melamine N N H 2 N N NH 2 HPLC system : Agilent 1200 RRLC Column : Agilent ZORBAX Rx-Sil, mm, 5 um Injection Volume :10μL Temp : 40 Flow rate : 0.2 ml/min Mobile phase : A - 5 mm Ammonium acetate in Water : B - 5 mm Ammonium acetate in ACN Isocratic : 95%B 30

31 @ACN 1uL Effect of Injection Solvent & Volume on Melamine uL Don t inject in strong solvent!!!! 31

32 Mobile Phase Page 32

33 Effect of Increasing % ACN on Cyanuric Acid Retention 60% Decreasing strong solvent (increasing AACN) increases retention. 70% 80% Same behavior as reversed-phase but strong and weak solvents are opposite! 90% 95% 33

34 Retention Time Changes with Changes in Mobile Phase Composition me, min ention Tim Rete Mobile phase: A= 5 mm NH 4 OAc/H20 B= 5 mm NH 4 OAc/ACN 4 % Acetonitrile 34

35 Effect of Ammonium Acetate Concentration on Cyanuric Acid Retention Buffer strength can dramatically 5 mm affect retention and resolution and should be a key parameter in method development and optimization on HILIC columns. 10 mm 20 mm 35

36 16.0 Effect of ph, Buffer on Retention on Silica 14.0 Strong Acids 12.0 Ret'n Tim me Amines Neutral 2-nsa p-xsa phenol caffeine nortrip diphhyd procain Ref: McCalley, J. Chromatogr. A 1171 (2007) mM 1.52mM 153mM.. 0.1% 4 ph 3.5 ph 3.0 ph 3.0 TFA Page 36

37 16.0 Effect of ph, Buffer on Retention on Silica procainamide Ret'n Tim me Amines benzylamine diphenhydramine nortrip diphhyd procain A, B, C - HCOONH4/85% ACN; D 95% ACN Ref: McCalley, J. Chromatogr. A 1171 (2007) 46 0 A mm B mm C - 153mM.. D - 0.1% 4 ph 3.5 ph 3.0 ph 3.0 TFA Page 37

38 Effect of ph, Buffer on Retention on Silica p-xylenesulfonic acid A, B, C - HCOONH4/85% ACN; D 95% ACN 2-naphthalenesulfonic acid Strong Acids Ret'n Tim me caffeine phenol Neutral 2-nsa p-xsa phenol caffeine 0.0 Ref: McCalley, J. Chromatogr. A 1171 (2007) 46 0 A mm B mm C mm.. D -40.1% ph 3.5 ph 3.0 ph 3.0 TFA Page 38

39 Effect of Buffer, ph Controls Ionization of Silica Controls Ionization of Analyte Organic solvent affects the actual [H + ] Acids can be repulsed (reduces ret n time) by anionic silica Buffer can mask anionic sites (reduce ret n time of amines, increase for acids) 39

40 Effect of Temperature Higher temperature improves kinetics sharper peaks, shorter ret n times Lower temperature improves selectivity Affects buffer equilibrium (effective ph) Page 40

41 Columns Page 41

42 Comparison of Silica and Amino HILIC Columns HPLC system Agilent 1200 Column 1 Agilent Zorbax-NH2, mm, 5 um Column 2 Agilent Zorbax-Rx Sil, mm, 5 um Injection Volumn 2 ul Temp 25 Flow rate 0.4 ml/min Mobile phase A - 5 mm Ammonium acetate in Water Isocratic 95%B B - 5 mm Ammonium acetate in ACN 42

43 HILIC Separation Using Zorbax NH 2 5 mm Ammonium acetate in 5% H 2 O/ACN x MRM ( > ) C_M_-mrm-reversed-NH2-11.d Melamine Cyanuric Acid Counts (%) vs. Acquisition Time (min) 43

44 HILIC Separation Using Zorbax Rx-Sil 5 mm Ammonium acetate in 5% H 2 O/ACN x MRM ( > ) C_M_Si_01.d Cyanuric Acid Melamine Counts (%) vs. Acquisition Time (min) 44

45 HILIC Separation of Sugars Using Zorbax Rx-NH 2 Comparison 70/30 and 75/25 ACN/H 2 O p 2 glucose 45

46 Method Development Page 46

47 Method Development/Optimization Systematic Approach to Method Development Stationary phase Mobile Phase Buffer, Buffer concentration ph 47

48 Method Development/Optimization Systematic Approach to Method Development Separations are too complicated to expect to find optimum conditions using Random Walk Investigate t effects of buffer, ph, buffer concentration ti Recommend use of experimental design cf: B. Dejaegher, D. Mangelings, Y. Vander Heyden, J. Sep. Sci., 31 (2008)

49 Method Development/Optimization Starting Conditions Silica Ammonium formate, 5, 10, 20 mm ph 3.0, 3.5, 4.0 (5, 6?) ACN/H 2 O 97%, 95%, 90%, 85%, 75% TFA? 49

50 Typical Stationary Phases - Polar Silica Amine Diol Amide Zwitterionic Page 50

51 Elution Order Inversely Related to RPLC, BUT Not Opposite Mechanism, Retention ti is not exactly opposite of RPLC RPLC cannot be used as predictive model Not truly orthogonal, but dissimilar separation, complementary technique Acids often poorly retained Page 51

52 Summary - Advantages of HILIC Retention of polar compounds Good peak shape for basic compounds where RPC may give tailing and/or low efficiency Higher flow rates and/or long columns can be used due to low viscosity mobile phases with high organic content; greater efficiency Enhanced detection sensitivity with MS Efficient spraying and de-solvation in electrospray MS As much as 3X sensitivity Can directly inject extracts from C18 SPE cartridges or acetonitrile precipitated plasma supernatant Dissimilar to RPLC (2D separations); elution order reversal, Complementary separations 52

53 References A.J. Alpert, J. Chromatogr. 499 (1990) 177 BA B.A. Bidlingmeyer, et. al., Anal. Chem. 54 (1982) 442 D.V. McCalley, J. Chromatogr. A 1171 (2007) 46 D.V. McCalley, J. Chromatogr. A 1217 (2010) 858 B. Dejaegher, D. Mangelings, Y. Vander Heyden, J. Sep. Sci., 31 (2008) 1438 B. A. Olsen, J. Chromatogr. A 913 (2001) 113 R.Li, Junxiong Huang, J. Chromatogr. A 1041 (2004) 163 T. Langrock, P. Czihal, R. Hoffmann, Amino Acids 30 (2006) 291 Page 53

Too Polar for Reversed Phase What Do You Do?

Too Polar for Reversed Phase What Do You Do? Too Polar for Reversed Phase What Do You Do? June 20, 2013 Mark Powell Columns and Consumables Technical Support Page 1 C8 or C18 Doesn t Always Do the Job Typical reversed phase conditions involve water/buffer

More information

penta-hilic UHPLC COLUMNS

penta-hilic UHPLC COLUMNS penta-hilic UHPLC COLUMNS penta-hilic Highly retentive, proprietary penta-hydroxy-ligand Excellent peak shape for polar compounds with a variety of functional groups: acids, bases, zwitterions strong and

More information

penta-hilic UHPLC COLUMNS

penta-hilic UHPLC COLUMNS penta-hilic UHPLC COLUMNS Highly retentive, proprietary penta-hydroxy-ligand penta-hilic Excellent peak shape for polar compounds with a variety of functional groups: acids, bases, zwitterions strong and

More information

LC and LC/MS Column Selection Flow Chart

LC and LC/MS Column Selection Flow Chart LC and LC/MS Column Selection Flow Chart To use the column selection diagram below, simply follow the path for your analyte and mobile phase. At the far right, follow your final column selection to the

More information

HILIC Method Development in a Few Simple Steps

HILIC Method Development in a Few Simple Steps HILIC Method Development in a Few Simple Steps Monica Dolci, Luisa Pereira, Dafydd Milton and Tony Edge Thermo Fisher Scientific, Runcorn, Cheshire, UK Overview This poster presents a systematic approach

More information

Hydrophilic Interaction Liquid Chromatography: Some Aspects of Solvent and Column Selectivity

Hydrophilic Interaction Liquid Chromatography: Some Aspects of Solvent and Column Selectivity Hydrophilic Interaction Liquid Chromatography: Some Aspects of Solvent and Column Selectivity Monica Dolci, Thermo Fisher Scientific, Runcorn, Cheshire, UK Technical Note 20544 Key Words Hydrophilic, HILIC,

More information

Fast Separation of Vastly Different Compounds by Isocratic HPLC

Fast Separation of Vastly Different Compounds by Isocratic HPLC Fast Separation of Vastly Different Compounds by Isocratic HPLC Aimee N. Heyrman and Yury Zelechonok Resolution Systems, Inc. 590 E. 32nd St. Holland. MI, 49423 SIELC Technologies, 15 E. Palatine Rd, Suite

More information

Does one-size-fit-all?

Does one-size-fit-all? A Look at Column Choices: Does one-size-fit-all? Bill Champion Agilent Technologies, Inc. 800-227-9770 opt 3/opt3/opt 2 william_champion@agilent.com Page 1 Separation Techniques Hydro - philic Volatile

More information

Packings for HPLC. Packings for HPLC

Packings for HPLC. Packings for HPLC Summary of packings for HPLC In analytical HPLC, packings with particle sizes of 3 to 10 µm are preferred. For preparative separation tasks, also particles with diameters larger than 10 µm are applied.

More information

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

HICHROM. Chromatography Columns and Supplies. LC COLUMNS SeQuant ZIC-HILIC. Catalogue 9. Hichrom Limited HICHROM Chromatography Columns and Supplies LC COLUMNS SeQuant ZIC-HILIC Catalogue 9 The Markham Centre, Station Road Theale, Reading, Berks, RG7 PE, UK Tel: + (0)8 90 660 Fax: + (0)8 9 8 Email: sales@hichrom.co.uk

More information

Comparison of different aqueous mobile phase HPLC techniques

Comparison of different aqueous mobile phase HPLC techniques June 009 ewsletter Pharmaceutical Analysis: What is Your Problem? SIELC Technologies, Inc., Prospect Heights, IL 0070 Pharmaceutical analysis involves liquid chromatography of various compounds, from active

More information

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

William E. Barber, Ph.D. Applications Chemist October William E. Barber, Ph.D. Applications Chemist ctober 0 00 Secrets of Good Peak Shape in HPLC Time: :00 p.m. CET Telephone Number: + 44 0 76 088 Chairperson: John Vis The Secrets of Good Peak Shape in HPLC

More information

Agilent s New Weak Anion Exchange (WAX) Solid Phase Extraction Cartridges: SampliQ WAX

Agilent s New Weak Anion Exchange (WAX) Solid Phase Extraction Cartridges: SampliQ WAX Agilent s New Weak Anion Exchange (WAX) Solid Phase Extraction Cartridges: SampliQ WAX Technical Note Agilent s SampliQ WAX provides Applications for strongly acidic, acidic and neutral compounds Excellent

More information

Dilution(*) Chromatography

Dilution(*) Chromatography WA20264 Poster # 184, HPLC 2002, Montreal, 4-5 June 2002 At-Column Column-Dilution for Preparative Dilution(*) Chromatography Cecilia Mazza, Jie Cavanaugh, Ziling Lu,Tom Sirard,Tom Wheat and Uwe Neue Waters

More information

and water enriched layer at the stationary phase particle-eluent interface [9,11].

and water enriched layer at the stationary phase particle-eluent interface [9,11]. 8 A Simple, Generally Applicable HILIC Method Development Platform Based Upon Selectivity by Alan P McKeown, Advanced Chromatography Technologies Ltd, 1 Berry Street, Aberdeen, Scotland AB25 1HF, UK amckeown@ace-hplc.com

More information

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

Analyze Nucleotides, Nucleosides, Purine, and Pyrimidine Bases Simultaneously with the Ultra IBD Column pharmaceutical #9 Applications note Analyze Nucleotides, Nucleosides, Purine, and Pyrimidine Bases Simultaneously with the Ultra IBD Column Mixtures of nucleotides, nucleosides, and their respective purine

More information

Advantages of polymerbased. an alternative for ODS

Advantages of polymerbased. an alternative for ODS Advantages of polymerbased HPLC columns an alternative for ODS Yukiko Higai Shodex, 1 Content (1) Comparison between Silica and Polymer-based RP columns (2) Characteristics of Polymer RP columns - an example:

More information

It s hot! ZIC -chilic

It s hot! ZIC -chilic It s hot! ZIC -chilic Complementary selectivity for HPLC and LC-MS separation of polar hydrophilic compounds EMD Millipore Corp. is a subsidiary of Merck KGaA, Darmstadt, Germany ZIC -chilic Your benefits

More information

State-of-the-art C18 HPLC Columns

State-of-the-art C18 HPLC Columns An HPLC GL Sciences Newest and Most Advanced ODS Phase-New For 00 State-of-the-art C HPLC s Improved Peak Shapes and Heights Enhancing Sensitivity High Resolution Fast Equilibration Compatible with 00%

More information

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

The Secrets of Rapid HPLC Method Development. Choosing Columns for Rapid Method Development and Short Analysis Times The Secrets of Rapid HPLC Method Development Choosing Columns for Rapid Method Development and Short Analysis Times Rapid Analysis Is More Than Run Time It is developing a method to meet a goal and developing

More information

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

Welcome to our E-Seminar: Choosing HPLC Columns for Faster Analysis Smaller and Faster Welcome to our E-Sear: Choosing HPLC Columns for Faster Analysis Smaller and Faster High Throughput/Fast LC Requires. Short columns 0 mm or shorter Small particle sizes. µm Rapid Resolution or new.8 µm

More information

Reversed Phase Solvents

Reversed Phase Solvents Part 1. General Chromatographic Theory Part 2. verview of HPLC Media Part 3. The Role of the Mobile Phase in Selectivity Part 4. Column Care and Use Reversed Phase Solvents 2 Solvents for RP Chromatography

More information

C18 Column. Care & Use Sheet

C18 Column. Care & Use Sheet C18 Column Care & Use Sheet HALO Description HALO C18 is a high-speed, high-performance liquid chromatography column based on a new Fused-CoreTM particle design. The Fused-Core particle provides a thin

More information

How proteins separate on reverse-phase HPLC

How proteins separate on reverse-phase HPLC 1 Reverse Phase How proteins separate on reverse-phase HPLC RP chromatography separates proteins through the interaction of the hydrophobic foot of the protein with a nonpolar surface of the particle RP

More information

Choosing Columns and Conditions for the Best Peak Shape

Choosing Columns and Conditions for the Best Peak Shape Choosing Columns and Conditions for the Best Peak Shape What is Good Peak Shape and Why is it Important? Good peak shape can be defined as a symmetrical or gaussian peak and poor peak shape can include

More information

for Acclaim Mixed-Mode HILIC-1 Column

for Acclaim Mixed-Mode HILIC-1 Column for Acclaim Mixed-Mode HILIC-1 Column Product Manual for ACCLAIM Mixed-Mode HILIC-1 Page 1 of 17 Product Manual for ACCLAIM Mixed-Mode HILIC-1 Column 5µm, 4.6 x 250mm, P/N 066844 5µm, 4.6 x 150mm, P/N

More information

Acclaim Mixed-Mode WCX-1

Acclaim Mixed-Mode WCX-1 Acclaim Mixed-Mode WCX-1 Product Manual for the Acclaim Mixed-Mode WCX-1 Column Page 1 of 28 PRODUCT MANUAL for the Acclaim Mixed-Mode WCX-1 Columns 4.6 x 150 mm, P/N (068353) 4.6 x 250 mm, P/N (068352)

More information

Simultaneous Determination of Paraquat and Diquat in Environmental Water Samples by HPLC-MS/MS

Simultaneous Determination of Paraquat and Diquat in Environmental Water Samples by HPLC-MS/MS Simultaneous Determination of Paraquat and Diquat in Environmental Water Samples by HPLC-MS/MS Richard Jack, Xiaodong Liu, Leo Wang, and Chris Pohl OT70806_E 08/13S 1 The world leader in serving science

More information

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

Product Bulletin. ACE LC/MS and Rapid Analysis HPLC Columns. HPLC Columns Product Bulletin HPLC Columns ACE LC/MS and Rapid Analysis HPLC Columns 0 mm, 30 mm, 35 mm and 50 mm column lengths.0,., 3.0, 4.0 and 4.6 mm column diameters Configured for High Sample Throughput Specially

More information

Analysis of Stachydrine in Leonurus japonicus Using an Agilent ZORBAX RRHD HILIC Plus Column with LC/ELSD and LC/MS/MS

Analysis of Stachydrine in Leonurus japonicus Using an Agilent ZORBAX RRHD HILIC Plus Column with LC/ELSD and LC/MS/MS Analysis of Stachydrine in Leonurus japonicus Using an Agilent ZORBAX RRHD HILIC Plus Column with LC/ELSD and LC/MS/MS Application Note Traditional Chinese Medicine Author Rongjie Fu Agilent Technologies

More information

Inertsil Technical Library

Inertsil Technical Library HPLC COLUMN INERTSIL TECHNICAL LIBRARY H P L C C o l u m n Inertsil Technical Library Inertsil ODS-4, C8-4 Comparison of Performance List of Compared Columns Experimental Explanation, Analytical Conditions

More information

Xiqin Yang, Jun Dai, Peter W. Carr* Chemistry Department University of Minnesota

Xiqin Yang, Jun Dai, Peter W. Carr* Chemistry Department University of Minnesota Comparison of the Reversed-Phase and Ion- Exchange Contributions to Retention on Polybutadiene Coated Zirconia and Octadecyl Silane Bonded Silica Phases Xiqin Yang, Jun Dai, Peter W. Carr* Chemistry Department

More information

High Performance Liquid Chromatography

High Performance Liquid Chromatography Updated: 3 November 2014 Print version High Performance Liquid Chromatography David Reckhow CEE 772 #18 1 HPLC System David Reckhow CEE 772 #18 2 Instrument Basics PUMP INJECTION POINT DETECTOR COLUMN

More information

High Performance Liquid Chromatography

High Performance Liquid Chromatography Updated: 3 November 2014 Print version High Performance Liquid Chromatography David Reckhow CEE 772 #18 1 HPLC System David Reckhow CEE 772 #18 2 1 Instrument Basics PUMP INJECTION POINT DETECTOR COLUMN

More information

Liquid Chromatography

Liquid Chromatography Liquid Chromatography 1. Introduction and Column Packing Material 2. Retention Mechanisms in Liquid Chromatography 3. Method Development 4. Column Preparation 5. General Instrumental aspects 6. Detectors

More information

HPLC Background Chem 250 F 2008 Page 1 of 24

HPLC Background Chem 250 F 2008 Page 1 of 24 HPLC Background Chem 250 F 2008 Page 1 of 24 Outline: General and descriptive aspects of chromatographic retention and separation: phenomenological k, efficiency, selectivity. Quantitative description

More information

for Acclaim Mixed-Mode WCX-1

for Acclaim Mixed-Mode WCX-1 for Acclaim Mixed-Mode WCX- Product Manual for the Acclaim Mixed-Mode WCX- Column Page of 8 Product Manual for Acclaim Mixed-Mode WCX- Columns Acclaim Mixed-Mode WCX-, 3µm, Analytical column, 3.0 x 50mm

More information

Method Development Kits

Method Development Kits Method Development Kits sales representative for ordering information or contact...77 Method Development Kits First column choice for Pharma Development USP L C Method Development Kits Maximize retention

More information

Chemistry Instrumental Analysis Lecture 28. Chem 4631

Chemistry Instrumental Analysis Lecture 28. Chem 4631 Chemistry 4631 Instrumental Analysis Lecture 28 High Performance Liquid Chromatography () Instrumentation Normal Phase Chromatography Normal Phase - a polar stationary phase with a less polar mobile phase.

More information

Small Molecule Selectivity Sampler

Small Molecule Selectivity Sampler Small Molecule Selectivity Sampler HALO 90 Å C18, AQ-C18, BIPHENYL Fused-Core Particle Technology Why Choose Fused-Core Particle Technology? Fused-Core Out-Performs Totally Porous Particles. HALO particles

More information

Shodex TM ODP2 HP series columns

Shodex TM ODP2 HP series columns HPLC Columns Shodex TM ODP2 HP series columns Better retention of highly polar substances Technical notebook No. 6 Contents 1. Introduction 1-1. Specifications 1-2. Eluent Compatibility of ODP2 HP Series

More information

UPLC Method Development and Validation

UPLC Method Development and Validation UPLC Method Development and Validation Rev. 2 2008 Waters Corporation Challenges of Method Development Methods are developed throughout the drug development process Samples vary in complexity Redundancy

More information

Part 2. Overview of HPLC Media

Part 2. Overview of HPLC Media Part 1. General Chromatographic Theory Part 2. Overview of HPLC Media Part 3. The Role of the Mobile Phase in Selectivity Part 4. Column Care and Use 1 HPLC Particle Technology Core-Shell Particle Fully

More information

LC Columns with Liquid Separation Cell Technology

LC Columns with Liquid Separation Cell Technology Obelisc LC Columns with Liquid Separation Cell Technology Obelisc HPLC Columns - Liquid Separation Cell Technology Introduction Obelisc HPLC columns are the latest, innovative columns from SIELC Technologies,

More information

8. Methods in Developing Mobile Phase Condition for C18 Column

8. Methods in Developing Mobile Phase Condition for C18 Column I. HPLC Columns Technical Information 8. Methods in Developing Mobile Phase Condition for C18 Column Introduction In reversed phase HPLC, octadecyl group bonded silica columns (C18, ODS) are the most widely

More information

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

Ion Exchange Columns ---- Inertsil AX 078 Inertsil CX 080. HPLC Column Catalog Ion Exchange ---- Inertsil AX 078 Inertsil CX 080 HPLC Column Catalog 077 Inertsil AX Physical Properties Silica : Series High Purity Silica Gel : μm Surface Area : 0 m /g Pore Size : 00 Å (0 nm) Pore

More information

"Theory and Practice of High Speed Chromatography for Bioanalysis" Stuart Coleman March 20, 2007

Theory and Practice of High Speed Chromatography for Bioanalysis Stuart Coleman March 20, 2007 "Theory and Practice of High Speed Chromatography for Bioanalysis" Stuart Coleman March 20, 2007 General Observations about HPLC Separations A good separation is necessary for good bioanalytical quantitation.

More information

Investigation of retention mechanisms in HILIC chromatography: Important considerations for robust method development

Investigation of retention mechanisms in HILIC chromatography: Important considerations for robust method development Investigation of retention mechanisms in HILIC chromatography: Important considerations for robust method development Anders Fridström Sigma- Aldrich GMBH Abstract Investigation of Retention Mechanisms

More information

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

columns Acclaim Mixed-Mode WCX-1 for Separating Basic Molecules columns Acclaim Mixed-Mode WCX- for Separating Basic Molecules The Acclaim Mixed-Mode WCX- is a novel, high-efficiency, silica-based column specially designed for separating various basic analytes. This

More information

Hydrophilic Interaction Liquid Chromatography: Method Development Approaches

Hydrophilic Interaction Liquid Chromatography: Method Development Approaches Hydrophilic Interaction Liquid Chromatography: Method Development Approaches Monica Dolci, Luisa Pereira, and Tony Edge Thermo Fisher Scientific, Runcorn, Cheshire, UK Abstract This poster summarizes the

More information

Improve Peak Shape and Productivity in HPLC Analysis of Pharmaceutical Compounds with Eclipse Plus C8 Columns Application

Improve Peak Shape and Productivity in HPLC Analysis of Pharmaceutical Compounds with Eclipse Plus C8 Columns Application Improve Peak Shape and Productivity in HPLC Analysis of Pharmaceutical Compounds with Eclipse Plus C8 Columns Application Pharmaceuticals Authors John W. Henderson Jr., Nona Martone, and Cliff Woodward

More information

HICHROM. Chromatography Columns and Supplies. LC COLUMNS SIELC Primesep. Catalogue 9. Hichrom Limited

HICHROM. Chromatography Columns and Supplies. LC COLUMNS SIELC Primesep. Catalogue 9. Hichrom Limited HICHROM Chromatography Columns and Supplies LC COLUMNS SIELC Primesep Catalogue 9 The Markham Centre, Station Road Theale, Reading, Berks, RG7 4PE, UK Tel: +44 (0)8 90 660 Fax: +44 (0)8 9 484 Email: sales@hichrom.co.uk

More information

Method Development with ZirChrom's Ion Exchange Phases

Method Development with ZirChrom's Ion Exchange Phases For Peak Performance ZIRCHROM ION EXCHANGERS Phases for Sugars and Proteins Wide Range of Ion Exchange Selectivity No Shrinking or Swelling Use Any Organic Solvent Significantly Higher Efficiency than

More information

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

An HPLC column offering moderate retentivity with superb peak shape Inertsil ODS Sprint An HPLC column offering moderate retentivity with superb peak shape Inertsil ODS Sprint An important new column in the Inertsil series Bonded-Phase Structure Perfect balance of retention for both polar

More information

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

Barry E. Boyes, Ph.D. Consumables and Accessories Business Unit May 10, 2000 Barry E. Boyes, Ph.D. Consumables and Accessories Business Unit May 0, 000 HPLC Column Troubleshooting What Every HPLC User Should Know :00 a.m. EST Telephone Number: 86-650-06 Chair Person: Tim Spaeder

More information

Optimisil HPLC & UPLC Column catalogue

Optimisil HPLC & UPLC Column catalogue Optimisil HPLC & UPLC Column catalogue Analytical Hub Plot No: 37, P & T Colony, Vikrampuri, Secunderabad 500009 Tel: 040-64621313, Mobile: +91 7799931313 Website: www.analyticalhub.com email: contact@analyticalhub.com

More information

Novel LC/MS-Compatible Stationary Phase with Polar Selectivity

Novel LC/MS-Compatible Stationary Phase with Polar Selectivity Novel LC/MS-Compatible Stationary Phase with Polar Selectivity Carmen T. Santasania and David S. Bell Supelco 595 N. Harrison Rd., Bellefonte, PA 16823 T403168 GJU Introduction Stationary phases based

More information

LC Column Troubleshooting. Do you have an equivalent column for my LC column?

LC Column Troubleshooting. Do you have an equivalent column for my LC column? LC Column Troubleshooting Do you have an equivalent column for my LC column? LC columns from different suppliers may have very different retention properties even if the bonding is the same. Although there

More information

The new Agilent 1290 Infinity Quaternary LC

The new Agilent 1290 Infinity Quaternary LC The new Agilent 1290 Infinity Quaternary LC How binary pump-like performance can enhance your lab efficiency Sebastian Krahe Produktspezialist HPLC Agilent Technologies Page 1 The 1290 Infinity Quaternary

More information

Method Development Guide. A Step by Step Approach to Developing Reproducible HILIC Methods + + ACE HILIC Method Development Guide

Method Development Guide. A Step by Step Approach to Developing Reproducible HILIC Methods + + ACE HILIC Method Development Guide + ACE HILIC Method Development Guide A Step by Step Approach to Developing Reproducible HILIC Methods + ++ + ACE HILIC Method Development Guide ACE Ultra-Inert, Base Deactivated HPLC and UHPLC columns

More information

The Theory of HPLC. Ion Pair Chromatography

The Theory of HPLC. Ion Pair Chromatography The Theory of HPLC Ion Pair Chromatography i Wherever you see this symbol, it is important to access the on-line course as there is interactive material that cannot be fully shown in this reference manual.

More information

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

ACE SuperC18. ACE UHPLC and HPLC Columns. Ultra-Inert UHPLC and HPLC Columns with Extended ph Stability ACE SuperC8 TM Ultra-Inert UHPLC and HPLC Columns with Extended ph Stability Exploit selectivity at low, intermediate and high ph Use with LC/MS compatible buffers between ph.. High efficiency µm, µm,

More information

Silanol. Groups Liquid Chromatography

Silanol. Groups Liquid Chromatography Abstract umber: 0-8P Study of Secondary Interaction Based on Residual lanol Groups for Reversed-Phase Liquid Chromatography II orikazu agae Ph.D, Kouji Yamamoto, Chiaki Kadota Chromaik Technologies Inc.

More information

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

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 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 HALO column packings are not made the typical way. Instead, the particles packed into HALO columns

More information

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

Analysis - HPLC A.136. Primesep 5 µm columns B.136 Primesep 5 µm columns Primesep columns feature double functionality of the bonding i.e : alkyl chain with anionic or cationic group, chelating group. This feature creates unique selectivities when using

More information

Chromatographic Methods: Basics, Advanced HPLC Methods

Chromatographic Methods: Basics, Advanced HPLC Methods Chromatographic Methods: Basics, Advanced HPLC Methods Hendrik Küpper, Advanced Course on Bioinorganic Chemistry & Biophysics of Plants, summer semester 2018 Chromatography: Basics Chromatography a physical

More information

LC Technical Information

LC Technical Information LC Technical Information Method Transfer to Accucore.6 μm Columns Containing solid core particles, which are engineered to a diameter of.6μm and a very narrow particle size distribution; Accucore HPLC

More information

Yuhui Yang, An Trinh, Michael Ye, and Tom Henderson 595 North Harrison Road, Bellefonte, PA T HKB

Yuhui Yang, An Trinh, Michael Ye, and Tom Henderson 595 North Harrison Road, Bellefonte, PA T HKB Use of An Improved Version of C8+SCX Mixed-Mode Solid Phase Extraction Material for Clean Extraction and Recovery of Basic, Zwitterionic, Neutral and Acidic Compounds from Biological Fluids Yuhui Yang,

More information

Acclaim HILIC-10 Columns

Acclaim HILIC-10 Columns Thermo Scientific Acclaim HILIC- Columns Product Manual P/N: January 3 Part of Thermo Fisher Scientific Product Manual for Acclaim HILIC- Analytical Column Acclaim HILIC-, 3 µm, Analytical, 4.6 x 5 mm

More information

Advanced HPLC Method Development Using Quality by Design

Advanced HPLC Method Development Using Quality by Design Advanced HPLC Method Development Using Quality by Design A comprehensive course in the systematic development of liquid chromatography separations using QbD principles COURSE HANDOUT: [SECTION 2] Separation

More information

RPLC/HILIC/API-MS: polarity extended analysis for organic molecules in water bodies

RPLC/HILIC/API-MS: polarity extended analysis for organic molecules in water bodies RPLC/HILIC/API-MS: polarity extended analysis for organic molecules in water bodies Sylvia Grosse and Thomas Letzel Analytical Research Group Ixia, Rhodes, 1st September, 2015 content polarity extension

More information

Acclaim Mixed-Mode WAX-1 Columns

Acclaim Mixed-Mode WAX-1 Columns User Manual Acclaim Mixed-Mode WAX- Columns 06565 Revision 03 October 05 Product Manual for the Acclaim Mixed-Mode WAX- Column Page of 3 Product Manual for Acclaim Mixed-Mode WAX- Columns 5µm, 0 x 50 mm,

More information

Analysis of Beer by Comprehensive 2D-LC with the Agilent 1290 Infinity 2D-LC system

Analysis of Beer by Comprehensive 2D-LC with the Agilent 1290 Infinity 2D-LC system Analysis of Beer by Comprehensive 2D-LC with the Agilent 1290 Infinity 2D-LC system Application Note Food Testing & Agriculture Authors Edgar Naegele Agilent Technologies, Inc. Waldbronn, Germany Keiko

More information

ProntoSIL C18-EPS Reversed-Phase HPLC Columns

ProntoSIL C18-EPS Reversed-Phase HPLC Columns ProntoSIL C8-EPS Reversed-Phase HPLC Columns Provides excellent separation of polar compounds Better peak shape for acids and bases Stabilized bonded phase for rugged, robust HPLC methods More retentive

More information

Quantitative analysis of small molecules in biological samples. Jeevan Prasain, Ph.D. Department of Pharmacology & Toxicology, UAB.

Quantitative analysis of small molecules in biological samples. Jeevan Prasain, Ph.D. Department of Pharmacology & Toxicology, UAB. Quantitative analysis of small molecules in biological samples 100 Jeevan Prasain, Ph.D. Department of Pharmacology & Toxicology, UAB % 0 300 500 700 900 1100 1300 1500 1700 m/z Class Overview Introduction

More information

Harris: Quantitative Chemical Analysis, Eight Edition CHAPTER 25: CHROMATOGRAPHIC METHODS AND CAPILLARY ELECTROPHORESIS

Harris: Quantitative Chemical Analysis, Eight Edition CHAPTER 25: CHROMATOGRAPHIC METHODS AND CAPILLARY ELECTROPHORESIS Harris: Quantitative Chemical Analysis, Eight Edition CHAPTER 25: CHROMATOGRAPHIC METHODS AND CAPILLARY ELECTROPHORESIS CHAPTER 25: Opener Aa CHAPTER 25: Opener Ab CHAPTER 25: Opener B 25-1 Ion-Exchange

More information

Technical Guide Thermo Scientific Syncronis HPLC Columns

Technical Guide Thermo Scientific Syncronis HPLC Columns Technical Guide Thermo Scientific Syncronis HPLC Columns Consistent, predictable separations, Column after column, time after time Thermo Scientific Syncronis Columns When developing a new method, one

More information

PolyHYDROXYETHYL Aspartamide for Hydrophilic Interaction Liquid Chromatography(HILIC)

PolyHYDROXYETHYL Aspartamide for Hydrophilic Interaction Liquid Chromatography(HILIC) PolyHYDROXYETHYL Aspartamide for Hydrophilic Interaction Liquid Chromatography(HILIC) Hydrophilic Interaction Liquid Chromatography(HILIC) is a variant of normal-phase chromatography which can be performed

More information

The Effect of Contact Angle and Wetting on Performance in Solid Phase Extraction

The Effect of Contact Angle and Wetting on Performance in Solid Phase Extraction The Effect of Contact Angle and Wetting on Performance in Solid Phase Extraction Edouard S. P. Bouvier, Randy E. Meirowitz and Uwe D. Neue Waters Corporation, 34 Maple Street, Milford, MA 01757 USA Presented

More information

Fluophase and Fluofix Columns

Fluophase and Fluofix Columns TG 0-0 Fluophase and Columns Introduction Fluorinated packings exhibit extra retention and selectivity for compounds that have fluorine and chlorine substituents. They also exhibit shape selectivity for

More information

Uptisphere CS Evolution Core Shell columns for fast & highly efficient identification & quantification of small molecules

Uptisphere CS Evolution Core Shell columns for fast & highly efficient identification & quantification of small molecules Uptisphere CS Evolution Core Shell columns for fast & highly efficient identification & quantification of small molecules www.interchim.com www.blog.interchim.com www.forum.interchim.com www.facebook.com/interchim

More information

Rapid Screening and Confirmation of Melamine Residues in Milk and Its Products by Liquid Chromatography Tandem Mass Spectrometry

Rapid Screening and Confirmation of Melamine Residues in Milk and Its Products by Liquid Chromatography Tandem Mass Spectrometry Rapid Screening and Confirmation of Melamine Residues in Milk and Its Products by Liquid Chromatography Tandem Mass Spectrometry Application Note Food Authors Jianqiu Mi, Zhengxiang Zhang, Zhixu Zhang,

More information

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

ACE Excel SuperC18. ACE UHPLC and HPLC Columns. Ultra-Inert UHPLC and HPLC Columns with Extended ph Stability ACE Excel SuperC8 TM Ultra-Inert UHPLC and HPLC s with Extended ph Stability Exploit selectivity at low, intermediate and high ph Use with LC/MS-compatible buffers between ph.. High efficiency µm, µm,

More information

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

QUALITY HPLC COLUMNS PARTICLE PORE PAGE SIZE SIZE. C18 Bidentate C18 Extremely long column life and rugged 4um 100Å 3 Cogent HPLC Columns Manufactured by Table of Contents QUALITY HPLC COLUMNS PARTICLE PORE PAGE SIZE SIZE C18 Bidentate C18 Extremely long column life and rugged 4um 100Å 3 C8 Bidentate C8 Extremely long

More information

Mechanisms of retention in HPLC

Mechanisms of retention in HPLC Mechanisms of retention in HPLC María Celia García-Álvarez-Coque Department of Analytical Chemistry University of Valencia Valencia, Spain https://sites.google.com/site/fuschrom/ 1 Part 3 Mechanisms of

More information

Fast and Flexible Optimization of Modifier Concentrations Using an Agilent 1290 Infinity LC System with Blend Assist

Fast and Flexible Optimization of Modifier Concentrations Using an Agilent 1290 Infinity LC System with Blend Assist Fast and Flexible Optimization of Modifier Concentrations Using an Agilent 129 Infinity LC System with Technical Overview Author A.G.Huesgen Agilent Technologies, Inc. Waldbronn, Germany Abstract During

More information

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

Zirconia-Based Phases as a Powerful Complement to Silica-Based Phases for LC and LC-MS under Non-extreme Mobile Phase Conditions Zirconia-Based Phases as a Powerful Complement to Silica-Based Phases for LC and LC-MS under on-extreme Mobile Phase Conditions Richard A. Henry, Shawn R. Wyatt, Carmen T. Santasania and David S. Bell

More information

Ch.28 HPLC. Basic types of Liquid Chromatography Partition (LLC) Adsorption (LSC) Ion Exchange (IC) Size Exclusion (SEC or Gel Chromatography)

Ch.28 HPLC. Basic types of Liquid Chromatography Partition (LLC) Adsorption (LSC) Ion Exchange (IC) Size Exclusion (SEC or Gel Chromatography) Ch.28 HPLC 28.1 Basic types of Liquid Chromatography Partition (LLC) Adsorption (LSC) Ion Exchange (IC) Size Exclusion (SEC or Gel Chromatography) High Performance (Pressure) LC Glass column st.steel (high

More information

InertSustainSwift C8

InertSustainSwift C8 Physical Properties Silica Particle Size Surface Area Pore Size Pore Volume Bonded Phase End-capping Carbon Loading ph Range USP Code :ES (Evolved Surface) Silica Gel :.9 μm, μm, μm :00 m /g :00 Å (0 nm)

More information

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

Method Development in Solid Phase Extraction using Non-Polar ISOLUTE SPE Columns for the Extraction of Aqueous Samples Technical Note 101 Method Development in Solid Phase Extraction using Non-Polar ISOLUTE SPE Columns for the Extraction of Aqueous Samples This technical note includes by specific information on the extraction

More information

Reversed Phase Chromatography: Mobile phase Considerations

Reversed Phase Chromatography: Mobile phase Considerations Reversed Phase: Mobile Phase Considerations Dr. www.forumsci.co.il/hplc Example of a Reversed Phase Method with UV Detection 1 Chromatographic Process B+A Mobile phase Elution through the Column-movie

More information

Alltech Alltima HP Introduction

Alltech Alltima HP Introduction Alltech Introduction High-Stability, High-Purity, High-Performance, Low-Bleed Columns for Demanding Applications Better Peak Symmetry high-purity silica eliminates peak tailing problems Long Column Life

More information

We recommend you cite the published version. The publisher s URL is:

We recommend you cite the published version. The publisher s URL is: McCalley, D. V. (2015) Study of retention and peak shape in hydrophilic interaction chromatography over a wide ph range. Journal of Chromatography A, 1411. pp. 41-49. ISSN 0021-9673 Available from: http://eprints.uwe.ac.uk/26428

More information

Introduction to Chromatographic Separations (Chapter 1) Many determinations involve separation followed by analysis chromatography electrophoresis

Introduction to Chromatographic Separations (Chapter 1) Many determinations involve separation followed by analysis chromatography electrophoresis Introduction to Chromatographic Separations (Chapter 1) Many determinations involve separation followed by analysis chromatography electrophoresis Chromatography: sample transported by mobile phase electrostatic

More information

Integrated approach to HPLC method development: Using all the tools in the chromatographer s toolbox

Integrated approach to HPLC method development: Using all the tools in the chromatographer s toolbox Integrated approach to HPLC method development: Using all the tools in the chromatographer s toolbox Rosario LoBrutto Pharmaceutical and Analytical Development Novartis Pharmaceuticals Overall Strategy

More information

High Resolution Fast LC

High Resolution Fast LC High Resolution Fast LC Easier Than You Think Rita Steed LC Columns Application Engineer May 9, 2013 What is High Resolution Fast LC? Maintain Resolution with Faster Run Time Increased Resolution with

More information

SEPARATIONS ESSENTIALS IN MODERN HPLC. 2University of Bucharest, Bucharest, Romania

SEPARATIONS ESSENTIALS IN MODERN HPLC. 2University of Bucharest, Bucharest, Romania ESSENTIALS IN MODERN HPLC SEPARATIONS Serban C. Moldoveanu1, Victor David2 'R.J. Reynolds Tobacco Co., Winston-Salem, NC, USA 2University of Bucharest, Bucharest, Romania ELSEVIER AMSTERDAM BOSTON HEIDELBERG

More information

Solid Phase Extraction Method Development Tips and Tricks

Solid Phase Extraction Method Development Tips and Tricks Solid Phase Extraction Method Development Tips and Tricks Carol Haney Ball, Ph.D. Application Scientist Agilent Technologies, Inc. February 3, 2009 Sources of Error Generated and Time Spent During a Typical

More information

Evaluation of ZIC-HILIC columns for the analysis of flavonols

Evaluation of ZIC-HILIC columns for the analysis of flavonols Current Topics in Analytical Chemistry Vol. 9, 212 Evaluation of ZIC-HILIC columns for the analysis of flavonols Aleksandra Sentkowska, Magdalena Biesaga and Krystyna Pyrzynska* Department of Chemistry,

More information

A C18 Silica Column With Exceptional Temperature and ph Stability

A C18 Silica Column With Exceptional Temperature and ph Stability A C18 lica Column With Exceptional Temperature and ph Stability Brian A. Jones 1, Stephanie J. Marin 1, Jody Clark 1, Nathan L. Porter 1, J. Andreas Lippert 2, and Todd M. Johnson 2 1. Selerity Technologies,

More information