G OU IN R 3 AT 0 R tth R B Y E E L L A

E R R

C C



 



 

     1987-2017

April 2017 Volume 30 Number 4 www.chromatographyonline.com

Characterizing Complex Polymer Formulations The benefi ts of coupling SEC with MS to investigate polyether polyol formulations

LC TROUBLESHOOTING PERSPECTIVES IN COLUMN WATCH Increasing resolution by MODERN HPLC New chromatography columns increasing retention New HPLC systems and and accessories for 2017 related products

G OU IN R 3 AT 0 R tth R B Y E E L L A

E R R

C C



 



 

     1987-2017

April 2017 Volume 30 Number 4 www.chromatographyonline.com

Characterizing Complex Polymer Formulations The benefi ts of coupling SEC with MS to investigate polyether polyol formulations

LC TROUBLESHOOTING PERSPECTIVES IN COLUMN WATCH Increasing resolution by MODERN HPLC New chromatography columns increasing retention New HPLC systems and and accessories for 2017 related products All other trademarks are the property of their respective owners. WHOSE TRIPLE QUAD CAN GIVE YOU 15% MORE TIME? Copyright © 2016 PerkinElmer, Inc. 400358A_03. All rights reserved. PerkinElmer® is a registered trademark of SIMPLE: PERKINELMER.

Whether you’re performing environmental, food, or industrial testing, your lab needs to analyze more and increasingly complex samples under tighter regulatory standards. Because our new QSight™ triple-quad LC/MS/MS needs no routine cleaning and maintenance, you get up to 15% more uptime (that’s up to 35 more days per year) to take on your increased testing workload. Plus, our patented technology delivers exceptional sensitivity and reliability along with all that productivity. QSight: A triple quad that simply knows the value of time.

Learn more at www.perkinelmer.com/QSight Published by UBM Americas Editorial Advisory Board

Daniel W. Armstrong Hian Kee Lee University of Texas, Arlington, Texas, USA National University of Singapore, Vice President/Group Digital Production Manager Günther K. Bonn Singapore Publisher Sabina Advani Institute of Analytical Chemistry and Wolfgang Lindner Radiochemistry, University of Innsbruck, Institute of Analytical Chemistry, Mike Tessalone [email protected] Austria University of Vienna, Austria [email protected] Deirdre Cabooter Henk Lingeman Managing Editor Special Department of Pharmaceutical and Faculteit der Scheikunde, Free University, Editorial Director Projects Pharmacological Sciences, University of Amsterdam, The Netherlands Leuven, Belgium Tom Lynch Laura Bush Kaylynn Chiarello-Ebner Peter Carr BP Technology Centre, Pangbourne, UK [email protected] kaylynn.chiarello.ebner@ Department of Chemistry, University Ronald E. Majors of Minnesota, Minneapolis, Minnesota, USA ubm.com Analytical consultant, West Chester, Editor-in-Chief Jean-Pierre Chervet Pennsylvania, USA Antec Leyden, Zoeterwoude, The Alasdair Matheson Art Director Debby Mangelings Netherlands Department of Analytical Chemistry [email protected] Dan Ward Jan H. Christensen and Pharmaceutical Technology, Vrije [email protected] Department of Plant and Environmental Universiteit, Brussels, Belgium Managing Editor Sciences, University of Copenhagen, Phillip Marriot Kate Mosford Senior Director of Copenhagen, Denmark Monash University, School of Chemistry, Danilo Corradini Victoria, Australia [email protected] Marketing and Audience Istituto di Cromatografia del CNR, Rome, David McCalley Data Italy Department of Applied Sciences, Assistant Editor Ronda Hughes-Stovern Hernan J. Cortes University of West of England, Bristol, UK Lewis Botcherby H.J. Cortes Consulting, Robert D. McDowall ronda.hughes-stovern@ubm. Midland, Michigan, USA [email protected] McDowall Consulting, Bromley, Kent, UK com Gert Desmet Mary Ellen McNally Transport Modelling and Analytical Sales Manager DuPont Crop Protection,Newark, Separation Science, Vrije Universiteit, Delaware, USA Oliver Waters Brussels, Belgium Imre Molnár Subscriber Customer Service [email protected] John W. Dolan Molnar Research Institute, Berlin, Germany Visit (chromatographyonline.com) LC Resources, McMinnville, Oregon, Luigi Mondello USA Sales Executive to request or change a Dipartimento Farmaco-chimico, Facoltà Anthony F. Fell di Farmacia, Università di Messina, Liz Mclean subscription or call our customer Pharmaceutical Chemistry, Messina, Italy University of Bradford, Bradford, UK [email protected] service department on Peter Myers Attila Felinger Department of Chemistry, Senior Director, Digital +001 218 740 6877 Professor of Chemistry, Department of University of Liverpool, Liverpool, UK Analytical and Environmental Chemistry, Janusz Pawliszyn Media University of Pécs, Pécs, Hungary Hinderton Point, Department of Chemistry, University of Michael Kushner Francesco Gasparrini Waterloo, Ontario, Canada [email protected] Lloyd Drive, Dipartimento di Studi di Chimica Colin Poole Cheshire Oaks, e Tecnologia delle Sostanze Wayne State University, Detroit, Biologicamente Attive, Università “La Michigan, USA Webcast Operations Cheshire, Sapienza”, Rome, Italy Fred E. Regnier Manager CH65 9HQ, UK Joseph L. Glajch Department of Biochemistry, Purdue Kristen Moore Tel. +44 (0)151 353 3500 Momenta Pharmaceuticals, Cambridge, University, West Lafayette, Indiana, USA [email protected] Massachusetts, USA Harald Ritchie Fax +44 (0)151 353 3601 Jun Haginaka Trajan Scientific and Medical, Milton Project Manager School of Pharmacy and Pharmaceutical Keynes, UK Sciences, Mukogawa Women’s Koen Sandra Vania Oliveira University, Nishinomiya, Japan Research Institute for Chromatography, [email protected] Javier Hernández-Borges Kortrijk, Belgium Department of Analytical Chemistry, Pat Sandra Nutrition and Food Science University of Research Institute for Chromatography, Subscribe on-line at Laguna, Canary Islands, Spain Kortrijk, Belgium www.chromatographyonline.com John V. Hinshaw Peter Schoenmakers Serveron Corp., Hillsboro, Oregon, USA Department of Chemical Engineering, Tuulia Hyötyläinen Universiteit van Amsterdam, Amsterdam, VVT Technical Research of Finland, The Netherlands Finland Robert Shellie 46#4$3*15*0/4
-$r($
&VSPQF
JT
GSFF
UP
RVBMJàFE
SFBEFST
JO
&VSPQF Hans-Gerd Janssen Trajan Scientific and Medical, To apply for a free subscription, or to change your name or address, go to Van’t Hoff Institute for the Molecular Ringwood, Victoria, Australia www.chromatographyonline.com, click on Subscribe, and follow the prompts. Sciences, Amsterdam, The Netherlands Yvan Vander Heyden To cancel your subscription or to order back issues, please email your request to Kiyokatsu Jinno Vrije Universiteit Brussel, [email protected], putting LCE in the subject line. School of Materials Sciences, Toyohasi Brussels, Belgium Please quote your subscription number if you have it. University of Technology, Japan MANUSCRIPTS: For manuscript preparation guidelines, visit www.chromatographyonline.com or call the Editor, +44 (0)151 353 3500. All submissions will be handled with reasonable care, but Huba Kalász the publisher assumes no responsibility for safety of artwork, photographs or manuscripts. Every Semmelweis University of Medicine, precaution is taken to ensure accuracy, but the publisher cannot accept responsibility for the Budapest, Hungary accuracy of information supplied herein or for any opinion expressed. DIRECT MAIL LIST: Telephone: +44 (0)151 353 3500. Reprints: Reprints of all articles in this issue and past issues of this publication are available (250 minimum). Contact Brian Kolb at Wright’s Media, 2407 Timberloch Place, The Woodlands, TX 77380. Telephone: 877-652-5295 ext. 121. Email: [email protected]. © 2017 Advanstar Communications (UK) Ltd. All rights reserved. No part of this publication may be reproduced in any material form (including photocopying or storing it in any Follow us @ LC_GC ‘Like’ our page LCGC Join the LCGC LinkedIn group medium by electronic means and whether or not transiently or incidentally to some other use of this publication) without the written permission of the copyright owner except in accordance with the provisions of the Copyright, Designs & Patents Act 5IF
1VCMJTIFST
PG
-$t($
&VSPQF
XPVME
MJLF
UP
UIBOL
UIF
NFNCFST
PG
UIF
&EJUPSJBM
"EWJTPSZ
#PBSE
(UK) 1988 or under the terms of a licence issued by the Copyright Licensing for their continuing support and expert advice. The high standards and editorial quality associ- Agency, 90 Tottenham Court Road, London W1P 0LP, UK. Applications for the BUFE
XJUI
-$t($
&VSPQF
BSF
NBJOUBJOFE
MBSHFMZ
UISPVHI
UIF
UJSFMFTT
FGGPSUT
PG
UIFTF
JOEJWJEVBMT copyright owner’s permission to reproduce any part of this publication should be LCGC Europe provides troubleshooting information and application solutions on all aspects of forwarded in writing to Permissions Dept, Hinderton Point, Lloyd Drive, Ellesmere separation science so that laboratory-based analytical chemists can enhance their practical Port, Cheshire, CH65 9HQ. Warning: The doing of an unauthorized act in relation to a 10% Post knowledge to gain competitive advantage. Our scientific quality and commercial objectivity copyright work may result in both a civil claim for damages and criminal prosecution. Consumer provide readers with the tools necessary to deal with real-world analysis issues, thereby Waste increasing their efficiency, productivity and value to their employer. www.chromatographyonline.com 175 April | 2017 COVER STORY Volume 30 Number 4 178 Coupling Size-Exclusion Chromatography to Mass Spectrometry for the Analysis of Low-Molecular-Weight Polymers: A Versatile Tool to Study Complex Polyether Polyol Formulations Marcel C. van Engelen, Ron A. Salome, Hamed Eghbali, Melissa N. Dunkle, John R. Stutzman, and Edwin P.C. Mes Complete characterization of low-molecular-weight polyether polyol formulations that are often simultaneously heterogeneous in composition, molecular weight, and architecture can be a daunting task. This article describes the hyphenation of size-exclusion chromatography (SEC) to mass spectrometry (MS) as a promising approach to characterize complex polyether polyol formulated systems.

Columns 190 LC TROUBLESHOOTING Count the Cost, Part 2: Increasing Resolution by Increasing Retention John W. Dolan We will discover how to fi nd the “sweet spot” in terms of retention for a liquid chromatographic separation as well as how much retention change is to be expected for a selected change in organic mobile-phase percentage or column temperature.

196 COLUMN WATCH New Chromatography Columns and Accessories for 2017 David S. Bell Our annual review of new liquid chromatography columns and accessories, introduced at Pittcon and other events.

208 PERSPECTIVES IN MODERN HPLC New HPLC Systems and Related Products: A Brief Review Michael W. Dong This instalment highlights some of the new HPLC systems, modules, accessories, and related technology introduced at Pittcon this year.

Departments 219 Products 222 Events Editorial Policy: All articles submitted to -$t($
&VSPQF are subject to a peer-review process in association with the magazine’s Editorial Advisory Board.

Cover: Original materials courtesy: Timofey_123/ Shutterstock.com

176 -$r($
&VSPQF April 2017

178 techniques such as nuclear magnetic resonance magnetic resonance asnuclear techniques such investigation. To formulations, investigate polyol polyether are under the sample followedschemes on depending Typical tools. ofanalytical deformulations combination and theproper selection formulated systems on depends of analysis markets. Successful profileand competitor provestudy failure analysis, mode patent infringement, developtune product newapplications, performance, weight and composition molecular ofdifferent a blend frequently usedinformulated systems, where they exist as ofthepolyether polyol. Polyether are polyols composition ( asethylene oxideethers such cyclic are sucrose. Oxides typically or assorbitol such pentaerythritol or Polyether PolyolPolyether Formulations A Versatile Tool to Study Complex Polymers: Low-Molecular-Weight Spectrometry for the Analysis of Mass to Chromatography Coupling Size-Exclusion SEC separation and reduction of multiple charging effects. charging ofmultiple reduction and separation SEC efficient and tofast given willbe Attention routines. processing todata approaches best and optimization, method techniques, the interfacing including methods, ofSEC–MS development the with made advances and considerations experimental ofthe some details work This mixtures. complex tounravel these able signifi techniques these Hyphenating systems. polyol formulated polyether complex tocharacterize approach as apromising chromatography ofsize-exclusion hyphenation the describes article This task. adaunting be can architecture and weight, molecular incomposition, heterogeneous simultaneously often are that polyol formulations polyether oflow-molecular-weight characterization Complete Engelen van C. Marcel R f weight. Typical includewater initiators in thepresence ofacatalyst to thedesired molecular active hydrogen one organic atleast oxide an bearing with arePolyether by polyols produced reacting initiator an grow average atan rate annual of4% during2014–2019. will polyols ofpolyether projected thattheconsumption fluids.Itis functional and lubricants include synthetic polyols ofpolyether applications important Other plastics. of consumption oftheglobal 6% makeand upabout blocks for polyurethanes. Polyurethanes are widespread high industrialrelevance. usedasbuilding are They mostly with polyols arePolyether polyols ofpolymeric aclass Mes PO) = & Knowledge of the exact composition is required oftheexact to composition Knowledge D – Analytical Sciences, Midland, Michigan, USA Michigan, Midland, Sciences, D –Analytical 2) or polyols such asglycerin 2) such polyols or 1 , (

Figure 1). are by influenced the properties Product 1 Dow Benelux B.V.–Core Benelux Dow R ( f cantly reduces ion overlap in MS spectra and results in a versatile and powerful tool better better tool powerful and in aversatile results and spectra inMS overlap ion reduces cantly = 4), initiators functional higher or 1 , Ron A. Salome A. , Ron ( EO) propylene and oxide & D – Analytical Sciences, Terneuzen, Sciences, Netherlands, D –Analytical The ( trifunctional, f trifunctional, ( difunctional, 1 , Hamed Eghbali , Hamed = ( NMR), 3) ( 1). 1 , Melissa N. Dunkle N. , Melissa ( ( spectrometry mass time-of-flight desorption–ionization liquid chromatography structure of the formulation structure oftheformulation regarding providecan quantitative information theoxide weight weight ofnumber-average thedetermination allows SEC molecular by may analyses from SEC. beobtained weight information Conversely, remains unknown. the formulation molecular the oxide structure within polyols oftheindividualpolyether weights andHowever, molecular on information detailed type the on information SEC) are often used. When setupcorrectly,SEC) used. are When often MALDI-TOF-MS), and size-exclusion chromatography size-exclusion and MALDI-TOF-MS), • By installing a installing By • results was exploited FastSEC to UHPSEC–MS obtain • was MS to high-resolution SEC nonaqueous Coupling • KEY POINTS experiment would take more than 40 min. 40 take than more would experiment than in less systems. formulated toestablished characterize polyol polyether complex over 1000 Da. for simplifiedspectra polyols for polyether allowing charging ofmultiple was reduced, effect problematic ( M n ) and weight-average molecular weight weight-average) and molecular 1 5 , John R. Stutzman R. , John min, whereas a conventional SEC min,whereas SEC aconventional 210 ( SEC) to mass spectrometry spectrometry SEC) tomass Po source chamber, intheESI the 2 ( ( s) ofinitiators. amount and The Dow Chemical Company–Core Company–Core Chemical Dow The LC), matrix-assisted laser ( 2), to inaddition obtaining 2 , and Edwin P.C. Edwin , and LC•GC Europe LC•GC 13 C NMR C NMR ( April 2017 M w ( MS) )

Photo Credit: Timofey_123/Shutterstock.com Combined forces Nexera MX and LCMS-8060: ultra-fast multiplexing UHPLC meets ultra-trace level detection

The Nexera MX ultra-fast multiplexed UHPLC system combined with Unmatched speed the LCMS-8060 triple quadrupole mass spectrometer provides routine with fastest sample injection, data acquisition and polarity switching high performance LC-MS/MS analysis that makes a real difference in time increasing laboratory efficiency. Nexera MX features two analytical flow lines in a single UHPLC system; this doubles sample processing Dedicated software packages capability when compared to the conventional single channel approach. for automated method and batch creation as well as simplified data The LCMS-8060 adds ultra-fast polarity switching and scanning speed, evaluation highest sensitivity and robustness to push the limits of LC-MS/MS quantitation.

Boosted operating efficiency through multiplexing technology

www.shimadzu.eu/nexera-mx Nexera MX and LCMS-8060 van Engelen et al.

based on an external calibration curve. In addition, the dispersity and the overall functionality (in cases where Figure 1: Reaction of an initiator, where R could be a di, tri, or higher functional species with propylene oxide or the -OH number has been determined separately) can be ethylene oxide to form a polyether polyol. obtained (3). Detailed structural information on the nature of the formulation, however, is limited. Off-line coupling of Catalyst LC or SEC with MALDI-TOF-MS may provide more detailed R OH n m O O OOR O H information regarding the distributions observed. However, T m this is labour intensive because off-line coupling is not n Initiator Propylene oxide Ethylene oxide Polyether straightforward because of the need for spotting the SEC (PO) (EO) Polyol eluent on the MALDI plate, evaporating the solvent, and derivatizing the spots prior to laser ablation (4). Figure 2: Schematic layout of the analytical system with LC coupled to MS is a well-established tool in structure post-column addition of organic modifi er to enhance elucidation. Conversely, SEC directly coupled to MS is ionization and parallel UV-RI and MS detection. less common as is clear from the number of publications 1 mL/min SEC System DAD RID available. Whereas nearly 6000 publications are available 0.6 mL/min

on the topic of LC–MS (2016), a Scifinder search lists a 0.4 mL/min Post column 0.1 mL/min total of less than 50 publications on the topic of SEC–MS MS ADC Data Modifier 0.5 mL/min (2001–YTD). Although practiced since the early 1990s (5), today one of the most active groups in SEC–MS is the Barner-Kowollik group (6). Figure 3: Example of a SEC–RI–MS chromatogram of = = A workflow was established that benefits from both the a polyether polyol mixture (Red TIC; Blue RI). The spectra at the top of the distributions are shown as (a), separation power of SEC and the identification strength (b) (Homopolymer M <2500 Da) and (c), (d) (Copolymer of high-resolution mass spectrometry. A regular SEC w M > 4000 Da). system coupled to an ultraviolet refractive index detector w (UV-RI) setup was extended by introducing MS detection parallel to the optical detectors. As such, MS offered the possibility of identification next to the quantitation by RI. As the optical cell of an RI detector is pressure sensitive (d) (c) (b) (a) while MS is a destructive technique, both detectors need to be the last in line of the detection scheme. To have two

final detectors, the flow after the columns was split and 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 one part was directed towards the UV and RI detector in Response Units (%) vs. Acquisition Time (min)

series (Figure 2) and the other part was directed towards z=1 (a)z=5 (c) 1096.7927 821.5844 z=3 z=3 the MS. Although other modifiers exist, ammonium 557.4137 z=4 1371.9578 1022.4728 formate was chosen because polyether polyols easily form z=2 z=2 2012.3968 ammonium adducts. As ammonium formate is not soluble in tetrahydrofuran (THF, typical solvent for polyether polyol 200 400 600 800 1000 1200 1400 1600 1800 2000 2200 2400 2600 2800 200 400 600 800 1000 1200 1400 1600 1800 2000 2200 2400 2600 2800 SEC) and the polystyrene/divinylbenzene columns used Counts (%) vs. Mass-to-Charge (m/z) Counts (%) vs. Mass-to-Charge (m/z)

are not compatible with water, the modifier was added z=2 (b)z=7 (d) 1014.2348 836.4928 post-column before entering the source of the mass z=2 682.1690 z=1 spectrometer. As the RI detector is not supported by the 2009.4691 z=6z=5 z=4 z=8 1344.9768 z=3 software version used, the RI data was acquired via an AD 2175.9084 Converter (ADC). 200 400 600 800 1000 1200 1400 1600 1800 2000 2200 2400 2600 2800 3000 200 400 600 800 1000 1200 1400 1600 1800 2000 2200 2400 2600 2800 Counts (%) vs. Mass-to-Charge (m/z) Counts (%) vs. Mass-to-Charge (m/z) Experimental Chemicals: Samples were prepared at an approximate with two optical detectors in series; a diode array detector concentration of 1 mg/mL in THF. Prior to separation, the scanning from 190 nm to 400 nm followed by a differential samples were filtered over a PTFE 0.45 μm Millex LCR refractive index detector set at 35 °C. syringe-driven filter unit (EMD Millipore). SEC separation The LC–MS and LC–charge reduction–MS (LC–CR–MS) was performed in THF (J.T. Baker; non-stabilized, HPLC experiments were performed on an Agilent 1290 Infinity grade). Ammonium formate (Fluka; LC–MS grade) was II LC system. PEG3800 analyzed under reversed-phase used as 0.2% solution in water (Millipore Milli-Q grade). separation conditions was used to demonstrate the The MS tuning solution was obtained from Agilent (ESI-L technique. The LC–MS and LC–CR–MS conditions and Low concentration tuning mix G1969). experimental design have been previously described (7). Chromatographic System: All SEC–UV-RI–MS Columns: Injections ranging from 1 to 20 μL were made separations were performed on an Agilent Technologies on four 7.5 mm × 30 cm, 5-μm PS/DVB packed columns 1260 infinity system. The system consisted of an (Agilent) connected in series. The columns were coupled autosampler, a binary pump delivering 1 mL/min of THF, in decreasing porosity: 103 Å, 500 Å, 100 Å, and 50 Å. For and an isocratic pump set at a flow rate of 0.1 mL/min to the ultrahigh-performance size-exclusion chromatography introduce the organic modifier. Solvents were degassed (UHPSEC) experiments, two 4.6 mm × 15 cm, 1.7-μm using a degasser. Columns were placed in a thermostatic Acquity APC XT 45 columns (Waters) were connected column compartment at 35 °C. The system was equipped in series. These specific type of columns display a

180 LC•GC Europe April 2017 Thinking Forward. GPC/SEC Theory or practice? PSS are world worldld lead lleaders ders ini mmacromolecularacromolellhcular chcharacteri-aractet rii- zation and have the expertise to help you with your analysis requirements. We offer a range of products and services from instruction courses and training If there is one thru contract analysis, consulting, method develop- ment and qualifi cation services all the way up to thing we can supplying turnkey GPC/SEC and LC/2D systems. do, it’s both. All this comes with the personal and direct support of our dedicated team of innovative and pioneering specialists. Is there any better way of achieving your analysis goals?

Driving GPC/SEC forward

Phone + 49 6131 962390 You‘ll fi nd the ideal GPC/SEC solution at PSS under: www.pss-polymer.com [email protected] van Engelen et al.

Figure 4: Overlay of a typical SEC–MS chromatogram Figure 5: van Deemter plot (H vs. u0) obtained with a with a heat map of the observed m/z values. The heat single APC XT 45 column for benzene as test component map shows a clear separation of the oxide structure with THF as the mobile phase. Each data point was where the x-axis shows an increase in EO and the y-axis measured in triplicate and averaged (RSD <3%) (black visualizes the increase in PO. circles). Pressure vs. u0 plot where the pressure values were corrected for the system pressure (empty squares).

1200 25 600 1100

1000 500 20 900

800 400 2 15 700

Mass (Da) 1 600 300 H (μm) 10

500 (bar) Pressure 200 400

300 5 100 13 14 15 16 17 18 19 20 21 Retention Time (min) 0 0 0.0 0.5 1.0 1.5 2.0 2.5 3.0 u (mm/s) linear separation range between 200 Da and 5000 Da 0 (polystyrene-based). Mass Spectrometer: Parallel to the optical detectors Figure 6: BPC and TIC showing the analysis of a typical an Agilent accurate-mass quadrupole time-of-flight polyether polyol. Two APC XT 45 (L = 15 cm) columns (QTOF) mass spectrometer was used for detection. The were coupled in series in combination with QTOF-MS TOF-MS was tuned to 12k resolution over the m/z range detection. 50–3000 in positive electrospray. Stable spray conditions were obtained at a nebulizer pressure of 50 psi and a drying gas at 10 L/min at 350 °C. The capillary voltage 1.4 was set to 4500 V with the fragmentor and skimmer at 150 V and 65 V, respectively. Mass correction was 1.2 BPC

performed against THF (m/z 73.06479; eluent) and HP-921 1 DEG

(m/z 922.009798; hexakis[1H,1H,3H-tetrafluoropropoxy] 2 (mw = 106) 0.8 phosphazine delivered by the instrument calibrant delivery x10 system). 0.6 For the LC–MS and LC–CR–MS measurements the 0.4 H20 + 27 EO = chromatographic system was coupled to a G6538 UHD 0.2 (mw 1206) accurate mass QTOF mass spectrometer (Agilent) (7). 0 Data System: Both LC and MS instrumentation were 1.4 1.6 1.82 2.2 2.4 2.6 2.8 33.23.4 3.6 3.8 4 4.2 4.4 4.6 controlled by Mass Hunter software (Agilent). The RI Counts (%) vs. Acquisition Time (min) detector was controlled by a 1200 instant pilot (Agilent) and the RI analog signal was fed into the Mass Hunter acquisition system via an AD converter. suggesting the polymer is based on propylene oxide 6 = + ( 58 Da). The charge adduct in this series is NH4 . The Results and Discussion monoisotopic mass of 1096.7927 Da suggests a formula Homopolymer: Figure 3 displays a typical SEC–RI– of C54H110O20 (error 0.16 ppm). Calculation of the residual MS chromatogram. The figure shows an overlay of mass results in a residual mass of 34, suggesting the the RI and the total ion count (TIC), blue and red line polyether polyol is a glycerin-initiated PO polyether polyol respectively, for an artificial mixture of polyether polyols. (Table 1). The accompanying mass spectra are shown below the chromatogram. MOD(Neutral mass,mass repeating unit) = Spectrum A is a typical spectrum for a homopolymer. MOD(1096.7927-18.03383,58.04186) = 34 [1] Identification of a homopolymer is straightforward because the oxide structure follows from the spacing of the Similarly, Figure 3(b) exhibits a spacing of 58, which individual oligomers (EO: 44 Da; PO: 58 Da). The initiator suggests the distribution centred around m/z 2009.4691 is can easily be determined from the residual mass of the most likely a water-initiated PO polyether polyol. polymer, that is, the mass remaining after subtraction of the As seen from Table 1, the residual mass can provide maximum integer number of monomers. The residual mass an indication on the initiator of the homopolymer. It can be determined by using the simple Excel function is, however, not a unique number and in some cases “MOD”. additional information—from, for example, NMR—may be For example, Figure 3(a) shows a typical spectrum for required to fully establish the polyether polyol structure. In a homopolymer, with the spacing of the isotope clusters the m/z 2009 example, an alternate suggestion may have

182 LC•GC Europe April 2017 van Engelen et al.

Figure 7: Diagram of the 210Po α-particle source in the ESI source of the MS. The α-particle source is positioned within a few millimetres on the electrospray ionization capillary. Emitted α-particles interact with ambient gases and solvent molecules, forming bipolar reagent ions that undergo further reactions with the analyte ions. These reactions result in charged particle transfer from the analyte + ions to the reagent ions (that is, NH4 transfer), generating charged-reduced analyte ions prior to entering the mass spectrometer. World leaders in

LC thermal desorption for environmental air monitoring

Ion source

MS

Tailor your TD system to your requirements – canisters, 210Po source on-line and sorbent tubes been a trimethylolpropane-initiated often leads to overlapping isotope polyether polyol because that would clusters (Figure 3[c] and [d]) and other result in the same m/z and residual measures need to be taken (vide infra). mass. Copolymer: The identity of Once the initiator and the oxide copolymers by SEC–RI–MS is more structure are established, a formula difficult and additional information database can be created, which from other sources may be needed. allows a targeted approach to fully Whereas the oxide structure can assign the polymer distribution. readily be determined by the spacing When assuming the response of of the isotope clusters, for example, the individual oligomers within the in the case of an EO–PO mixture, 6 = 6 = distribution being equal over the PO-PO 58, EO-EO 44, and 6 = distribution, a series of extracted ion EO-PO 14, the identity of the chromatograms or single oligomer initiator can be obtained from NMR. ɵ Maximise efficiency by running profiles can be generated and Alternatively, if the number of potential VVOC to SVOC on one platform integrated to determine the M and initiators is small, its identity may n ɵ Sample splitting and re-collection Mw of the polymer using the formula be calculated using the molecular below (8): formulas obtained from the accurate for easier validation and method masses. compliance N M 2 = i i NiMi Based on the calculated formula Mn Mw= [2] ɵ Reduce your costs – avoiding Ni NiMi the EO–PO ratio per peak can use of liquid cryogen Ni is the ion count of the single readily be calculated and assigned. oligomer profile as determined from For example, in Figure 4 a SEC– the total ion chromatogram and Mi MS chromatogram is shown where the molecular weight of that oligomer. the dotted overlay is a graphical Identification of the distributions representation of the observed Find out more becomes more cumbersome when masses versus retention time forming http://chem.markes.com/XR moving to higher molecular weights a heat map. As is clear from the and copolymers. Once the molecular figure, there is a distinct ordering in weight starts to be higher than the observed masses. For example, 1000 Da, multiple charging starts to when looking at the blue dot assigned occur. For homopolymers this still “1” a m/z 673 was observed. Accurate results in reasonably well interpretable mass calculations revealed an spectra, but for copolymers this elemental composition C33H68O13, www.chromatographyonline.com 183 van Engelen et al.

while from NMR it was known that no specific initiators Figure 8: (a) Total ion chromatogram obtained under besides H2O were present. Therefore: positive ionization from the LC–MS analysis of PEG3800. C33 H68 O13 = H2O + n C2H4O + m C3H6O (b) Summed spectra of the peak eluting from 10–12.5 min. (c) Summed CR spectra from 11–12.5 min. (d) Summed C : n 2 + m * 3 = 33 * [3] CR spectra from 10–11 min front shoulder. O : n + m = 12 = = n 3; m 9 1.1 (a) 1 0.9 When moving up along the blue dots, an increase of m/z 0.8 0.7 H O with spacing of 44 can be observed indicating an increase in 2 0.6 O H

x10 0.5 0.4 EO. The entire “blue” distribution as such can be assigned as 0.3 0.2 a water-initiated (PO)9 with 3–15 EO. Similarly, the “red” series, 0.1 0 starting with 2, can be assigned as series based on 10 PO -0.1 1234567891011121314 and 3–14 EO. As such, only a few of the oligomers have to be Counts (%) vs. Acquisition Time (min) assigned to identify the entire distribution because one can (b) 1.8 6+ observe an increase in EO loading along the y-axis, while the 1.6 5+ 1.4 + n+ 7 [M+nNH4] x-axis shows an increase in PO. These plots can be generated 1.2 3 1 4+ without difficulty using commercial software or open source x10 0.8 tools such as MZMine or OpenChrom (9,10). When moving 0.6 0.4 3+ from left to right through the chromatogram the PO loading 0.2 2+ 0 increases observed by a 58 Da spacing, whereas the y 500 750 1000 1250 1500 1750 2000 2250 25000 2750 3000 3250 3500 3750 4000 4250 4500 4750 5000 direction sees an increase in EO with a 44 Da spacing. So Counts (%) vs. Mass-to-Charge (m/z) once a few peaks have been identified, one is able to “walk” (c) 3 + + through the distribution. 2.5 [M NH4] Ultrahigh-Performance SEC–MS for Fast Identifi cation: 2 1

Over the last couple of years there has been a trend towards x10 1.5 miniaturization in SEC. Columns with smaller inner diameter 1 0.5 (i.d. = 4.6 mm) and particle sizes (

x10 3 time can be obtained at the cost of higher back pressures. 2.5 2 1.5 This miniaturization trend is analogous to the trend that 1 0.5 adsorption chromatography has undergone already with 0 500 750 1000 1250 1500 1750 2000 2250 25000 2750 3000 3250 3500 3750 4000 4250 4500 4750 5000 the introduction of ultrahigh-performance chromatography Counts (%) vs. Mass-to-Charge (m/z) (UHPLC), which is in a more mature stage. Various manufacturers have recently started the commercialization of products following the aforementioned miniaturization separation window of the analytical method because of the trend (14,15). A recent system for polymer analysis that was pronounced polydispersity of certain samples. Ultimately, introduced allows sized-based separations with high resolution all of the above indicates that advanced instrumentation to be achieved in very short analysis times (14). The column allowing high pressures (>1000 bar) with minimized external stationary phase used is based on sub-3-μm ethylene-bridged band broadening is required to run these columns at hybrid (BEH) silica particle technology, which reportedly optimal conditions. Since the application of these columns provides an increased mechanical strength (16). can shorten the analysis time significantly with respect Figure 5 shows a typical van Deemter plot obtained to conventional SEC, the combination with MS detection with such a sub-3-μm particle column where the flow rate was investigated to explore the possibility of fast polymer was varied between 0.1 mL/min and 1.7 mL/min. It can identification. be observed that the shape of the curve is very flat in the Figure 6 shows a UHPSEC–MS chromatogram (BPC high-velocity region (C-term region) where plate height values and TIC) of a typical polyether polyol. A fast separation = as low as Hmin 3.9 μm are reached. It clearly illustrates that was achieved in less than 5 min, which is in contrast to this type of column can be operated at high flow rates without conventional SEC where a separation run can easily take sacrificing column efficiency. To operate at the optimum more than 40 min. The BPC signal shows that the sample conditions (best performance [Hmin] and shortest analysis contains multiple components with different molecular weights. = time) one should work at a linear velocity of u0 2.5 mm/s, Despite the fact that some external band broadening in the which corresponds with a column pressure close to 500 bar. MS source is inevitable, the good separation resolution of this Extrapolating the van Deemter plot in Figure 5 towards higher type of stationary phase is still reflected. Identification with mobile phase velocities even projects that working at higher MS revealed that the various separated components were pressures could bring additional gain in separation speed polyether oligomers formed by polymerization of EO where without affecting the efficiency. In SEC it is also very common water served as initiator. Oligomers with a Mw up to 1200 Da to couple different columns in series to extend the overall could be identified.

184 LC•GC Europe April 2017 ® Valves, fittings, detectors, and more for chromatography and liquid handling Generation of Calibration Gas Standards

Dynacalibrator® Model 505 s4OUCHSCREENCONTROL s4WOCHAMBERSWITHINDEPENDENT temperature control s%XTREMELYSTABLETEMPERATURECONTROLn deviation of <0.002°C at set point / 1°C change in ambient temperature s5TILIZESSAFE CONVENIENT$YNACAL® permeation devices

Dynacal®0ERMEATION4UBES s00"TOHIGH00-RANGE s!CCURATE STABLECONCENTRATIONS s%CONOMICAL mEXIBLEALTERNATIVETO BULKYBOTTLEDMIXTURES s.)34TRACEABLE

North America, South America, and Australia/Oceania contact: Europe, Asia, and Africa contact: ® Valco Instruments Co. Inc. ® VICI AG International tel: 800 367-8424 tel: Int + 41 41 925-6200 fax: 713 688-8106 fax: Int + 41 41 925-6201 [email protected] [email protected] van Engelen et al.

carriers superimposed on each other). The multiple charging Table 1: Examples of residual masses. The residual mass phenomenon can easily be observed in Figure 3(c) and (d); as is obtained by subtracting the maximum integer number the molecular weight of the polymer increases, an increase in monomer units from the observed neutral mass. For example, glycerin (M = 92) would give a residual mass of multiple charging occurs. 34 on a propylene oxide structure (92 – 58 = 34) whereas Different strategies exist for overcoming the challenges sucrose: 342 – 5*58 = 52. of multiple charging. Charge reduction mass spectrometry (CRMS) is a chemical approach to minimize the complexity of Initiator EO PO mass spectra by stripping or removing charge from gaseous

H2O 18 18 ions (18). In the current study, CRMS was performed to reduce the multiple charging of poly(ethylene) glycol. This technique Glycerin 434 has been widely studied for the direct infusion of biopolymers TMP 218 (19–22); however, the coupling of liquid chromatography to Pentaerythritol 420charge reduction mass spectrometry (LC–CRMS) has only Sucrose 34 52 recently been applied (7). Stutzman and coauthors were able Sorbitol 68to accomplish the coupling of LC to charge reduction MS by introducing a polonium-210 α-particle source into the ESI UHPSEC with MS Detection: When coupling with MS, source (Figure 7). Briefly, the α-particle source is positioned one needs to take into account that a regular electrospray within a few millimetres of the electrospray ionization capillary. source cannot handle more than 1 mL/min of column effluent Emitted α-particles interact with ambient gases and solvent without compromising instrument performance. When setting molecules, forming bipolar reagent ions that undergo further up UHPSEC in combination with MS detection, several reactions with the analyte ions. These reactions result in considerations related to data acquisition need to be taken charged particle transfer from the analyte ions to the reagent + into account. In a regular SEC separation, where peaks of the ions (NH4 transfer), generating charge-reduced analyte ions individual oligomers can easily be a half a minute (FWHM) prior to entering the mass spectrometer. wide, an acquisition rate of 1 spectrum per second is typically In Figure 8 the charge reduction effect is clearly used with RI and UV detectors and even a triple quad may demonstrated for the LC–CRMS analysis of a relatively simple be used. With such an aquisition rate, sufficient points are 3800 molecular weight poly(ethylene glycol) (PEG3800) collected to properly describe the chromatographic peak and sample. Figure 8(a) shows first the positive mode total to allow for integration. In the example shown in Figure 6, a ion chromatogram (TIC) from the reversed-phase LC–MS single oligomer elutes in a peak only 2 s wide. Using a typical analysis of the PEG standard. Ion signal was observed from acquisition rate would be far too slow to properly analyze approximately 10–12.5 min. Mass analysis of the PEG3800 such a sample. Fortunately, the acquisition rate of a TOF response generated an ion signal consistent with multiply + 2+ instrument can be readily adjusted because each individual ammoniated PEG cations (Figure 8[b]), that is, [M 2NH4] + 7+ 210 mass spectrum is composed of thousands of transients over through [M 7NH4] . With the introduction of the Po the entire mass range. By decreasing the number of transients α-particle emitter into the ion source of the mass spectrometer, for each spectrum, the acquisition rate of the instrument can be CRMS was performed following liquid separation. Mass increased. The increased acquisition rate, however, invariably analysis of the PEG3800 response generated a distribution leads to a decrease in the signal-to-noise ratio (S/N) because of singly charged ions centred at approximately m/z 4000, less transients are used to build the spectrum. Depending which was consistent with the water-initiated PEG3800 on the mass range of interest, the loss in S/N can (partly) be standard (Figure 8[c]). Notably, on the front shoulder of the compensated by changing the pusher frequency and limiting PEG TIC response, a distribution ranging from approximately the acquired mass range. With the decreased mass range the m/z 1200 to 2800 was also observed, likely arising from an number of transients per spectrum is increased (typically 14 k impurity (Figure 8[d]). The ion signal from the lower molecular per second for m/z 20–1700). One has to be aware that species weight PEG distribution was later identified and confirmed beyond the set mass range may appear as low-molecular-weight in the multiply-charged mass spectrum; however, the low artefacts in the spectrum because the pusher may operate faster relative abundance compared to PEG3800 and overlapping than the time required to “empty” the flight tube of the instrument. distributions initially limited its identification. Overall, LC–CRMS Multiple Charging: Multiple charging is a phenomenon of PEG3800 exhibited simplified mass spectra and enabled of electrospray ionization that occurs when molecules enhanced characterization of the complex polyether polyol containing several charge-carrying functional groups ionize mixture. By combining LC–MS with LC–CRMS data, one has (17). Multiple charging of polyether polyols typically occurs an additional powerful method to decipher complex polyether starting from a molecular weight of approximately 1000 Da polyol formulations containing material > 1000 Da. and ultimately adds to the complexity of the spectrum. An interesting observation is that the multiple charging occurs Conclusions by the presence of different combinations of charge carriers Detailed analysis of polyether polyol formulations was + + + + + (n*H , n*NH4 , and n*NH4 (m-n)*H ). Multiple charging achieved by coupling SEC to RI–MS. The oligomer is useful in the identification of proteins because it shifts the distributions (in terms of molecular weight and oxide structure) charge state distribution to lower values, thereby allowing as well as initiator type could be determined. Homopolymer the detection of higher molecular weight species. However, systems can sometimes be fully unravelled by SEC–RI–MS, for polydisperse samples such as polyether polyols, it is whereas more complex copolymer polyether polyol systems detrimental for the identification because several occurrences often require additional data from other techniques, such as overlap (different oligomers, charge states, and charge NMR. UHPSEC coupled to RI–MS can significantly increase

186 LC•GC Europe April 2017 van Engelen et al. resolution and simultaneously reduce the analysis time by a factor of 5 to 6. Multiple charging of higher Mw species significantly hampers the correct deformulation of complex polyether polyol systems. Installation of a 210Po source close to the ESI spray in the MS source reduces the average charge Brighter state to singly charged species and thereby significantly Separations simplifies the spectra. We have shown that LC–CRMS technology is particularly useful for the characterization of State-of-the-art HPLC complex polyether polyols containing material > 1000 Da. Columns for small mole- References cules bringing you reliable, (1) M.F. Sonnenschein, Polyurethanes: Science, Technology, Markets and Trends (John Wiley & Sons, Inc., Hoboken, New Jersey, USA, 2015). brilliant results (2) ASTM, D4875 - 11: Standard Test Methods of Polyurethane Raw Materials: Determination of the Polymerized Ethylene Oxide Content of Polyether Polyols. 2011, ASTM International. (3) ASTM, D4274 - 16: Standard Test Methods for Testing Polyurethane Raw Materials: Determination of Hydroxyl Numbers of Polyols. 2016, ASTM International. (4) S.M. Weidner, J. Falkenhagen, and I. Bressler, Macromol. Chem. Phys. 213, 2404–2411 (2012). (5) W.J. Simonsick and L. Prokai, Adv. Chem. Ser. 247, 41–56 (2009). (6) T. Gruendling, S. Weidner, J. Falkenhagen, and C. Barner-Kowollik, Polym. Chem. 1, 599–617 (2010). (7) J.R. Stutzman, M.C. Crowe, J.N. Alexander, B.M. Bell, and M.N. Dunkle, Anal. Chem. 88, 4130–4139 (2016). (8) Agilent, Polymer Molecular Weight Distribution and Definitions of MW Averages, 5990-7890EN (2015). (9) T. Pluskal, S. Castillo, A. Villar-Briones, and M. Orešic񡚕 , BMC Bioinformatics 11, 1–11 (2010). (10) M. Sturm, A. Bertsch, C. Gröpl, A. Hildebrandt, R. Hussong, E. Lange, N. Pfeifer, O. Schulz-Trieglaff, A. Zerck, K. Reinert, and O. Kohlbacher, BMC Bioinformatics 9, 1–11 (2008). (11) M. Janco, J.N. Alexander, E.S.P. Bouvier, and D. Morrison, J. Sep. Sci. 36, 2718–2727 (2013). (12) E. Uliyanchenko, P.J. Schoenmakers, and S. van der Wal, J. Chrom. A 1218, 1509–1518 (2011). (13) E.S.P. Bouvier and S.M. Koza, TrAC Trend. Anal. Chem. 63, 85–94 (2014). (14) Waters, ACQUITY Advanced Polymer Chromatography System, LITR134729285 (2015). (15) S. Luke, P. Cooke, and G. Cleaver, GPC/SEC miniaturization delivers lower solvent costs, higher throughput, and excellent separations. Access Agilent eNewsletter, (2015). (16) Waters, ACQUITY APC Columns, LITR134729585 (2014). (17) J. Pitt, Clin. Biochem. Rev. 30, 19–34 (2009). • Monolithic - Chromolith® (18) L. Gong and J.S.O. McCullagh, Rapid Commun. Mass Spectrom. 28, 339–350 (2014). Low backpressure – Long lifetime - (19) D.D. Ebeling, M.S. Westphall, M. Scalf, and L.M. Smith, Anal. Chem. Suitable for dirty samples – Available 72, 5158–5161 (2000). in micro format (20) W.J. Herror, D.E. Groeringer, and S.A. McLuckey, Anal. Chem. 68, ® 257–262 (1996). • Fused core particles - Ascentis (21) M. Scalf, M.S. Westphall, J. Krause, S.L. Kaufman, and L.M. Smith, Express – Efficiency – Peak symme- Science 283, 194–197 (1999). try – Wide range of phases (22) J.L. Stephenson and S.A. McLuckey, J. Am. Chem. Soc. 118, 7390–7397 (1996). • Fully porous particles - Purospher® STAR – 100% aqu- Marcel van Engelen is a research scientist focusing on NMR ous phase compatibility – pH stability 1.5 – 10.5 – Column/batch reproduc- and MS techniques. ibility – High loadability Ron Salome is a senior chemist and is an expert in liquid chromatography. sigma-aldrich.com/hplc-columns Hamed Eghbali is a senior chemist specialized in advanced LC separation and detection techniques. Melissa Dunkle is a senior chemist focusing on UHPLC–MS and advanced GC techniques. Edwin P.C. Mes is a senior R&D manager of Core R&D specialized in advanced separation and detection techniques. Marcel, Ron, Hamed, Melissa, and Edwin are all members The life science business of Merck operates of Core R&D - Analytical Sciences at Dow Benelux B.V. in as MilliporeSigma in the U.S. and Canada. Terneuzen, The Netherlands. Copyright © 2017 Merck KGaA. All Rights Reserved. Merck and the vibrant M are trademarks and Chromolith and Purospher are registered trademarks of Merck. Ascentis is a registered trademark of Sigma-Aldrich John Stutzman is a senior chemist focused on advanced Co. LLC. or its affiliates. MS techniques. He is a member of Core R&D - Analytical Sciences at The Dow Chemical Company in Midland, USA. www.chromatographyonline.com 187 MarvelXACT: A Worry-Free Fitting System for Liquid A Q&A Chromatography

hromatographers need and want We also wanted to create a fitting that is Crepeatable, reproducible results. A highly re-usable. Ultra high-performance key aspect of ensuring such repro- liquid chromatography (UHPLC) is typically ducibility is avoiding guess-work when tight- run at very high pressures, and there’s very ening fittings. To learn more about a recent few tubing material and fitting technology innovation in liquid chromatography con- out there that lends itself to such pressure. If nections called MarvelXACT, LCGC recently fittings are highly re-usable, labs don’t have Eric Beemer spoke with Eric Beemer, senior development to discard items every time they change Senior Development Engineer engineer at IDEX Health & Science. columns or connections, thereby increasing and Inventor of MarvelXACT IDEX Health & Science the value of our new technology. LCGC: Why did IDEX develop MarvelXACT and what kind of field feedback led to it? Ease of use is also important. Although we have some existing finger-tight con- Beemer: At IDEX, we’ve done a lot of nections that don’t require tools, concerns Voice of the Customer (VOC) analysis in about whether they are properly assembled the field and have had close communica- still exist. With MarvelXACT, the connection tion with customers to understand the pain is not a concern anymore because it enables points of chromatography users and to de- an exact, proper connection every time. velop a set of criteria from which to work. We also have significant chromatography LCGC: What are the key features of experience from our internal lab work MarvelXACT? and incorporated that knowledge into the MarvelXACT specifications. Beemer: One key feature is its torque- limiting mechanism. Torque is important We wanted to develop a “worry-free” fit- because if you don’t have enough torque, ting. By “worry-free,” I mean no carryover, no leaks can develop—and with leaks come peak broadening, or any kind of tailing—just bad chromatography. repeatable chromatography. I think that’s what all chromatographers want. The leaks By applying enough torque, you maximize that we tend to see with fittings currently the fitting’s sealing potential. If you have available on the market have a big impact too much torque, you can damage the on troubleshooting time and chromatogra- port or the tubing itself. Once the tubing phy results. is damaged, an increase in back pressure usually occurs, which can be detrimental for achieving the flow rates that might be required by the analysis.

SPONSORED BY MARVELXACT: A WORRY-FREE FITTING SYSTEM FOR LIQUID CHROMATOGRAPHY

With MarvelXACT, by having a torque-limiting “click” element, but MarvelXACT builds upon MarvelX by adding mechanism, the proper amount of torque is provided to the torque-limiting mechanism to make it easier to use. maximize the seal while preventing too much torque, which Nonetheless, much of the mechanism for creating a seal can limit the tubing’s re-usability and the functionality. The and the resulting chromatographic results are the same as click also provides good feedback; you know it has been those previously obtained on the proven MarvelX technology. properly assembled. It’s like your automobile’s gas cap. MarvelXACT has been extensively tested to verify a superior When you hear the click, you know that you’ve assembled product incorporating these new elements. it properly and that it will seal every time. LCGC: Is MarvelXACT a patented technology? Another key feature is face-sealing. Face-sealing is basi- cally how the end of the tubing seals at the bottom of the Beemer: Yes; both the torque mechanism and the fitting port. No cones, ferrules, or other components are needed. have been granted patents. The tubing itself and the face A load is applied to the back of the seal to create the seal sealing element is patent pending. on the front end. MarvelXACT connections are available in either stainless steel or all PEEK flow path options, the latter LCGC: What is IDEX’s vision on MarvelX technology? providing a flow path that is smooth and very inert, which is What should users expect for the near future? important for “sticky” applications like protein separations. Beemer: IDEX Health & Science is committed to the growth LCGC: IDEX launched a new connection about a year of the MarvelX product family as evidenced by the expansion ago called MarvelX. What are the similarities and dif- of the face-sealing MarvelX technology used in MarvelXACT. ferences between MarvelXACT and MarvelX? We’re looking at ways we can use different tubing internal diameters and are experimenting with different materials Beemer: MarvelXACT is really just the advanced version such as a new fused silica flow path to try and open up other of MarvelX. It still utilizes the same tubing and face-sealing applications of the technology.

IDEX Health & Science is the global authority in fluidics and optics, bringing life to advanced optofluidic technologies with products, people, and engineering expertise. The company is respected worldwide for solving complex problems and de- livering complete path innovation for analytical, diagnostic, and biotechnology applications for the life sciences market. As a genuine and trustworthy partner, IDEX Health & Science solves its customers’ most demanding challenges with the industry’s most extensive portfolio of state-of-the-art components and capabilities that are unrivaled in breadth, quality, performance, and design. IDEX’s vision of the complete path goes far beyond just meeting its customers’ needs—it anticipates them, with intelligent solutions for life.

Product offerings include: connections, valves, pumps, degassers, column hardware, manifolds, flow cells, microfluidics consumables, sensors, integrated fluidic systems, optical filters, lenses, shutters, laser & light engines, and integrated optical systems. For more information, visit: www.idex-hs.com LC TROUBLESHOOTING Count the Cost, Part 2: Increasing Resolution by Increasing Retention

John W. Dolan, LC Troubleshooting Editor

We will discover how to fi nd the “sweet spot” in terms of retention for a liquid chromatographic separation as well as how much retention change can be expected for a selected change in organic mobile-phase percentage or column temperature.

R = N0.5 α k + k k This is the second instalment s ¼ ( -1)( /[1 ]) [1] some increase in , the cost of i ii iii k R in a series about how to use ( ) ( ) ( ) an increase in to increase s, and chromatography fundamentals the stability of the separation as it to estimate the impact of various where α is the selectivity between relates to k. k k k parameter changes on liquid two peaks with -values of 1 and 2: The Advantages of Increased chromatography (LC) separations. k-Values: The plot of Figure 1 α = k k k In the first discussion (1) we looked 2/ 1 [2] shows us that for small values of , R at the influence of the column s increases rapidly with increased plate number (N) and saw that for Although we could use retention k. For example, a change of k from t most situations, starting method time ( R), it is usually simpler to use 0.5 to 1.0 (black dots in Figure 1) development with a 10,000-plate the retention factor in discussions increases the relative resolution by column makes the most sense. This like the present one. Recall that the 50% from 0.35 to 0.53. A further t k is a good compromise between retention factor is calculated from R doubling of from 1.0 to 2.0 gives t separation power, pressure, and and from the column dead time ( 0 or only a 33% increase in resolution. t k run time. The effect of further M) as: A fivefold increase of from 2 to 10 changes in column length (L) or increases resolution by another third, d k = t t t k particle diameter ( p) can be easily ( R – 0)/ 0 [3] but a change of from 10 to 20 only estimated without doing the actual changes resolution by 5%. This tells experiments. From equation 1, we can see that us that if we want to use k as a lever k + k R This month, we’ll continue the resolution is a function of /(1 ) to improve s, it will be most effective iii R k k discussion with an emphasis on the (term ). Thus, s will increase with , at small values of . influence of the retention factor (k) but not linearly. For example, if k = 1, The Cost of Increased k-Values: on the separation, expressed as k/(1 + k) = 0.5, whereas if k = 100, Another way to look at the data of R k + k resolution ( s). We’ll consider the /(1 ) ≈ 1.0. I find that this Figure 1 is to consider how much general case, specific cases, and relationship is easier to understand in it costs for an increase in k and how to estimate the influence of a a graphic representation, such as in compare this to the gain in resolution change in mobile phase percent Figure 1, where retention, expressed we might get for that cost. Because organic (%B) or temperature on as k, is plotted against resolution, most LC methods are run in an retention. As with the prior instalment expressed as k/(1 + k). Initially automated mode, the cost of analysis of this series, we’ll limit ourselves resolution increases rapidly with time is usually not significant as to isocratic separations and increases in k, then flattens out as k long as a batch of samples can be reversed-phase conditions to simplify exceeds ~5. run in a reasonable time, such as a the discussion, but much of this 14-h (840-min) overnight run. As an instalment will apply to gradients and Target Ranges for k example, let’s consider a method other separation modes. The relationship between k and run on a 150 mm × 4.6 mm column R s shown in Figure 1 can help us at 2.0 mL/min and a sample batch The Influence of Retention on to decide in advance what is the size (comprising both samples and Resolution desired range of k-values we would calibration standards) of 100 injections. t Once again, we’ll use the like to aim for in a separation. Let’s Under these conditions, 0 ≈ 0.75 min; fundamental resolution equation to look at three different aspects of this: we can rearrange equation 3 and R guide the discussion: the improvement in s we get for solve for retention time:

190 LC•GC Europe April 2017

magentablackcyanyellow ES913817_LCE0417_190.pgs 04.06.2017 20:54 ADV LC
5306#-&4)005*/(

'JHVSF
 Plot of retention factor versus resolution (expressed as k/[1 + k]) based on equation 1. Black dots represent results for k = 0.5, 1, 2, 10, and 20.

1.0

0.8

0.6

0.4 Relative resolution 0.2

0.0 0 5 10 15 20 Retention (k/[1+k])

above with k = 0.5, 1, 2, 10, and 16 min, respectively. (I’ve rounded = + tR t0(1 k) [4] 20 (black dots on Figure 1), we can my numbers here, so your results use equation 4 to calculate retention may vary a bit if you repeat these If we use the same examples as times of 1.3, 1.5, 2.3, 8.3, and calculations.) These translate to

GO WITH A HIGH FLOW INTRODUCING THE ALL NEW HIGH DEGASI PREP+ FLOW RATE 75 ml/min Per channel The world’s first high flow rate, in-line degasser, for organic solvents Biotech in co-operation with IDEX H&S are proud to announce the world’s first in-line, membrane degasser ready to use with aggressive media and organic solvents, while maintaining flow-rates up to 150ml/min and above.* Available configura- tions: Stand alone, OEM open frame and 1-4 channels.

* 150ml/min is only a guideline value, the degasser will in many application work with flow rates well above this guideline. The degasser comes with 2-channels, where each channels recom- mended flow rate is 75 ml/min, connecting these chambers in a series will allow for a combined flow rate of 150 ml/min.

INNOVATIVE PRODUCTS FOR FLUIDIC SYSTEMS

HEADQUARTER ° BIOTECH AB ° SWEDEN ° TEL +46 (0)300-56 91 80 ° [email protected] USA ° BIOTECH USA LLC ° TEL 612-703-5718 ° [email protected] www.biotech.se JAPAN ° BIONIK INC. ° +81 545-38-9125 ° [email protected]

XXXDISPNBUPHSBQIZPOMJOFDPN 191 LC
5306#-&4)005*/(

batch run times 100 times larger of 113, 150, 225, 825, and 1575 min. 'JHVSF
 Plot of log k versus %B according to equation 5. Data for a “regular” You can see that methods where the sample from reference 5 are triazine herbicides separated on a C18 column in mixtures of methanol–water as the mobile phase. last peak is eluted with k > 10 would exceed the allotted 14-h window. Another potential cost of adjusting k has to do with detection limits for a method. Longer retention times 3 mean broader peaks (peak width is directly proportional to retention time under isocratic conditions). If peak area is assumed constant, peak 2

height will be inversely proportional 0 to peak width (or retention time). This k

relationship means that doubling log retention, as when k is increased 1 from 10 to 20, will halve the peak height. For trace analysis, peak height, not peak area, is the limiting factor for detection limits. Clearly smaller k-values are desired for taller 0 10 20 30 40 50 60 peaks. %B ( x 100) A final cost-related factor is that of solvent costs. The purchase price, preparation costs, and disposal mobile-phase preparation is certainly k
*T
/PU
UIF
8IPMF
4UPSZ costs all add up. Shorter run times within the normal expectations of The data in Figure 1 implies that an (smaller k-values) will reduce laboratory variation. Because the increase of k will always increase solvent costs. curve of Figure 1 is quite steep at resolution, but this conclusion is a low k-values compared to higher rather broad generality that tends to 5IF
SFMBUJPOTIJQ
CFUXFFO
ones, small k-value samples will fail in the case of specific samples. k R be more susceptible to significant To understand this relationship
BOE
T
DBO
IFMQ
VT
UP
changes in resolution caused by better, we can classify samples EFDJEF
JO
BEWBODF
XIBU
normal fluctuations in mobile-phase as “regular” and “irregular” (see JT
UIF
EFTJSFE
SBOHF
PG
composition. For this reason, it is best discussion of reference 2 for more kWBMVFT
XF
XPVME
MJLF
UP
to avoid conditions that give k < 2 for details). It has been shown (3,4) maximum method robustness. that the relationship between k BJN
GPS
JO
B
TFQBSBUJPO 4FMFDUJPO
PG
k3BOHF
*T
B
and the mobile-phase percent $PNQSPNJTF
After considering organic (%B, expressed as Φ, where 4UBCJMJUZ
PG
4FQBSBUJPO
The general the above discussion, it is clear that Φ = %B × 0.01): curve shape for Figure 1 is one selection of an ideal value of k has = Φ we often see in chemistry, such as tradeoffs—robustness, resolution, log k log k0 – S [5] when tracking reaction completion. run time, detection limits, solvent It is common to see a rapid rise in expense, and so forth. Another Where k0 is the k-value of a solute at the dependent variable with a small complicating factor is that the typical 0% B and S is the slope of the plot, change in the independent variable, sample contains components that and is a characteristic of the solute. then with continuing increases in the vary somewhat in retention time, so we S can be estimated as follows: independent variable, the change would like to fit the sample into a range in the dependent one levels off. The so that the k-value of the first peak is S ≈ 0.25 MW0.5 [6] same happens here for changes not too small and the k-value of the in resolution as k is increased. last one is not too large. Based on the where MW is the molecular weight in For values of k > 5, there is only a above discussion, a range of 2 ≤ k ≤ 10 daltons. For some types of samples, small increase in resolution for each is ideal, being a good compromise plots of equation 5 appear as in successive increase in k, whereas between robustness and run time. Figure 2. Here it can be seen that for k < 2, the changes are quite However, some or many samples, the log k versus S plots are parallel dramatic. Values of k can change depending on the application, have or tend to fan out from each other as because of normal laboratory too broad a polarity range to fit within %B is reduced. In a case like this, variation. For example, as discussed this k-range. In such cases, extending the chromatogram would spread below (Rule of Three), a 1% change the range to 1 ≤ k ≤ 20 is reasonable. out, with increasing resolution at in %B (the organic content of the When the ratio of the k-values of the smaller values of %B, as might mobile phase) can be expected to first and last peaks exceeds 20–40, be expected from the behaviour change k by approximately 10–15% a gradient method will usually be observed in Figure 1. This kind of for a 500-Da sample. A 1% error in favoured over isocratic. sample has been referred to as a

 -$r($
&VSPQF

April 2017 'JHVSF
 Plot of log k versus %B according to equation 5. Data for an “irregular” sample from references 6 and 7 are substituted aromatics separated on a C18 column with mixtures of acetonitrile–water as the mobile phase.

3

2

0 k log

1

0 10 20 30 40 50 60 %B ( x 100)

“regular” sample. Regular samples and at %B < 40% B, it comes out tend to be those closely related after the black one. In contrast to in structure, such as homologs or the expected behaviour of Figure 2, isomers. The sample of Figure 2 is a the compounds represented by the group of structurally related triazine black and blue lines converge as %B herbicides. is reduced, so their resolution gets worse, not better, with increased .BOZ
JG
OPU
NPTU
TBNQMFT
k. Samples in which the slopes of JODMVEF
DPNQPOFOUT
UIBU
the log k versus %B plots differ significantly are referred to as BSF
OPU
BT
DMPTFMZ
SFMBUFE
“irregular” samples. BT
UIPTF
KVTU
EJTDVTTFE
The information we gather from 4VDI
TBNQMFT
WBSZ
JO
Figures 1 and 3 can be valuable guides for LC method development. DPSF
TUSVDUVSF
BT
XFMM
BT
Figure 1 tells us that we’ll get the JO
UIF
UZQF
BOE
OVNCFS
PG
“best” chromatographic behaviour GVODUJPOBM
HSPVQT
with 2 ≤ k ≤ 10 (or 1 ≤ k ≤ 20 if the sample won’t fit within this restricted Many, if not most, samples include retention range). This is a good components that are not as closely general guideline. However, samples related as those just discussed. that behave as the one in Figure 3 Such samples vary in core structure are common enough that we can’t as well as in the type and number assume that larger k-values (longer of functional groups. It is common retention) will always improve the for plots of equation 5 for these separation. Sometimes α (equation samples to appear more like that of 2) will remain relatively constant as Figure 3. In Figure 3, compounds %B is changed and sometimes it will represented by the black, blue, and vary significantly. This tells us that green lines behave much like those fine-tuning k can be a powerful tool of Figure 2 in that they are roughly to fine-tune the resolution. parallel. However, the compound represented by the red line moves 1SFEJDUJOH
3FUFOUJPO relative to the others. At ~45% B, the Another useful tool that we can red and black lines cross, so the two use to save experimental runs is compounds would be coeluted. This to estimate the change in retention is also the crossover point for the red (expressed as k) for a selected and black compounds; at %B > 40%, change in %B. We can do this the red compound comes out first estimation quite accurately based

XXXDISPNBUPHSBQIZPOMJOFDPN LC
5306#-&4)005*/(

on two experimental runs that allow assumptions we make). Equation 6 Thus for S = 5 and Δ%B = 10%, us to make plots as in Figures 2 and tells us that a 400-Da compound will we get Δk = 10(0.1 × 5) ≈ 3. This is 3. For example, chromatograms have an S-value of approximately what is often referred to as “The run at 30% B and 50% B would 5, which is a convenient number Rule of Three”, where a 10% change allow us to measure retention for to use as an average or typical in the organic solvent concentration each compound, convert it to k S-value for small molecule samples of the mobile phase is expected and plot the results as in Figure 2 (for example, <1000 Da). From to change k by approximately or 3. Alternatively, we could use equation 5, we can construct an threefold. This is a very handy tool equations 5 and 6, along with estimate of the change in k (Δk) for estimating how much retention a single experimental run and for a given change in %B (Δ%B, will change when the mobile knowledge (or guess) of the analyte where Δ%B = 10% is equivalent phase is modified. I used a similar molecular weights to construct to ΔΦ = 0.1): estimate for the earlier discussion similar plots with a little less of the effect of a 1% change in accuracy (depending on how many Δk = 10(ΔΦ S) [7] %B due to normal variation in the laboratory.

8F
BMTP
TBX
UIBU
XIJMF
JU
JT
B
HFOFSBM
QBUUFSO
UIBU
SFTPMVUJPO
JODSFBTFT
XJUI
We’re making BO
JODSFBTF
JO
k
UIFSF
BSF
NBOZ
DBTFT
JO
XIJDI
UIF
hybrid pump design TBNQMF
DPNQPOFOUT
WBSZ
FOPVHI
UIBU
DIBOHFT
JO
elementary. SFMBUJWF
SFUFOUJPO
BOE
FWFO
QFBL
SFWFSTBMT
DBO
CF
TFFO
XJUI
FJUIFS
JODSFBTFT
PS
EFDSFBTFT
JO
# That’s because our Bal Seal® spring- energized seal, which combines proven Although I won’t go into detail here canted coil spring energizer technology with (see discussion in Section 2.3.2.2 of advanced polymer formulations and profiles, reference 2 for more information), we provides unparalleled performance and can get plots similar to Figure 2 or 3 flexibility for a new generation of if we plot log k versus 1/T, where T is equipment designed to handle both temperature in kelvins. From such SFC and UHPLC processes. data, we can generate a rule of thumb that an increase in temperature of 1 °C will decrease k by 1–2%. This rule of thumb can be useful to help The Bal Seal® estimate the effect of a change in spring-energized seal column temperature on retention and also alerts us to the importance of column temperature control if we want stable retention times. For pressures starting at $PODMVTJPOT aspiration to 15 kpsi and above, We have examined the influence of at temperatures up to 80 °C, it seals the retention factor, k, on resolution. consistently for millions of cycles, making After we considered the risks (costs) more accurate sample resolution and faster and benefits of various values of throughput a reality. k, using Figure 1 as a guide, we Call or click today, and start designing concluded that we are likely to get for tomorrow. the “best” chromatography and a good balance of costs (time, solvent costs, resolution, and so forth) if we can adjust the chromatographic +31 20 638 6523 conditions to achieve 2 ≤ k ≤ 10, www.balseal.com or if the sample won’t fit into this retention range, 1 ≤ k ≤ 20. When k < 1, retention tends to be less

194 -$r($
&VSPQF

April 2017 LC
5306#-&4)005*/( stable, resolution is very sensitive accurate estimates can be made for 3FGFSFODFT to small changes in k, and there is the change in retention that can be (1) J.W. Dolan, LCGC Europe (3), more chance of interferences from expected with a specific change 138–142 (2017). (2) L.R. Snyder, J.J. Kirkland, and J.W. the unretained materials at t0. When in %B. Dolan, Introduction to Modern Liquid k > 20, run times become excessive Chromatography, 3rd edition (Wiley, and peak heights suffer. When 2010), section 2.5.2.1. 3FHVMBS
TBNQMFT
UFOE

(3) K. Valkó, L.R. Snyder, and J.L. Glajch, samples won’t fit in a retention range J. Chromatogr. A 656, 501–520 (1993). of 1 ≤ k ≤ 20, a gradient is likely a UP
CF
UIPTF
DMPTFMZ
(4) L.R. Snyder, J.W. Dolan, and J.R. Gant, better choice. SFMBUFE
JO
TUSVDUVSF

J. Chromatogr. A 165, 3–30 (1979). We also saw that while it is a (5) T. Braumann, G. Weber, and L.H. TVDI
BT
IPNPMPHT
PS
Grimme, J. Chromatogr. A , general pattern that resolution JTPNFST 329–343 (1983). increases with an increase in k (as (6) N.S. Wilson, M.D. Nelson, J.W. Dolan, in Figure 1 or 2), there are many L.R. Snyder, R.G. Wolcott, and P.W. cases in which the sample Often if we start method Carr, J. Chromatogr. A 961, 171–193 (2002). components vary enough that development with a column that (7) N.S. Wilson, M.D. Nelson, J.W. changes in relative retention and generates N ≈ 10,000 plates (1), Dolan, L.R. Snyder, and P.W. Carr, J. even peak reversals can be seen find conditions that generate Chromatogr. A 961, 195–215 (2002). with either increases or decreases 1 ≤ k ≤ 20 for our sample in %B. This means that even if the components, and then fine-tune “LC Troubleshooting” Editor +PIO
sample is eluted in the 1 ≤ k ≤ 20 %B for the best retention, we will %PMBO has been writing “LC range, fine-tuning %B can be a obtain a satisfactory separation. Troubleshooting” for LCGC for more useful tool to move peaks around If this is the case, method than 30 years. One of the industry’s relative to each other. development can be a fairly most respected professionals, John Finally, we saw that we can further simple process. If we have is currently a principal instructor save the cost of experimental runs optimized these two factors (terms for LC Resources in McMinnville, by estimating the change in retention i and iii of equation 1) and are not Oregon, USA. He is also a member or k for a specific change in %B or successful, it is time to look for help of LCGC Europe’s editorial advisory temperature. If two experimental in term ii of equation 1, as will be board. Direct correspondence about runs are made, such as at 30% B discussed in the next instalment of this column via e-mail to LCGCedit@ and 50% B for Figures 2 and 3, quite this series. ubm.com

HDHT. First cement-free syringe for PAL Combi-xt headspace autosamplers

Chemically inert Temperature stable up to 200ºC

Hamilton Bonaduz AG Switzerland [email protected] www.hamiltoncompany.com

XXXDISPNBUPHSBQIZPOMJOFDPN 195 COLUMN WATCH

New Chromatography Columns and Accessories for 2017

David S. Bell, Column Watch Editor

Our annual review of new liquid chromatography (LC) columns and accessories introduced at Pittcon and throughout the previous year.

In accordance with recent tradition, the small‑molecule realm three improved performance, inertness, this article is intended to cover subcategories could be established: and robustness as compared to chromatography columns and reversed‑phase, hydrophilic conventional FPP products. Another accessories commercially released interaction liquid chromatography theme consistent with 2016 is the after Pittcon 2016 through this (HILIC), and chiral offerings. Each of continuing effort to increase column year’s conference held in Chicago these will be discussed separately. stability for wider pH range utility. 5–9 March 2017. LCGC once again For large‑molecule products, Agilent Technologies (HPH‑C18), sent out a survey in early 2017 subareas involving reversed‑phase ChromaNik Technologies (SunShell asking vendors to supply them with mode and size‑exclusion C18), and Imtakt Corporation any products launched after Pittcon chromatography are identified and (DZ‑C18) all launched phases 2016. Instrumentation is covered discussed. Columns commercialized with enhanced pH stability. in this issue of LCGC (1), and using ion‑exchange chromatography Separation Methods Technologies gas chromatography and sample for both small and large molecules commercialized an interesting set preparation will be covered in the will be discussed separately. of short‑chain reversed‑phase May issue. Because information for Furthermore, there were products chemistries (C1, C2, and C4). this instalment of “Column Watch” launched for supercritical fluid The company contends that their is obtained sporadically over the chromatography (SFC) that deserve proprietary bonding strategy course of many months, it is very some discussion. An interesting and provides “proper deactivation” of possible that some information potentially game‑changing microchip silanols as well as resistance to has been missed. Readers are chromatography device based acid hydrolysis that plagues many encouraged to check with specific on pillar array technology is also short‑chain surface chemistries. vendor sites for additional products featured. Lastly, several accessories Another interesting trend from this as well as more detailed information relating to HPLC are noted. year’s offerings is the presence of regarding what is covered here. several “polar C18” chemistries. The vendors that responded to Small-Molecule Separations Phenomenex launched a polar the survey with high performance Reversed-Phase Chromatography: C18 stationary phase in both the liquid chromatography (HPLC) The product offerings assigned to Kinetex and Luna Omega lines and and ultrahigh‑pressure liquid the small‑molecule, reversed‑phase Waters added its T3 chemistry to chromatography (UHPLC) columns category are listed in Table 2. A total the Cortecs brand. In each case and accessories are listed in of 22 new surface chemistries are the surface modification is noted Table 1. The list contains very noted from the list. Continuing recent to impart a balanced retention well‑known vendors such as Waters trends, many of the new columns are for hydrophilic and hydrophobic and Agilent Technologies, but also intended to round out selectivity or molecules. Phenomenex also came several lesser‑known vendors. The particle size offerings within existing out with a chemistry denoted as PS product range is also highly varied lines. Of the 22 columns listed, nine C18 which is reported to include a from traditional reversed‑phase are manufactured using superficially positively charged modification to columns using fully porous materials porous particles (SPP), indicating the surface that aids in the retention to specialty columns. As in years that the particle architecture of acidic analytes. More complete, past, with such a varied list, further continues to garner attention. full line offerings were launched by discussion requires some breakdown Enhancements of fully porous Agilent (InfinityLab Poroshell) and of the different offerings. Similar particles (FPP) are also noteworthy. Restek (Force Performance). to 2016, one differentiation can be For example, Phenomenex’s Luna Hydrophilic Interaction made based on offerings that target Omega phase reportedly includes Chromatography: HILIC, which is small or large molecules. Within a thermal modification that provides often employed for the retention and

196 LC•GC Europe April 2017

magentablackcyanyellow ES913725_LCE0417_196.pgs 04.06.2017 20:31 ADV HILICON

L+,/,&‹ )XVLRQ DGYDQFHV+,/,&VHSDUDWLRQVLQ+3/&DQG8+3/&

L+,/,&‹ )XVLRQ L+,/,&‹ )XVLRQ  L+,/,&‹ )XVLRQ 3 6LOLFDEDVHG 6LOLFDEDVHG 3RO\PHUEDVHG

 &KDUJHPRGXODWHGK\GUR[\HWK\ODPLGHDPLGH+,/,&FROXPQV  &RPSOHPHQWDU\VHOHFWLYLWLHVIRUSRODUK\GURSKLOLFDQDO\WHV  ([FHOOHQWGXUDELOLW\DQGXOWUDORZEOHHGLQJIRU/&06  3HUIHFWFROXPQVIRUμ2PLFVμVWXGLHV  L+,/,&‹‹ )XVLRQDQGL+,/,& )XVLRQ  S+DQG˜P  L+,/,&‹ )XVLRQ 3 S+˜P

 $03 $'3  $73

 W∓



  5HWHQWLRQ7LPH PLQ

)RUPRUHLQIRUPDWLRQ (PDLOLQIR#KLOLFRQFRP_:HEVLWHZZZKLOLFRQFRP Œ+,/,&21$%$OOULJKWVUHVHUYHG_L+,/,&‹ LVDUHJLVWHUHGWUDGHPDUNRI+,/,&21$%6ZHGHQ COLUMN
8"5$)

separation of very polar compounds, 5BCMF
 2017 LCGC new product survey responding vendors continues to show growth. Table 3 lists product offerings in the HILIC Company Product category. The seven new entries Advanced Chromatography ACE Amino columns in this category match the number Technologies Limited of new columns reported in 2016. Advanced Materials Technology, HALO 1000Å C4, 2.7-μm columns It is interesting to note that three Inc. of the seven entries are built InfinityLab Poroshell 120 1.9-μm columns on polymeric phases. HILICON Six different chemistries: introduced a polymeric version InfinityLab Poroshell 120 EC-C18 1.9-μm columns in its iHILIC-Fusion line. Shodex InfinityLab Poroshell 120 EC-C8 1.9-μm columns commercialized two polymerically based HILIC phases. The VN-50 Agilent Technologies InfinityLab Poroshell 120 Phenyl-Hexyl 1.9-μm columns 2D diol phase is designed for oligosaccharide analysis, and the InfinityLab Poroshell 120 PFP 1.9-μm columns VC-50 2D carboxyl chemistry is InfinityLab Poroshell 120 HPH-C18 1.9-μm reportedly applicable to monoamine columns type neurotransmitters and oral InfinityLab Poroshell 120 HILIC 1.9-μm columns anti-diabetic drugs. The company Analytical Sales & Services, Inc. MonoSLEEVE single-column heater controller also notes that the PEEK housing Chiralpak IA-U columns reduces undesired interactions with Chiral Technologies Chiralpak IC-U columns the column hardware. MicroSolv Technology continues to fill out Chiralpak IG columns its line of silica hydride based Chromanik Inc. Japan SunShell C18, 2-μm columns phases with a new amide chemistry. ChromegaChiral CCU columns Fully porous– and superficially ChromegaChiral CCX columns ES Industries porous–based HILIC phases Epic SCX columns continue to be promoted. Advanced Chromatography Technologies MacroSep Bio Gold C18, C8, C4, PFP, napthyl, biphenyl, diphenyl, and HPR columns added an amino chemistry to its FPP ACE offering, and both Agilent SpeedCore BIO C18 Peptide columns Technologies (bare silica) and Fortis Technologies SpeedCore BIO Protein C18, C8, and C4 ChromaNik Technologies (amide) columns added to their respective SPP lines. Hilicon iHILIC-Fusion(P) column $IJSBM
$ISPNBUPHSBQIZ
Eight new chiral phases were introduced Imtakt Corporation Dacapo DX-C18 column after Pittcon 2016. This number Jordi Labs LLC Jordi Resolve GPC columns is a significant increase over the Nucleodur C18 Gravity-SB columns same time period from Pittcon 2015 Macherey-Nagel Nucleodur π2 columns through the “New Chromatography Microsolv Technology Columns and Accessories for 2016” Cogent Amide HPLC column Corporation column instalment where only three Mott Corporation PerfectPeak HPLC–UHPLC mixers entries were noted (2). Cellulose- and amylose-based chiral phases Optimize Technologies EXP Trap columns dominate the list provided in Table 4. PharmaFluidics μPAC chip separation device Chiral Technologies extended its line Clarity Oligo-SAX columns of meta-immobilized 5-μm phases Clarity Oligo-XT C18 columns through the introduction of Chiralpak Kinetex Polar C18 columns IG. The company also introduced amylosic and cellulosic phases on Luna Omega C18 columns 2-μm FPP particles designed for Phenomenex Luna Omega Polar C18 columns UHPLC. ES Industries continued to Luna Omega PS C18 columns fill out its ChromegaChiral line with Lux AMP columns two new chemical modifications of amylose that provide what Lux i-Cellulose-5 columns the company refers to as unique Yarra 1.8u SEC-X300 columns enantiomeric separation behaviour. PolyHeptyl A columns Phenomenex introduced an PolyLC Inc. PolyHexyl A columns immobilized cellulosic phase, Lux i-Cellulose-5, which is designed to PolyPentyl A columns ZircoFit UHPLC fitting system for fused-silica be more robust for strong organic Prolab Instruments GmbH solvents than coated polysaccharide capillaries

198 -$r($
&VSPQF

April 2017 Advertisement

'VMMZ
"VUPNBUFE
*OTUSVNFOU
GPS
Solution Viscosity in Polymeric Materials

The determination of the solution viscosity of polymeric materials is very important to both the research and manufacturing industries because it can be used to estimate molar mass, thereby providing important information relating to the physical properties of polymers. The Intrinsic Viscosity Analyzer, IVA, was developed in response to the market’s need for a reliable and automated instrument for intrinsic viscosity analysis of polymeric materials, which sometimes demand temperatures of up to 200 ºC for dissolution. A wide range of polymers, even the most crystalline ones, can be analyzed with convenience and safety in this instrument. The IVA presents a precise and convenient approach to IV measurement as a result of the automation of all the analytical procedures, from filling of the vials to the reporting of the results. Samples are put in solid form into 20-mL vials and brought to the instrument’s autosampler tray. Under software control, the instrument takes care of adding solvent to the vials, controlling the exact dissolution time for each vial, injecting each solution, and rinsing the capillary lines. No solvent needs to be handled by the analyst at any time and no vapours are released to the atmosphere because the system is airtight at all times. Accuracy is also achieved with sample care protocols implemented to maintain the sample’s integrity throughout the dissolution and measurement processes. Before dissolution starts, a purge of the vial’s atmosphere with an inert gas (nitrogen) is performed to minimize oxidation. The vials then remain in an external tray at room temperature before the software schedules their transfer to the dissolution oven, where the time spent at high temperature is kept only at the necessary length for dissolution. In addition, efficient heat transfer to the vial, together with a gentle shaking without magnets, and preheating the solvent prior to its delivery to the vial, help to shorten the time required for dissolution. This minimizes oxidative and thermal degradation ensuring that accurate IV is measured, even for the most challenging ultrahigh molar mass materials and oxidation-prone polymers, such as polypropylene. The viscosity measurement is performed by the built-in two capillary viscometer placed inside an oven with extremely precise and stable temperature control (0.01 ºC). The instrument fluidic design is rather simple, having a single valve inside the oven, which results in high reliability. Moreover, the IV results obtained by IVA are in agreement with the ISO 1628-3:2010 method. A new run can be immediately started after finishing the previous one, achieving a throughput of 40 samples a day in standard operation conditions. Unlike the glass capillaries used in Ubbelohde viscometers, the IVA stainless steel capillaries and tubing do not require additional washing or rinsing, and provide robust and precise viscosity values over time. The IVA can be used with different solvents for analyzing a wide variety of polymers. When analyzing polyolefins or other polymers containing significant C-H content, the instrument can incorporate the Infrared detector IR4. With this on-line detector, the injected polymer mass is accurately quantified, which results in improved precision and accuracy of IV results.

Key Features: t
)JHIUFNQFSBUVSF
BVUPTBNQMFS
GPS
EJTTPMVUJPO
PG
TBNQMFT
JO
UIF
TBNF
JOTUSVNFOU t
'VMM
BVUPNBUJPO
PG
EJTTPMVUJPO
BOE
BOBMZTJT
QSPDFTTFT t
4FMGDMFBOJOH
EFTJHO
CBTFE
PO
B
EVBM
DBQJMMBSZ
SFMBUJWF
WJTDPNFUFS t
)JHI
QSFDJTJPO
BDIJFWFE
CZ
BVUPNBUJPO t
/P
OFFE
GPS
BDDVSBUF
XFJHIJOH
XIFO
UIF
PQUJPOBM
*3
EFUFDUPS
JT
VTFE t
4JNQMF
UP
PQFSBUF
BOE
SFMJBCMF
JOTUSVNFOU t
6Q
UP

TBNQMFT
DBO
CF
BOBMZ[FE
TFRVFOUJBMMZ
XJUIPVU
VTFS
JOUFSWFOUJPO t
"OBMZTJT
PG
IJHI
BOE
VMUSBIJHI
NPMBS
NBTT
QPMZNFST t
-PX
TPMWFOU
DPOTVNQUJPO

Polymer Char Website: www.polymerchar.com/intrinsic_viscosity_analyzer www.chromatographyonline.com 199 COLUMN
8"5$)

phases. The company also 5BCMF
 2017 LCGC new product survey responding vendors Contd.... launched the Lux-AMP phase. The proprietary chemistry is specifically Company Product designed for the enantiomeric Force Performance Biphenyl columns separation of amphetamine and Restek Force Performance C18 columns methamphetamine. Separation Force Performance Fluorophenyl columns Methods Technologies reported the SMT CIB-TE - Specialty Teicoplanin columns Separation Methods development of a teicoplanin-based SMT MEB C1, C2, and C4 columns chiral stationary phase. The Technologies SMT PAH1 columns company reports that the column is HK-404L columns useful for performing a wide variety LW-803 columns of chiral separations with amino Shodex acids, carboxylic acids, and neutral VC-50 2D columns compounds. Like 2016, it is clear VN-50 2D columns from these new product introductions IonPac AS23 columns Thermo Fisher Scientific that amylose and cellulose phases IonPac CS20 columns based on FPP particles continue to Vici VICI Jour JR-BPR3 back pressure regulator dominate the field. New this year is CORTECS Shield RP18 columns the introduction of sub-2-μm chiral Waters CORTECS T3 columns columns (Chiral Technologies); Torus SFC columns however, SPP technologies have still Welch Materials Inc. Ghost-Buster column not entered this realm. YMC Co. Ltd. YMC-Pack Diol SEC columns

5BCMF
 Reversed-phase columns for small-molecule separations Company Product Name Stationary Phases Particle Sizes (μm) Particle Type

EC-C18, EC-C8, phenyl- Agilent Technologies InfinityLab Poroshell 120 1.9 SPP hexyl, PFP, HPH-C18

ChromaNik Technologies Inc SunShell C18 2 SPP

Imtakt Corporation Dacapo DX-C18 2.5 FPP Macherey-Nagel GmbH & Co. Nucleodur C18 Gravity-SB 1.8, 3, and 5 FPP

Macherey-Nagel GmbH & Co. Nucleodur π2 5 FPP

Kinetex Polar C18 2.6 SPP

Phenomenex Luna Omega PS C18, polar C18 1.6, 3, and 5 FPP

Luna Omega C18 1.6 FPP

Force Performance LC C18, biphenyl, and Restek Corporation 1.8, 3, and 5 FPP Columns fluorophenyl

Separation Methods SMT PAH1 Proprietary 1.7, 3, 5, and 10 FPP Technologies

Separation Methods SMT-MEB C1, C2, and C4 1.7, 3, 5, and 10 FPP Technologies

Waters CORTECS Shield RP18, T3 1.6, 2.7 SPP

 -$r($
&VSPQF

April 2017 COLUMN
8"5$)

-BSHF.PMFDVMF
4FQBSBUJPOT BIO-Gold) demonstrates that new (160 Å) and SpeedCore BIO Protein The separation of large molecules lines based on fully porous particles (300 Å) columns. Also based on such as peptides, proteins, and are still being developed. New wider pore SPP technology, these oligonucleotides often requires columns intended for reversed-phase offerings are intended for improved larger pore sizes than are typically chromatography of biomolecules are separations of peptides and proteins, used in small molecule applications. presented in Table 5. Also included respectively. Oligonucleotides Full characterization of these in the table is a new set of offerings also fall into this category of large complex molecules often requires from PolyLC, Inc. for HIC. molecules and offer their own set multiple modes of separation, Advanced Materials Technologies of chromatographic challenges. including reversed-phase introduced a 1000-Å C4 column Phenomenex recently launched chromatography, size-exclusion intended for very large molecules Clarity Oligo-XT, an organo-silica chromatography (SEC), ion-exchange such as monoclonal antibodies SPP phase intended for the chromatography, and hydrophobic (mAbs). The larger pore size as reversed-phase characterization interaction chromatography (HIC). compared to many “large-molecule” and purification of synthetic 3FWFSTFE1IBTF
$ISPNBUPHSBQIZ columns (typically 160–400 Å) oligonucleotides. The column is In last year’s report, columns provides less restriction into the reported to be stable at high pH, intended for reversed-phase analysis pores and thus can provide higher which is often used in oligonucleotide of large molecules was dominated resolution of minor variants of separations. by chemical modification of SPP mAbs. The column is based on SPP HIC likely deserves its own particles. That trend continues this technology. Fortis Technologies category within the large-molecule year. However, a new line of columns reports the expansion of its SPP realm. However, for the sake of from ES Industries (MacroSep portfolio with SpeedCore BIO Peptide space it is lumped in here with

Dimensions (mm) Comments Robust superficially porous 1.9-μm particles and optimized column loading give manageable pressure, which allow columns to be run at optimum linear velocities or the use of longer columns without exceeding column or instrument pressure rating. This feature leads to faster run times or improved resolution. Benefits are decreased costs from higher throughput or improved data quality and decreased costs from reduced 50 × 2.1 to 150 × 3.0 re-work. This particle and loading technology also provides long column lifetimes under UHPLC conditions. Benefits are decreased costs because of fewer column purchases, reduced workflow disruption, and less re-work. For the HPH-C18 phase, an organic compound is integrated into the porous layer during particle manufacture to give stability at extremes of pH. Stable over a pH range from 1 to 10. More than 300,000 plates per metre. Symmetrical peak shape for basic, 50 × 2.1 to 150 × 2.1 acidic, and metal chelating compounds. Not specified Hybrid silica and polymeric particle dual-matrix technology provides high pH stability. 30 × 2 to 250 × 4.6 Hydrophobic C18 phase with distinct polar selectivity for improved resolution of early eluted compounds. Hydrophobic phase with alternative selectivity compared to classical C18 modifications. The biphenylpropyl 50 × 2 to 250 × 4.6 ligand provides π-π and hydrophobic interactions. Common analytical lengths for 2.1, 3.0, and Balanced retention of polar and nonpolar compounds under reversed-phase or 100% aqueous conditions. 4.6 mm i.d. 1 mm to 2.1 mm i.d. (1.6 μm) , 2.1 mm to Polar C18: Balanced retention of nonpolar compounds alongside enhanced retention of polar compounds 4.6 mm i.d. (3 and 5 under reversed phase or 100% aqueous conditions. PS C18: Positively charged ligands on surface aid μm) common analytical in the retention of acidic compounds through ionic interactions, while the C18 ligand promotes general lengths. Preparative reversed-phase retention. dimensions also available 1 mm to 2.1 mm i.d. Thermally modified fully porous particles provide greater performance, inertness, robustness, and performance common analytical compared to conventional fully porous products. lengths 30 × 2.1 to 100 × 3.0 (1.8 μm), 30 × 2.1 to 150 Fully scalable from 1.8 μm to 3 and 5 μm for easy method transfer. Long-lasting and reproducible even under × 4.6 (3 μm), and 50 × the stress of elevated UHPLC pressures and rapid cycling. 2.1 to 250 × 4.6 (5 μm)

50 × 2.1 to 300 × 50 For the separation of structurally similar isomers of polyaromatic hydrocarbons (PAHs).

Columns with short carbon chains for rapid analysis of basic, neutral, and mildly acidic drugs and small molecules and biomolecules. These columns provide excellent baseline resolution for small basic molecules, 50 × 2.1 to 300 × 50 including therapeutic drugs. The bonding scheme ensures proper deactivation substrate silanols and resistance to acid hydrolysis. 30 ×2.1 to 150 × 3.0 Shield RP18: The embedded polar group helps to “shield” residual surface silanols, reducing peak tailing for basic (1.6 μm) and 30 × 2.1 to analytes and providing alternative selectivity for polyphenolic compounds such as flavanoids. T3: 100% aqueous 150 × 4.6 (2.7 μm) compatible reversed-phase column designed to give balanced retention to both polar and nonpolar compounds.

XXXDISPNBUPHSBQIZPOMJOFDPN  COLUMN
8"5$)

the reversed-phase offerings. HIC at HPLC 2016 (pentyl, hexyl, and solvents such as tetrahydrofuran is often a valuable tool used to heptyl) that permit MS-compatible is normally referred to as GPC. separate polar variants of proteins. HIC for proteomics and quality Columns used in SEC and GPC are HIC, however, traditionally uses control of biotherapeutics. often characterized by strict control high concentrations of nonvolatile 4J[F&YDMVTJPO
$ISPNBUPHSBQIZm
of pore size and by inert surface buffers, rendering it incompatible (FM
1FSNFBUJPO
$ISPNBUPHSBQIZ
chemistry. Control of the pore size with mass spectrometry (MS) SEC continues to witness resurgence dictates how molecules of various detection. A presentation by Andrew largely as a result of the focus sizes can diffuse into the pores, Alpert of PolyLC, Inc., at HPLC 2016 on large-molecule therapeutics. while the inertness of the surface described efforts to render HIC MS SEC resolves analytes based on limits secondary interactions that friendly (3). Using more-hydrophobic molecular size and is often used may interfere with molecular size analogs of a poly(propyl as a complementary technique to separation and determination. aspartamide) stationary phase, Alpert reversed-phase analysis of protein. Table 6 lists the SEC and GPC demonstrated that proteins could be Gel permeation chromatography column offerings launched since denatured in MS-friendly(er) mobile (GPC) uses the same process Pittcon 2016. The number of new phases that may lead to a possible of separation by molecular size. offerings is similar to those reported marriage of the two techniques Separation by molecular size last year. Jordi Labs released a (4). Since that time, PolyLC, Inc., in aqueous mobile phases is divinylbenzene GPC phase in both has launched three versions of the commonly referred to as SEC, 3- and 13-μm particle sizes that polyaspartamide phases discussed whereas separation in organic promises high efficiency, high

5BCMF
 HILIC columns for small-molecule separations Company Product Name Stationary Phases Particle Sizes (μm) Particle Type Advanced Chromatography ACE Amino Amino 1.7, 3, and 5 FPP Technologies InfinityLab Poroshell 120 Agilent Technologies Unbonded 1.9 SPP HILIC Amide, phosphate, quaternary HILICON AB iHILIC-Fusion(P) 5 Polymeric ammonium MicroSolv Technology Cogent Amide Amide 4 Silica hydride Corporation VN-50 2D Diol 5 Polymeric Shodex VC-50 2D Carboxyl 5 Polymeric

5BCMF
 Chiral columns for small-molecule separations Company Product Name Stationary Phases Particle Sizes (μm) Particle Type Amylose tris(3-chloro-5- Chiralpak IG 5 FPP methylphenylcarbamate) Chiral Technologies, Inc Amylose tris(3-chloro-5- (Daicel Corporation) Chiralpak IA-U and methylphenylcarbamate) and <2 FPP Chiralpak IC-U Cellulose tris(3-chloro-5- methylphenylcarbamate) Methylbenzylcarbamate ChromegaChiral CCU and 3-chloro-4- 3, 5, 10, and 20 FPP methylphenylcarbamate–amylose ES Industries Methylbenzylcarbamate and ChromegaChiral CCX 3,5-dimethylphenylcarbamate– 3, 5, 10, and 21 FPP amylose

Lux AMP Proprietary 3 FPP

Phenomenex Cellulose tris(3,5-dichlorophenyl- Lux i-Cellulose-5 3, 5 FPP carbamate)

Separation Methods SMT CIB-TE Teicoplanin 5 FPP Technologies, Inc.

 -$r($
&VSPQF

April 2017 COLUMN
8"5$) capacity, and lower back pressure. YMC Co. Ltd. launched a 2-μm diol Table 7, however, the product Shodex continued to expand its phase that reportedly exhibits greater launches this year are dominated by offering with the release of two new resolution than standard 3- and 5-μm columns intended for small-molecule chemistries on polymeric supports. materials. analysis. ES Industries released The HK-404L phase is intended for *PO&YDIBOHF
$ISPNBUPHSBQIZ
the Epic SCX column, which the rapid GPC analysis, and the LW-803 Ion-exchange chromatography company claims provides superior product is designed for antibody is a separation technique that cation loading and stability that and antibody aggregate separations. exploits strong interactions between results from a high density bonding Smaller-particle designs continue opposite charges of a surface technology. Thermo Fisher Scientific to infiltrate the SEC–GPC market in and an analyte. Various surface also released two new chemistries an attempt to enhance resolution supports may be modified to carry in its IonPac line. According to the and speed. Phenomenex extended permanent (strong cation and anion company, the AS23 phase provides its line of sub-2-μm SEC columns exchange) or variable (weak cation advanced capabilities to detect with the release of the Yarra 1.8μ and anion exchange) charge that oxyhalides among other inorganic SEC-X300 column. The column’s can be used to interact with and anions in various water matrices 300-Å pore size is intended for the separate analytes with the opposite of interest and the CS20 product characterization of mAb aggregates, charge. As with SEC, ion-exchange provides enhanced performance for antibody–drug conjugates (ADCs) chromatography is often used as a the analysis of inorganic cations and and biosimilars with a separation complementary tool for the analysis amines. The lone “large-molecule” range of 10–700 kDa. Similarly, of large molecules. As shown in ion-exchange product release came

Dimensions (mm) Comments Highly reproducible and ultrainert amino phase for reversed-phase, normal-phase, weak anion-exchange, Not disclosed and HILIC modes. Proprietary bonding technology provides robustness with excellent column lifetimes.

50 × 2.1 to 150 × 3.0 Robust superficially porous 1.9-μm particles and optimized column loading.

Unique selectivity from enhanced hydrophilic partitioning, weak electrostatic interaction, and hydrogen 20 × 2.1 to 250 x 4.6 bonding capacity; HILIC applications at high pH. 100 × 2.1 plus additional Stability of column with greater column lifetime and unique selectivity. dimensions 150 × 2.0 Diol-functionalized PVA-based HILIC column designed for oligosaccharide analyses. Packed in PEEK, this column is designed for monoamine type neurotransmitters and oral anti-diabetic 150 × 2.0 drugs. The polymer column housing decreases interactions between compounds and column hardware.

Dimensions (mm) Comments Meta immobilized columns offer mechanistic advantages in chiral recognition, higher success rates 100 × 4.6 to 250 × 10 using fewer columns. The columns are designed for robustness and extended column life.

These columns are designed to be used with UHPLC systems to achieve ultrafast and high-resolution 50 × 3.0 and 100 × 3.0 separation of enantiomers.

Capillary to preparative Packed in PEEK, this column is especially designed for monoamine type neurotransmitters and oral anti-diabetic drugs. The polymer column housing decreases interactions between compounds and column hardware. Capillary to preparative

Common analytical lengths for Designed and tested for chiral separation of amphetamine and methamphetamine enantiomers. 3.0- and 4.6-mm i.d. columns Common analytical lengths Immobilized portion of this new chiral stationary phase greatly increases column robustness by tolerating for 2.1-, 3.0-, and 4.6-mm strong organic solvents such as DMSO, DCM, ethyl acetate, MTBE, and tetrahydrofuran to be injected i.d. columns; preparative onto the column. dimensions available SMT CIB-TE column is made through bonding of the antibiotic, teicoplanin with 1,6-diisocyanatohexane, using SAM Total Coverage technology, both patented and proprietary. This covalently bonded 50 × 2.1 to 300 × 22.1 teicoplanin column is extremely useful for performing a wide variety of chiral separations with amino acids, carboxylic acids, and neutral compounds.

XXXDISPNBUPHSBQIZPOMJOFDPN  COLUMN
8"5$)

5BCMF
 Reversed-phase separations for large-molecule separations Company Product Name Stationary Phases Particle Sizes (μm) Particle Type Advanced Materials HALO 1000Å C4 C4 2.7 SPP Technology

MacroSep BIO-Gold HPLC C18, C8, C4, PFP, napthyl, ES Industries 1.9, 3, 5, and 10 FPP columns biphenyl, diphenyl, HPR

SpeedCore BIO Peptide C18 2.6 SPP Fortis Technologies SpeedCore BIO Protein C18, C8, C4 3.5 SPP

Phenomenex Clarity Oligo-XTC181.7, 2.6, and 5 SPP

Polypentylaspartamide, PolyPentyl A, PolyHexyl PolyLC, Inc. polyhexylaspartamide, and 2, 3, and 5 FPP A, and PolyHeptyl A polyheptylaspartamide

5BCMF
 SEC columns for large-molecule separations Company Product Name Stationary Phases Particle Sizes (μm) Particle Type Jordi Resolve GPC Monodisperse Jordi Labs LLC Divinylbenzene 5 and 13 columns polymeric

Phenomenex Yarra 1.8u SEC-X300 Proprietary 1.8 FPP

Shodex HK-404L Styrene–divinylbenzene 3.5 Polymeric

Shodex LW-803 Not disclosed 3 FPP

YMC Co. Ltd. YMC-Pack Diol SEC Dihydroxypropyl 2 FPP

5BCMF
 Strong cation-exchange columns for large-molecule separations

Company Product Name Stationary Phases Particle Sizes (μm) Particle Type

ES Industries Epic SCX Aromatic sulfonic acid 1.8, 3, 5, and 10 FPP

Phenomenex Clarity Oligo-SAX Quatenary amine 5 Nonporous

IonPac AS23-4μm Not disclosed 4 Not disclosed Thermo Fisher Scientific IonPac CS20 Not disclosed Not disclosed Not disclosed

from Phenomenex. The Clarity and speed the technique often is designed especially for SFC Oligo-SAX phase is interestingly built provides. The lone line of new SFC and is enhanced by a two-stage on a nonporous, 5-μm particle and phases reported since Pittcon 2016 bonding process that provides stable is intended for the high-resolution comes from Waters Corporation. retention and unique selectivity. analysis of synthetic oligonucleotides. Several surface modifications, 1JMMBS"SSBZ
4FQBSBUJPO
%FWJDF 4VQFSDSJUJDBM
'MVJE
including 2-picolamine, diethylamine, An interesting and potentially $ISPNBUPHSBQIZ
SFC continues diol, and 1-aminoanthracene built on game-changing separation device to receive interest because of Water’s bridged ethyl hybrid (BEH) was featured at Pittcon 2017. improvements in instrument design, particle technology were released PharmaFluidics launched the first the desire to perform greener under the brand name Torus. The microchip chromatography device that separations, and the high resolution company claims that the particle is manufactured using lithographic

 -$r($
&VSPQF

April 2017 COLUMN
8"5$)

Dimensions (mm) Comments Higher resolution of minor variants of mAbs and narrower peak widths by 12–40% compared to current 20 × 2.1 to 250 × 4.6 products. MacroSep BIO-Gold packings are based on ultrahigh-purity spherical silica, state-of-the-art high density 50 × 2.1 to 300 × 4.6 bonding, and full endcapping intended for the separation of high-molecular-weight compounds such as proteins and peptides.

Not disclosed An addition to the SpeedCore line of columns intended for peptide analysis.

Not disclosed Additional phases to the SpeedCore line of columns intended for protein analysis. Common lengths with 2.1 mm i.d. (1.7 μm), common lengths for 2.1 and 4.6 mm i.d. (2.6 and Designed to provide rugged high performance and high pH stability for the LC–MS characterization of 5 μm); semipreparative and synthetic DNA and RNA samples as well as purification of these target oligos. preparative dimensions also available

These materials are unique in permitting HIC of proteins in native structure with volatile (MS-compatible) Capillary to process scale mobile phases for top-down proteomics and quality control.

Dimensions (mm) Comments

300 × 7.8 High efficiency, high separation capacity, and lower back pressure with greater bed stability.

Common analytical lengths For separation and characterization of mAb aggregates, antibody–drug conjugates (ADCs) and for 4.6 mm biosimilars. Biomolecule separation range 10–700 kDa complements existing Yarra 1.8 μm SEC-X150. Monodisperse styrene–divinylbenzene particles create an environment for rapid analyses on both traditional GPC systems as well as micro-GPC and UHPLC (APC) systems. The HK-404L is a 300 × 8 mixed-bed linear type column for separations ranging from 100–1,000,000 Da. No need to purchase expensive APC XT columns. LW-800 series is specially designed for antibody and antibody aggregates separations. The ultralow 300 × 8 shed makes this series ideal for multiangle light scattering systems. The 2-μm particle exhibits greater resolution than standard 3- and 5-μm materials, which allows smaller 150 × 4.6 and 300 × 4.6 diameter and shorter length columns to be used. This increases throughput while decreasing mobile-phase consumption.

Dimensions (mm) Comments

Epic SCX uses a high density bonding technology that provides for superioer cation loading and Not disclosed robustness. Common analytical lengths Innovative high capacity strong anion-exchange column delivers high-resolution analysis of synthetic for 4.6 mm i.d. oligonucleotides. Advanced capabilities to detect oxyhalides and common inorganic anions in drinking water, Not disclosed groundwater, wastewater, and other diverse sample matrices. Designed to offer choice performance for the analysis of inorganic cations and amines. The columns use Not disclosed gradient ion chromatography to offer high selectivity for detecting a variety of alkyl and alkanol amines. techniques. The device features a -JRVJE
$ISPNBUPHSBQIZ
of the MonoSLEEVE column heater perfectly ordered backbone that is "DDFTTPSJFT device, which features advanced formed by etching interstitial volume out Although most of the attention in “plug-and-play” technology. Mott of a silicon wafer. The company reports this series of column instalments Corporation launched HPLC–UHPLC that the device produces highly reliable goes to the development of new static mixers intended to supply and robust reversed-phase, nano-LC columns, accessories play a vital better sensitivity by greatly reducing separations with unprecedented role in enabling liquid separations. baseline noise and improving lower peak capacity. It is certainly possible Table 8 lists several LC-related limits of detection. According to that this development may provide accessories that have been recently the company, the mixers are easy a revolutionary advance for liquid developed. Analytical Sales and to install, are pressure rated up chromatography. Service, Inc., reported the release to 20,000 psi, and can maintain

XXXDISPNBUPHSBQIZPOMJOFDPN  COLUMN
8"5$)

5BCMF
 Accessories for liquid chromatography Company Product Name Comments MonoSLEEVE is the next generation of the original HotSleeve controller. With its new advanced Analytical Sales and MonoSLEEVE "plug-and-play" technology, the controller can accommodate single HotSleeve Plus or AgileSleeve Service, Inc. Plus column heaters of different sizes without having to recalibrate to each individual sleeve. PerfectPeak HPLC–UHPLC mixers are designed to provide better sensitivity by greatly reducing baseline Mott Corporation HPLC–UHPLC noise and maximizing lower limits of detection. The mixers are pressure rated up to 20,000 psig mixers and reportedly maintain high efficiency over a wide range of flows. The EXP Trap column system enables chemists to quickly remove detergents or salts that can affect the ionization process in MS work. This trapping technique can concentrate the sample Optimize EXP Trap directly on-line thus allowing for increased recovery of precious sample material compared Technologies, Inc. columns to off-line techniques. On-line trapping readily lends itself to automation for high-throughput analysis in UHPLC–MS applications. The company’s Free-Turn architecture allows users to change cartridges by hand without breaking fluid connections on the holder inlet–outlet. ZircoFit is a novel capillary fittings system that allows users to assemble their custom-length or Prolab Instruments custom-volume connections within minutes. Using a simple technique, a special tool presses GmbH Zircofit the composite sleeves onto the bare ends of fused-silica capillaries with 10–75 μm i.d. They then fit into 1/16-in. HPLC ports, achieving a pressure rating of 20,000 psi (patent pending). As an extension in the line of the VICI Jour back-pressure regulators, the new JR-BPR3 offers an VICI Jour extended application range. Pressures from 90 to 300 bar (1300–4300 psi) are easily manually VICI AG JR-BPR3 adjustable. The stainless steel made body and the inert membrane enables the usage of any International back-pressure HPLC mobile phase and of supercritical carbon dioxide. The back-pressure regulator can also regulator be mounted into any column oven that maintains temperatures up to 80 °C. Additionally an installation using a 17-mm panel hole is possible. This column is designed to be installed between the gradient mixer and the sample injector for removal of impurities in the mobile phase that contribute to ghost peaks. Without any contact Ghost-Buster with the sample and an easy installation, the column provides superior performance in removing Welch Materials, Inc. column ghost peaks from the baseline, leading to significantly enhanced resolution and minimal impact to established chromatography methods. Because of these unique features, the Ghost-Buster column can be easily adopted in a broad spectrum of application methods.

high efficiency over a wide range of flow rates. Optimize automation for high-throughput analysis and can be Technologies released a series of trap columns under used for multidimensional chromatography processes. the brand name EXP. On-line trapping facilitates High-pressure capillary connections often frustrate the practitioner. Prolab Instruments has developed a novel capillary fittings system called Zircofit that is designed to allow users to assemble connections within minutes. The company reports the achievement of 20,000-psi pressure ratings with this design. VICI AG International launched a new back-pressure regulator, JR-BPR3, which reportedly extends the application range of the company’s existing line. The company claims the new regulator is manually adjustable between 90 bar and 300 bar and can be mounted into any column oven that maintains temperatures up to 80 °C. Lastly, ghost peaks in gradient LC can not only be a nuisance, but may also interfere with detection and quantitation of target analytes. The source of ghost peaks is often impurities in solvents used in mobile phases. Welch Materials has developed what it calls the Ghost-Buster column to help alleviate the issue. The column, packed with proprietary material, is installed between the gradient mixer and the sample injector and is intended to remove contaminants from the mobile phase and thus reduce or eliminate undesired responses in gradient analyses.

$PODMVTJPOT Overall, the products released over the past year represent evolutionary developments. The greatest number of new columns falls into the category of line extensions or “gap-filling” new product lines for individual companies based on existing accepted technologies. New known chemical modifications of both SPPs and FPPs of various particle sizes continue to be developed, and these modern particles continue to find utility in niche separation modes such as HILIC and chiral. Several reported treatments of FPP surfaces may lead

 -$r($
&VSPQF

April 2017 COLUMN
8"5$) to more revolutionary products, and %BWJE
4
#FMM is senior manager directly in the chromatography several new chemistries, especially of separations technologies industry, Dave has focused his efforts hydrophobic–hydrophilic balanced and workflow development on the design, development, and phases, may offer unique and at MilliporeSigma (formerly application of stationary phases powerful selectivities. Sigma-Aldrich/Supelco). With a B.S. for use in HPLC and hyphenated As opposed to last year, there degree from SUNY Plattsburgh and a techniques. In his current role at seems to be less emphasis on Ph.D. in analytical chemistry from The MilliporeSigma, Dr. Bell’s main focus large-molecule separations in terms Pennsylvania State University, Dave has been to research, publish, and of the number of phases that have spent the first decade of his career present on the topic of molecular been launched. That said, several within the pharmaceutical industry interactions that contribute to developments in large-molecule performing analytical method retention and selectivity in an array separations could play a key role in development using various forms of of chromatographic processes. the future of HPLC. An emphasis on chromatography and electrophoresis. Direct correspondence to: dave. even larger-pore-size particles may, During the past 15 years, working [email protected] for example, enable improved analysis of mAbs and other biotherapeutics. Techniques complementary to reversed-phase chromatography such as SEC–GPC and ion-exchange chromatography continue to grow at a CHIRAL steady pace. It is noteworthy, however, HPLC & SFC that no affinity chromatography columns were reported to be released over the past year. Is it possible that the emphasis on large-molecule separations is slowing? The commercialization of pillar-array nano-LC devices may be the most revolutionary development over the past year. It will be interesting to see if the device leads us to more widespread acceptance of chip-based separations.

"DLOPXMFEHFNFOU Product reviews would not be possible without the contributions and cooperation of the manufacturers that have responded to the LCGC survey. Their effort is greatly appreciated. Although LCGC has made every attempt to include every submission TIME TO REFLECT ON YOUR in the series of review articles, it is possible that some have been missed. CHIRAL COLUMN! Free If there have been omissions or if you testing want to be sure to be included in the and 2018 review series, please contact scouting Laura Bush, Editorial Director, LCGC Coated and Immobilised Europe, at [email protected]. Polysaccharides 3FGFSFODFT (1) M.W. Dong, LCGC Europe (4), 208–218 (2017). (2) A. Alpert, A Series of New Materials for Direct HIC-MS Analysis of Proteins in Top-Down Proteomics, presented at the 44th International Symposium Full scalability of High performance Liquid Phase Stable in NP, RP 3-5-10-20 μm Wide application Separations and Related Techniques (HPLC 2016), San Francisco, California, and SFC Bulk available range USA, 2016. (3) D.S. Bell, LCGC Europe (9), 506–515 (2016). (4) D.S. Bell, LCGC Europe (4), 214–224 (2016). discover at www.ymc.de XXXDISPNBUPHSBQIZPOMJOFDPN More than an alternative… PERSPECTIVES IN MODERN HPLC New HPLC Systems and Related Products: "
#SJFG
3FWJFX

.JDIBFM
8
%POH
Perspectives in Modern HPLC Editor

5IJT
JOTUBMNFOU
IJHIMJHIUT
OFX
IJHI
QFSGPSNBODF
MJRVJE
DISPNBUPHSBQIZ
)1-$
TZTUFNT
NPEVMFT
BOE
TPGUXBSF
UIBU
XFSF
JOUSPEVDFE
BU
1JUUDPO

BOE
EVSJOH
UIF
ZFBS
QSJPS
BOE
EFTDSJCFT
UIFJS
JOOPWBUJWF
GFBUVSFT
BOE
TJHOJà
DBOU
CFOFà
UT

This annual review of new high 13,300 after three days. With open /FX
)1-$
BOE
6)1-$
performance liquid chromatography admission on Thursday, a total 4ZTUFNT
BOE
-JOF
&YUFOTJPOT
systems and related products covers of above 14,100 was expected, New HPLC, UHPLC, supercritical products introduced during the which is very good news for the fluid chromatography (SFC), ion last 12 months. The majority of the conference and an indication of chromatography (IC), modules, products were launched at Pittcon a healthier economy and positive chromatography data systems (CDS), 2017. outlook for instrumentation. This and related software are summarized The Pittsburgh Conference on year’s conference featured a plenary in Table 1 and described with more Analytical Chemistry and Applied lecture by Dr. Karl Deisseroth, a details below. Spectroscopy (Pittcon) is one of professor of bioengineering at Agilent 1260 and 1220 Infinity II the world’s largest conferences on Stanford University, on insights systems—Two years ago Agilent laboratory science. The 69th Pittcon on optogenetic proteins using introduced the 1290 Infinity II was held at McCormick Place in electron microscopy and mass LC systems. This year, Agilent Chicago, the largest conference spectrometry (MS). Once again, completed the InfinityLab LC Series centre in the United States, from Pittcon 2017 provided a “mecca” with the 1260 and 1220 Infinity II 5–9 March 2017. The last time for analytical chemists and a systems, all to deliver full method Pittcon was held in Chicago was premier venue for instrument compatibility. Innovative 1290 Infinity in 2014 (and 2011 prior), and it will manufacturers. II LC technology is leveraged with be there again in 2020. Note that the 1260 Infinity II Multisampler Chicago, Orlando, Philadelphia, .FHBUSFOET
JO
)1-$
with dual-needle sampling and New Orleans, and Atlanta are the &RVJQNFOU Vialsampler. A larger column oven, cities selected for hosting Pittcon The perfect marriage of which fits 30-cm columns and can with Chicago in the rotation every ultrahigh-pressure liquid accommodate four columns with a 3 years and the others about every chromatography (UHPLC) and MS single switching valve, and a range 6 years. This frequency makes sense instruments continues to be the of detectors are common to both since Chicago is the third largest city megatrend in chromatography in systems. The 1260 Infinity II pumps in the U.S. and is also an industrial, recent years (1–5), even though have a 600-bar pressure limit and agricultural, financial, transportation, this is another off year for new are available in either stainless steel and communication centre. We UHPLC system introductions. With or bioinert format. The 1220 Infinity had great weather this year at the many second-generation UHPLCs II system remains an integrated conference with many sunny days debuted by all major manufacturers 600-bar system available with an and spring-like temperature in the in prior years (2,3), this year we isocratic or a dual-channel gradient 50s. are seeing mostly line extensions, pump that can be paired with a Pittcon is impressive with ~2000 application-specific systems, manual injector or an autosampler, presentations, 100+ short courses, modules (particularly new MS or MS a variable-wavelength UV detector and a huge exposition with over add-ons), and software products. or a diode-array detector, and 800 exhibitors, including over 120 Table 1 lists the new product external special detectors including new manufacturers displaying for introductions (arranged a mass-selective detector. All the first time. Although the number alphabetically by vendor name), systems can be operated with of attendees has been dropping followed by more detailed Agilent OpenLab CDS Software as in recent years, registration hit descriptions or commentaries. well as third-party software with the

208 -$r($
&VSPQF

April 2017 www.sedere.comwww.sedere.com LowLow T Temperatureemperature EvaporativeEvaporative L ight-Light-ScatteringScattering D Detectorsetectors

BECAUSE YOUR LC DESERVES A UNIVERSAL DETECTION

SEDEX LT-ELSD™ model 100 is the new low temperature evap- orative light scattering detector from SEDERE. This new state- of-the-art compact LT-ELSD™ provides the best detection for liquid chromatography or SFC. This detector presents a num- ber of outstanding innovations thereby providing the best re- sults at a competitive price for pharmaceutical, food, chemical or cosmetics applications. A complete range of detectors Sensitivity, versatility, unri- to fulfill your needs: valled detection of semi volatiles r
Quantitative results and innovative SAGA* (providing more than five decades of dynamic range) makes SEDEX r
Easy optimization LT-ELSD™ model 100 a new breakthrough r
Compatible with in the category of universal detectors. your system

Introducing the new SEDEX LT-ELSD™ model 100 at http://www.sedere.com

S E N S I T I V I T Y SEDERE CONTACT [email protected] BP 10027 - Parc Volta - 9, rue Parmentier TO FIND YOUR CUSTOM SOLUTION. FLEXIBILI T Y 94141 Alfortville Cedex - FRANCE Tel: +33 (0)1 45 18 05 18 -Fax +33 (0)1 45 18 05 25 *Sedex Automated Gain Adjustment E X P E RIE N C E Email: [email protected] PERSPECTIVES IN MODERN
)1-$

5BCMF
 New HPLC product introductions at Pittcon 2017 or the prior year

7FOEPS 1SPEVDU%FTDSJQUJPO
BOE
$PNNFOUT Software includes overlaid logD curves, enhanced column databases, and better ACD/Labs AutoChrom 2016 version connectivity to Chromeleon 7, Empower, and other MS data systems. LUMINATA is a new multitechnique software product that offers comprehensive data ACD/Labs LUMINATA aggregation for impurity characterization. MetaSense is an LC–MS software solution to facilitate identification of predicted and ACD/Labs MetaSense unexpected metabolites. 1260 and 1220 Infinity II Agilent Updated platforms for Agilent’s Infinity II family of HPLC and UHPLC systems. systems Offers an improved autosampler and an updated control module with an optional Agilent 1260 Infinity II SFC binary pump. 1290 Infinity II preparative Enables collection of up to 432 fractions or volumes up to 5.9 L triggered by time-, Agilent fraction collector peak- or mass-based events. This version of OpenLAB CDS provides a networked or standalone environment that Agilent OpenLab CDS 2.1 combines ease of use and regulatory compliance. Analytical Sales Sentinel A fluid-level indicator for monitoring up to three bottles of waste or mobile phases. and Services DEGASi Compact and DEGASi Compact with small internal membrane volume and DEGASi Prep+ Biotech AB Prep+ degassers supports preparative HPLC and flash chromatography. OCTOPUS purification A flexible modular purification system for both flash and preparative HPLC with Bonna-Agela system automatic fraction collection and many detection modes. Bruker Elute LC series UHPLC systems provide pressures from 700 to 1300 bar for LC–MS analysis. Cecil Electrochemical Detection A modular HPLC system with electrochemical detection that supports Instruments Adept HPLC DC-amperometric, scanning, and pulsed modes. ChromPerfect for small laboratories that controls up to four chromatographs and is ChromPerfect ChromPerfect SL CDS compatible to tablets control and data analysis. A portable, battery operated HPLC–UV system with novel approaches to column Cromite A portable HPLC/UV and sample probe design. An updated software version operating under Windows 10 with enhanced instrument DataApex Clarity v.7.2 CDS connectivity and MS extensions. ECOM ECF 2096 fraction collector An automated collector that collects up to 48 tubes of 8-mL volumes. Very small photometric, fluorimetric, and conductivity HPLC detectors based on LED FF Runge GmbH Mikron detector series technology. JASCO offers both LC-4000 Series HPLC–MS and SFC–MS systems with an add-on JASCO HPLC–MS and SFC–MS of Advion’s Expression SQ MS system. JASCO offers method scouting software for automated solvent and column JASCO Method scouting software screening (up to 10 solvents and 10 columns). A monitoring system for HPLC reservoirs and waste containers using sonic detection JM Science Sonic reservoir system and LED display. Azura Analytical and Semi- A family of analytical and semi-preparative HPLC systems (200, 400, 700, and Knauer prep HPLC 1000 bar) in stainless and biocompatible formats. The Eco IC is a new ion chromatograph designed for routine analysis of anions, Metrohm Eco IC cations, and polar compounds in environmental testing and education. The QSight triple-quadrupole system is a self-cleaning analyzer for testing food PerkinElmer QSight MS/MS System contaminants in compliance with regulatory standards. Prolab ZirconiumHT UHPLC The Zirconium High-Throughput UHPLC system is a micro-UHPLC system with novel Instruments system trapping configuration for 2D applications. Sciex X500B QTOF MS A compact QTOF system for standardized biotherapeutics characterization. Shimadzu Cannabis Analyzer for The Shimadzu Cannabis analyzer is an HPLC–UV system designed specifically for Potency quantitative determination of cannabinoid content. Shimadzu CLAM-2000 automation CLAM-2000 clinical laboratory automation module automatically performs the module for sample preparation necessary for analyzing blood and other biological samples by LC–MS/MS LC–MS/MS. Shimadzu LCMS-8045 This system performs demanding routine quantitative analyses in food safety and environmental testing.

 -$r($
&VSPQF

April 2017 PERSPECTIVES IN MODERN
)1-$

7FOEPS 1SPEVDU%FTDSJQUJPO
BOE
$PNNFOUT A version of Fusion QbD software for HPLC method development using design of S-Matrix Fusion QbD v. 9.8.0 experiments with interfaces to most HPLC systems. Thermo Vanquish Flex binary A biocompatible binary pump for the Vanquish Flex UHPLC system rated at Scientific pump 1000 bar (15,000 psi). Thermo Q Exactive HF LC–MS/MS A compact orbital trap hybrid LC–MS/MS system capable of a maximum resolution Scientific system of 240,000 FWHM. Thermo This version of Chromeleon is a multitechnique, multivendor scalable platform for Chromeleon 7.2 SR5 CDS Scientific GC, LC, IC, and MS applications with many enhancements. The latest AppsLab library is a searchable on-line repository for access to Thermo AppsLab methods library chromatography and MS applications including analytical methods, chromatograms, Scientific and related compound information. Acquity H-Class UPLC for Waters offers an automated injector for dissolution based on Sotax’s Xtend Waters/Sotax dissolution testing Dissolution Line. Waters announced cloud-based Empower 3 software available through Amazon Waters Empower Cloud Web Services. Wyatt ViscoStar III An on-line viscometer detector offers sensitive measurements of intrinsic viscosity for macromolecular size and polymer branching. universal Agilent Instrument Control Cromite portable HPLC—Cromite The system is designed with novel Framework. introduced a handheld, portable approaches to mobile-phase Bruker Elute LC series—Bruker HPLC system that consists of an storage and sample introduction introduces the Elute LC series isocratic pump with a 275-bar or and preparation, and it includes a systems for LC–MS analysis: SP 4000-psi pressure limit and a max single-ended leak-proof column. (700 bar), UHPLC (1300 bar for flow rate of 4 mL/min, and a UV–vis The system weighs 2.5 lb (1 kg) and ultrafast analysis), OLE (for on-line detector with a light-emitting diode has overall dimensions of 6.8 × 3.2 extraction), and HT (high-throughput (LED) source that provides up to × 3.4 in. (173 × 81 × 86 mm). The with the new PAL3 autosampler). four customizable wavelengths. device can be controlled from a PC,

Leverage the outstanding inertness, low bleed, and high reproducibility of Rxi® 3-in-1 technology to gain: 
  
ata



 
 

 
 
 Put Rxi® columns to work in your lab. Visit www.restek.com/rxi and order yours today.

Rxi® columns come with an unbeatable guarantee. That is Restek® Pure Satisfaction.

Pure Chromatography www.restek.com/rxi

XXXDISPNBUPHSBQIZPOMJOFDPN  PERSPECTIVES IN MODERN
)1-$

smartphone, or tablet via Bluetooth and can be powered off a 12 VDC 'JHVSF
 An example chromatogram obtained using Cromite’s portable HPLC system. Column: 100 mm × 3 mm, 5-μm d Cromite C18; mobile phase: 70:30:0.5 power supply or external battery. The p (v/v/v) water–acetonitrile–acetic acid; detection: UV absorbance at 275 nm; flow rate: company is targeting applications 1 mL/min. in education, environmental, and production QC testing with this Benzoic acid offering. Figure 1 shows an example chromatogram obtained using the Cromite portable HPLC system. 50 8JUI
PQFO
BENJTTJPO

PO
5IVSTEBZ
B
UPUBM
40 PG
BCPWF
 
XBT
FYQFDUFE
XIJDI
JT
WFSZ
HPPE
OFXT
GPS
30

UIF
DPOGFSFODF
BOE
Benzonitrile BO
JOEJDBUJPO
PG
B
20 IFBMUIJFS
FDPOPNZ
BOE
Absorbance (mAU) 2-Aminopyridine QPTJUJWF
PVUMPPL
GPS
JOTUSVNFOUBUJPO
10

Knauer Azura Analytical and Semi-Prep HPLC—Knauer offers the Azura as a system solution for 0 analytical to preparative HPLC. The analytical systems are available with 0123 binary or quaternary pumps (stainless Time (min)

HPLC GC MS SPE IR BLS BIO powered by We have 1000’s of eLearning topics CHROMacademy is the world’s largest eLearning website for analytical scientists, containing 1000’s of interactive learning topics. Lite members have access to less than 5% of our content. Premier members get so much more!

Find out more about CHROMacademy Premier membership contact: Glen Murry on +1 732.346.3056 | e-mail: [email protected] Peter Romillo: +1 732.346.3074 | e-mail: [email protected] www.chromacademy.com PERSPECTIVES IN MODERN
)1-$

'JHVSF
 An example chromatogram of a 10.0-mg/L standard mixture of 11 cannabinoids separated on the Shimadzu Cannabis analyzer using the instrument’s high-sensitivity method. Gradient elution conditions with acid-modified water and acetonitrile were used with a C18 column.

60

50 CBN

40 CBDV CBGA ∆9-THC CBC THCA CBDA

30 CBD CBG THCV ∆8-THC

Response (mV) 20

10

0

2.0 3.0 4.0 5.0 6.0 7.0 Time (min) steel at 700 and 1000 bar as well as is a HPLC–UV system designed dissolution analyzer, the wait is a biocompatible, ceramic version specifically for quantitative over. Waters, in conjunction with at 400 bar). The semipreparative determination of cannabinoid Sotax, now offers an automated 200-bar systems are capable of content for potency with three UHPLC system based on the Acquity flow rates as high as 50 mL/min and method packages optimized for H-Class UPLC system and Sotax’s are available in both stainless and high-throughput, high sensitivity, and dissolution testing apparatus with biocompatible formats. These systems high resolution. Figure 2 shows an Sotax’s sample manager (SAM) to can be configured with appropriate example chromatogram for potency allow sample withdrawals, fractions autosamplers, column ovens, testing of 11 cannabinoid standards collection, and autoinjections from and UV–vis, refractive-index, and on the system. USP dissolution systems. diode-array detectors. The systems Bonna-Agela OCTOPUS purification can be controlled by a controller, a 1JUUDPO
JT
JNQSFTTJWF
XJUI
system—Bonna-Agela Technologies mobile tablet, or a chromatography _
QSFTFOUBUJPOT
+
(acquired by Danaher last year) data system (CDS) such as OpenLAB introduced a flexible and easy-to-use EZChrom, ClarityChrom, and TIPSU
DPVSTFT
BOE
B
IVHF
modular purification system for both Chromeleon. FYQPTJUJPO
XJUI
PWFS

flash and preparative HPLC with a Prolab Instruments ZirconiumHT— FYIJCJUPST
JODMVEJOH
PWFS
family of four pumps (1–200 mL/min The Zirconium High-Throughput and pressure limits of 14–200 bar UHPLC system is a micro-UHPLC 
OFX
NBOVGBDUVSFST
[200–3000 psi]) that can be equipped system with a novel trapping EJTQMBZJOH
GPS
UIF
àSTU
with automatic fraction collection, an column configuration for loading UJNF autosampler, a column oven/switcher, and analyzing samples at the same and UV–vis, photodiode-array, time using only one binary pump. Sotax Xtend On-Line Dissolution evaporative light scattering, Suggested applications are 2D micro with Waters Acquity H-Class UPLC— refractive-index, or MS detection. and nano-LC analyses. For those who have been waiting for Agilent 1260 Infinity II SFC—The Shimadzu Cannabis analyzer— a modernized replacement of the Agilent 1260 Infinity II SFC system The Shimadzu Cannabis analyzer Alliance-based 2695D automated now offers an improved Multisampler

XXXDISPNBUPHSBQIZPOMJOFDPN  PERSPECTIVES IN MODERN
)1-$

autosampler, which uses an innovative feed injection principle 'JHVSF
 A screen shot of the Agilent OpenLab 2.1 CDS showing four tiled panels on that allows 0.1–90 μL of injection data processing, chromatograms, peak analysis, and processing methods. volume range with the higher sample capacity of 432 2-mL vials. The 1260 Infinity II SFC binary pump can provide pressures of 600 bar at up to 5 mL/min. The Infinity II SFC control module now offers full flow into evaporative light scattering or MS detection systems. A novel low dispersion nozzle with variable back-pressure settings enables outstanding peak shapes in this case. Metrohm Eco IC—The Eco IC ion chromatograph is designed for routine analysis of anions, cations, and polar compounds in environmental testing and education applications. The system is complete with a suppressor, a conductivity detector, software, and an optional autosampler with up to 36 samples.

)1-$
.PEVMFT

Cecil Instruments Electrochemical Detection Adept HPLC system— Cecil Instruments offers a modular

FUN HYP APP YOU FUNDAMENTALS HYPHENATIONS APPLICATIONS YOUNG & TUTORIALS

Key Dates Registration Fees March 6, 2017 Early Regular Abstract deadline for Best Poster Award Academic 650 € 790 € March 20, 2017 Industry 900 € 1 080 € Deadline for early registration payment Student 240 € 340 € April 17, 2017 Abstract deadline for poster presentations Student + 2 Short Courses 360 € 460 € [email protected] www.hplc2017-prague.org PERSPECTIVES IN MODERN
)1-$

HPLC system with electrochemical detection that Shimadzu LCMS-8045—Shimadzu’s LC-MS/MS-8045 supports DC-amperometric, scanning, and pulsed system is a mid-range triple-quadrupole MS system that modes. performs routine quantitative analyses in food safety and FF Runge Mikron detector series—These very compact environmental testing. The instrument features a scan photometric, fluorimetric, and conductivity HPLC speed of 30,000 u/s without the loss of mass accuracy detectors are based on LED technology. and a polarity switching time of 5 ms. It is operated using Thermo Scientific Vanquish Flex binary pump—The Shimadzu’s LabSolutions software. A dual electrospray Vanquish Flex binary system now features a binary ionization (ESI) and atmospheric-pressure chemical high-pressure gradient pump with 2 × 3 solvent channels ionization (APCI) source is available. (1000 bar or 15,000 psi) and low gradient delay volume Shimadzu CLAM-2000 clinical laboratory automation capable of delivering flow rates of up to 8 mL/min. The module for LC–MS—The CLAM-2000 system automatically entire system provides a biocompatible flow path. performs all of the sample preparation procedures Wyatt ViscoStar III—The Wyatt ViscoStar III viscometer necessary for analyzing blood (from blood collection tubes) detector offers measurements of intrinsic viscosity and other biological samples by LC–MS/MS for therapeutic for sensitive macromolecular size and conformation drug monitoring and testing of abused drugs. for polymer branching analysis. The detector uses Thermo Scientific Q Exactive HF—This compact Mark-Houwink-Sakurada parameters and hydrodynamic quadrupole-orbitrap hybrid LC–MS/MS system designed radii intrinsic viscosity calculations using Wyatt’s ASTRA for rapid quantitation of proteins, peptides, lipids, software package in size-exclusion chromatography. metabolites, and contaminants is capable of scan rates up to 18 Hz and a maximum resolution of 240,000 FWHM. 5IJT
ZFBST
DPOGFSFODF
GFBUVSFE
B
QMFOBSZ
MFDUVSF
CZ
%S
,BSM
%FJTTFSPUI
)1-$
"DDFTTPSJFT
Agilent 1290 Infinity II preparative fraction collector—This B
QSPGFTTPS
PG
CJPFOHJOFFSJOH
BU
automated open-bed fraction collector allows efficient 4UBOGPSE
6OJWFSTJUZ
PO
JOTJHIUT
PO
and flexible automated collection of up to 432 fractions PQUPHFOFUJD
QSPUFJOT
VTJOH
FMFDUSPO
or volumes up to 5.9 L triggered by time-, peak-, or mass-based mode. NJDSPTDPQZ
BOE
NBTT
TQFDUSPNFUSZ
Analytical Sales and Services Sentinel—The Sentinel .4 
is a fluid-level indicator for monitoring up to three bottles

.BTT
4QFDUSPNFUSZ

JASCO HPLC–MS and SFC–MS—JASCO offers the Advion Expression compact single-quadrupole MS system for its HPLC–MS (LC-4000 Series) and SFC–MS (SFC-400 Series SFC) systems with mass-triggered fraction collection and integrated system control using the ChromNAV CDS. ADVANCING BIOPHARMACEUTICAL DEVELOPMENT PerkinElmer QSight MS/MS system—The QSight triple-quadrupole system is a highly sensitive dual source self-cleaning LC–MS/MS analyzer that enables laboratories to test complex samples for food contaminants (such as pesticides) for regulatory compliance. QSight provides a complete pesticide solution from sample preparation and extraction to analysis and reporting. Sciex X500B QTOF—Sciex launches the compact X500B QTOF system designed to bring simplicity, performance, and robustness for biotherapeutic analyses. The new SCIEX OS software interface and BioPharmaView 2.0 processing software provide an integrated LC–MS solution for experts or novices performing standard biotherapeutic 22-24 May 2017 characterization. Sciex QTRAP 6500+ LC–MS/MS—This combined The Malton Hotel, Killarney, Ireland quadrupole–ion trap system includes the highly sensitive triple-quadrupole 6500+ MS/MS system for high-throughput LC–MS/MS analysis for both small- and large-molecule quantitation. The MS system is Scan here or visit equipped with the multicomponent IonDrive technology www.casss.org for for enhancing sensitivity and is capable of six orders of program updates. magnitude of linearity, up to 20,000-u/s scan rates, and 5-ms polarity switching.

XXXDISPNBUPHSBQIZPOMJOFDPN  PERSPECTIVES IN MODERN
)1-$

'JHVSF
 A screen shot of ACD/Labs LUMINATA software displaying multiple panels of an overall synthetic scheme of a drug substance, a table of potential impurities, and the data associated with each impurity (HPLC chromatogram, UV, and mass spectra).

of waste or mobile-phase containers $ISPNBUPHSBQIZ
%BUB
4ZTUFN
under Windows 10 and can be and is equipped with status display BOE
0UIFS
)1-$3FMBUFE

connected to four instruments and and alarm functions. 4PGUXBSF
up to 32 data channels. Its optional Biotech AB DEGASi Compact Agilent OpenLAB CDS 2.1—This control modules provide integrated and Prep+ degassers—Two new newest version of OpenLAB CDS control of selected instruments. degassers were introduced: The provides a networked or standalone Extensions are available for DEGASi Compact degasser with environment that combines ease diode-array detectors, gel permeation a small internal volume and the of use with rigorous regulatory chromatography (GPC) analysis, MS DEGASi Prep+ degasser that compliance. This latest platform (single quadrupole and time of flight), supports preparative HPLC, is an LC-, GC-, LC–MS-, and and system suitability testing. flash chromatography, and GC–MS-capable CDS for the Waters Empower Cloud— high-throughput applications. QC laboratory with strong data Waters announced a release of integrity and audit trail review cloud-based Empower 3 (feature 5IF
QFSGFDU
NBSSJBHF
capabilities. Version 2.1 contains release 4) available through Amazon PG
VMUSBIJHIQSFTTVSF
many new features including an Web Services in late 2017. This extended support for non-Agilent cloud-based CDS platform is MJRVJE
DISPNBUPHSBQIZ
instruments and improvements in particularly suited for pharmaceutical 6)1-$
BOE
.4
MS quantitation, peak integration, companies to reduce costs for JOTUSVNFOUT
DPOUJOVFT
and reporting. Figure 3 shows a implementation and maintenance and UP
CF
UIF
NFHBUSFOE
JO
screen shot of the Agilent OpenLAB can globally support all subsidiaries CDS with four tiled panels for data and contract organizations. DISPNBUPHSBQIZ
JO

processing, chromatograms, peak ACD/Labs AutoChrom 2016 SFDFOU
ZFBST analysis, and processing methods. version—This automated HPLC ChromPerfect SL—ChromPerfect method development software ECOM ECF 2096 fraction introduces a CDS for small or mobile uses predictive simulation and collector—The ECF 2096 automated laboratories that controls up to four can interface with both UV and MS fraction collector collects up to 48 chromatographs with web-based and detection. This version includes tubes of 8-mL volumes. Windows 10 operating systems. It overlaid logD curves, better JM Science Sonic reservoir is compatible with tablets with data peak tracking, and an enhanced system—A monitoring system for up processing, reporting, and analysis column data base with Tanaka to three HPLC reservoirs and waste functions. parameters and superficially porous containers using sonic detection and DataApex Clarity v.7.2 CDS—The particle columns. It also has better an LED display. Clarity v.7.2 advanced CDS operates connectivity to Chromeleon 7,

 -$r($
&VSPQF

April 2017 FREE CHROMACADEMY MEMBERSHIP

Agilent Technologies is offering five years complimentary access to CHROMacademy for all university students and staff. CHROMacademy is an intuitive, comprehensive e-learning and trouble- shooting platform with more than 3,000 pages of content for HPLC, GC, sample preparation, and hyphenated techniques. No other online resource offers separation scientists more live streaming events, a knowledge base, practical solutions, and new technologies in one easy to navigate website. Get your free five year membership worth US $1,995* by submitting the form at www.chromacademy.com/agilent.

* Five years free access to CHROMacademy only available to customers affiliated with an academic or research institution, conditions apply. A valid university e-mail address if required.

© Agilent Technologies, Inc. 2017 PERSPECTIVES IN MODERN
)1-$

Empower, and other MS data unique workflow automation called restaurants and the warmer weather— systems. eWorkflows. The new platform is and I plan to visit both the exposition ACD/Labs LUMINATA—LUMINATA optimized for large-scale installation and the theme parks. multitechnique software offers for thousands of users and This instalment provides a brief comprehensive data aggregation instruments with data integrity features and nonexhaustive overview of new for impurity characterization and for regulatory compliance. This latest HPLC product introductions (systems, knowledge management applied platform is designed particularly for modules, MS, CDS, software, and to pharmaceutical development MS analysis including biotherapeutics related products) at Pittcon 2017 and applications. Figure 4 shows a and is equipped with capabilities for during the prior year. Readers should screen shot from this software automated target mass analysis for contact manufacturers for additional displaying multiple panels including both LC–MS and GC–MS. technical details. Note that new HPLC the overall synthetic scheme of Thermo Scientific AppsLab methods columns are covered separately in the drug substance, a table of library—The latest AppsLab library LCGC ’s “Column Watch” annual review potential impurities, and the data is a searchable on-line repository for by David Bell. The opinions expressed associated with each impurity easy access to chromatography and in this instalment are the author’s (HPLC chromatogram, UV, and mass MS application databases including own and bear no reflections on those spectra). analytical methods, chromatograms, of LCGC magazine, the Pittsburgh ACD/Labs MetaSense—MetaSense and related compound information Conference, or any other organizations. software uses LC–MS data as (1886 applications and 380 part of its workflow in metabolite ready-to-use eWorkflows for "DLOPXMFEHFNFOUT identification. It facilitates faster, Chromeleon CDS). I wish to give thanks to the marketing easier, and more accurate detection and editorial staff of LCGC and all and identification of predicted and $PODMVEJOH
BOE
1FSTPOBM
HPLC manufacturers for their timely unexpected metabolites. Built on the 3FNBSLT
responses to my numerous requests ACD/Spectrus platform, MetaSense Most analytical chemists come to for information. I am particularly offers functionalities including the Pittcon for professional development grateful to Davy Guillarme and facilitation of decision-support and networking. My schedule this Szabolcs Fekete of U. Geneva, Harika and effective knowledge sharing year included teaching several short Vemula of U. Missouri at Kansas throughout organizations as well courses on HPLC, UHPLC, and City, David Van Meter from EAG as the ability to support LC–MS method development, and chairing an Laboratories, Hernan Fuertes from analytical data from most instrument invited symposium on UHPLC method Ardelyx, and Matt Mullaney from vendors. development in pharmaceutical Pentec Health for their invaluable JASCO method scouting software— analysis. My networking events editorial and technical input. JASCO offers a method scouting included a Chrom Mixer sponsored software for automated solvent and by Chrom Forum Delaware Valley in 3FGFSFODFT column screening with up to 10 memory of Jack Kirkland (more than (1) C.H. Arnaud, Chem. & Eng. News solvents and 10 columns. It also 500 attendees), a networking dinner (24), 29–35 (2016). (2) M.W. Dong, LCGC North Amer. (4), supports multiple detectors and can event with the Chinese American 262–273 (2016). be configured for use with up to 30 Chromatography Association (CACA) (3) M.W. Dong, LCGC Europe 29(4), columns and parallel screening. attended by more than 120 conferees 223–231 (2015). S-Matrix Fusion QbD method (featuring a lecture by Dr. Perry (4) D. Guillarme and M.W. Dong, Amer. Pharm. Rev. (4), 36–43 (2013). development software v. 9.8.0—The Wang of the US FDA on the perfect (5) D. Guillarme and M.W. Dong (Eds.), Fusion QbD software for automated marriage of HPLC and MS), and the UHPLC: Where We Are Ten Years After LC method development using annual Pittcon presidential reception, Its Commercial Introduction, Trends in Anal. Chem. , 1–188 (2014). principles of quality by design where I personally thanked many (QbD) and design of experiments Pittcon staff and programme chairs. .JDIBFM
8
%POH is a principal of (DOE) has been updated to include I spent considerable time on the MWD Consulting, which provides an enhanced user interface with exposition floor to verify information training and consulting services in an integrated chromatogram on new HPLC products for this column HPLC and UHPLC, pharmaceutical viewer, new project management instalment. I returned home Thursday analysis, and drug quality. He capabilities with extended data night, slept late, and woke up to find was formerly a Senior Scientist at security, additional support for GC 4 inches of snow on the ground in Genentech, Research Fellow at Purdue and capillary electrophoresis (CE) Connecticut. We were lucky as the Pharma, and Senior Staff Scientist at method development, and increased weather can be unpredictable in Applied Biosystems/PerkinElmer. He instrument support for most HPLC early March in Chicago, as well as holds a Ph.D. in analytical chemistry and UHPLC systems, including hybrid the Northeast. Rumour says that next from City University of New York. He systems. year will be the last year for Pittcon has more than 100 publications and a Thermo Scientific Chromeleon 7.2 in Orlando because the vendors best-selling book in chromatography. SR5 CDS—The Chromeleon CDS complained about conference He is an editorial advisory board is a multitechnique, multivendor attendees visiting Disney World member of LCGC North America. scalable platform for GC, LC, IC, instead of the exposition. I love Direct correspondence to: LCGCedit@ and MS applications and includes a Orlando because of the excellent ubm.com

 -$r($
&VSPQF

April 2017 PRODUCTS MALS detector ICP-MS system The μDAWN is, The Thermo Scientifi c iCAP TQ according to the ICP-MS system reportedly provides company, the world’s scientists with access to a triple fi rst multi-angle light quadropole mass spectrometer scattering (MALS) that offers advanced interference detector that can be removal and lower detection limits coupled to any UHPLC for challenging sample matrices, system to determine while offering a sample introduction absolute molecular weights and sizes of polymers, peptides, component that bolsters experiment and proteins or other biopolymers directly, without resorting reproducibility. The detection of metals and several to column calibration or reference standards. The WyattQELS nonmetals is essential to evaluate clinical samples for Dynamic Light Scattering (DLS) module, which measures disease, contamination on industrial sites, and the safety hydrodynamic radii “on-the-fl y”, reportedly expands the of food for human consumption. versatility of the μDAWN. www.thermoscientifi c.com www.wyatt.com Thermo Fisher Scientifi c, California, USA. Wyatt Technology, Santa Barbara, California, USA.

Thermal desorption consumables Hydrogen gas for GC Markes International has released its 2017–2018 catalogue for thermal The Vici DBS range of FID gas desorption tubes, accessories, and stations with sophisticated software spares. According to the company, the control and alarm capability present catalogue contains everything the TD the GC user with the opportunity analyst needs for successful VOC and to reap all the benefi ts offered SVOC monitoring and analysis. This by hydrogen carrier gas, whilst edition also contains Markes’s HiSorb overcoming the safety concerns. products for sorptive extraction in According to the company, these addition to an extended section on thermal desorption sorbent unique instruments combine the sampling tubes. The catalogue is available as an interactive reliability of the Vici DBS hydrogen and zero air generators on-line version (chem.markes.com/TDCat) or as hardcopy from into one compact and convenient FID package. [email protected]. http://www.dbsinstruments.com/en/prodotti/fi d_ www.markes.com tower_plus/ Markes International Ltd., Llantrisant, UK. Vici AG International, Schenkon, Switzerland.

Solvent transportation Chromatography software To facilitate laboratory operations, Clarity is modular; users can Polymer Char has designed a convenient confi gure Clarity to fi t their tool to transfer fresh and waste solvents, needs using optional application such as TCB and DCB, between a fi lling extensions. According to the station and different instruments without company, Clarity can grow coming into direct contact with either from a simple solution for one solvents or vapours. Solvents can be chromatograph into a complex transferred in a safe and convenient way solution for laboratories with multiple chromatographs around the laboratory using the Solvent operating in a regulated environment. It enables control Handling Trolley, according to the company. of hundreds instruments from one place and has six http://polymerchar.com/solvent_handling_trolley language localizations. The company offer a free demo Polymer Char, Valencia, Spain. version. www.dataapex.com DataApex, Prague, Czech Republic.

www.chromatographyonline.com 219 PRODUCTS

Preparative system LC/SFC system Quattro countercurrent and centrifugal Shimadzu’s Nexera UC/s partition chromatographs and (SFC/UHPLC switching extractors are designed to work with, system) allows measurements and complement standard fl ash by liquid chromatography and HPLC laboratory and process (LC) and supercritical fl uid instrumentation. When appropriate, chromatography (SFC) on replacing the solid–liquid columns with a single system. Switching unique liquid–liquid instrumentation between SFC and LC enables allows preparations from milligram to tonnes per annum. No rapid screening for optimum on-column adsorption or degradation will occur, according to the separation conditions resulting in improved analytical company. A mass-balance is the norm for CCC/CPC. Typically efficiency. Through a newly released upgrade kit, a 50–80% solvent saving occurs. Standard biphasic solvents, UHPLC units already installed can be upgraded to the ionic liquids, liquid chiral selectors, and ion exchangers may Nexera UC/s, thereby decreasing investment cost for the all be utilized. According to the company, crude material that additional SFC system. would poison standard columns can be injected without causing www.shimadzu.eu contamination. 4IJNBE[V
&VSPQB
(NC)
%VJTCVSH
(FSNBOZ www.quattroprep.com "&$42VJL1SFQ
-UE
-POEPO
6, Degasser Ion chromatography The white Biotech Degasi Compact is a new line of standalone degassers Modular, versatile, effortless, combining the cutting-edge Systec reliable, and sensitive, IonQuest technology with a very small footprint Ion Chromatography, with at an affordable cost. It is available automation options, can reportedly with 2-, 4-, or 6 degassing channels be used in water, environmental, in a housing with only a 167 × 56 mm cosmetics, electroplating, mining, footprint. Closed-loop control with a fracking, semi-conductor, effluent, continuously running vacuum pump biotechnology, clinical chemistry, gives a smooth baseline. petrochemical, pharmaceutical, and https://www.biotech.se/ food and beverage laboratories. Biotech AB, Onsala, Sweden. The low drift and low noise conductivity detector may be purchased separately for use with third-party chromatography systems. www.cecilinstruments.com Cecil Instruments Limited, Cambridge, UK.

LC–MS/MS instrument HPLC solvents PerkinElmer, Inc., has announced the launch Carlo Erba’s solvents for HPLC of its QSight Triple Quadrupole LC–MS/MS meet the requirements of HPLC instrument with patented fl ow-based mass by guaranteeing the optimal spectrometry that enables laboratories to specifi cations on purity, volatile test highly complex samples and experience residue content, and UV transmission, increased throughput, according to the according to the company. The company. Combined with PerkinElmer’s product line includes solvents for Altus UPLC instrument, the QSight system UHPLC–MS, solvents, additives and reportedly offers a complete solution from blends for LC–MS, solvents for HPLC sample preparation to results and reporting for gradient, solvents for HPLC isocratic food, industrial, and environmental applications. For regulatory food and solvents for HPLC preparative, ion pair reagents, safety purposes, the QSight instrument specializes in detecting a syringe fi lter, vials, micro-inserts, and much more. wide range of pesticides that are increasingly found in crops. It can http://www.carloerbareagents.com also test foods for mycotoxins and antibiotics as well as analyze $BSMP
&SCB
3FBHFOUT
4"4
7BM
EF
3FVJM
'SBODF veterinary drugs and nutritional components. www.perkinelmer.com/Product/qsight-220-multi-opt-dual- source-system-bc003382 1FSLJO&MNFS
*OD
.BTTBDIVTFUUT
64"

220 -$r($
&VSPQF
April 2017 PRODUCTS

Sample preparation system LC system The Microlab 600 is a Agilent has launched its new highly precise syringe InfinityLab product family, pump designed to quickly including the Agilent 1260 dilute liquids. According to Infinity II liquid chromatography the company, this positive system. Integrating the displacement system high-end technology of provides better than 99% Agilent’s flagship 1290 LC accuracy, independent system into the company’s core platform, the 1260 Infinity of a liquid’s viscosity, system offers an ergonomic design that aims to increase vapour pressure, and customer efficiency, reduce costs, and improve overall temperature. The inert fluid path minimizes sample carryover usability. According to the company, the InfinityLab family and improves compatibility with harsh chemicals. provides the end-to-end solution laboratories need to http://www.ml600.com/ ensure analytical excellence. Hamilton Bonaduz AG, Bonaduz, Switzerland. www.agilent.com Agilent Technologies, Inc., California, USA.

HPLC columns Mobile MS benches Phenomenex Inc., has introduced a 5-μm particle size to its family of Kinetex F5 Manufacturer of innovative pentafluorophenyl propyl (PFP) core– mobile benches for LC/GC/ shell columns. The Kinetex F5 is a robust MS. IonBench for mass PFP core–shell phase that, according to spectrometry reportedly the company, overcomes reproducibility removes 75% of the and performance limitations of other noise, eliminates 99% of PFP and F5 products on the market the vibration, and saves and significantly reduces method up to 30% of floor space. development time with its dynamic According to the company, IonBench for HPLC improves and responsive chemical functionality. system performance, enhances laboratory safety, and With five retention mechanisms and contributes to productivity. five separation modes, it is an effective www.ionbench.com orthogonal alternative to the widely used C18 and C8 phases. IonBench, Joigny, Burgundy, France. www.phenomenex.com Phenomenex Inc., California, USA.

Customizable assemblies UHPLC columns Merck has launched the Mobius Versatility, robustness, and MyWay Portfolio, a program scalability from UHPLC to prep: that allows more flexibility, YMC-Triart. “Hybrid-style” phase better supply predictability, offer a wide range of stability and shorter lead times for towards pH, temperature, and more efficient and safer drug 100% aqueous eluents. Six manufacture, according to the bonded phases provide a broad company. Merck’s portfolio offers selectivity range: C18, C18 a choice of off-the-shelf and configured-to-order assemblies, ExRS, C8, phenyl, PFP, and as well as fully customizable configurations. Assemblies are diol-HILIC. Scalable from 1.9 μm segmented into three categories, each offering a different level of to 20 μm—from UHPLC to prep. configuration flexibility. http://ymc.de/ymc-triart.html www.merckmillipore.com :.$
&VSPQF
(NCI
%JOTMBLFO
(FSNBOZ Merck Group, Darmstadt, Germany.

www.chromatographyonline.com 221 &7&/5
NEWS

*41"$

UI
*OUFSOBUJPOBM
4ZNQPTJVN
PO m
.BZ
 1PMZNFS
"OBMZTJT
BOE
$IBSBDUFSJ[BUJPO $ISPN4PDT
4QSJOH
4ZNQPTJVN 4VQFSDSJUJDBM
'MVJE

6MUSB)JHI
The UI
*OUFSOBUJPOBM
4ZNQPTJVN
PO
1FSGPSNBODF
$ISPNBUPHSBQIZ 1PMZNFS
"OBMZTJT
BOE
$IBSBDUFSJ[BUJPO
Innovation House, Kent, UK *41"$ will take place in -JO[
"VTUSJB, on &NBJM
chromsoc@ m
+VOF
. meetingmakers.co.uk *41"$ is a non-profit scientific 8FCTJUF
www.chromsoc.com/ organization formed to provide an chromsocevents international forum for the presentation of recent advances in the field of polymer analysis and characterization methodologies. This unique m
+VOF
 symposium brings together analytical chemists and polymer scientists )1-$
 who are involved in the analysis and characterization of polymeric Prague, Czech Republic materials. Meetings are held annually, rotating to venues in the USA, &NBJM
[email protected] Europe, and Asia, with ISPAC 2016 taking place in Singapore. 8FCTJUF
www.hplc2017-prague.org *41"$
sessions comprise a three-day programme and feature invited lectures, poster sessions, and discussion and information exchange m
+VMZ
 sessions, all focused towards polymer analysis and characterization 4BNQMF
1SFQBSBUJPO
4VNNFS
approaches, techniques, and applications. Each session features multiple $PVSTF



lectures and a one-hour open discussion period where participants from Chania, Crete, Greece academic, industrial, and government settings can discuss different &NBJM
sampleprep2017@ aspects of polymer analysis and characterization approaches, techniques, enveng.tuc.gr and applications. The symposium offers a perfect networking platform, 8FCTJUF www.sampleprep2017.tuc.gr enabling the free exchange of ideas and tips about different techniques, as well as an opportunity to learn about the latest developments m
+VMZ
 first-hand. *OUFSOBUJPOBM
4ZNQPTJVN
GPS
*41"$
 will focus on several topics including a broad spectrum )JHI
1FSGPSNBODF
5IJO-BZFS
of polyolefin-related themes, specific topics on advanced fractionation, $ISPNBUPHSBQIZ
)15-$
 natural polymers, solar applications of polymers, and electrical properties Berlin, Germany of polymers. Esteemed scholars from different institutions will give plenary &NBJM
[email protected] lectures that will be followed by invited and contributing papers. The first 8FCTJUF www.hptlc.com day of the conference will be a workshop on advanced characterization methods and is specifically designed for graduate students and young researchers. The topics and speakers of the workshops are as follows: m
+VMZ
 r
"OESFBT
4DIPFOIBMT
#".
#FSMJO
(FSNBOZ *OUFSOBUJPOBM
4ZNQPTJVN
&YIJCJU
Broadband Dielectric Spectroscopy as a Complementary Analytical 
8PSLTIPQT
PO
1SFQBSBUJWF
BOE
Tool to Characterize Polymer Systems 1SPDFTT
$ISPNBUPHSBQIZ
r
.BOGSFE
8JMIFMN
,BSMTSVIF
*OTUJUVUF
PG
5FDIOPMPHZ
(FSNBOZ Philadelphia, Pennsylvania, USA Rheology: From the Basics up to Complex Polymers 8FCTJUF www.prepsymposium.org r
4UFQBO
1PE[JNFL
6OJWFSTJUZ
PG
1BSEVCJDF
$[FDI
3FQVCMJD
Determination of Molar Mass Distribution and Branching of Synthetic m
+VMZ
 and Natural Polymers Using Separation Techniques with Advanced UI
*OUFSOBUJPOBM
4ZNQPTJVN
Detection BOE
&YIJCJU
PO
UIF
4FQBSBUJPO
r
"OUKF
1PUUIBTU
6OJWFSTJUÅU
GÛS
#PEFOLVMUVS
5VMMO
"VTUSJB 1VSJà
DBUJPO
BOE
$IBSBDUFSJ[BUJPO
Analyzing Lignocellulosic Biopolymers—Challenges, Requirements, PG
#JPMPHJDBMMZ
*NQPSUBOU
and (some) Solutions .PMFDVMFT
*4111
 r
$MFNFOT
4DIXBS[JOHFS
+PIBOOFT
,FQMFS
6OJWFSTJUZ
-JO[
"VTUSJB
Philadelphia, Pennsylvania, USA Advanced Mass Spectrometry of Polymers &NBJM [email protected] 8FCTJUF www.ISPPP.org A number of invited speakers have already confirmed their attendance from institutions across the world as well as a number of sponsors who m
0DUPCFS
 are supporting the conference and will be providing an accompanying UI
*OUFSOBUJPOBM
/VUSJUJPO

instrument exhibition. %JBHOPTUJDT
$POGFSFODF


More details on the ISPAC 2017 symposium including registration and Hotel Duo, Prague, Czech Republic abstract submission can be found at XXXJTQBDDPOGFSFODFTPSH
&NBJM
[email protected] As the organizers of this conference we hope to see you there and look 8FCTJUF www.indc.cz forward to celebrating the 30th anniversary of ISPAC with you. 1SPG
)BSBME
1BTDI
6OJWFSTJUZ
PG
4UFMMFOCPTDI
4PVUI
"GSJDB
1MFBTF
TFOE
BOZ
VQDPNJOH
FWFOU
1SPG
$MFNFOT
4DIXBS[JOHFS
+PIBOOFT
,FQMFS
6OJWFSTJUZ
-JO[
JOGPSNBUJPO
UP
-FXJT
#PUDIFSCZ
"VTUSJB MFXJTCPUDIFSCZ!VCNDPN

222 -$r($
&VSPQF April 2017 THE ESSENTIAL PRINT & DIGITAL RESOURCE FOR SEPARATION SCIENCE IN PRACTICE

March 2016 Volume 29 Number 3 www.chromatographyonline.com

SUPPLEMENT TO June 2015 www.chromatographyonline.com A Numbers Game How suitable is the column plate number for system suitability testing?

MS — THE PRACTICAL ART GC CONNECTIONS SAMPLE PREPARATION PERSPECTIVES Nontargeted metabolite Carrier-gas flow or velocity? profiling Trends in sample prep

Recent Developments in HPLC and UHPLC print

SUPPLEMENT TO October 2015 www.chromatographyonline.com

Advances in Biopharmaceutical Analysis

February 2016 Volume 29 Number 2 www.chromatographyonline.com powered by E-APP NOTE

The Rise of HILIC ALERTS In untargeted clinical metabolomics

LC TROUBLESHOOTING COLUMN WATCH QUESTIONS OF QUALITY Chemical attack The differences between alkyl Do you understand CSV? E-NEWSLETTERS and PFP phases digital DIGITAL EDITIONS & APPS WEBINARS

CHROMATOGRAPHYONLINE.COM E-BOOKS WE LIVE EFFICIENCY

Automated LC Method Development

Focus is key to efficiency, in sports and in science. At Agilent, we are passionate about helping you to maximize the efficiency of your LC workflows.

The Agilent InfinityLab LC Method Development Solution gives you automated access to hundreds of unique sets of analytical separation conditions – without changing a single column or solvent bottle.

Keep focused, live efficiency.

www.agilent.com/chem/livelcmetdev #WeLiveEfficiency #EfficientUHPLC

© Agilent Technologies, Inc. 2017