from Discovery to Recovery

2006

The Golden Jubilee year of
The Chromatographic Society
Recognising half a century of supporting progress in separation science.
sigma-aldrich.com/supelco

International Labmate Ltd

Environmental Technology (Publications) Ltd

ALL TRADEMARKS ARE THE PROPERTY OF THERMO ELECTRON CORPORATION AND ITS SUBSIDIARIES. ©2006 THERMO ELECTRON CORPORATION. ALL RIGHTS RESERVED.

The Perfect Fit

Thermo Electron’s innovative
GC and MS technologies
deliver superior performance,
reliability, and value.
Visit our website at www.thermo.com/gc
or contact us for more information.
Tel: +1 800-532-4752 • Email: analyze@thermo.com

Look closer for answers

DSQ™ GC/MS

Analyze • Detect • Measure • Control™

Chromsoc Final doc

24/4/06

3:55 pm

Page 1

Contents
The Golden Jubilee year of
The Chromatographic Society

Foreword to the Golden Jubilee Book

2

by Dr. Chris D. Bevan

Recognising half a century of supporting progress in separation science.

50 years of the Chromatographic Society,

The Chromatographic Society would like to thank everyone
who has contributed to this celebratory journal.

The Pye 104 Club and the disappearing heritage

Chris Bevan
President The Chromatographic Societyof instrumental chromatography
by Ian Wilson

Denise Wallworth
CE Executive Committee Honourary Secretary Member

James Waters and his Liquid Chromatography

Ted Adlard
Retired Long Serving CS Executive Founder Member andFormer Chairman

10

People: A Personal Perspective
by Patrick D. McDonald

Bernie Monaghan
Chromatography Consulting International Ltd.
Consulting Editor-Separations Science and Spectroscopy
International Labmate

The Gas Chromatography Discussion Group20

by Ted Adlard

Helen Sweetland
Senior Designer - Designs for You Ltd

A Historical Overview of Size Exclusion

Jessamy Fox
Designer - Designs for You Ltd

22

Chromatography
by Greg Saunders

Russell Purvis
Managing Director - Designs for You Ltd

The Future of Chromatography

Michael Pattison
Chairman - Designs for You Ltd

30

by Peter Myers

Everything

Presentation is

4

10%

disco
u
all de nt on
sigmemb for Chro n workmers compan socies

we offer the complete
design service...

We are...a fast, affordable and effective solution

confident we cancut your printing

Designs for You has the resources to produce any type of corporate design. This can include:• advertisement design• exhibition display graphics• company posters• stationery• company brochures• promotional literature• leaflets or newsletters• corporate identity

costs significantly -ask for a quote

Designs for You Ltd, Oak Court Business Centre, Sandridge Park, Porters Wood, St Albans, Herts, AL3 6PH, United Kingdom
tel. +44 (0)1727 731819 • info@designs4you.biz • www.designs4you.biz

Designed and printed by Designs for You Ltd

2006 The Golden Jubilee year of The Chromatographic Society

1Chromsoc Final doc

24/4/06

3:55 pm

Page 2

editorial
Foreword to the Golden Jubilee Book
by Dr. Chris D. Bevan, President of the Chromatographic Society
If you are reading this book you are

the Society. The Chromatographic Society’s bursaries have enabled many

perhaps have a career involving

symposia and make important contributions by presentations of their work.

probably a chromatographer andseparation science. If not, you willlearn of a branch of science towhich many industries owe a
Dr. Chris D. Bevanconsiderable debt of gratitude.

Much of our modern world relies on the action of complex mixturesof substances combined together, often in a unique way. Drugs areprobably the most obvious of such formulations and the ability toseparate and analyse these is essential to the pharmaceutical and finechemical industries. Chromatography is probably the best known of
the branches of separation science.

In 1954 a meeting on gas chromatography convened by A.F. Williams andW. Murray was held in Ardeer, Scotland at ICI Nobel Division. In 1956

the emergence of GC as a useful technique for the petroleum industry

created a nucleus of scientists with a common interest in discovering its
scope and developing it further. The first international symposium onchromatography was held in London in 1956 with the inspired supportof Denis H. Desty, and his colleagues in the Hydrocarbon ResearchGroup of the Institute of Petroleum.

The past 50 years has seen the introduction and development of gas,liquid, supercritical fluid, capillary, preparative and numerous other variantsof separation techniques that have become an indispensable part of theanalytical chemist’s toolkit.

When a technique is born, scientists want to discuss it, test it, apply it andmake money from it. In order to encourage such discourses scientists
crystallise into specialist interest groups and if sufficiently popular anduseful these develop further into scientific societies. The
Chromatographic Society has evolved from such roots.

The Gas Chromatography Discussion Group was formed to organise,
disseminate and promote interest and information exchange in this

rapidly developing science. As the various methods of chromatography

were discovered and developed so the Discussion Group matured into
what we now know as the Chromatographic Society.

The influence of the Society as a nucleus for discourse and a source offunding to inspire and assist progression in this area shouldn’t be

underestimated. The committee formed to run the society has been

populated by specialist innovative practitioners and commercially inspired

technologists to ensure a balanced representation across the whole field of

The Society’s Martin and Jubilee medals are awarded to recognise
particularly significant contributors to separation science.

The key and significant influence of the CS must be in hosting andmanaging international symposia. In the past 50 years the Society has
been the principal organiser of eight International Symposia on

Chromatography in the UK and Ireland. In addition to these home-

based symposia the committee has assisted and cooperated with their

European partners to host a further eighteen, of which the current one
is in August this year in Copenhagen. The number of lectures andposters given must run into thousands and the dissemination ofknowledge and know-how greatly facilitated by the Society.

Nationally the Society has been even more active with selective

specialised seminars, workshops and symposia all designed to assist thediscussion and adoption of knowledge, techniques and instrumentation
used in chromatography.

This book’s object is to enlighten you in the heritage of chromatography.
Chromatography is largely an instrumentally-based science. Instrumental
developments have enabled the pursuance and realisation of theseparations devised by the experimentalist. In the same way as one
may reminisce over cars one has driven, or holiday venues one has

visited, the content and intention of this book is to allow you to indulge
yourself in nostalgia by revisiting and sharing your memories ofequipment with others. Amateur museum collections coupled withmemories of using these instruments have been commissioned forinclusion here. A trip down memory lane for the experienced and arealisation of the evolution of instrumentation for the newcomer iscatered for here. Memories from the past and visions of the futureserve to commemorate the pivotal position the CS has played over the
past half-century.

Photos of some members of the Executive Committees over the years
have been included as a tribute to their voluntary contributions to themission of our society. If the Society had never existed many of theadvances in chromatography would still have been made but due tothe activities of the Society they have been more rapidly disseminatedand more widely known. Opportunities to hear, meet and interact with

the innovators are truly facilitated by the society and its meetings, acatalyst for the advancement of separation science.

separation science. Important connections and alliances between the UK-

As President of the Chromatographic Society I hope you will enjoy

counterparts were, and still are, promoted and realised by the activities ofand knowledge of our science with others.

based scientists and their European, American and former Soviet block2

students and scientists without sufficient means, to attend international

2006 The Golden Jubilee year of The Chromatographic Society

reading this book and that it will inspire you to share your experiences

Chromsoc Final doc

24/4/06

3:55 pm

1200 RapidRes_EURO_180x254

Page 328-03-2006

13:34

Pagina 1

We could have called it:

The Mega Fast Ultra Resolution
Hyper Precise Super Scalable
LC System...

…but we think numbers speak louder than words
Numbers define performance, and performance is what

All the Performance. All the Time.

the new Agilent 1200 Series Rapid Resolution LC system isall about. In combination with Agilent’s second genertion
ZORBAX RRHT 1.8 µm LC columns, you can analyze
samples faster and with higher resolution than ever before
without compromising your data quality _ and still run

The Agilent 1200 Series Rapid Resolution LC System• Up to 15 times faster than conventional HPLC
• 60% higher resolution• 30-second cycle time
• 30,000 plates efficiency• > 600-peak capacity

traditional HPLC methods. Plus you have the added
confidence that comes from flexible and comprehensive
services that keep your operations at peak performance.
After all, the Agilent 1200 Series builds on the 1100 Series
platform, with the proven reliability counted upon by

www.agilent.com/chem/1200RRE6

60,000 system installations worldwide.

©Agilent Technologies, Inc. 2006. MC 15177

2006 The Golden Jubilee year of The Chromatographic Society

3Chromsoc Final doc

24/4/06

3:55 pm

Page 4

editorial
50 years of the Chromatographic Society,

The Pye 104 Club and thedisappearing heritage of
instrumental chromatography
by Ian Wilson
Department of Drug Metabolism and Pharmacokinetics, AstraZeneca
Mereside, Alderley Park, Macclesfield, Cheshire SK10 4TG, UK.
Ian Wilson

Instrumental chromatography is a relatively recent innovation, andcollection is, unfortunately, slightly damaged but Roly had apparently

many amongst us who can remember when HPLC was not routine, but

was intact.

commercial instrumentation even more recent. Indeed, there are still
a new and untried technique, expensive, temperamental and frankly,

when compared to packed column GC, pretty low resolution with lousy
sensitivity.There are a diminishing number of pioneers who canremember the era before instrumental GC, when you made your own
kit out of glass and the whole concept was completely new, where

every result was novel and new discoveries were being made on a daily
basis. If you want to see how exciting chromatography could be, thephotograph in Figure 1 shows a commercial, all glass, gas

chromatograph. It was given to me by the late Dr Roly Jones, (Martin

Medallist and a former Chairman of the Chromatographic Society) andwas manufactured in London by a firm

However, fortunately most of us never experienced this type ofequipment and increasingly, in the world of pharmaceutical analysis at least,
our laboratories are filled with eager young scientists whose only

exposure to chromatographic analysis is via HPLC-MS-MS.This wonderful
technology is accepted as a commonplace rather than the miracle ofmodern analytical chemistry that it is. Indeed those of us who remember

the moving belt or direct liquid interfaces had thought that, if this was thebest that could be done in linking LC and MS, we could forget routine
LC-MS in our lifetimes (if you are too young to remember theseinterfaces, be grateful).

trading under the imaginative name of Gas

Anyway, at some point, and I cannot really remember when it was, I

much easier in those days, you had aactually use!) were disappearing from the lab. If they were lucky they were

Chromatography Ltd (marketing was so

product, you knew what it did and you soldit as that!).The firm was based at Number

176 Old Brompton Rd, SW5. Roly told me(amongst other things) that their demo lab

was in a cellar that backed on to a Tube line.
When trains went past they caused somuch vibration that the chart recorders

couldn’t function properly so, as a result,demos were run according to the train
timetable. Prices started from £20

according to a price list in my possession
and, if you wanted extra sensitivity you
could also buy a “Lovelock Ionisation

Detector”. The gas chromatograph in myFig 1. The all glass chromatograph from Gas Chromatography Ltd, circa
1960. This is very difficult to photograph because it’s 1.4 meters high but only
a few cm wide. The device is shown with the boiler pipe insulation thatsurrounded the whole thing opened up to show the l.3m long “U” shaped, fixed,
glass columns contained within the outer glass tube that acted as the oven.

4

managed to donate another to the Science Museum in London that2006 The Golden Jubilee year of The Chromatographic Society

noticed that the instruments I had been trained on (i.e those I could stillgoing to some deserving, but under-funded university

department, but often they were heading straight into the skip.That’s what
happens to instruments, they become obsolete or wear out, better kit
comes along, and the old stuff is replaced. And no bad thing either

because, despite what people may say, in general the instruments are
better these days. However, it was slightly sad to think of all of this“wonderful” old instrumentation being lost forever. After all, when it went
into the skip a small part of my life went with it. Also there was some kit
that was so badly designed that it deserved to be remembered so thatthe same mistakes would NEVER be repeated. So it was with a vague
feeling of regret that I realised that I would never again have theopportunity again to burn my fingers trying to change a GC column in ahurry in a Perkin Elmer F17 (Figure 2) without breaking the column(actually it wasn’t possible to change a column in a hurry in this

instrument, but burning your fingers was guaranteed). Worse, it was

becoming impossible to do chromatography without a computer tocontrol the pumps and detector etc.

At some point this vague feeling of regret crystallised into a conversation
on a bus (at the 20th ISC in Bournemouth) with Prof. Mike Cooke that

Chromsoc Final doc

24/4/06

3:56 pm

Page 5Chromatographic Society’s Bulletin (Number 40). Naturally it just aboutsank without trace but, for those of you who really need to be a member,the 104 Club is still going and anyone who wants to join can do so by aone-off payment (for lifetime membership) of the princely sum of £15 forfellowship (where ideally you should have actually USED a 104, but we are
flexible….) or membership at a trifling £10.There will only ever be 104

members in each category, so membership is much more exclusive thane.g., the Royal Society.

Fig. 2 A Perkin Elmer F17 GC once owned by the late Dr Roly Jones.

concluded that “something had to be done” (there may have been beer
involved, I don’t remember, though it is highly likely, but no minutes orother formal record of the “meeting” were taken). One of the things we
decided to do as a result of the conversation was to form the Pye 104

Club as a special interest group of the Chromatographic Society. So why

the Pye 104 Club and not the National Society For The Preservation And
Celebration Of Historic Chromatographs? Well “Pye 104 Club” is shorter(and less pretentious) for a start, and also we rather thought that thepeople who would be interested in this sort of thing would be the sort ofFig 4. A Waters HPLC system comprised of two M6000 A Pumps, the Model
660 gradient controller, a UV detector, and a U6K injector (lying on its side in
front of the pumps). I’m not sure that the UV detector is quite right for thisparticular model, and Ive been told that the “Rolls Royce” system came in acustom built cabinet (presumably with a proper mounting for the U6K). The 660
controlled the pumps via a pair of cables of which I have one. So anyone who
can help with a contemporary UV detector, cabinet or controller cable…. I’d love
to hear from you. I do have an authentic Waters refractive index detector which
does fit with this set up.

chromatographers who knew about this iconic instrument. A photograph

What you get for this is a certificate that says you’re in, which thereby

courtesy of Dr Derek Stevenson, is shown in Figure 3 (next to an almost

NO benefits. However, the certificates are signed by a variety of greatof a 104 from my collection, obtained from the University of Surrey

identical looking Pye 105, which was the prep version).The 104 was built
in the 1960s, and was built to last (indeed there are still some in use), it
weighs a ton and is practically indestructible. It was such a popular

instrument in the UK that it could be found, often in large numbers, inmost industrial and university analytical labs for many, many years, andgenerations of PhD students learnt their craft on them. It also had thebenefit of a capacious oven that you could heat your pies up in so that,even if the separation didn’t work, you were at least set up for lunch. A
classic. Anyway the Pye 104 Club was launched in October 1994 to ahuge fanfare of publicity in the widely read and ever popular

implies that you have more money than you need as membership has
chromatographers including for example John Knox or James Lovelock so
if you’re an autograph hunter….The money goes to the Chrom. Soc. andevery now and again (i.e. when there is enough) is used to provide
sponsorship for students to attend meetings.

Sometime after the 104 club was formed I started to acquire
instruments. Some of these I sought out, because they meant

something to me, and some just turned up.They still do.They included
not just GCs but also HPLC systems as well. So one of my first HPLC
systems was the Waters M6000A setup shown in Figure 4. A

wonderful machine, it could do gradients (ofvarious shapes) and had a good UV detector andthe U6K injector, which was good compared toe.g the Spec Ac, but lost out to Rheodyne in later
years. Like the 104, the M6000 series dominatedthe HPLC landscape for some time before nimble
predators like the Constametric pumps fromMilton Roy began to bite back. I have an early

Milton Roy in the collection (Figure 5) which I
obtained from Klaus Borner in Germany.The

instrument was delivered with a manual that said
Fig 3. A pair of Pyes!. The Pye 104 GC on the right was obtained from Dr Derek Stevenson of the Universityof Surrey whilst the (almost identical looking to the non-enthusiast!) Pye 105 on the left (the prep. version of
the 104 was found by Dr Mark Powell (at the time of donation of Liverpool John Moores University).

“give this to the man or men who will install the
instrument”, not something you are likely to see
on any document these days. Next to it are a2006 The Golden Jubilee year of The Chromatographic Society

5Chromsoc Final doc

24/4/06

3:56 pm

Page 6

Now, by this point many of you are probably wondering
how I get away with filling the house with all of thishistoric equipment (or junk as it has been called by lesscharitable souls). Well, I’m lucky in that I have a large, andreasonably dry, cellar. Not an ideal place to store
equipment, but better than a skip. I also have an

understanding and long suffering wife (who doesn’t yet
have a use for the cellar!!!). However, if I can find a

museum that will take the whole collection and lookafter it I will gladly pass it on (well most of it, it will takea lot to get take my last 104 and the M6000 system).
But that is not as easy as you might think. For a startmuseums are not, as many people suppose, in the
Fig 5. An old Milton Roy HPLC pump (on the right) from Klaus Borner together with a ex
ICI/Zeneca Constametric III HPLC pump and UV detector. The upright device between theConstametric and Milton Roy pumps is a primitive a computing integrator. No peaks, just a roll of
paper giving retention times and peak areas. Things HAVE got better.

Constametric III pump (with a UV detector sitting on top) and a very
user-unfriendly integrator.

As well as kit I also have acquired a whole range of interesting “bits”
including some old bottles of stationary phases, columns and otherephemera.

At this point I have a total of 17 GCs in the collection (with a further 2
on long term loan to Schimadzu), I also have some 12 LC pumps and avariety of detectors. In the collection I also have the odd mystery

instrument, where I don’t even have the name of the manufacturer.
Does anyone recognise the portable GC shown in Figure 6?

Unfortunately, I have fewer manuals than instruments, and manuals arequite important. Manuals however, do become detached from systems,
and worse, thanks to GLP, are often archived when the instrument isdisposed of. So if anyone as a manual for a HP5890, another iconic

instrument, that they don’t want I would be very pleased to receive it.

business of preserving everything that you or I might
think was important.There is not enough money orspace in the world for that, so they have to collect anddisplay what will appeal to the public. At any one timevery little of any museum’s collection can be displayedand storage space costs. At this point ask yourself how many people in

the whole of Britain are that interested in chromatography and analysis?
(even with the benefit of CSI Miami etc,) and would therefore actually

be interested in looking at a collection of old chromatographs and you
will see the problem. Well, the group possibly includes you and me(obviously), but its not going to be a large number. Also, and this is nota criticism but a fact, museums are run by professional curators, notenthusiasts (like libraries are run by librarians and not authors).

Enthusiasts run steam railways and love them, curators don’t. Curators
are professionals with a lot to think about and if, at some point, anartefact no longer fits in with the collection they have no compunction
in disposing of it. Also, in moving things from one temporary storage
space to another, things get “lost”. For example I know of one major

museum that cannot find a particularly large 1970s mass spectrometer(a Kratos MS8) that it had in storage and now wants. Now its not easy
to “lose” something of this size (think “that’s a big bit of kit”, and thendouble it) and in all probability it was “cleared out” when the space

was needed by someone who did not realise what it was, but did seethat it was taking up an awful lot of room. As an aside, if you want toannoy a curator at a dinner party ask him if he knows the sad story of
the last stuffed Dodo in the world. A genuine curator will.This unique
exhibit, which if I remember correctly was in the Ashmolean Museumin Oxford, was in a fairly decayed state and was tossed on a bonfire bya curator who probably thought it “didn’t fit the collection any more”.
Another curator was passing by and saw it burning. He did know its

value, managed to retrieve a foot and the beak, and that is all we haveleft. What no museum will do these days is take an instrument on

loan, you have to give it to them and, if they decide that they don’t

want it any more, they wont give it back (they will offer it around toother museums but if none of them want it, it’s disposal time again!).

So whats to be done? Well the museums will conserve some of these
instruments (just perhaps not the ones that you or I would havechosen) and not necessarily for ever. For example, the Science

Museum in London has a very large collection but obviously, becauseof space constraints, most of it is not on display.Then there are a

Fig 6. A mystery portable GC. I know nothing about this other than that I
obtained it for Dr Derek Stevenson of the University of Surrey, no manual, noidea who the manufacturer was/is. Any help much appreciated on either front

6

2006 The Golden Jubilee year of The Chromatographic Society

couple of positive strategies that could easily be adopted by thechromatographic community if preserving instrumentation was seen as
being worthwhile. Firstly the instrument manufacturers could be asked

Chromsoc Final doc

24/4/06

3:56 pm

Page 7

to keep an eye on their heritage. Many of them already dothis and, as mentioned above, two of the instruments in
my care are going home to Schimadzu UK in Milton

Keynes on long term loan. This strikes me as one of the

HPLC, GC & MS Support

best options but, that said, takeovers happen, times change

Agilent/Hewlett Packard•Shimadzu•Waters/Micromass

buying an analytical chromatograph from WG Pye Ltd of

Dionex/Gynkotek•Beckman•Kontron•Rheodyne•Valcoand companies go bust etc., (if you don’t believe me try

Gilson•Jasco•Merck-Hitachi•Perkin Elmer•ThermoSeparation•Varian

Cambridge).You can save the instruments yourself, if you• Preventative Maintenance Agreements• Validation / Calibration / OQ / PQ Agreements• Engineer Callout or Workshop Support• Reconditioned and Nearly New Instrument Sales• Instrument Rental and Leasing Schemes• Original Manufacturers and quality alternative Spare Parts• Wide Range of Consumables and Accessories

have the space, and let the Pye 104 Club know about it(via the Chrom. Soc. or direct to me atian.Wilson@astrazeneca.com). Perhaps in this way we canestablish a extended “virtual museum” and whilst its hard

to see it ever attaining the same degree of interest as you

Jaytee Biosciences Ltd
171-173 John Wilson Business Park, Whitstable, Kent, CT5 3RB
Tel: 01227 265333 Fax: 01227 265331
this is a good long term strategy (I shudder to thinkofthe1076 ChromSociety50th 2/13/06 10:10 AM Page 1
Email sales@jaytee.com www.jaytee.com
problem my heirs will have when they

see at a steam engine rally it might still be fun. I’m not surecome to empty the cellar) but an

uncertain future is better than no future
at all. Then there is the option that

preserves the memory but not theartefact - images on the net (not somuch the ghost in the machine as theghost of the machine!). Having justacquired a digital camera I’m in theprocess of photographing the instruments
in my collection from all angles, andhopefully the Chrom. Soc. will put themon its web site. It would be a fitting thing
to do in the Society’s 50th year. Perhaps

we could all do that?, its not the same ashaving the instrument to hand but, itsbetter than nothing.

Finally if there is anyone out there who
knows of a Pye Pan Chromatograph, aVarian Aerograph or a Perkin Elmer
Model 154 Vapour Fractometer, or

anything else of a historic nature that’s stillintact and just about to be disposed of,

Finally, the best of all worlds
in one HPLC detector.
With performanceexceeding RI, ELSD,
and often UV, theCorona is ideal for:and 2) please don’t get in touch to say,

The promise is now delivered. Corona™
Charged Aerosol Detection (CAD™) from
ESA has redefined the standard for universal
HPLC detection in regard to sensitivity,
dynamic range, and reproducibility.
Which explains why Corona CAD won
the 2005 R&D 100 Award (R&D Magazine)
and the 2005 Silver Pittcon Editor’s Awardfor best new product – for out-of-this-world
performance, right here at home.

week, last month or last year but we

The Corona Charged Aerosol Detector

well I still have space in the cellar for a

few more classic instruments ( though at
this point I need to make two important
points here, 1) I save them, I don’t buy

them, so please don’t ask me how much I
will give you for an old chromatograph,“we had one of those until yesterday, lastchucked it away, if only we had known” I• Pharmaceutical R&D• Sugars and lipids• Industrial chemicals

would prefer not to know). Manuals
would also be of interest.

Those of you who are still in the earlystages of your career probably can’t see
what all the fuss is about, but trust me,

978.250.7000 www.coronacad.com

one day you will.

2006 The Golden Jubilee year of The Chromatographic Society

7Chromsoc Final doc

24/4/06

3:56 pm

Page 8We’ve got it COVERED!
International Labmate Ltd are worldwide publishers of Laboratory,

International Labmate

Environmental and Petrochemical publications for scientists working bothInternational Environmental Technology

inside and outside the laboratory. Titles include International Labmate, Lab

Petro Industry News

Asia, Labmate UK & Ireland, International Environmental Technology, AsianLabmate UK & Ireland

Environmental Technology, Petro Industry News and Measurement

Lab Asia

Analysis China. Copies of our publications are available free of charge to

Asian Environmental Technology

qualified readers.

Measurement Analysis China

For prices on advertising or to request a copy, please contact:

International Labmate Ltd. Oak Court Business Centre, Sandridge Park, Porters Wood

International Labmate Ltd

Environmental Technology (Publications) Ltd

St Albans, Hertfordshire, AL3 6PH, United Kingdom
Tel. +44 (0)1727 855 574 • Fax +44 (0)1727 841 694 • info@intlabmate.com • www.internationallabmate.com

www.searchaproduct.com

HPLC is not only Technology . . .

Find products
and suppliers
quickly andeasily.

:
US AT
MEET
ean
Europ osium,pme Sy
Peptid k / Poland
Gdans r 3-8, 2006
embe
Sempt
or
P,
/ ISPP ia
A
C
I
P
S
tr
s
u
A
uck /
Innsbr 7-20, 2006
er 1
Oktob

No more wasted
time trawlingthe internet!
This on-line
directory features
OVER 4,000

Laboratory,
Environmental& Petrochemical
products andsuppliers, on
one easy to
use web-site.

Please contact: Carla Sweeney,

Guide Manager International
Labmate, Sandridge Park,
Porters Wood, St.Albans, Herts,
AL3 6PH, United Kingdom... but also Arts, Attitude and Spirit. YMC analytical and preparative high purity
silicas for sophisticated normal and reversed phase and HILIC chromatography,
high performance analytical columns from capillary to process-scaledimensions (> 1 meter diameter), column tool kits for method development,
validation, combinatorial chemistry, LC-MS and scale-up, multi-ton
capacities for industrial supply of silica and bonded phases (10-150 µm),
methods development and custom purification services / toll manufacturing,seminars and workshops.
Please inquire for YMC’s general catalogue.

The Selectivity Company

Tel: +44(0)1727 855574
carla.sweeney@intlabmate.com

8

2006 The Golden Jubilee year of The Chromatographic Society

Schöttmannshof 19 · D-46539 Dinslaken · Phone: +49 (0) 20 64 / 427-0 · Fax: +49 (0) 20 64 / 427-222
E-mail: info@ymc.de · Website: www.ymc.deLife sccie
and Dranienpm
earch harmaceutical Life sc nce Food d Drin Research
ce FPage9
tical Lif 3:56
s
Pharmaceu24/4/06
e
o
R
o
k
d
i
P
n
i
and Drenes
and inkhhPharmaceutical Life cience Food anesearc Pharmaceutical Life scie ce Food an Dr k Researc
R
k
n
i
r
d
D
n
h
h
d r inPharmaceutical Life science Food anesearc Pharmaceutical Life science Food an D k Researc
R
k
n
i
r
d
D
h
s
n
earc Pharmaceutical Life sci ce Food d Dri Research
Pharmaceutical Life cience Food a
s
e
R
k
n
i
nd Dr
arch armaceutical Life s ence Foo and Drink esearchesPharmaceutical Life science Food aneRhd Drinksrch P rmaceutical Life cience Food and Drink R searchaes
Pharmaceutical Life cience Food aneesciencdd Drink R arch Pha aceuticale Foo and Drink R searcheLsi
Pharmaceutical Life science Food an
m
f
e
r
e
a
scien
d an
d Drink R
h Phshink RePharmaceutical Life cience Food and Drink Researc Pharmaceutical Life scie ce Food a d Dr k Researcnnhd r inh
Pharmaceutical Life science Food anesearc Pharmaceutical Life science Food an D k Researc
R
k
n
i
r
d
D
h
s
n
i
d Dr
arch
earc harmaceutical Life sc ce FChromacoles
Pharmaceutical Life cience Food an
o
s
e
o
R
e
d
k
R
Ltd
n
i
P
i
ence
and Drinkd DrhhFooIndustrial
Pharmaceutical Life science Food anCentre,searc
esearc Pharmaceutical Life scie3nMundells
R
e
d
k
R
n
i
r
a
k
d
D
ndAL7DIEWhce City,r in
rch
uti Welwyn Garden
Pharmaceutical Life science Food a
ooHerts,
searc
Resea ch Pharmace cal Life scienc Tel:F+44
e
d
k
R
n
i
r
a
n
k
d
D
n
s
h
n
i
c
d
Dr
e oo (0)1707 394949
arceutical Life s
arcPharmaceutical Life ience Food and(0)1707
ciencFax:F+44
k Rese rch Pharma
Rese rch
391311
n
i
r
a
k
d
D
n
s
n
i
d
c
r
D
i
e
e
a
nce Fooceutical Life semail: enquiries@chromacol.com
Pharmaceutical Lifecienc Food and Drink Resea chd and Drink Rese rch Pharmascie Fwww.chromacol.com
e
t
a
c
u
i
c
e
e
m
a
a
n
l Life sc Website:utical L
ac
Phar
ood a
esear hR
ife scie ce Food and Drink Rese rch Pharm
i
e
k
n
n
n
i
d
c
r
D
e
a
nce FooarctPharmaceutical Lifed and Drink Rese ch Pharmaceu ical Life scienc Food and Drink Rese h
s
c
c
r
e
Pharmaceutical Life ience Food an
Resea h Pharmaceutical Life scienc Food and ink Resear h
k
n
i
r
d
D
s
c
c
r
D
i
r
e
Food
Pharmaceutical Life ence Food an
searcesea Pharmaceutical Life scie
R
e
k
R
n
i
r
a
k
d
D
n
n
h
d Dr i ncechrcPharmaceutical Life science Food an
Resea h Pharmaceutical Life scienc Food and ink Resear h
k
n
i
r
d
D
s
c
c
Dr
e
rc
ar
aceutical
Pharmaceutical Life ience Food ak Rese rch Pharm utic Life science Food and Drink Research
n
i
r
n
d
D
s
c
i
e
a
c
a
nce Fooce al Life s
Pharm eutical Lifecienc Food and Drink Resea chd and Drink Rese rch Pharma
s
c
i
e Foo
e
t
a
c
u
i
c
e
a
e
m
a
n
c
r
l
u
ce Food
Life sctical Lif
Pha
Rese h Pharma
esear h
d
k
R
n
i
i
a
e
r
e
a
n
k
d
D
n
n
n
s
i
d
c
r
c
c
D
Pharmaceutical Life ience Food an
searc
esear Pharmaceutical Life scien e Food an
R
e
k
R
n
i
r
k
d
D
h
s
h
in
Pharmaceutical Life cience Food and Drink Researc Pharmaceutical Life scie ce Food a d Dr k Researc
n
n
n
h
i
d
searc
Pharmaceutical Life science Food an
maceutical Life scie ce Food a Dr Researchd Drink Re earch Pharnce Fnsciencenkhood a d Dri Researc
Food and Drink Res
knd D r i n

Chromsoc Final doc

Chromatography vials andconsumables for all markets

WWW.CHROMACOL.COM

ClarityTM

Bio Sol uti

ons

patent pending

Pharmaceutical • Research • Food and Drink • Life Science

2006 The Golden Jubilee year of The Chromatographic Society

9Chromsoc Final doc

24/4/06

3:56 pm

Page 10

editorial
James Waters and his
Liquid Chromatography People:
A Personal Perspective
by Patrick D. McDonald
Patrick D. McDonald

A golden jubilee, a celebratory

young U.S. Navy veteran, armed with a degree in physics and engineering

every 50 years — what betterand enthusiasm, was building instruments in the basement of his parents'

period provided by ancient law
time to stand in the thresholdof our discipline, like Janus,

looking at once to both thepast and the future.We are

fortunate, as was Newton, tostand on the shoulders ofgiants whose perception,

perspiration, and perseverance

prompted a cycle of revolution
James L. Waters, ca. 1965

and evolution in the practice ofchromatography, leading it to its present position as a central science[1]. I am pleased to have this opportunity to review the role of

Waters, both the man and his company [2], in some of the mostfundamental developments that have determined how we practiceliquid chromatography today, and that are leading the way toward
solving the significant separation problems of tomorrow.

So many of the important achievements in chromatography, unlike in othermodest colonial home in Framingham, Massachusetts. His mother, notwishing to have strangers in her house, decided when he hired his first
employee that it was time to move his fledgling business elsewhere. In

1958, after selling his first company, James Logan Waters incorporated J.L.
Waters, Inc., and moved from a spare room in a local hardware store to
the basement of the former police station.

Early on, Jim developed instruments on a contract basis. But when his

design for an on-line refractometer in process monitoring applications

proved popular, it became a successful product for his five-man company.

Engineers at Dow Chemical, Freeport,Texas, skilled at keeping current withthe latest technology, spotted Jim's detector and alerted one of theirpolymer-chemist colleagues, Dr. John Moore. John called Jim in 1962 andasked for a custom unit with a 10x-smaller-volume flow cell. At first, Jim,

unaware of what John was doing, argued that this was not necessary forprocess control. But, when he learned that John would pay for such a
modification, Jim agreed to make it.

Several months passed. Finally, curiosity got the best of Jim. He sent his

scientific fields, have been accomplished by industrial scientists andenterprising sales manager, Larry Maley, down to Texas to see how John

boundaries of physics, chemistry, mathematics, and engineering, is perfectly[4], so John was free to show Larry his new, homemade instrument

engineers, not university professors. Separation science, crossing themated to the interdisciplinary teamwork so strongly rooted in companies

focused on uniting requisite resources to solve problems, create products,
and partner with customers to meet their needs. Unfortunately, industrial
inventors, unlike their academic colleagues, may perish if they publish, sothey often labor in relative obscurity and disappear into the baseline ofchromatographic history. A true entrepreneurial spirit—and a desire to
make the world a better, safer place—pervades their best attempts toprovide solutions that add value to, and have impact on, laboratory

practice. In a sense, we in the analytical instrument industry are toolmakers.
But revolutionary tools enable and challenge users to do what they wereunable to do before.This activity then advances science in ways that truly
improve our world.

Before Chairman Dr.Tony James [3] and Secretary Denis Desty convened,in 1956, the earliest GC practitioners and pioneers to share tips andtechniques in the Gas Chromatography Discussion Group [GCDG], a10from Columbia University and his characteristic boundless determination

2006 The Golden Jubilee year of The Chromatographic Society

was using his refractometer. Fortuitously, patent applications had been filed
designed to pass solvent through a column filled with controlled-porosity,
cross-linked polystyrene-divinylbenzene beads and determine themolecular weight distribution of synthetic polymer samples [5]. John called
his process gel permeation chromatography [GPC]. His concept had been

inspired by the seminal work of Porath and Flodin who used dextran gelsto separate biomolecules by size [6].

Larry took the decisive step on the path to the first commercial HPLC,
concluding John's demonstration by boldly stating, "Waters would like tobuild this for you!" John had to convince his superiors in Midland that thebest option would be to contract this dynamic, caring, but tiny, company in
Massachusetts to commercialize his system, and not some giant analytical

instrument concern like Beckman. In January 1963, Jim's hand trembled as

he wrote a check in the daunting amount of $10,000 as a down payment
on royalties for his exclusive license to GPC. He traveled to Freeport,

spent three weeks learning every aspect of the process, including how toChromsoc Final doc

24/4/06

3:56 pm

Page 11synthesize the polymer beads, later to be sold under the mark Styragel, and

Dr. James Little, told him that Woodward was perhaps one niche below

improved version of John's instrument. John, recalls Jim now, was a true

concluded on the following Monday morning that he should tackle thisreturned to Framingham filled with ideas on how to construct an

genius who not only envisioned the mechanism, means of execution, andmanifold applications for GPC but also assembled by himself all the pieces
needed to practice it.

Within three months, Jim and his team had built five GPC-100 prototypes,
using plywood for the cabinets to save time; three were sold to Dow, andone each to Dr. Jack Cazes at Mobil Chemical, and Dr. Dale Harmon at B.F.
Goodrich.Though Jim is the first to credit the Nobel work of Professors
Moore and Stein at Rockefeller University, after WWII, on systems foramino acid analysis via partition on potato starch [suggested to them by

Dr. Synge], then ion exchange [7], it can be argued that these GPC-100s,
delivered in March, 1963, were the first commercial HPLC systems.

Professor Josef Huber in Vienna and, a bit later, Professor Csaba Horváth in
New Haven, by their own admission [8], built their initial bench-top

breadboard HPLC systems in 1963 and 1964, respectively. Csaba coined
the name HPLC for a PittCon 1970 oral presentation [9]. And both heand Josef Huber used silica-based particles, rather than organic polymers.
However, all the components of an HPLC instrument were embodied in
Jim's GPC-100. In that system's manual, which Jim wrote himself, he evenlabeled its flow schematic Waters Liquid Chromatography Assembly.

It did not take long for polymer chemists to learn that a determination

taking weeks could now be done in a few hours. Another brilliant move by
Larry Maley was the creation of an annual GPC Symposium where

successful early adopters and struggling users could learn from each other,
thereby fostering best practices, not unlike the early days of the GCDG.

First held in the cold of a Cleveland, Ohio, winter, 1965, it moved eventuallyto the warmth of Miami Beach, Florida [10], and, no doubt, like the GCDG,

Sir Robert Robinson atop the pantheon of organic synthesis gods, Jim

problem himself. After all, he had wanted to penetrate this market eversince being told in 1968 by a Mobil colleague of Jack Cazes that everyorganic chemist ought to have an HPLC on his or her bench. So, hepacked some columns, trundled an ALC-100 to the lab on Oxford

Street in Cambridge, Massachusetts, and proceeded to solve separation
problems for Helmut and his fellow postdoc, Dr. John McCall, one after
another, while the master was traveling.

I was in the audience in Boston on Wednesday afternoon, July 28, 1971,
when Woodward concluded an afternoon memorial symposium on

Natural Product Synthesis [XXIIIrd IUPAC Congress], following suchluminaries as Sir Derek Barton, Sir Alan Battersby, Professor Karl Folkers,

and Professor Gilbert Stork. He began the B12 story at four and stopped

at 7:45 p.m., pausing every 45 minutes or so to declare "we could nothave done this without liquid chromatography." Every organic chemist ofnote was in the audience that had swelled to nearly 5000 after theparallel sessions ended at 5 p.m. No one left!

Woodward never mentioned who did these HPLC separations, or

which company's products were used, but cognoscente spread theword like wildfire [12]. Jim Waters was not present at this seminal

lecture. He was busy compiling a list of 900 names and addresses froman American Chemical Society directory of chemistry faculties. He
mailed to each a package, including a glossy photo of himself with

Helmut, Woodward, and, his new friend, Josef Huber [chromatography

consultant to Woodward's B12-race rival, Professor Albert Eschenmoser[13] at the ETH in Zürich], with the message: "Look what we did for

Woodward. We can do the same for you!" Woodward later echoed: "Theinspired many other user forums in our discipline — and elsewhere.

power of these high pressure liquid chromatographic methods hardly canbuilding nearby which once housed the Bay State Car Company factory. In

represent relatively simple instrumentation, and I am certain that they will

Jim's young company soon grew out of the police station and into a large
1964, the second-generation GPC-200 debuted, and, the next year, Jim
licensed some technology from Shell Development for liquid-liquid

be imagined by the chemist who has not had experience with them; they

be indispensable in the laboratory of every organic chemist in the very near
future [14]." Following his example, a single HPLC peak soon

chromatography. Its implementation was unsuccessful, but two years later, in

supplanted a single TLC spot as a criterion for chemical purity in

and RI detectors for executing all modes of Analytical Liquid

While following up the 100 replies to his marketing campaign catalyzed

1967,Waters introduced the ALC-100, a versatile system with both UV
Chromatography.Then, in 1969, there followed the Chromatoprep™, a

dedicated, open-frame, phone-booth-sized, preparative HPLC system withintegral fraction collector.

Dr. Karl Bombaugh,Waters R&D Director, in the late 60s, initiated a fruitfulconsulting relationship with Professor István Halász, Csaba's mentor inFrankfurt [11], who guided Waters through the development of the firstcommercial GC and LC chemically bonded silica phases and the firstpellicular silica packings for HPLC [trademarked Durapak and Corasil,
respectively].These signaled the ongoing commitment of the young

instrument manufacturer to materials science, spanning both inorganic andorganic substrates and surfaces, that continues to this day. However, thepersonal dedication Jim Waters instilled in his colleagues to make every
customer successful is what carried his business to the next plateau.

Frustrated by failure on a Friday in late 1970, Dr. Helmut Hamburger,

the Swiss-trained, chief postdoctoral fellow in Nobel-laureate Professor
Robert Woodward's lab at Harvard, called Waters to ask for help in

separating isomers of key intermediates in the total synthesis of Vitamin
B12. Jim had no idea who Woodward was, but when his marketing VP,

organic synthesis [15].

the onset of phenomenal growth for Waters, Jim, meanwhile, wassupervising his last instrument design, one that inspired a succession of
innovations destined to alter the HPLC landscape forever. He teamedwith a brilliant engineer, Burleigh Hutchins, then VP of R&D, and an

equally insightful, UK-born craftsman and machinist, Louis Abrahams, toinvent the first dual-reciprocating-piston, 6000-PSI pump for HPLC, theModel 6000 [16]. Utilizing non-circular gears, a step motor, and feedback

control circuitry to drive small-diameter parallel plungers sequentially fornearly pulse-less flow, this precision-volume solvent delivery device, firstshown at PittCon 1972, was a true revolution — and so ruggedly
reliable that many continued to be used for decades!

Competitors soon retired pressurized vessels and other primitive

pumping means and sought to imitate the wildly successful M6000.

Cottage industries sprang up to supply sapphire plungers, high-pressure
seals, check valve assemblies, pulse dampeners, and other replacement
parts. All this activity spurred rapid acceptance of HPLC in labs

worldwide and accelerated the growth of the nascent HPLC industry.
Burleigh and Jim, together with further improvements by Dr. William

2006 The Golden Jubilee year of The Chromatographic Society

11Chromsoc Final doc

24/4/06

3:56 pm

Page 12

Carson and John Roe [17], redefined the state-of-the-art forarena where it rose to predominance as an analytical, and ultimately, a

the benchmark Model 401 detector.To complete a nearly simultaneous

A major component outlier in the early drive by Waters to control its

temperature stability and sensitivity of the differential refractometer intriple play, Lou also invented a novel valve and fluid circuit [18] that heand Burleigh used as the core of the first 6000-PSI-capable, septum-less
HPLC injector, the Model U6K.The success of these products funded

the construction of, and a move in the fall of 1973 to Waters present
corporate headquarters and manufacturing location in the Bear Hill
woods of Milford, Massachusetts.

This M6000/U6K combination was not designed merely to meetcustomer needs. It was necessary to fulfill the prophecy of Dr. Martin
and Dr. Synge who wrote in 1941: "the smallest H.E.T.P. should beobtainable by using very small particles and a high pressure difference
across the length of the column [19]." For, in a parallel Waters R&D
program, Dr. Richard Vivilecchia, with Norma Thimot and the late

Richard Cotter, created the first commercial 10-micron silica columns for
HPLC: µ Porasil™ silica announced in December 1972, andµBondapak® C18 in early 1973. Using reactions invented by John Speierat Dow Corning [20], Dick Vivilecchia synthesizedoctadecyldimethylchlorosilane and therewith created the firstcommercial monofunctionally bonded C18 column.Though his

proprietary process for silica treatment, surface bonding, and end

capping prompted a host of attempts at imitation, it remains unique tothis day. Later in 1974, based upon my work in preparative LC, I

suggested that Dick switch to a superior silica substrate.This improvedµBondapa® k C18 packing produced the best-selling HPLC column inhistory. Waters chose not to patent the silane or disclose the process

details. Five years later, when Waters subcontracted the silane synthesis

on a non-exclusive basis, it became commercially available for anyone touse, and nearly everyone did!

A column's physicochemical and performance properties may ultimately
lead to successful separations in the hands of experienced practitioners,

but in the earliest days of HPLC, novices needed guidance and feasibility
demonstrations to induce them to use this novel technique. Out of theranks of pharmaceutical chemists, a prophet emerged to preach thegospel of reversed-phase HPLC. Charles Pidacks left his position as LC
guru at Lederle Laboratories to join Waters in mid-1971 andproceeded systematically to inject every known therapeutic compound

into a Corasil®/C18, or later, a µBondapak™ C18, column. He compiled abible of pharmaceutical separations and, for a decade, traveled

extensively to every corporate lab and industry forum to convert hisformer colleagues. James Clifford, an artist with a pen as well as aprotocol, who personally packed more than 100,000 columns in his

three-decade career at Waters, at Charlie's request, drew a cartoon
showing a column flying on angel's wings.To combat the despair of

those yet uninitiated in the intricacies of HPLC, Charlie used this image
to show that when every good column dies, as it must, it goes toheaven, not hell. Charlie, more than any other single person, wasresponsible for the rapid acceptance of HPLC in the pharmaceuticalpreparative separation tool.

HPLC system destiny was the outsourced UV detector. Enter Ken

Nelson, another interdisciplinary wizard, who conceived the taper cell[21] and designed the optics bench, as well as the circuitry, for the

Model 440 detector, creating a benchmark for sensitivity and stability.
Meanwhile, Burleigh and Lou were busy inventing another pumpconcept, a simpler design using only two check valves and a single

motor to drive separate primary and accumulator pistons in series [22].
While the ensuing M45 product did not generate the sales of theM6000, the concepts embodied in this seminal design were used togreat advantage by major competitors.

Often overlooked is the catalytic role Waters played in the emergenceof LC-grade solvents for mobile phase and sample preparation. In thelate 1960s, a Waters applications chemist, William Dark, asked suppliers

to find a way to stabilize chloroform with something other than ethanol.
Nominally 0.75%, the actual ethanol concentration varied so widely thatit wreaked havoc on retention-time reproducibility in normal-phaseseparations on silica gel.The first to solve this problem was Mallinckrodt.
They created a special grade of chloroform containing a percent of

mixed amylenes, non-polar inhibitors of phosgene formation. About thesame time, a young company in Michigan began to sell a range of useful,high-purity, organic solvents distilled in glass. When customers asked
Waters chemists which brand of solvents they used, theirrecommendation was Burdick & Jackson. Word spread, and B&J's
business boomed.

For a few years, before 1980, Waters forged a foray into the LC-solvent
business. Specifications and analytical protocols using LC and GC that I
helped to develop for our suppliers led to major improvements in thequality of methanol, acetonitrile, tetrahydrofuran, and other key mobile
phase components, especially water. Even before our merger with
Millipore in 1980, we taught them how to use gradient HPLC to

determine trace organic levels in high-purity water.Their MILLI-Q®

System, with an integral organic-removal cartridge, ultimately achievedbenchmark status for water quality. When we exited the business, weshared all our test procedures and demanding requirements with theindustry, thereby creating a competitive environment from which allusers ultimately derived significant quality and performance benefits.
In retrospect, instrumentation expressly designed to operate newcolumn technology, as with the GPC-100 or the M6000/U6K, has beena sure signal of separation power with the potential to alter the courseof science. Waters did this a third time in March, 1976, with theintroduction of the PrepLC™/System 500, incorporating a special dualradial-compression chamber to hold dry, disposable, thin-plastic-walledcolumn cartridges filled with nominal 75µ-silica packing. When I joined
Waters in 1974, Jim Waters challenged me to prove his theory thatlarger particle columns, in series or via recycle, could equal the

Figure 1: Clockwise from top left: One of the first five GPC-100 Prototypes in its plywood cabinet; Parents' home in Framingham, MA, where Jim Waters began his firstbusiness; Flow schematic on page 4 of 1963 Instruction Manual for first GPC-100 [personal copy from Dr. Jack Cazes]; 1965 Waters First Gel Permeation
Chromatography Symposium in Cleveland: left to right, holding model of a high-molecular-weight polymer chain: Jim Waters, Dr. John C. Moore [Dow Freeport], plenary
lecturer Prof. Fred W. Billmeyer, Jr. [R.P.I.], and Waters sales manager Larry Maley; Nicky Anastis tends a Styragel® particle synthesis [reactor made from a 55-gallon
metal drum] in former ladies' cell on second floor of police station; Cover of 1969 brochure for Waters Chromato-Prep preparative chromatograph [note then new 'batwings'logo]; Second generation GPC-200 [1964]. [© 2006 Waters Corporation, used with permission]

122006 The Golden Jubilee year of The Chromatographic SocietyChromsoc Final doc

24/4/06

3:56 pm

Page 132006 The Golden Jubilee year of The Chromatographic Society

13Chromsoc Final doc

24/4/06

3:56 pm

Page 14

separation power of small particles but with orders of magnitude higher

He and Raymond Fisk even applied organic polymerization principles topacking columns in the then 75-year history of chromatography [24].

Nova-Pak, 1982], for which they won the Grand Annual Millipore Award

load capacity.This I did [23] while also finding the first new way of

Operating under non-linear-adsorption isotherm conditions, flying in theface of conventional wisdom, we routinely purified ten grams of sample
in ten minutes, at flow rates up to 0.5 L/min.

Our best serviceman, Richard Turner, and I installed the third

PrepLC™/System 500 instrument in the chromatography lab at Harvard
shared by Woodward and Professor Elias James 'EJ' Corey.That day, stillon a wooden wall rack in the corner, I was excited to find the long steel

columns, with tags in Jim's handwriting, that he had used in the ALC-100
there six years earlier to do the B separations. EJ, who was awarded a12Nobel Prize in 1990, was so impressed with our system's performancethat he later called it, in a New York Times interview: "a revolution inliquid chromatography." He cited it and an Exxon catalyst as: "goodexamples of the fine technological work in American industry." [25]Following this success, Burleigh sought to divide the radial compression
team and conquer new worlds. Carl Rausch was teamed with Dr.Yuri

Tuvim, Dick Vivilecchia, and a new hire fresh from graduate school, Dr.
Uwe Neue.They invented Radial-Pak™ Cartridges and the Radial
Compression Separation System for analytical HPLC [26].

I, on the other hand, was commissioned to "find new, faster, moreconvenient ways to do traditional sample preparation operations." In aninternal memo, dated June 13, 1977, I proposed a project to "solve a real

analytical problem using, whenever possible, our LC technology," and "get to

product design and development as quickly as possible." Together with Dick
Vivilecchia and a young chemist, David Lorenz, hired appropriately fromthe synthesis of an inorganic polymer [a sol-gel silica, trademarked
for Technical Innovation.This seminal groundwork led to several

subsequent materials-science blockbusters, beginning with Symmetry®

spherical-silica packings in 1994, synthesized by Dr. John Petersen's teamin high purity directly from tetraethoxysilane. While not the very first

HPLC silica made this way, it was, and remains, the most reproducible, inlarge measure because of Waters commitment to controlling every
aspect of its synthesis.

Commercially available bonded-phase functionalities [C18, C8, C4, phenyl,etc.] became generic in name, if not necessarily in quality or performance.
Dr. Uwe Neue and John directed a young Ph.D. candidate, Carsten

Niederländer to synthesize a new, long-chain alkyl silane containing anembedded-polar carbamate group.With this reagent, they made, in asingle step, a packing material exhibiting superior peak shape for basic

compounds [28].This shield chemistry, so called because of its apparent
ability to mitigate the effect of surface silanols, was used first to make

SymmetryShield™ RP18 columns. Later it was applied with even greater
success to silica-hybrid substrates [see later] to achieve the best peaksymmetry ever seen for basic compounds, as predicted by a radically new

theory about how reversed-phase separations function [29]. Shield phases
serve as a unique, and, in some cases [e.g., phenols], a dramatic [30],alternative to the traditional selectivity of strictly hydrophobic reversed
phases.To share his vast experience and insight, Uwe, the 2005 Halász
Medalist [31], literally, wrote the book on HPLC columns [32].

What Symmetry® columns did for retention times, Waters Alliance®

Dr. James Thean's Florida State Department of Agriculture lab in

HPLC Systems, introduced in 1996, did for flow rate and mobile phase

1977, the birthday of the SEP-PAK® Cartridge [27]. We used triaxial bed

changes in pumping practice were developed at Waters in the late 80s.

Tallahassee, we watched the fruit of our labor ripen on October 24,

compression and hermetically sealed, individual packaging to maintain

adsorbent activity, bed integrity, and performance uniformity. On January
9, 1978, we shipped the first commercial, disposable, silica-based-

adsorbent LC columns for Sample Enrichment and Purification via atechnique one of the myriad of competitors, J.T. Baker, years later

named, in a play on acronyms, solid phase extraction [SPE].Two years andeight months elapsed before a competitive product came to market.

What followed was explosive growth in the application of SPE to a fullcomposition, reproducibility and system reliability.Two fundamental

First,Theodore Dourdeville and David Trumper devised, for the 600

system, a means to use a microprocessor for real-time modulation of

motor speed to enhance the stability of short-term flow rate [33].Tadthen designed a pump, first used for capillary LC, with a one-motor-per-piston, linear drive and with one pressure transducer for each cylinder, toprovide independence between, and precise control over, the intake anddelivery strokes [34].This concept was refined into a two-motor, twopiston, serial-flow format for the Alliance® 2690 Separations Module.

spectrum of sample preparation problems in every lab around the

Accuracy in its low-pressure solvent-proportioning feature was thereby

stands as a predominant analytical technique.

as a single trace [35]! SDI termed this new yardstick by which HPLC

globe.Today, nearly three decades later, SPE is still growing rapidly andIn the 1980s, HPLC became the tool for applications in every field

where a sample could be dissolved in a mobile phase. Even through thisgreatly enhanced. An overlay of ten gradient chromatograms appeared
performance was measured "one of the most successful products in thehistory of analytical instruments" and "an important influence inevolutionary period, there were undercurrents of invention andfundamentally transforming the industry." [36]Schultz, a brilliant polymer chemist, and his team improved all ourleader in mass spectrometry by acquiring Micromass® Ltd., Manchester,technology developments in many areas. At Waters, the late Dr. Herman
polymer-synthesis processes while learning to tighten control ofimportant variables such as particle diameter and pore-size distribution.

That same year, Waters morphed from a minor player to a major

England. And, further validating that year's slogan, "It's All Important," weintroduced the next revolution in SPE at ISC '96 in Stuttgart: Waters

Figure 2: Clockwise from top left: Publicity photo for organic synthesis marketing program: left to right, postdoctoral fellow Dr. Helmut Hamburger, Prof. Josef F.K. Huber [Vienna],
Jim Waters, and Prof. Robert B. Woodward [Harvard], taken in Woodward's chromatography lab [ALC-100 at right]; Three views of new 50,000-sq.-ft. Waters Milford facility
under construction in early 1973 [spring snow on the ground]; Charles Pidacks, holding new brochure for Waters PIC™ Reagents [about 1977]; First issue of Waters
Chromatography Notes, January 1970 [headline: The Renaissance in Liquid Chromatography]; Cover of first brochure for Model 6000 Solvent Delivery System [1973]; Waters
ALC-200 series cabinet fitted with R400 Refractive Index Detector, M6000 pump, and Model U6K injector [1973]; Separation of Vitamin B12 synthesis intermediate cobyrinate
isomers, differing only in conformation at position 13, on five 2' x 1/8" I.D. Corasil II silica columns in series, using recycle; Cover of Abstract Book from 1971 IUPAC Congress,
Boston, where Woodward described this work; Cover and illustration from brochure for ALC-100 [1967]. [© 2006 Waters Corporation, used with permission]

14

2006 The Golden Jubilee year of The Chromatographic SocietyChromsoc Final doc

24/4/06

3:56 pm

Page 152006 The Golden Jubilee year of The Chromatographic Society

15

Chromsoc Final doc

24/4/06

3:56 pm

Page 16

Oasis® HLB sample extraction products [37]. In the past decade, thisUPLC performance continues to be extraordinary [44] and award

polymer unique in its ability to remain wetted in aqueous phases, while

inventors [nearly 40 new patents filed to date] and principal

growing family of devices, containing adsorbents based upon a patented
retaining analytes by hydrophobic or mixed-mode interactions [38], hasrisen to a position of performance and market leadership, unmatchedin the industry.

At HPLC '99 in Granada, Spain, Waters announced its next materials-

science breakthrough: hybrid-particle technology [39]. Dr. Zhiping Jiang andhis colleagues blended two co-monomers, tetraethoxysilane andtriethoxymethylsilane, to create a porous spherical particle with organic
functionality distributed throughout its backbone, reducing by one-third
the residual population of surface silanol groups. Proprietary bonding
protocols using traditional silanes generated packings with a familiar

winning [45]. Even more amazing is the effort of more than six dozen
contributors from Waters R&D organization, supported by a team of
more than 400 who brought it to the marketplace. Unfortunately,
memory and space do not permit naming here the myriad ofcontributors to the successful development and launch of all the Waters
innovations in the last five decades.

On November 16, 2005, Jim Waters and his bride of 58 years, Faith,

visited Milford to celebrate his 80th birthday [October 7th] with about
thirty folks he had hired and hundreds he has inspired. He told us how
proud he was that we had used his simple formula for success—innovation, a good attitude, and hard work—to usher in the dawn of arange of selectivity, improved peak symmetry, and an order-of-magnitude

new era in separation science. I am humbled and honored by themaking them, by far, the fastest selling HPLC columns in history—until

time, the tale of Jim, his liquid chromatography people, and theirincrease in high pH stability. Sales of XTerra® columns sky-rocketed,even this steep trajectory was surpassed in 2005 by that of XBridge™
columns containing second-generation hybrid technology. Now

successfully armed with an ability to design and synthesize tailoredparticle surfaces and backbones, even deep within micropores, we may
consider experiments that can challenge or confirm novel notions of
how chromatographic separations occur [29].

Continuing our hybrid research program, Dr. Kevin Wyndham and his

co–workers made two exciting discoveries after preparing particles madeby incorporating an ethylene bridge into a silane co-monomer. Resistanceto hydrolysis at high pH was improved by another order of magnitude

beyond that of the first generation methyl hybrid. Even more important,their 1.7 µm-diameter particles were highly pressure-resistant [40].
This encouraged our engineers to design the ACQUITY Ultra

Performance LC™ System, capable of operating at 15,000 PSI. For the
fourth time, Waters created new instrumentation to accommodate
novel column technology [41]. Introduced at PittCon 2004 withACQUITY UPLC™ BEH Columns, this holistically designed systemfurther refined the 2690 pump concept into a two-motor, two-piston

serial-flow device for high-pressure gradient formation.The linear drive,
as well as brilliantly designed check valves, seals, transport tubing, andsoftware algorithms, are key for uniform delivery of compressible liquids

at very high pressure.

Using 1.7 µm particles at 15,000 PSI is something about which Martin
and Synge could only dream [19]. A key to minimizing band dispersion

opportunity to document in this golden jubilee volume, for the firstenormous impact on the practice of liquid chromatography.
Acknowledgments:

I wish to thank Jim Waters for selling me an ALC-100 during mypostdoctoral days in 1971, and Dick Vivilecchia [now at Novartis] andBurleigh Hutchins for hiring me in 1974.They introduced me to HPLC,
taught me how to invent, and enabled my career. Special thanks to Jim

also for his extended conversations with me in recent years, as well as

several long-time Waters employees, whether retired, or still active, whohave supplied many details, documents and images for this historical

record.Thanks to Jim and to my talented colleagues Brian J. Murphy, Dawn
Maheu,Tad Dourdeville, Uwe Neue,Tom Walter, and Mark Baynham fortheir critical review of this manuscript and assistance in its preparation.
References:

Note: Where indicated below, use search term following * to access PDF document in Waters
Library at http://www.waters.com
1. M.S. Lesney, Today's Chemist at Work 7(8): 67-68, 71-72 [1998]
2. B.J. Murphy, Waters History * WA66666; L.S. Ettre, LC-GC 23(8): 500-506 [2005]
Waters is a registered trademark of Waters Corporation.
3. A.T. James, A.J.P. Martin, Biochem. J. 50: 679-690 [1952]
4. J.C. Moore, U.S. Patent 3,326,875 [filed Jan. 1963; issued June 1967]
5. J.C. Moore, J. Polymer Sci. A-2: 835 [1964]
6. J. Porath, P. Flodin, Nature 183: 1657-1659 [1959]; see also:
http://www.bionity.com/articles/d/47068/
7.

http://nobelprize.org/chemistry/laureates/1952/synge-lecture.pdf

for highest efficiency, while maximizing sensitivity, in this UPLC™ system

8. J.F.K. Huber, Chap. 17 in: J. Chromatogr. Library 17: 159-166 [1979];

Ken Nelson's legacy by tapering the light beam, rather than the cell,

9. C. Horváth, op. cit. pp. 157-158

is detector flow cell design. In 1989, Dr. Anthony Gilby had transformed
used in Waters optical detectors in the early 90s [42].Two years later,
he and Bill Carson conceived a high-efficiency, light-guiding flow cell

C. Horváth, Chap. 16 in: J. Chromatogr. Library 17: 151-158 [1979]

10. Proceedings: Waters First International Seminar on Gel Permeation
Chromatography, Cleveland, Jan. 1965; Second: Boston, Sept., 1965;

approach ideal for reduced-volume separations. Ongoing refinement by

11. C. Horváth, op. cit. p. 152

capillary-scale LC and, eventually, in the UPLC system.

12. P.D. McDonald, Chap. 1 in: J. Chromatogr. Library 38: 1-103 [1987] and references
cited therein, esp., R.B. Woodward, Pure Appl. Chem. 333: 145-177 [1973]

Tony,Tad, and Dennis DellaRovere [43] led to the cell types used inThird: Geneva, May, 1966; Fourth: Miami, May, 1967

Figure 3: Clockwise from top left: Dr. Herman S. Schultz [1984]; William A. Dark [1984]; Norma Thimot [1984]; PrepLC™/System 500 with dual Prep-PAK™
Cartridge compression chamber at right—chemist pointing at retained sample debris on used cartridge [1976]; Sep-Pak® Silica Cartridges and original sealed foil pouches [Feb.
1978]; David R. Lorenz, left, and Dr. James Thean in Talahassee laboratory [1977]; Radial Compression Separation System and Radial-Pak™ Cartridges [1978]; Alliance®
HPLC System [2005]; ACQUITY UPLC™ System and its principal inventors and developers, holding 2005 R&D 100 Award plaque; Synthesis scheme for secondgeneration hybrid particles [2004]; R&D 100 Award winners for hybrid-particle technology: left to right: front: Dr. Zhiping Jiang, Pamela C. Iraneta, Raymond P. Fisk, rear:
Dr. John E. O'Gara, Christopher C. Benevides, Dr. Thomas H. Walter [2000]; left to right: Dr. William Carson, Theodore 'Tad' Dourdeville, and Dr. Richard M. King[1985]. [© 2006 Waters Corporation, used with permission]

16

2006 The Golden Jubilee year of The Chromatographic SocietyChromsoc Final doc

24/4/06

3:56 pm

Page 172006 The Golden Jubilee year of The Chromatographic Society

17Chromsoc Final doc

24/4/06

3:57 pm

Page 18

13. A. Eschenmoser, C. E. Wintner, Science 196: 1410-1420 [1977]
14. R.B. Woodward, op. cit. pp. 165-166
15. R.B. Woodward, op. cit. pp. 175-176
16. L. Abrahams, B.M. Hutchins, Jr., J.L. Waters, U.S. Patents 3,855,129 [Dec. 1974];
3,958,898 [May 1976]; 3,981,620 [Sept. 1976]

31. Halász Medal awarded by Hungarian Society for Separation Sciences to Dr. Uwe
Dieter Neue on September 9, 2005, 6th Balaton Symposium, Siófok, Hungary
32. U.D. Neue, HPLC Columns: Theory, Technology and Practice, Wiley-VCH, NewYork [1997]
33. T.A. Dourdeville, D.L. Trumper, U.S. Patent 4,767,279 [Aug. 1988]

17. W.W. Carson, J.S. Roe, U.S. Patent 4,284,352 [Aug. 1981]

34. T.A. Dourdeville, U.S. Patent 5,637,208 [June 1997]

18. L. Abrahams, U.S. Patent 4,027,983 [Jan. 1977]

35. Waters PerformancePerspective WPP33 [1998] – * 990018

19. A.J.P. Martin, R.L.M. Synge, Biochem. J. 35: 1358-1368 [1941]

36. Strategic Directions International, Instrument Business Outlook 6(23): 1, 6-7 [1998]

20. J. L. Speier, J. A. Webster, G. H. Barnes, J. Am. Chem. Soc. 79(4): 974-979 [1957];
B. Sharp Industrial Research 20: 65-66 [1978]; R.R. Jones, Ibid. 20: 9 [1978]; J.L.
Speier, Ibid. 20: 131-134 [1978]

37. P.D. McDonald et al., Poster #151, ISC '96, Stuttgart [1996] – * 970939

21. K.E. Nelson, U.S. Patents 4,011,451 [Mar. 1977]; 4,276,475 [June 1981]
22. B.M. Hutchins, L. Abrahams, U.S. Patent 4,245,963 [Jan. 1981]
23. P.D. McDonald, op. cit. pp. 29-30
24. P.D. McDonald, op. cit. pp. 81-90; P.D. McDonald, C.W. Rausch, U.S.
Patent 4,250,035 [filed Dec. 1975; issued Feb. 1981]
25. V.K. McElheny, New York Times, Col. 1, Pg. 7, Sec. 4 [March 8 1978]
26. C.W. Rausch, Y. Tuvim, U.D. Neue, U.S. Patent 4,228,007 [Oct. 1980]
27. P.D. McDonald, R.V. Vivilecchia, D.R. Lorenz, U.S. Patent 4,211,658[filed Jan. 1978; issued July 1980]; P.D. McDonald, Poster #P112, ISC '98,
Rome [1998] – * 981040

38. E.S.P. Bouvier, R.E. Meirowitz, P.D. McDonald, U.S. Patents 5,882,521 [filed
Apr. 1996; issued Mar. 1999], 5,976,367 [Nov. 1999], 6,106,721[Aug. 2000], 6,254,780 [July 2001], 6,468,422 [Oct. 2002], 6,726,842 [Apr.
2004], 6,773,583 [Aug. 2004]; P.J.J. Lee, J.E. O'Gara, U.S. Patent 6,322,695[Nov. 2001]
39. Z. Jiang, R.P. Fisk, J.E. O'Gara, T.H. Walter, K.D. Wyndham, U.S. Patent
6,686,035 [filed May 1999; issued Feb. 2004]; T.H. Walter, Waters
Whitepaper [1999] – * 990858
40. K.D. Wyndham, T.H. Walter, P.C. Iraneta, U.D. Neue, P.D. McDonald, D.
Morrison, M. Baynham, Waters Whitepaper [2005] – * 720001159EN
41. For detailed information, see http://www.waters.com/uplc
42. A.C. Gilby, U.S. Patent 5,153,679 [Oct. 1992]
43. A.C. Gilby, W.W. Carson, U.S. Patent 5,184,192 [Feb. 1993]; T.A. Dourdeville,

28. U.D. Neue, C. Niederländer, J.S. Petersen, U.S. Patent 5,374,755 [Dec. 1994]

A.C. Gilby, D. DellaRovere, U.S. Patent 6,188,813 [Feb. 2001]

29. P.D. McDonald, Chap. 7 in: Adv. Chromatogr 42: 323-375 [2003]; P.D.
McDonald et al., Lecture #154, ISC 2000, London [2000] – * WA09000;
P.D. McDonald et al., Poster #P0236, HPLC 2001, Maastricht[2001] – * WA10200; R. Stevenson, Am. Lab. 33(16): 50, 52-56 [2001]

44. UPLC™ Application Notebooks – * 720001136EN; * 720001417EN
45. Follow links at http://www.waters.com/uplc for press releases: PittCon 2004
Gold Editors' Award, 3/19/04; Frost & Sullivan Award, 3/14/05; R&D 100
Award, 7/25/05

30. U.D. Neue, Y-F. Cheng, Z. Lu, B.A. Alden, P.C. Iraneta, C.H. Phoebe, K.
Tran, Chromatographia 54: 169-177 [2001]

Discover a better alternative to helium
New UHP H2 Generator for carrier
gas applications
Call FREE on 0800 092 6202 or+44 (0) 191 402 9000

N
P
N
V
T
E
S

Quote ref. CS0406

scientific@domnickhunter.com

18

2006 The Golden Jubilee year of The Chromatographic Society

www.domnickhunter.com/scientificChromsoc Final doc

24/4/06

3:57 pm

Page 19

Chromatography Solutions for Polymer Characterization

GPC/SEC...Get it Right& Every
the First^Timewith High Performance Technology
from the Specialists in GPC
Robotic & ManualSample Prep Systems

Organic & Aqueous GPC/SEC Columns
PLgel, PlusPore, PL EnviroPrep, PL aquagel-OH

PL-SP 260VS, PL-XTR
Highly Characterized Polymer Calibrants

GPC-FTIR Characterizationof Polymers

PL Polymer Standards

GPC-FTIR Interface, SCB Software,
Integrated Systems for Ambient
Temperature GPC

GPC-FTIR Data Import Module

PL-GPC 50

ELSD for Polymer & Compositional
Analysis

Integrated Systems for Mediumand High Temperature GPC
PL-GPC 220, PL-GPC 120, PL-XT 220

PL-ELS 1000, PL-ELS 2100
Polymer Molecular Weight Monitoring Systems
PL-PMC Systems

29 Years of Experience and Expertize
High Level of Technical Support
Applications Service

For further information or to discuss a custom project, please contact us today.
North America
Polymer Laboratories
Now a part of Varian, Inc.
Varian, Inc.
Tel 800 767 3963
EmailSalesUS@polymerlabs.com

UK / International
Polymer Laboratories Ltd
Tel (+44) 01694 723581

Germany
Polymer Laboratories GmbHTel (+49) 06151 860690

Benelux
Polymer Laboratories BV
Tel (+31) 011 8671500

Fax (+44) 01694 722171
EmailSales@polymerlabs.com

Fax (+49) 06151 860670
Email
PLDeutsch@polymerlabs.com

Fax (+31) 011 8671502
Email
PLBenelux@polymerlabs.com

www.polymerlabs.com/gpc

FrancePolymer Laboratories SARL
Tel (+33) 04 91 17 64 00
Fax (+33) 04 91 17 64 01
EmailSupport@polymerlabs.fr
Website www.polymerlabs.fr

Chromsoc Final doc

24/4/06

3:57 pm

Page 20

editorial
The Gas Chromatography
Discussion Groupby Ted Adlard
Ted Adlard

In 1950,A.T.James was working with A.J.P.Martin at the National
Institute for Medical Research at Mill Hill on the outskirts of London.
This was a remarkable institution where scientists were given a
considerable degree of freedom in their investigations so that a
research project which departed from its original goal was notuncommon.An account of this remarkable institution can be found in
Jim Lovelock’s autobiography “Homage to Gaia.”

Tony James was trying to develop an automated method for fractional
crystallisation (which later became zone melting) and when he became
discouraged at his lack of progress, Martin suggested that he should try
gas-liquid partition chromatography (because, according to James, he,Martin, knew that it would work).The possibility of gas-liquid partition
chromatography had been clearly stated by Martin and Synge in theirseminal paper on partition chromatography in 1941 but no one had
followed up the suggestion in the intervening nine years although several
groups in Europe and the USA had developed gas-solid chromatography
with limited success.

After trying over 30 columns, James and Martin were sufficiently
successful to describe their work to small meetings of the Biochemical
Society in the next two years. As biochemists, they were interested in
the separation of fatty acids and amines which had the advantage thatthese compounds could be detected and measured by manual titrationof the column effluent. However they realised that the technique shouldhave wider general applicability and they decided to try to separatehydrocarbon mixtures. Martin contacted the Anglo-Iranian (now BP)
Research Centre at Sunbury-on-Thames with a request for somesample hydrocarbons and D.H. (Denis) Desty was sent from the
Research Centre to find out exactly what was required. Desty
immediately realised that significance of James and Martin’s work andpersuaded his management that the method should be investigated at
Sunbury. Somewhat earlier, an ICI research worker, N.H.Ray, had visited
Mill Hill and had initiated work on gas chromatography at ICI. Ray had
also suggested to Martin that a thermal conductivity device might beused as a detector. For some reason, Martin did not favour the idea andso, when he and James started to separate hydrocarbons which were
not amenable to titration, Martin went on to develop the elegant, butultimately doomed, gas density balance. Ray went on to develop the
now ubiquitous method of sample injection through a rubber septum
and ICI suggested to Shell that gas-liquid chromatography would be asuitable method for the analysis of commercial xylene mixtures.
By 1952 James and Martin’s work was becoming more widely known and
Courtenay Phillips at Oxford who had been investigating gas-solid

20

2006 The Golden Jubilee year of The Chromatographic Society

chromatography switched to gas–liquid chromatography with considerable
success.The real breakthrough occurred at a meeting of the Society for
Analytical Chemistry held in Oxford in September of that year, attended bysome 400 people from industry. Martin and James presented a paper
which, among other things, described an elegant automatic titration
detector which they had designed and built.The enormous leap forwardthat their equipment represented can be best appreciated by comparing itto the apparatus described by Claesson in his work published in 1945.
Phillips also gave a paper that included the first example of column
temperature programming.The excitement generated by thesepresentations was intense and many industrial organisations, particularly
Shell, BP, ICI and Distillers instituted major development programmes,
Advances followed rapidly and Desty persuaded his immediate supervisor,
Dr S.F.Birch, to obtain the backing of the Hydrocarbon Research Group to
sponsor a symposium which was held in London in May/June 1956. (The
Hydrocarbon Research Group had been established during the war by anumber of major UK companies to sponsor research on the synthesis ofpure hydrocarbons required for the IR analysis of enemy fuels).This
meeting was an outstanding success and it was agreed that a discussion
group should be formed to disseminate the wealth of ideas that werebeing generated by the new technique on an almost daily basis. Again
Denis Desty was the instigator and champion of the idea and so the Gas
Chromatography Discussion Group was formed.The second international
symposium took place in Amsterdam in 1958 and in retrospect it is clearthat this meeting was almost the most important in the history ofchromatography since Ian McWilliam described the flame ionisation
detector and Marcel Golay described capillary columns. Perhaps the only
comparable advances were the development of HPLC in the early 1970s
and of silica capillary columns in 1979. Golay worked for Perkin Elmer in
the USA and it is somewhat ironic that whereas capillary columns were
eagerly seized upon in Europe (especially in the petroleum industry) it was
not until the development of silica capillaries that such columns were muchused in the USA.
From 1957 the Gas Chromatography Discussion Group held 3-4
meetings a year where people such as Ray Scott, Howard Purnell, Ervin
Kováts and Jim Lovelock gave papers on their work. It was at one of thesemeetings that Lovelock first described the argon ionisation detector, theforerunner of the electron capture detector. From 1957 until 1972 theGas Chromatography Discussion Group was run under the auspices of
the UK Institute of Petroleum which provided secretarial help and a venue
for committee meetings.This was a mutually advantageous arrangement,bearing in mind the very large part in the development of GC played by
the petroleum companies but by 1972 this arrangement had outlived its

Chromsoc Final doc

24/4/06

3:57 pm

Page 21

usefulness and the Group became an independent organisation based at
Nottingham Trent Polytechnic.This came about through Ralph Stock, amember of the Group for many years and the head of the Science
Department at Trent Polytechnic. At this time liquid chromatography was
beginning to appropriate the instrumental developments of GC andbecame HPLC, so it was decided to change the name to the
Chromatography Discussion Group. In1984 the name was finally changed
to the Chromatographic Society, the Chairman became President and thesecretariat moved to offices in the centre of Nottingham. C.E.R.(Roly)
Jones was the first President under this new title.

The Gas Chromatography Discussion Group was the model for similarorganisations in a number of European countries such as France, Germany
and Denmark and for similar bodies in the USA. By the late 1970s themembership was well over 1000 but gradually declined from this number
as GC and HPLC became “mature” techniques..The strength ofchromatography was greatly enhanced when first GC and then HPLC
were successfully combined with mass spectrometry (a combination
predicted by Howard Purnell in the late 1950s). GC-MS instruments werefirst manufactured in the early 1960s but it was not until the advent of thequadrupole mass spectrometer and computer data handling that the
combination was anything but a research tool.We have now reached thestage of HPLC-UV-MS-MS-NMR and who knows what the future holds.
The next advances are likely to be in the realm of microtechnology soelegantly predicted by Martin in 1962. It is hard to imagine what theanalyst would do if chromatographic methods did not exist and no doubt
Tswett and Martin would be amazed to see how their infant ideas havegrown to the overwhelming importance they have today.

A Century of Separation Science. Celebrating the Past: Predicting the Future
separation science from its earliest

beginnings to possible future prospects.
The meeting was attended by 130

delegates from a number of countries
and also hosted an exhibition by the

major instrument and supply companies.
The Chromatographic Society, one of

the sponsors of the meeting, invited anumber of it past Chairmen/Presidents
and the group photograph of all those
who attended shows a line up of a

On 1st and 2nd April 2004, the Chromatography and Electrophoresis

number of ‘‘old timers’’ who thoroughly enjoyed the reunion. Sadly Dr

on the above topic in York, to mark the 100 years that have passed

following week.Tom Lynch, the Chairman of the C & E Group, is(C & E) Group of the UK Royal Society of Chemistry held a meeting

since Michael Tswett’s first publication on chromatography.Twenty-two
papers were presented covering practically the whole range of

C.E.R. (Roly) Jones, shown fifth right on the front row, died theslightly to the left in the middle ground, immediately behind Professors
John Knox and Keith Bartle.

The Specialists in Separation
Science Consultancy with over
25 years experience
Do any of these apply to you?
• Up to date Market Intelligence data required?
• Need to Rebrand products or company?
• Reduce Product-to-Market cycles?

Need to turn Problems into Opportunities?

Chromatography Consulting Internationalcan bring fresh perspective using proven solutions to theseand many more problems.

• Missing opportunities in India, China andAsia generally?

Join our expanding client list including many market leaders• Distributor chain in need of motivation?

Contact: Bernie Monaghan• Prelaunch, launch and post launch
management a worry?

Telephone: +44 (0) 1695 51973 • Fax: +44 (0) 871 242 3886
e-mail: bm@chromatographyconsulting.com

2006 The Golden Jubilee year of The Chromatographic Society

21

Chromsoc Final doc

24/4/06

3:57 pm

Page 22

editorial
A Historical Overview of Size
Exclusion Chromatography
by Greg Saunders, Polymer Laboratories
Dr Greg Saunders

Introduction

experiments may have taken several hours, analysis times of less thanmodern world.These versatile materials are used in an almost

achievable for specialist applications.

The development of polymers has had a profound effect on thelimitless array of products, ranging from drinks bottles and compact
discs to shampoo additives and prosthetic hip joints.The firstmodern truly synthetic polymer was ‘Bakelite’, developed by
Baekeland in 1907 from the condensation of phenol andformaldehyde. During the 1950s, advances in polymer science led tothe development of many of the now common synthetic polymers,such as polystyrene and poly(vinyl chloride). Engineering polymers

such as polyethylene, polypropylene and polyurethane followed in the
1950-60s, and have become extremely important commodities.
Recent developments have been in the area of speciality hightechnology materials, such as polymers displaying high thermal andchemical stabilities like the polyaryletherketones, polyimides andpolytetrafluoroethylenes. Over the years, many thousands of different
classes of polymer have been developed from a range of different
chemistries, making plastics the most versatile of materials.

The increase in the popularity of plastics has necessitated the need todevelop methods to fully characterise these materials.There are many
parameters that affect the performance of the polymer, including melt
viscosity, toughness and glass transition temperature. However, one

property that strongly influences many other physical parameters is themolecular weight distribution or the distribution of chain lengths in thesample.This article discusses the development of size exclusion

chromatography (SEC) as a technique for analysing polymer molecularweight distributions. (Note:The term gel permeation chromatography(GPC) is also a widely used term to describe this separation.) The

Chromatographic techniques for separating polymeric materials on thebasis of size in solution were first developed in the late 1950s and early
1960s using polydextran and polyacrylamide ‘soft gel’ materials.The

technique was named ‘gel filtration chromatography (GFC)’ and was used
to separate water soluble polymers.These soft gels had a ‘microporous’
structure, in which a low degree of crosslinking (typically under 10%)yielded a non-rigid material with the pore structure introduced by swellingin a solvent. Unfortunately, the low mechanical stability of these materials

restricted their use to applications involving low flow rates and long
analysis times and they were also intolerant of extremes of pH andorganic solvents.These materials are still employed for the analysis ofproteins and polyacrylamide, agarose and polystyrene/divinyl benzene

softgels are readily available. However, it was the limitations inherent inthese materials that led to the development of highly crosslinked
macroporous materials.

Macroporous semi-rigid particles for SEC were first developed by Moore.
Synthesised by suspension polymerisation, these materials had far higher
crosslinking than the microporous gels, resulting in rigid structure withlimited swelling.The pore structure was introduced by the use of

porogens during synthesis and was well defined compared to the softgels. As a result, these semi-rigid particles were mechanically strong andcould be used at higher flow rates.The pore structure could also be

tailored during synthesis to generate materials with a range of pore sizes.
Figure 1 compares and contrasts micro and macroporous materials.

development of hardware will be considered but the discussion of

As a result of the controlled pore sizes and mechanical stability ofthat often are used for the separation of proteins and other biologicalthan microporous gels.They are available in a wide variety of chemistries

columns shall be restricted to polymeric SEC gels, not the silica materials
macromolecules.The development of SEC methodologies for the rapid
determination of molecular weight employed in high throughput
experiments are also outside the scope of this article.

Column Technologies for SEC

Generally speaking, there has been a significant advancement in thecolumns used for SEC. With improvements in the manufacturing
processes used for the preparation of SEC media, higher column

efficiencies are now achievable compared to the materials originallymacroporous materials they are far more widely used for size separations
from (relatively) polar to non-polar, the most popular being copolymers ofstyrene and divinyl benzene. Early suspension polymerisations produced

macroporous gels with a narrow pore size distribution and a fairly narrow
particle size distribution. A range of manufacturers commercially produces
these ‘individual pore size’ gels.

SEC columns packed with individual pore size materials are quite versatile
in application, being robust enough for use with a wide variety of organic
solvents.The limitation of individual pore size materials is that they only

employed for separations. Consequently, the number and length ofgive resolution over a short range of molecular weight. Therefore, toconcurrent reduction in typical analysis times. Whereas early SEC

pore size must be combined together in series, for example, a column set

columns used in a typical analysis have decreased significantly, with a22thirty minutes are now commonplace and much faster chromatography is2006 The Golden Jubilee year of The Chromatographic Society

increase the resolving range of the separation, several columns of different

Chromsoc Final doc

24/4/06

3:57 pm

Page 23

Column type

Column Range

Organic or Aqueous

MultiPore

TSK-GEL HXL

Organic

PLgel

Organic

Mixed /

Individual

PlusPore

Shodex K-series
PL aquagel-OH

Aqueous

TSK-GEL PW

Aqueous

Shodex OH pak

Aqueous

Ultrastyragel

Aqueous

TSK-GEL HXL

Shodex K-series
Styragel

Figure 1. Comparison of micro and macroporous materials for size
exclusion separations.

comprising a PLgel 5µm 106Å, 104Å and 500Å resolves from over

10,000,000 g/mol down to 500 g/mol (polystyrene equivalent in THF).

Manufacturer

Polymer Laboratories
TosoBiosep

Showa Denko
Polymer Laboratories

Organic

Organic

TosoBiosep
Showa Denko

Organic

Waters Corp.

Table 1. Examples of column ranges from various manufacturers (the list
should be in no way considered exhaustive).

A further limitation of individual pore size materials is the artefacts that canoccur when combining columns together due to the overlap of theresolving range of the columns. Mismatches in pore volume in theseregions can cause shoulders on chromatograms known as ‘dislocations’.To
combat this effect, a new column type was introduced in the early 1980s,

the ‘linear’ or ‘mixed bed’ column. In these columns, a homogenised mixtureof individual pore size gels is introduced to cover the desired resolving

range, with the proportion of each gel carefully selected to ensure that theresulting calibration curve is linear and that no dislocations occur.The

majority of SEC applications are now performed on columns of this type.
Recently there has been a new development in organic SEC with theintroduction of ‘multipore’ gels.These materials are composed of a single
gel type (like individual pore size materials) but each polymeric particle

Figure 2. Overlaid calibration curves of typical individual, mixed-bed andpacked with a single type of material have very wide resolving ranges,

5µm MIXED-C and PolyPore columns).

contains a wide range of pore sizes. Consequently,‘multipore’ columns
removing the possibility of dislocations and the necessity to blend

materials (as in mixed bed or linear columns). A further advantage of themethods used to manufacture these gels is that the pore volume is very
high, leading to an increase in resolution compared to traditional SEC

materials.These ‘multipore’ materials represent the cutting edge in SEC
column technology.

Aqueous SEC columns have been developed in parallel with the products
for organic solvents, but the range of columns available is much smaller
due to the limited market for water soluble polymers. Of the threecolumn types discussed above, aqueous columns based on ‘individual poresize’ and ‘mixed bed’ gels are available, but there are no currently available‘multipore materials. For SEC columns to work successfully in water, thehydrophillicity of the packing material must be increased significantly.
Polymeric-based gels are typically highly non-polar and so somemodification of the gel must be performed, either in terms of secondary
treatment of the surface or alteration at the synthesis stage to utilise

different monomers. However, for many aqueous separations silica basedmaterials are still popular.

Table 1 shows examples of the three classes of columns available frommultipore SEC columns (Polymer Laboratories’ PLgel 5µm 500Å, PLgel
Hardware Technologies for SEC

Like other high performance liquid chromatography (HPLC) systems ofthe time, the first SEC systems were home-made instruments.Thesesystems were followed by ‘modular’ SEC equipment comprised of

separate pump, injection and detector modules, with various ancillarydevices such as autosamplers and column ovens.The advantage ofthese modular systems was that components could be mixed andmatched between manufacturers, however, there was nocommunication between instruments and the user had to learn tocontrol the interface of each device. For this reason, the use of modular
systems required skilled operators who were familiar with eachcomponent.

The emergence of integrated systems specifically designed for SEC has
been the trend over the last ten years.These instruments contain all
the components required for the analysis (pump, injection system,

column compartment, detectors and ancillary devices) within a single
unit.The benefit of such devices is that the positioning of thecomponents can be optimised, various safety features can be

introduced and custom computer software can be developed tovarious manufacturers. Figure 2 shows an overlay of calibration curves

control all parts of the system. Such instruments are therefore easier to(Polymer Laboratories’ PLgel 5µm 500Å, PLgel 5µm MIXED-C andconsiderably more safety features.They can also be tailored to suitfor typical individual pore size, mixed bed and multipore columns
PolyPore columns).

use (requiring less training) than modular systems, and containparticular analyses, such as high temperature work, much more easily
2006 The Golden Jubilee year of The Chromatographic Society

23

Chromsoc Final doc

24/4/06

3:57 pm

than modular systems. An

The growing popularity of SEC and the development of integrated

system expressly designed forcollection in early SEC systems was through chart recorders, with data

example of an integrated

SEC is the PL-GPC 50 shownin Figure 3.This instrument canbe operated from ambient to
50°C and is designed toaccommodate a range of
detector options.

SEC employing a single

concentration detector is often
Figure 3. The PL-GPC 50

3109 hiPage 24termed conventional SEC. In

this analysis, molecular weightaverages are calculated relative to a series of standard polymers of known

molecular weight.The disadvantage of this method is that the SEC column
separates on the basis of size not molecular weight, which leads todiscrepancies if the standards used for calibration are a different chemistry(and therefore size in solution) to the sample. Of increasing interest overthe last ten years has been so-called ‘multi detector’ SEC, employing one
or more detectors that respond to molecular properties other thanconcentration. Light scattering detectors and viscometers are the mostcommon detectors used in these applications, with the use of bothdevices (along with the required concentration detector) termed ‘triple

instrumentation have led to increased software requirements. Data

handling either by hand or through custom programming.These earlysystems were slow and difficult to use, and the calculations performed on
the data were cumbersome, greatly hindering the versatility of SEC andmaking these methods clearly not suitable for widespread use. However,as SEC grew in popularity, specialised data collection hardware andhandling software was developed. Many companies with an interest in

SEC have released software that is modern, powerful and versatile, andsoftware now represents a major component of the modern SEC system.
The majority of these SEC software packages are intuitive to use andWindows (TM) based, with some companies offering to fill complianceand regulatory requirements where necessary.

Table 2 shows a brief summary of some of the commercially available

integrated systems, light scattering and viscometry detectors and softwaredeveloped specifically for gel permeation chromatography.
Product type

PL-GPC 50, 120, 220

Instrument

detection’. These devices further aid the analytical scientist in that they

reveal other parameters relating to the behaviour of polymer molecules insolution. Light scattering detectors can be used to estimate the size ofmolecules in solution (in terms of Rg, the radius of gyration), andviscometers give an indication of the molecular density of polymer

Name
PL-BV 400, PL-ELS 1000

can then be exploited to investigate the morphology of polymer

molecules, an important property for many industrial applications.

SEC detectors

Viscotek

Alliance

Integrated SEC systems

Waters Corp.

Cirrus

SEC and Multidetector

Polymer Laboratories

Millenium

SEC and Multidetector

Waters Corp.

OmniSEC

SEC detectors

SEC detectors

SEC and Multidetector

Wyatt

Viscotek

Table 2. Examples of commercially available integrated systems, light scatteringand viscometry detectors and software developed specifically for gel permeation

Static light scattering detectors respond directly to molecular weight andchromatography (the list should be in no way considered exhaustive).

regarding the size of the polymer molecules in solution.The development

Initially introduced for the separation of water soluble polymers by size

so do not rely on a column calibration, and can also give informationof light scattering theory is well established, but the first commercial

Conclusions

in aqueous solution, the technique of gel permeation chromatography

instrument was only available from the 1960s. Since then, manyhas been greatly developed to allow the analysis of a huge variety ofcovering the range of designs and prices. As a consequence, light scattering

replaced in the majority of applications with more robust semi-rigid

instruments have been developed specifically for coupling to SEC systems,
has grown from a niche area of academic interest to a very popular

technique.The relatively high cost of light scattering instrumentation andthe expertise required for operation can, however, prove prohibitive.

Viscometry detectors can be employed to improve the accuracy of SEC

molecular weight calculations by removing the dependence of the calculated
molecular weights on the chemistry of the standards and sample.This isachieved using the Mark-Houwink relationship and the Universal Calibration.
Viscometry gives the intrinsic viscosity of the sample under investigation, aproperty that can be related to the molecular density of the polymer
molecules. Flow through (i.e. compatible with liquid chromatography)

viscometry detectors were developed in the 1960s, and although many

types exist, those which employ liquid flow-through capillaries are by far themost common.The advantage of viscometry is that the technique can greatly
improve the accuracy of a nSEC analysis compared to a conventional

experiment, and the instruments tend to be robust and reliable, although,historically the cost of such devices had limited their appeal. Recently, newand more cost effective designs have become available and viscometry is
becoming increasingly common in the SEC laboratory.

24

Integrated SEC systems Polymer Laboratories
Integrated SEC system

DAWN EOS,ViscoStar

molecules (a property related to the intrinsic viscosity). These parameters

Manufacturer

TDA

LALS,Viscometer

SoftwareApplication

2006 The Golden Jubilee year of The Chromatographic Society

synthetic and natural polymers. Early microporous soft gels have been
macroporous polymeric materials that allow analysis in a wide range ofsolvents.The development of mixed bed or linear columns and now
multiporous high pore volume materials has greatly increased theversatility of SEC. Progress in the design and technology of SEC

instruments has occurred to match this growth in interest. Integrated
instruments specifically designed for SEC have superseded the early

modular systems, and several companies now produce light scatteringand viscometry detectors, instruments once only available in dedicatedresearch laboratories. Specialised SEC software is available that greatly
enhances the ease-of-use of SEC.

Since its inception, SEC has grown steadily in popularity to become amainstay of the analytical laboratory.The growing number of new

developments in SEC demonstrates the popularity and versatility of thetechnique, and bodes well for those looking to separate
macromolecules on the basis of size in solution.

3109
hichrom
3:34 pm
Page
Chromsoc
Final24/4/06
doc 24/4/06
3:57
pm 1 Page 25

The Evolution of HPLC Column Packings
Nicola Vosloo, Stuart McKay, Hichrom Limited, Reading, UK

Since the introduction of liquid chromatography by Tswett in 1903, thefield of separation science has expanded to encompass all branches ofchromatography.
The packing materials for HPLC columns, primarily silica based, have
evolved from pellicular and irregular porous bead types of support in
the early 1970s to the spherical, chemically bonded ultra high purity
phases used today.
The materials used in the early commercially available HPLC columns,
now commonly referred to as conventional or type A silicas, wererecognised as having limitations for the chromatography of basic
analytes (figure 1 (i)). Breakthrough spherical materials of that era, whilst
offering improved chromatography over the irregular materials, still
exhibited variations in batch to batch reproducibility and the addition of
an ion-pair reagent was commonplace to improve the chromatographyof basic analytes.
Figure 1: Chromatography of basic analytes on (i) conventional silica,(ii) ultra high purity silica.

emergence of sub-2µm particles, whilst offering a slight increase inefficiency, requires investment into new instrumentation and the higher
packing pressures involved may effect column bed stability.
The nature of samples analysed today is changing, resulting in an
increased demand for the analysis of more polar, more hydrophilic andionisable compounds. Using standard reversed-phase columns, such
samples are often poorly retained, eluting close to the void volume.
Chromatographers can consider an AQ or polar embedded column(with an amide functionality for example within the alkyl chain) whichallow 100% aqueous mobile phases to be used without any drop in
retention. However such phases often suffer from increased column
bleed, and for very polar compounds retention is still unacceptable.
Moving to an alternative column, such as HILIC4 or mixed mode phase,
can solve the problem of poor retention of polar compounds.
Unique HILIC columns, such as the ZIC-HILIC phase, figure 3, employ
a hydrophilic zwitterion as the functional group, which offers
complementary selectivity to reversed-phase and the added benefit
of improved MS sensitivity.
Figure 3: Structure of the ZIC-HILIC phase.

(i)

(ii)

minutesminutes1) Norephedrine 2) Nortriptyline 3) Toluene 4) Imipramine 5) Amitriptyline, 22°C, 215nm
Mobile Phase: 80:20 MeOH/25mM KH2PO4 pH6.0, 250 x 4.6mm, 1.0ml/min

The purity and the bonding technology of the silica phases improved,with pioneers such as Jack Kirkland at the forefront of phasedevelopment. Column selectivity and peak shape are influenced by thecharacteristics of the underlying silica. Thus the resulting ultra high purity
phases, also referred to as type B silicas, give excellent chromatographyof not only neutral and acidic compounds, but also basic compounds(figure 1 (ii)). These ultra pure silica materials have a low metal ion
content, a uniform silica surface and very low silanol activity, which inturn offer enhanced stability at low pH1 and also at elevated pH2.
Silica based alkyl bonded phases have shown to be the workhorse formany pharmaceutical separations. Alternative chemistries developed,(e.g. CN, Ph and PFP) bonded to ultra high purity silica bases, now
offer alternative selectivity3, (figure 2) whilst maintaining excellent batch
to batch reproducibility and exhibiting improved column lifetime
compared to their conventional counterparts.

A novel range of mixed mode phases, Primesep, has emerged which
combines both ion-exchange and hydrophobic interactions. The
Primesep columns have an embedded ion-exchange group within thealkyl chain and by controlling the pH of the mobile phase, the extent ofionisation of the embedded group can be altered to tune the separation
resulting in the desired retention. Now the retention of polar
compounds can be readily achieved, and as figure 4 illustrates, thePrimesep 200 column offers superior retention of a range of polar
pharmaceutical entities.
Figure 4: Comparison of Pseudo k’ figures: Primesep 200 vs. ‘ReversedPhase polar column’, data courtesy of GSK.
k’ (Primesep 200)k’ (Reverse Phase column)
Note: Only UV visible compounds run on RP Column

Figure 2: Separation of vitamins: changing selectivity with C18 and phenyl
bonded phases.
1
5
1

4

2

C18

5
2

Phenyl

4

1

67

3

7 6
3

0

2

4

6

minutes

8

10

12

0

2

4

6

8

10

12

3

4

5

6

7

8

9

10

A

B

C

minutes

Column: 150 x 4.6mm 5µm ultra high purity silica Mobile phase: MeOH/20mM KH2PO4 (pH2.9),
linear gradient, 5 to 95% MeOH in 20 mins Flow: 1.50ml/min
Components; 1: Pyridoxine, 2: Pantothenic acid, 3: p-Aminobenzoic acid, 4: Cyanocobalamine,
5: Folic acid, 6: d-Biotin, 7: Riboflavin, i: impurity

The growth in LC/MS analysis has driven down the column dimensions
used in some application areas, but more traditional columns sizes (e.g.
250x4.6mm), are still ubiquitous. There has also been a trend towards
smaller particle sizes, from 10µm in the 1970s to 5µm, established as
the analytical standard, and down to 3µm in the 1990s. The recent

CHROM

2

Polar test compounds: A=Procainamide B=Cephaloridine C=4-Guanidinobenzoic acid,
Compounds 1-10, GSK polar compounds. 100x2.1mm column Mobile Phase A: 0.05%
TFA in Water Mobile Phase B: 0.20% TFA in MeCN Flow = 0.4ml/min direct into MS.
Gradient 0 to 100%B in 6 mins, hold for 3mins, back to 0% in 0.5 mins.

Despite the development of novel phases, the preferred starting point
for method development for many remains the tried and tested C18
column chemistry, at low pH, which offers longevity of column lifetime
coupled with excellent column and batch reproducibility.
REFERENCES

1: L.R. Synder, J.L. Glajch, J.J. Kirkland, Practical HPLC Method Development, Wiley-Interscience,
New York, 1996. 2: J.J. Kirkland, J.Chromatographic Sci., 34, 308-313 (1996).
3: M. Przybyciel, LCGC, January 2006, 19-27. 4: J.A. Alpert, J. Chromatography, 499, 17 (1990).

Hichrom Limited, 1 The Markham Centre, Station Road, Theale, Reading, Berkshire RG7 4PE
Tel: +44 (0)118 930 3660 Fax: +44 (0)118 932 3484 nvosloo@hichrom.co.uk www.hichrom.co.uk

CHROM

Chromsoc Final doc

24/4/06

3:57 pm

WWEM (Laboratory) ILM Half A4Page 26

20/4/06

12:37 pm

Page 1

WWEM 2006

Conference,

Water, Wastewater& Environmental Monitoring

Workshops.

1st-2nd November

Exhibition andTelford International Centre, Telford, England.
Are you responsible for monitoring andtesting Water or Wastewater?
The Ideal Event for Analytical Chemists and Scientists

Save £20 Pre-register today
to gain free access to theexhibition and all workshop
sessions. Please log on towww.wwem.uk.com

WWEM is a Conference, Exhibition and Workshops run in conjunction with the Environment
Agency and SWIG. Visitors will be able to discuss product and service needs with leading
manufacturers and service providers. Visitors will be able to participate/attend over 60 informative
walk-in and walk-out workshops free of charge on standards, legislation, mcerts and technology.
Avoid Disappointment Register Today!

WWEM is ideal for process operators, regulators, water authorities, scientists, end users andconsultants who want to learn about the latest product and technology innovations in the field,in the laboratory or in the plant.

Tel: +44 (0)1727 858840 Fax: +44 (0)1727 840310 email: info@wwem.uk.com www.wwem.uk.com

CONGRATULATIONS
to the Chromatographic Societyon their 50th Anniversary!

from Discovery to Recovery

Prep and Process Columns and Media, including easy-to-packhighly efficient SPRING™ Columns.
HPLC Instruments, including pumps, autosamplers, and the Alltech®
ELSD, the most sensitive ELSD from the leader in Evaporative Light
Scattering Detection technology.
Analytical HPLC Columns and Accessories for all applications,
including Alltima™ HP Columns for low-bleed, high-resolution LC/MS
and Everest® Columns for high quality protein separations.
GC Columns and Accessories for all applications, including highperformance individually tested Heliflex® AT™ Capillary Columns,
available in a wide variety of phases.

One Company - Integrated Solutions - From Discovery to Recovery
BE Tel: 09 340 65 65 • Email: info@alltech.be

FR Tel: 03 20 62 96 00 • Email: info@alltech.fr

DE Tel: (0)7457/9493-0 • Email: info.grom@grace.com

IT Tel: (02)-901 10 150 • Email: info@alltechweb.it

NL Tel: 076-571 75 76 • Email: info@alltech.nl

GB Tel: 01524 734451 • Email: info-uk.alltech@grace.com

ALLTECH®, ALLTIMA™, AT™, EVEREST®, FLEXIT™, GROM™, HELIFLEX®, JONES CHROMATOGRAPHY™, MODCOL®, SPRING™, and VYDAC® are trademarks of Alltech Associates, Inc. GRACE®, GRACE DAVISON®, DAVISON® and DAVISIL® are registered trademarks of W. R. Grace & Co.-Conn.

Grace Davison Discovery Sciences

26

EUR_ChromSoc_Ad.indd 1
2006
The Golden Jubilee year of The Chromatographic Society

2/7/06, 8:49:09 AM

Chromsoc Final doc

24/4/06

3:57 pm

Page 27

The First Ultra-Low Bleed
Polar Phase for LC-MS
Ascentis RP-Amide HPLC Column

NOTE: Fewer and lower intensity responses on Ascentis RP-Amide than oncompetitive embedded polar phase HPLC columns.
Separation of Anesthetics
The Ascentis RP-Amide HPLC column is the first amide-based embedded polar phase to exhibit ultra-low bleed for LC-MS. This virtuallyeliminates background mass response that can interfere with analyte identification and quantification.
This application demonstrates the suitability of Ascentis RP-Amide for the efficient separation of the anesthetics phencyclidine and
4-hydroxyphencyclidine by LC-MS.
LC-MS Analysis of Anesthetic Phencyclidine and Hydroxy Metabolitecolumn:mobile phase:flow rate:
temp.:
det.:
injection:
sample:Ascentis RP-Amide, 15 cm x 4.6 mm I.D., 5 µm particles (565324-U)
70:30, 10 mM ammonium acetate (pH 4.5 with acetic acid):acetonitrile
1 mL/min.
35 °C
ESI (+)
10 µL
1 µg/mL each in mobile phase

1

MOLECULAR BIOLOGYCELL CULTURE
CELL SIGNALINGPROTEOMICSANALYTICAL

2

L ABORATORY ESSENTIALSDRUG DISCOVERY

1. 4-hydroxyphencyclidine (m/z 260)
2. Phencyclidine (m/z 244)

For further technical information on Supelco products please email eurtechserv@europe.sial.com

C H R O M AT O G R A P H Y & A N A LY T I C A L P R O D U C T S F O R A N A LY S I S A N D P U R I F I C AT I O N

sigma-aldrich.com/supelco

Chromsoc Final doc

24/4/06

3:58 pm

Page 28

View articles
View thelatest products
Keep up-to-datewith all the newsand events

ALL ONLINE

Microscopy andMicrotechniques ...
Chromatography
and Spectroscopy ...
IT Solutions ...
Laboratory Products ...

Whatever yourspeciality they
are now all easily
available to viewand get quick
responses!
Intelligent Analytical Solutions
Your lab is smarter with the
NEW Agilent 5975 GC/MSD
International Labmate Ltd– We can make it
faster and deliver
even better
productivity!

To sign up and registerfor your newsletter go to
www.internationallabmate.com

Monthly newslettersstraight to your inbox…
28

To see how we can help, visit: www.anatune.co.uk
or email: info@anatune.co.uk

2006 The Golden Jubilee year of The Chromatographic SocietyChromsoc Final doc

24/4/06

3:58 pm

Page 29

The New 2006-2007 Waters Quality Parts®,
Chromatography Columns and Supplies Catalog
We are pleased to present the new 2006-2007 Waters Quality Parts®, Chromatography
Columns and Supplies Catalog. It contains many of today’s leading separation technology
products, such as the new Oasis® 2x4 method development tools, XBridge™ hybrid
particle technology columns, the paradigm changing ACQUITY UPLC™ columns andthe state-of-the-art SunFire™ Prep OBD™ column families.
No other company is as dedicated as Waters to ensuring customer commitment,
product innovation and performance. We invite you to learn more about us, ourtechnology, our resources and our capabilities at www.waters.com.

Waters Corporation U.S.A. 508 478 2000/800 252 4752

Request your copy atwww.waters.com/catalog©2006 Waters Corporation. Waters Quality Parts, Waters, Oasis, XBridge, ACQUITY UPLC, SunFire and OBD are trademarks of Waters Corporation.

Chromsoc Final doc

24/4/06

3:58 pm

Page 30

editorial
The Future of Chromatography
by Peter Myers
My great friend and mentor Denis Desty

will take us into this area. From these plots it is clear that this can be done

and writing about the future, be bold.

monolith or the particle size.

told me that when you are thinking of
Establish your thoughts from a soundbasis and develop them in all directions.

Don’t let conventional thinking restrain you. Denis practiced this in his
work, being very bold in his experimental work, but even bolder inapplying his thoughts to the future. Therefore, I have taken this as my startin writing this short paper on my personal views of the future. Denis got
some things very wrong,, mainly on time scales, but in concept, he got
many things correct. I hope I can come close to some of his visions.

The basis from where I start is the latest developments we have seen inliquid chromatography in the last five years. In this time we have seen the
introduction of monolithic columns offering a lower back pressure for thesame plate count as a classical particle-packed column. And opposing thiswe have seen the introduction of sub-two micron particles offering higher

plate counts but at the expense of far higher back pressures. Are either ofthese the future of chromatography, well let me make my first boldstatement and say no. My reasoning behind this comes from the strong

basic work that has recently (2005) been reported by Gert Desmit fromthe University of Gent who has shown how the classical van Deemter plot
has real limiations and maybe can even confuse us when we are looking at
the efficiencies of the small particle columns and comparing them toclassical 3 micron and above columns and to monoliths.

by increasing the order of the packing and decreasing the skeleton size of aIs all this possible?
Yes it is; today we have 1.5µm porous particles and instruments that willtake us to 1500 bar, but I really don’t see this as the way forward. If I lookat the past and all developments in technology, everything that has

moved forward has been made smaller and simplified. Look at thedevelopment in television; from the 60’s we have seen a change inelectric valve technology, high voltage CRT tubes and large boxes, totoday’s slimline, low voltage LCD TV’s. Another example is in therevolution we have seen in mobile telephone technology. In the late 80’s
I remember using a portable telephone that had a separate battery packweighing over 2Kg. Today most of us use a portable that is much smaller
and uses far less power so that it can operate for days on a 3.7V Li-ion
battery. In these comparisons I am comparing power requirements topressure. Although we can achieve the high pressures required for thepresent systems there are problems in total system requirements. The
alternative and what I think is the way forward into the future is theapproach that was started by Fred Reginer and now continued by many
including the work shown in Fig 2 by Peter Schoenmakers

In the kinetic plot approach, H and E are plotted as a function of thepressure drop-limited plate number (N). Such kinetic plots are as easy toestablish as a normal van Deemter plot and offer the advantage that theperformance of differently shaped and sized LC supports can be directlycompared. The kinetic plots can also be used to show areas of
improvement for LC columns as shown in fig 1.

Fig 2 Ordered pillared structures from the Schoenmakers group

Such structures give an increase in the order of the packing and bycontrolling the interstitial spacing of the columns also give therequired porosity. But the format of these new columns will also bevery different. Today we have not moved on in column design since
the first LC columns with compression end fittings. It still takes skill

to fit these columns into an instrument. After many years of trying I
today can just about connect a 4.6mm id 25cm column onto an
instrument. My bad fitting that introduces 2 microlitres of deadFig 1 Kinetic Plot as described by Desmit

The area to the right of the graph has been termed the forbidden area. To
develop chromatography further in terms of column efficiency andreduced separation time we need to develop supports and packings that30

2006 The Golden Jubilee year of The Chromatographic Society

volume can be accommodated. But I cannot connect a 150micron id
column that still uses compression fittings as, in connecting, thecolumn moves back and so introduces that dead volume and on suchsmall id capillary columns destroys the separation. So new column
connectors, maybe totally integrated column systems must be

Chromsoc Final doc

24/4/06

3:58 pm

Page 31developed. The way forward must be to build the injectors anddetectors into the column. Work on detectors using the total
column has been started by David Goodall.

But let’s go further forward. I do think that the column is the heartof any separation system, so in the future the columns used inchromatography will also be the centre of the system. They will carrya smart chip not only to identify the column but a chip that will carry
the method, such that no longer will we purchase a C18 column but
a column for PAH analysis or a column for basic drugs. The column

will be placed inside the instrument and it will be the column that will
set up the system parameters, perform QC procedures andcalibrations and finally decide when it requires replacing.

But this is only the first stage; there is no reason why using modern

micro and nano fabrication, techniques cannot be further developed
for columns such that a whole range of functions can be integrated
into a single column format. Such that at the front of a totally
integrated column there will be a solid

In fig 3 the transition from the first transistor into the integrated circuit is shownon the left. On the right the first experimental DFGF chips that could be
miniaturised and made into a DFGF separation array. To achieve such a device
a great deal of work needs to be done in the measurement and computer control,
but I did say this was ten years ahead.

phase extraction column, followed by a
pre and then main column with a postcolumn reactor before the analytes areplaced onto a Maldi target. Like in theprevious device, the separation chip willcontrol the instrument and as it will haveits own unique IP address, it will be able toreorder new reagents and then anothercolumn to replace itself when or before it
fails its internal QC.

But let me move forward into the real

future and I hope this is only about ten
years ahead, I believe we could see theend of the chromatography column foranalytical separations. One possibility is

the new developments we are seeing in
dynamic field gradient focusing. In this,separation is achieved by setting a

constant hydrodynamic flow velocity

against step changes in electrophoretic

velocity. Where these two velocities arebalanced for a given analyte, the analyte

focuses at that point because it is driven

to it from all points within the separation
channel. The position of the focused
analyte can be change dynamicallythrough changing the electric field

gradient. By dynamically changing theposition of the focused bands they canbe taken off the device or passed toanother dynamic field gradient device forfurther separation. Hence, individual

devices can be linked together to form aseparation array. I liken this to the way
the first transistors were used in the

manufacture of complex circuits, thesesmall DFGF devices could likewise belinked together.

2006 The Golden Jubilee year of The Chromatographic Society

31Chromsoc Final doc

24/4/06

3:59 pm

Page 32

Fifty years of The Chromatographic Society:
Past and present Chairmen/Presidents of the Society

Dr A.T.James FRS (1957-1960)

C.S.G. Phillips (1961-1965)

D.H.Desty (1966-1968)

A.T. (Tony) James and A.J.P.Martin were thefirst people to develop gas-liquid
chromatography in the early 1950s and Tony
became the first Chairman of the Gas
Chromatography Discussion Group. He went
on to occupy a senior position at Unilever
Research Centre at Colworth House, Bedford.
After retirement from Unilever, Dr Jamesbecame a director of the Wellcome Foundation.

Courtenay Phillips was a Fellow of
Merton College, Oxford. He started
research on GC in 1947 and was the first
person to publish the use of temperature
programmingin 1952.

Denis Desty was almost the first person outside
the biochemistry field to appreciate the enormous
importance of gas-liquid chromatography. Denis
worked tirelessly to advance the technique and toensure that information on developments should
be available to all. To this end he was theinstigator and prime mover of the Gas
Chromatography Discussion Group. Denis
spent his entire career at BP’s research centre at
Sunbury-on-Thames.

Ted Adlard (1970-1973 & 1979-1982)
Ted was for many years a Senior
Scientist at Shell Research Ltds,
Thornton Research Centre near Chester.
He was one of the first dozen people in
the world to make and use a GC. He
was a pioneer of the coupling of GC tomass spectrometry but in the early 1960s
the equipment was so unreliable that nowork on the topic was ever published!

C.L.A.Harbourne (1968-1970)“Happy” Harbourne was a colleague ofDenis Desty at B.P.s research centre at
Sunbury–on-Thames for many years. He
eventually left industry and took up ateaching career.

R.Stock (1974-1976)

C.E.R.Jones (1982 - 1985)

Ralph Stock was one of the very early workers
on gas-liquid chromatography and a memberof the Gas Chromatography Discussion Group
from its inception. In 1972 when the Groupdecided to part company with the Institute ofPetroleum and become an independent body,
Ralph was instrumental in finding a newhome for the Group at Trent Polytechnic (nowNottingham Trent University). He was also
responsible for finding the first two permanent
secretaries, both of whom gave sterling service to
the Group over many years.

C.R.R. (Roly) Jones was widely known asa unique, larger-than-life “character”. Roly
was a pioneer of pyrolysis GC for the
analysis of polymeric materials and did
much to standardise this technique. In his
latter years Roly was a Visiting Professor
in the archaeology department of a local
university and in addition to his scientificachievements, was an accomplishedorganist and musician.

G.A.P.Tuey (1976-1978)
Like Ralph Stock, Peter Tuey was afounder member of the Gas
Chromatography Discussion Group andplayed a key role when the Group decided
to become an independent body. The Rulesof the Society, to this day, are essentiallythe same as the ones written by Peter in the
1970s. Peter worked at May and Baker
for most of his career.

I.D.Wilson (1988-1990 & 2000-2001)
Dr John Dolphin (1985-1988)
John was an early pioneer of HPLC.
Until his recent retirement he worked inclose collaboration with Prof J.H.Knox
on the production of specialist column
packings for liquid chromatography.

32

Ian Wilson works for what used to be ICI,
Pharmaceuticals Divn. but is now
AstraZeneca at their research establishment at
Alderley Park, Cheshire. Ian has won a
number of awards including both the Silver
and Gold medals of the Analytical Division of
the Royal Society of Chemistry. He is, perhaps,
best known today as the exponent of the “bigblack boxes”, HPLC-UV-NMR-MS-MS andtheir application in drug research.

2006 The Golden Jubilee year of The Chromatographic Society

M. B Evans (1990-1992)
After starting work in the chemical industry,
Prof M.B.Evans was in turn a Lecturer,
Senior Lecturer and Professor at Hatfield
Technical College, now Hatfield University.
He supervised many Ph D students fromindustry in various applications of HPLC.

Tony Fell (1992-1994)
Tony Fell came to Bradford University fromthe Herriot-Watt College, Edinburgh. At
Bradford he is a Professor of Pharmaceutical
Chemistry and is now in charge of the
Graduate School of the University. Tony is best
known for his pioneering work on the use ofphotodiode array detection in HPLC. He has
close connections with scientists world-wide, inparticular those from Eastern Europe.

Chromsoc Final doc

24/4/06

3:59 pm

Page 33

Prof M.C.Cooke (1994-1996)

D.Stevenson (1996-1998)

Prof K.D. Bartle (1998-2000)

Dr. Chris D. Bevan (2001-2006)

After some time in industry Mike Cooke
was a Professor at Sheffield Hallam
University and then at the Royal
Holloway College, University of London.
Has now retired to country life.

Derek Stevenson started his career inindustry and then entered the academic
field. For the last ten years he has been
largely responsible for the very successful
Bioanalytical Forum meetings held at the
University of Surrey, Guildford. He also
initiated the first two meetings on chiralseparations run by the Society. Derek iscurrently a Senior Lecturer in Analytical
Chemistry at the University of Surrey,
specialising in environmental andbiological problems.

Keith Bartle was Professor of Chemistryat Leeds University until his recent
retirement. Until becoming an academic
Keith served his apprenticeship tochromatography under Dr David Grant.
Keith has carried out research on many
aspects of chromatography including
SFC, Comprehensive 2-D GC andUniversal chromatography. Nowadays
he can be found at Headingly or Trent
Bridge if it’s not raining.

Chris Bevan has been President of the CS for thelast 5 years which has been a particularly difficult
time for the Society. Chris works at the GSK
Medicinals Research Centre at Stevenage. He hasorganised three outstandingly successful meetings
at the Stevenage/Harlow sites of GSK on theapplications of large scale chromatography. This
is now a major technique for the preparation andpurification of pharmaceutical compounds.
Currently he is organising three meetings during
the year to celebrate the 50th anniversary of the
Gas Chromatography Discussion
Group/Chromatography Discussion Group/
Chromatographic Society.

LGC pharmaceutical services
With over 1,000 staff, working in awide range of scientific disciplines,
LGC is Europe’s leading independent
analytical laboratory. Equippedwith the latest in state-of-the-art
instrumentation, LGC is uniquely
positioned to meet the measurement
needs of the pharmaceutical sector.
LGC offers a range of complete
outsourced and specialised niche
services tailored to client needs,
which are supplied out of technicalcentres across Europe and in Asia.
As a supplier to the pharmaceuticalsector LGC has built on its industry
knowledge and the technicalexcellence developed from the roleof the UK’s designated national
laboratory for chemical and biochemical
metrology, to provide leading-edge
and cost-effective services in:• Leachables and extractables
LGC provides analysis andmethod development to determine
the leachable and extractablecomponents from delivery andpackaging systems to meet
regulatory requirements.
• Pharmaceutical reference materials
LGC supplies synthesis, production,
certification and distribution services
to meet the quality and measurement
requirements of quality control
and analytical development groups
within the pharmaceutical industry.
For further information on
LGC’s pharmaceutical services,
please contact
Neil Malone: +44 (0)1928 513986• Particle characterisation
LGC is highly experienced in thecharacterisation of pharmaceuticalparticulate materials using an
extensive array of leading edge
technologies and well establishedmaterials techniques to meet thedemands of QC testing, productand process development.

Innovative new technology
range from Glygen Corporation
for sample preparation andpurification of biologicalsamples at the micro- and nanolitre sample scale• Samples as small as 0.1µl handled• Ideal for MALDI and LC/MS• Fast, sample, easy-to-use withminimal sample loss
• Over 60 chemistry medias available• No contamination fromsupporting matrix• Enzyme Reactions, Affinity
Chromatography and Desalting
Applications• NEW – now available as
Lab-in-a-Plate (96, 384 or 1536)

www.lgc.co.uk© LGC Limited, 2006. All rights reserved

Lab-in-a-Tip™

For further details contact
Bernie Monaghan
Chromatography Consulting International Ltd.
115A Liverpool Road, Bickerstaffe, Lancashire, L39 0EQ, UK
Tel: +44 (0) 1695 51973 • Fax: +44 (0) 871 242 3886
e-mail: bm@chromatographyconsulting.com

1413/AS/0406

2006 The Golden Jubilee year of The Chromatographic Society

33

Chromsoc Final doc

24/4/06

3:59 pm

M.S.Tswett“The man who started it all”, Warsaw 1906

Archer John Porter Martin 1910-2002
With his colleague, R.L.M.Synge they canbe considered to be the founders of themodern chromatography era.

Page 34

R.L.M.Synge
Joint Nobel Prize Winner with A.J.P.Martin for
their 1941 paper on partition chromatography

Front row 2nd L, Dr R.P.W.Scott, Centre Prof. Phyllis Brown, 1st R Prof. Georges Guiochon, 2nd R Dr J. J.(Jack) Kirkland, ca. 1965. Ray
Scott was one of the pioneers of GC in the 1950s. Jack Kirkland was a pioneer in the production of HPLC packing materials and the co-authorof a well-known textbook. As a member of GAMS, Georges Guiochon was a close collaborator with the Society in the 1970s and 1980s.

Dr J.Janak,Dr R.P.W.Scott and Prof.Howard Purnell. Edinburgh 1960 Dr Janak was an earlypioneer of GC and inventor of a method of gas analysis using CO2 as the carrier gas. Howard
Purnell was the author of the definitive book on the theory of chromatography for many years.

Denis Desty, Martin and Victor Pretorius in South Africa ca. 1975

34

Martin with Synge in the background receiving their Nobel
Prizes from the King of Sweden,1952.

2006 The Golden Jubilee year of The Chromatographic Society

Dr S.Lipsky and Dr J.Janak
Sandy Lipsky of Yale University was an early exponent of capillary columns in the USA
and collaborated with Jim Lovelock on ionisation detectors.

Ted Adlard receiving the Martin Medal from A.J.P.Martin 1982.

Chromsoc Final doc

24/4/06

3:59 pm

Page 35

Professor L.S.Ettre, Martin Medallist, 1982. Well known for his many
original contributions and for his work on the history of chromatography

Dr G.A.P. Tuey , A.J.P. Martin, Dr R. Stock, E.R. Adlard, Dr C.E.R. Jones and D.H. Desty at the Science
Museum, London, 3May 1984.

E.Rees, E.R.Adlard and C.E.R.Jones with a Griffin and George GC presented to the
Science Museum, London 1987.

Back row: Dr J. Dolphin and Prof M.B. Evans. 2nd row: Dr G.A.P. Tuey, Mr I.W. Davis and Mr
M. Cooke. 3rd row: Dr P.A. Sewell, Mrs J.A. Challis and Mr M. Perry. Front row: Mr
N.G.McTaggart, Dr C.E.R. Jones, Mr E.R. Adlard at the Science Museum, 3 May 1984. These werethe members of the CS Committee at the time.

Dr John Dolphin, Dr Henri Colin, Prof J.H.Knox and Prof E.Sz Kováts. Prof. Ervin
Kováts was the inventor of the relative retention system that bears his name.

Professor J.E.Lovelock
and Mrs D.Desty at theRoyal Institution,
London, 2001. Jim
Lovelock is holding aminer’s lamp presented tohim by the
Chromatographic
Society.

Prof. John Knox receiving his Martin Medal from Dr John Dolphin.

2006 The Golden Jubilee year of The Chromatographic Society

35

Chromsoc Final doc

24/4/06

3:59 pm

Page 36

Prof. M.B.Evans, Mrs J.A.Challis and Dr C.E.R.Jones on the occasion of Mrs Challis’ retirement. Jennifer Challis was
the Secretary of the CS for 15 years. She now lives in Devon with her husband Prof Laurie Challis, former Professor of
Physics and Pro Vice-chancellor of Nottingham University.

Dr John Dolphin, Prof Ernst Bayer and Dr C.E.R.Jones. Prof Bayer, formerly Professor of Organic Chemisrty
at Tubingen University, died in 2002. Prof Bayer did his PhD under Prof Kuhn and can therefore be said to bea direct link, through Willstätter, to Michael Tswett. Prof Bayer was one of the earliest workers on
chromatography in Germany.

Alan Handley, current Vice-president of the CS. Alan formerly
worked for ICI but now works for LGC at the Heath in Runcorn.
His chromatography credentials go back many years.

Dr Michel Martin.
Dr Martin was a member of GAMS (the Frenchsociety that organised symposia on chromatography)
for many years and collaborated with the CS on anumber of meetings. He did research at the Ecole
Polytechnique, Paris under Prof Georges Guiochon.
(See group photo on page 34 ).

Prof. Walt Jennings, ca 2002, US pioneer of capillary GC. Until his recent
retirement Walt Jennings was known world-wide for his dynamic lecture
courses on GC.

Professor Peter Myers, former Vice-president of the CS, at a
Royal Society of Chemistry meeting in York in 2004
demonstrating particle packing with the aid of a packet of
M&Ms™.

Dr Ken Jones. Ken Jones was the Hon Treasurer of
the CS for a number of years. Among many otherclaims to fame, Ken was one of the founding
directors of Phase Separations Ltd.

2006 The Golden Jubilee year of The Chromatographic Society

36KROMASIL.® IT JUST GOES ON AND ON.
Kromasil HPLC columns are based on high purity
Kromasil silica. They give you symmetrical peaks evenfor basic compounds, just as you would expect fromother top performing analytical columns. The differenceis that Kromasil columns last much longer.
During the past ten years, Kromasil has been theleading brand for large-scale HPLC, where long lifetimeand thereby high chemical stability are crucial forminimizing costs.

Eka Chemicals AB. Bohus Sweden Tel +46 31 58 70 00
Eka Chemicals Inc. Marietta GA. USA Tel +1 800 2421 3900

Want to know how Kromasil gives you more analyses
from every column? You can find everything fromdetailed product descriptions to order information on
our website. Or why not contact your nearest Kromasil
distributor directly? It could be the beginning of a
relationship that just goes on and on.
Kromasil is available in particle sizes of 3.5, 5, 10,
13 and 16μm; pore sizes of 60, 100 and 300Å andseveral surface modifications.

The time has come for your lab to makethe step up to the Waters® ACQUITY UPLC™
system. With more data and unparalleledsensitivity, the ACQUITY UPLC system gives
you the results you’re searching for, withgreater efficiency and confidence. Whichmeans better method development. Morebreakthroughs. And greater success than you’ve
ever had before. Finally, technology is nolonger holding back progress. Instead, you
can push science forward. The future of
Liquid Chromatography is the ACQUITY UPLC
system. Visit www.waters.com/a1© 2006 Waters Corporation. Waters, ACQUITY Ultra Performance LC, UPLC, and ACQUITY UPLC are trademarks of Waters Corporation.