22 may 2014 volume 10 issue 9 cover story...

The Future of PharmaEmerging trends in pharmaceutical analysis

2 Trends in Pharmaceutical Analysis: A Technology Forum The Column spoke with experts in the pharmaceutical industry

about current and emerging trends in pharmaceutical analysis,

including the use of LC–MS instead of LC–UV for routine assays,

best practices for impurity profiling, and areas where commonly

used methods are likely to improve.

Cover Story

Features

17 The 2014 LCGC Awards (Part 2) Megan L’Heureux

The 2014 Emerging Leader in Chromatography Award was presented to André

de Villiers. Here we chart his career and accomplishments so far.

Regulars7 News

Potential biomarkers for larynx cancer, investigating the origins of metabolism,

and the latest apps for separation scientists are featured this week.

13 Evaluating the Temperature Shift in Analytical Temperature Rising Elution Fractionation Adrian Boborodea and André Luciani, Certech ASBL

This article presents a new method to evaluate the temperature shift observed

in analytical temperature rising elution fractionation (ATREF). The evaluation is

based on the dependence of the measured peak temperature as a function of

heating rates. Application of the proposed method does not require any

knowledge of the fluid circuit characteristics geometry and avoids the use of

narrow preparative TREF standards. The results are found to be more accurate

than the method that is usually applied.

10 Incognito 3D Printing — A Coming Revolution? Incognito speculates on how 3D printing could change life in the laboratory.

21 CHROMacademy

Update on what’s new on the professional site for chromatographers.

22 Training Courses and Events

24 Staff

22 May 2014 Volume 10 Issue 9

ES438346_LCTC052214_001.pgs 05.14.2014 22:47 ADV blackyellowmagentacyan

Trends in Pharmaceutical Analysis:A Technology ForumThe Column spoke with experts in the pharmaceutical industry about current and emerging trends in pharmaceutical analysis, including the use of LC–MS instead of LC–UV for routine assays, best practices for impurity prof ling, and areas where commonly used methods are likely to improve. Participants in this technology forum include Ann Van Schepdael, a professor at the KU Leuven in Leuven, Belgium, Tom van Wijk, a senior scientist at Abbott Healthcare BV in Weesp, the Netherlands, and Harm Niederlander, who was a project leader at Synthon Biopharmaceuticals in Nijmegen, the Netherlands, until August 2013.

Q: Has there been any signif cant

adoption of liquid chromatography

coupled to mass spectrometry (LC–MS)

for routine pharmaceutical analyses? Or is

liquid chromatography–ultraviolet

(LC–UV) still applied more often for

routine assays and quantitative analysis?

Ann Van Schepdael: Many monographs

still use UV as a detection technique and

LC–UV for assays and rela ted substances.

LC–UV equ ipment is affordable and robust,

and the available column chemistries allow

the analyst to play with the selectivity of the

system. LC–MS may also be in use in the

industry on a routine basis, but it appears less

in pharmacopoeial texts. It seems that LC–MS is

very important for the preparation of regulatory

f les for a new chemical entity (NCE): It is

signif cant for the structural characterization of

unknown impurities on the one hand, and for

quantitation of the drug and its metabolites in

biological samples on the other. This is because

of its better sensitivity and very good selectivity.

The study of a drug’s pharmacokinetics

(metabolite characterization, quantitation of

excretion, kinetics of metabolism, and drug

interactions) is quite well supported by LC–MS.

Tom van Wijk: LC–MS plays a crucial role

in pharmaceutical analysis, but in contrast

to biopharmaceutical analysis, LC–MS is

hardly used for routine analysis. Although it

is technically feasible to quantitate known

impurities with variations within current

requirements, the use of LC–MS for routine

testing in pharmaceutical analysis is generally

avoided. Aside from the fact that more

technical details need to be in place, in general

a number of non-technical issues, such as cost

of analysis, transferability, and obtaining and

maintaining suff cient knowledge levels, put off

running this type of method. Quantitative

LC–UV–MS methods are used in early

development in cases where specif c and

sensitive detection is required; for example, for

the analysis of low level genotoxic impurities

or impurities without a chromophore. For the

latter, often an alternative method will be

developed for quality control (QC) purposes. Ph

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Pharma Analysis Forum2 News7 Incognito10 Boborodea and Luciani137 100LCGC Awards 201417 CHROMacademy21 Training & Events22 Staff24212 2222


The Column www.chromatographyonline.com

Routine testing of genotoxic impurities in the

f nal product can often be avoided by

controlling impurities in the intermediate steps

of the process.

Harm Niederlander: To answer this, we

need to consider what is routine. LC–MS is

used in analytical method development in

pharmaceutical analysis more and more. As

method development is increasingly becoming

an automated task (though still requiring

case-by-case expert evaluation), LC–MS can

be considered an important tool in “routine

analysis”. Furthermore, though less routine, the

role of LC–MS in product characterization and

structure elucidation of “unknowns” in both

pharmaceutical and biopharmaceutical analysis

is indispensable. If, however, you are talking

about release and stability testing, LC–MS is

largely avoided because its application is still

not as straightforward as UV detection, for

example.

Q: What are the best practices for

conducting impurity prof ling of drugs?

What is the best approach to f nding

unknown impurities?

AVS: In order to safeguard the quality, safety,

and eff cacy of medicines, impurity prof ling

of drugs is paramount. The chemical structure

of these impurities is usually very much like

that of the API and therefore the separation

of API from impurities can be a challenge. As

a result, drug producers use methods with the

highest possible resolution to study the related

substances present in a drug.

When aiming to f nd all the impurities in

a drug, it is advisable to implement various

kinds of separation techniques. For instance,

LC can have a different separation selectivity

from capillary electrophoresis (CE) so applying

both techniques yields supplementary and

complementary information about a particular

sample. Moreover, within chromatography,

it is often advisable to use a combination of

columns with different selectivities, that is, to

apply orthogonal methods.

Another way of f nding all the impurities

in a sample is to combine a separation

technique with a variety of detectors. Each

type of detector can highlight different types

of compounds because the detector response

depends on the chemical structure of the

compound. Does it have a UV chromophore?

Does it exhibit good ionization in an MS probe?

Does it show good conductivity? The answers

to these questions point to different detectors.

TvW: The procedure applied strongly depends

on the level of knowledge of the drug

substance, the phase of development, and the

purpose of the impurity prof ling study. For

well def ned processes, applying the def ned

method of analysis for different batches

could be an acceptable prof ling approach.

However, state-of-the-art prof ling would

Pharma Analysis Forum

3


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require a different approach, starting with

performing a theoretical assessment based

on product knowledge and the literature on

the synthesis, degradation pathways, and

interaction with excipients. Based on the long

list of impurities from the assessment, selection

of the analytical techniques and methods will

be made, taking into account the physical and

chemical characteristics of the components,

such as presence of chromophores, (calculated)

pKa values, and (predicted) ionization in MS.

After tuning the methods based on a set

of representative components, the data are

processed, applying peak picking software

to enable any impurities to be found or to

compare (differences in) impurity levels. This

approach, combining prior knowledge, good

quality data, and suitable software, enhances

the chance of detecting unknowns and

increases the understanding of impurities

found. When the information is added to

a product knowledge document, it allows

the information level to be monitored and

estimations to be made at any point in time.

HN: It is not really possible to point out a

single procedure that is routinely applicable

for impurity profi ling. The very diverse nature

of (potential) impurities demands signifi cant

expertise in the selection of separation

and detection techniques to be included

in impurity profi ling studies. A theoretical

expert assessment of impurities that might

be expected should therefore always precede

any effort of practical profi ling. Based on

the estimated properties of these potential

impurities, a choice can be made for an array

of separation and detection techniques to

include in such studies. These separation

techniques should preferably be selected

so as to be orthogonal (that is, relying on

signifi cantly differing separation mechanisms;

for example, reversed-phase LC [various

modes], hydrophilic interaction chromatography

[HILIC], CE) to minimize the chances of missing

out on impurities that may potentially co-elute,

elute without retention, or not elute at all. In

addition, for detection, it is desirable to include

more than just a single technique (note that

no single detection technique is really generic).

Finally, after profi ling, investigation of mass

balance proves to be a versatile tool to estimate

if important impurities may have been missed.

Q: What are important topics for research

in pharmaceutical analysis of small

molecules?

AVS: For small molecules, analysts always

aim for an improvement in the sensitivity of

the analysis and an improvement in selectivity

of the separation technique, as well as an

improvement in effi ciency and speed. This

is why we will most probably witness in the

near future more and more methods using

ultrahigh-pressure liquid chromatography


4


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ES438479_LCTC052214_V4.pgs 05.14.2014 23:57 ADV blackyellowmagentacyan


(UHPLC), entering into monographs.

Companies can save a lot of time and

expense by adopting the newer miniaturized

separation techniques, and they have an

advantage in doing so from the start, that

is, when submitting the regulatory f le. They

can avoid the time-consuming method

transfer and adjustment process needed

to transfer a standard LC method to a

miniaturized one.

TvW: In general, activities that support impurity

prof ling offer opportunities for improvement;

for example, the improvement of method

development strategies, orthogonality of

methods and techniques, column selection,

prediction of degradation pathways, and

interaction with excipients. Analysis of polar

components is of special interest as these show

little retention in the classic LC–UV methods

on C18 columns; investigation into the use of

HILIC separation methods have grown in the

last few years as a result. Control of potential

genotoxic impurities is also an important area

for research. In contrast to impurity prof ling

of regular impurities, which focuses on the

detection of any unknowns above a specif c

threshold, the control of potential genotoxic

impurities today fully relies on assessments.

Although the impurity threshold for genotoxic

impurities is much lower, technical capabilities

allow screening for toxic impurities based

on their intrinsic reactivity, in addition to

the assessment. Although not required,

these screening methods have already been

developed for alkylation agents and a similar

approach would allow other classes of toxic

compounds to be screened for. With new

EMA (CHMP/SWP/4446/2000; 2013), USP

(232/233; 2014), and ICH (Q3D; 2013) guidance

on the horizon for heavy metals, there is a

strong increase in work performed in this f eld,

mainly using inductively coupled plasma–mass

spectrometry (ICP–MS).

HN: Important areas include: genotoxic

impurities; residues of (heavy) metal (catalysts);

and process analytical technology.

Q: How are methods used for quality

control and characterization of

biopharmaceuticals different from

those used with small-molecule

pharmaceuticals?

AVS: Biopharmaceuticals tend to be more

complex in primary and secondary structure.

In the past decades we have seen the arrival

of various forms of biopharmaceuticals, all

with their own specif city. Following the f rst

set of compounds made through genetic

engineering, we have seen the coming of

monoclonal antibodies and the conjugated

forms of biopharmaceuticals, made in order to

enhance their pharmacokinetic performance.

All of these biopharmaceuticals require proper

characterization such as a study of glycosylation

patterns and checking for the presence of

deamidated products. Thinking about nucleic

acid-based materials as oligonucleotides, the

determination of their sequence can be done

using MS coupled to a separation technique.

Purity testing can gain from the combination

of different orthogonal techniques, such as ion

exchange liquid chromatography and sieving

capillary electrophoresis.

For proteins, sequencing techniques and

tryptic maps can also perform structure

conf rmation. But the biopharmaceutical

f eld is in need of techniques that allow

quality assessment of intact proteins. The

latter are indeed the compounds that will be

administered to the patient, and their activity

and quality are determined by the structure of

the intact protein.

TvW: Biopharmaceuticals cover a wide range

of compound classes and, when compared to

small molecules, the classif cation of purity and

impurity is not that well def ned. Historically,

many people working in biopharmaceuticals

have a background in small molecules and

the ICH Q3A/B guidance may be followed

as a way of ensuring quality, however, this

may not always be feasible or required. Often

“f ngerprints” are used for characterization

purposes. For biopharmaceuticals, higher

reporting and identif cation levels of impurities

are acceptable because of the larger process

variation anticipated.

For both small-molecule pharmaceuticals and

biopharmaceuticals, high-end technology is

available and is more often applied as supportive

data for product characterization in regulatory

f lings. For routine quality control analysis,

however, the classic methods, such as ELISA and

SDS-page for antibodies, are still in place.

HN: Typically, the “purity” of

biopharmaceuticals extends beyond the level

of identifying or quantifying components

that are not the intended active ingredient.

Biopharmaceuticals may consist of mixtures

of iso-forms and slightly (differently) modif ed

proteins that can all represent (some)

activity. Therefore, prof ling the composition

of these mixtures is an important part of

biopharmaceutical analysis in characterization

and quality control. Parameters evaluated

often include: Folding and association using

spectroscopic techniques (circular dichroism,

f uorescence); oxidation, deamidation, and

N- and C- terminal heterogeneity using typtic

peptide mapping; charge heterogeneity using

cation-exchange chromatography (CEX)

or capillary isoelectric focusing (CIEF); and

glycosylation using digestion or deglycosylation

with reversed-phase LC, anion exchange

chromatography (AEC), or matrix-assisted

laser desorption–ionization time-of-f ight

(MALDI-TOF), and receptor assays.

Following on from the fact that drug activity

results from the combined effect of many


5




individual contributions, at least one (overall)

activity assay (often cell-based) is always

included.

Furthermore, the diversity of product-

and process-related impurities is generally

much wider for biopharmaceuticals than for

small-molecule pharmaceuticals. As a result,

the number of methods needed to cover all

of these is generally much wider too. These

include: Product-related impurities: Soluble

aggregation is tested using size-exclusion

chromatography (SEC); and cleavage,

decomposition, or proteolysis is tested

using SEC, SDS-page, or CE. Process-related

impurities: Host cell proteins are tested using

immunological techniques; DNA impurities

are analyzed using real-time polymerase chain

reaction (qPCR); and individual generally

xenobiotic process additives are analyzed

using immunological, chromatographic, or

spectroscopic techniques. Other: Bioburden

or virus-related testing is carried out using

compendial techniques; and general

parameters are also tested using compendial

techniques. Please note that my focus here has

primarily been on antibody biopharmaceuticals.

Q: Are spectroscopic techniques (without

chromatography) still important for

pharmaceutical analysis?

AVS: The answer to this question depends

on the purpose of the analysis. What type

of information does the analyst aim for?

A separation step might not be needed

when the compound is present as the only

active pharmaceutical ingredient (API) in a

pharmaceutical mixture, and is simply dissolved

in a simple matrix. In such cases, it may be

possible to apply a stand-alone spectroscopic

technique.

One example is identity testing of a bulk

pharmaceutical ingredient. This is most

conveniently performed by infrared (IR)

spectroscopy. Indeed, the typical f ngerprint

region in the IR spectrum allows conf rmation

of the identity of a compound after comparison

with the spectrum of a standard.

When the purpose of an analysis is assaying

the medicine, UV testing can be a good

choice. Formulations containing a single

API can be conveniently analyzed with UV

spectrophotometry if the excipients do not

interfere in the UV absorbance. It may or

may not involve a simple sample preparation

procedure, and the analysis could be applied

routinely because of its simplicity. It is also

possible to carry out UV analysis in the form of

a f ow injection analysis (FIA). In FIA the samples

are injected into a f owing stream of liquid that

continuously passes through a detector cell. UV

measurement in the cell allows very fast and

automated analysis of all the samples.

When people are testing for the presence of

impurities in medicines, in the majority of cases

evaluation or comparison of colour against

European Pharmacopoeia reference standards,

to make colour assessments more objective.

These examples show that new opportunities

can still be found for applying direct

spectroscopic techniques.

HN: Maybe less so in small molecule

pharmaceutical analysis, but in

biopharmaceutical analysis, spectroscopic

techniques (without chromatography) still

have a very important position in product

release, stability testing, or characterization.

A few examples include: UV absorbance

in the content analysis of protein products;

absorbance or f uorescence in immune- and

cell-based assays (and even in some chemical

assays like those that test for free SH groups);

and f uorescence and circular dichroism in

the characterization of secondary and tertiary

structure of proteins.

Ann Van Schepdael is a professor at the KU

Leuven in Leuven, Belgium.

Tom van Wijk is a senior scientist at Abbott

Healthcare BV in Weesp, the Netherlands.

Harm Niederlander was a project leader at

Synthon Biopharmaceuticals in Nijmegen, the

Netherlands, until August 2013.

a separation technique will be implemented. In

this case sample preparation may be needed

for the analysis of drug products containing

the formulated drug in the presence of

excipients.

There are also areas in which the use of

particular spectroscopic techniques (without

chromatography) is emerging, such as quick

and initial detection of a counterfeit drug in

suspicious medicines. Raman and near infrared

(NIR) spectroscopy have shown their strength in

this f eld. These spectroscopic techniques could

become as important as UV in the future.

TvW: Separation prior to detection generally

results in higher specif city. However, since this

is not always required, direct application of

spectroscopic techniques is therefore desired in

cases where timely and cost-effective analysis

is paramount. Direct UV measurement is the

preferred detection for dissolution testing.

Vibrational spectroscopic techniques such as

near infrared (NIR) and Raman are used in

process control and for anti-counterfeit analysis

where fast and nondestructive analyses are

required. Implementation of quality-by-design

(QBD) has resulted in a strong increase in

the use of these techniques. In addition,

spectroscopic techniques can replace visual

AVS: [email protected]: [email protected]: [email protected]


6



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Peak Scientific (Inchinnan, Scotland) has

been awarded a 2014 Queen’s Award for

Enterprise in International Trade decreed by

Her Majesty The Queen of Great Britain, on

the recommendation of the Prime Minister,

David Cameron. The awards are announced

each year on the Queen’s birthday, 21 April.

This is the fourth time that the company

has received this award in recognition of its

international trade and growth overseas.

Established in 1997, the company is now

supported by a network of distributors and

service partners in 70 countries worldwide.

Each award is valid for five years, and winners

are invited to a reception with HM The Queen

at Buckingham Palace in London, UK.

For more information please visit:

www.queensawards.org.uk

Over the last 30 years, the occurrence of head and neck cancer in Spain has been increasing, and now accounts for between 5–10%

of malignant tumours diagnosed each year.1 In a new study published in the journal Chromatographia, solid-phase microextraction

(SPME) with gas chromatography coupled to mass spectrometry (GC–MS) has been performed to identify two potential biomarkers of

epidermoid laryngeal cancer, that could be used in non-invasive diagnostic testing.

Adolfo Toledano, an otolaryngologist from the Alcorcón hospital (Madrid, Spain), approached Rafael García from Rey Juan Carlos

University (Madrid, Spain) with the idea of applying volatile compound analysis to the exhaled breath of patients as a method of

diagnosing head and neck cancers. García told The Column: “Previously, I had been working on the detection of volatile organic

compounds (VOCs) for environmental applications, and the chance to detect these organic compounds from the exhaled air of

patients suffering epidermoid laryngeal carcinomas, as non-invasive biomarkers to be used in early detection of this pathology,

was an outstanding challenge.”

A group of 31 volunteers were recruited for the study: 20 healthy volunteers, and 11 patients diagnosed with cancer of the

larynx undergoing treatment at Alcorcón Hospital. Samples were collected from patients exhaling

air after an 8 hr fast using gas sampling bags. SPME was performed to concentrate

compounds within the exhaled air, before analysis using GC–MS.

The method was able to detect seven potential biomarkers that were at different

concentrations between healthy and diagnosed patients. However, only two were

considered to be signif cantly higher in late-stage cancer patients (T3) when

compared with healthy or early-stage patients (T1) — ethanol and 2-butanone.

There is a note of caution from García who said: “The importance of the

work is quite relative and it is important to be cautious, since the evaluated

subjects are not statistically signif cant enough to extract def nitive conclusions,

and also because the more signif cant VOCs were detected in the later stages

of the disease. However, the importance of the investigation is the concept,

not only for this serious pathology but also from other neck and head

prevalent diseases. I am sure that the results are widely improvable,

and it will be improved.”— B.D.

Reference1. R. García, V. Morales, and Sergio Martín, Chromatographia 77, 501–509 (2014).

Peak Scientif c Win Queen’s Award for Enterprise: International Trade 2014

Larynx Cancer Biomarkers

7




Investigating the Origins of MetabolismWhat is the origin of life? Did organisms become metabolically active as they evolved or were these

reaction chains already in existence?

Reference

1. M.A. Keller, A.V. Turchyn, and M. Ralser,

Molecular Systems Biology DOI 10.1002/

msb.20145228 (2014).

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Bethany Degg, Assistant Editor, [email protected]

of energy, lipids, proteins, and more. The

reactions involved within the metabolism

network are largely conserved across all

organisms, suggesting a single origin from

which the network has evolved. Core

metabolism has been thought to have come

after the formation of proteins as enzymes are

key to many metabolic reactions.

The team reconstructed conditions of the

Archean Ocean (4 billion years ago) based

on the sediment composition proposed by

the literature. Metabolites key to central

metabolism (including glucose-6-phosphate

[G6P], fructose-6-phosphate [F6P], and fructose

1,6-bisphosphate [F16BP]) were incubated at

50–90 ºC in water, the temperature of water

around the hydrothermal vents of oceanic

volcanoes. The resulting compounds were

analyzed using LC–QTOF-MS.

The team was able to observe almost 29

spontaneous reactions, including the formation

and interconversion of glucose, pyruvate, the

amino acid precursor erythrose-4-phophate,

and the nucleic acid precursor ribose-5-

phosphate. By using single reaction monitoring

mode for the analysis, the team were able

to determine absolute quantities for 15

intermediates.

Markus Ralser of the University of

Cambridge and the National Institute for

Medical Research said: “Our results show

that reaction sequences that resemble two

essential reaction cascades of metabolism,

glycolysis and the pentose-phosphate

pathways, could have occurred spontaneously

in the earth’s ancient oceans.” He added:

“In our reconstructed version of the ancient

Archean ocean, these metabolic reactions

were particularly sensitive to the presence

of ferrous iron, which was abundant in the

early oceans, and accelerated many of the

chemical reactions that we observe. We were

surprised by how specif c these reactions

were.”

The paper demonstrates the

possibility that the iron-rich oceans

could have been suff cient to catalyze

the beginning of core metabolism,

without the presence of enzymes. It also

suggests that RNA molecules could have

been formed from a basic mix of sugar

metabolites. However, the origin of the

sugar metabolites is still in question and the

chance that a small solution of metabolites in

an ocean could trigger the beginnings of life

is questionable. — B.D.

Scientists at the University of Cambridge

(Cambridge, UK) have reconstructed the early

conditions of the Earth’s oceans to f nd that

spontaneous chemical reactions could have

generated the f rst biological molecules, before

the evolution of organisms or the existence of

enzymes.1 The study published in the journal

Molecular Systems Biology presents data

collected from liquid chromatography–triple

quadrupole mass spectrometry (LC–QTOF-MS)

suggesting that reactions central to our

core metabolism could have spontaneously

occurred.

Life in its primitive form is widely thought

to have begun around 4 billion years ago on

Earth. At this time, the oceans are believed

to have been iron-rich. As to how it began,

there are a number of theories. One of the

most prominent and well-known is that of

Miller and Urey in 1953 who demonstrated

that amino acids could be created by applying

an electrical charge to a “primordial soup” of

hydrogen, methane, water, and ammonia. The

hypothesis that the amino acids spontaneously

formed could be lthe basis of enzymatic

proteins required for metabolism.

Core metabolic processes are essential to

life, and are responsible for the production

News

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Platforms: iPad and iPhone (iOS 4.2 or later, 6.9 MB), Android (v2.1 and up, 336 k).Features: This app published by Sigma Aldrich calculates conditions for transfer of an isocratic or gradient method from one HPLC column to another; recommends f ow rates for analytical columns; allows method scaling from microbore through preparative column range based on two sets of column variables and current method conditions; and predicts the gradient delay that must be applied when transferring a gradient method, based on the user-entered value of dwell volume. Cost: Free

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10


3D Printing — A Coming Revolution?

I hate being in that intellectual hinterland

where I am on the fringe of conversations

without knowing enough to contribute in

any meaningful way. As a relatively “early

adopter” on the technology curve, I always

liked to keep up-to-date with the latest

advances, without straying into “geekdom”

too far! On a recent trip to the Science

Museum in London, UK, I felt very much

in the hinterland as I toured an excellent

exhibition on 3D printing. Working printers,

the varied products of these printers, and

some very stimulating pieces on what 3D

printing may mean for us in the future

were all on display.

In an attempt to get ahead of the curve,

I’ve read up a little and spoken to a few

very early adopters of the technology

about their experiences. This includes

my own son: His school’s technology

department has had one for absolutely

ages and in his words “Oh my goodness

Dad, where on earth have you been!” The

basics of 3D printing using the “additive

manufacturing” approach are reasonably

straightforward. In fused deposition

modelling (FDM), a thermoplastic

filament (wire) of poly lactic acid (PLA)

or acrylonitrile butadiene styrene (ABS) is

fed into a heated print head that extrudes

the molten plastic and deposits layers

or dots, according to a 3D printable

stereolithography (STL) file, which builds

up the object in layers from the fast

thermosetting plastic. Once completed,

the piece has any sprues (frames) removed

and is filed smooth and finished or is

finished by a higher resolution printing

device using a subtractive (ablation or

similar) process. The resolution of the

printer and the number of different print

heads and feedstock materials it can

handle are linked to cost. Lower resolution

printers are available for under $500 while

high-resolution, large format printers using

multiple feedstocks can cost hundreds

of thousands of dollars. Other printing

processes such as granular materials

binding (GMB) and print feedstock

materials are available, and all of the

variations can of course be studied in more

detail from the excellent Wikipedia article.1

The applications of the technology rather

than the technology itself are the most

fascinating. On display at the exhibition

at the Science Museum were running Ph

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Incognito speculates on how 3D printing could change life in the laboratory.



shoes, models of organs, foetuses, and

brains, prosthetics, “selfies” of loved ones

and pets, and a working hand-cranked

version of a ¼-scale rotary aeroplane

engine — all printed in a single session

with no further assembly required. More

serious and beneficial applications are

seen in bio-printing where living cells are

deposited onto a gel medium to build up

3D structures that can include intricate

vascular systems that have already been

used to research the potential manufacture

of trachea, major blood vessels, and liver

and kidney tissue. Prosthetic applications

have included pelvic reconstructions from

titanium, jaw bones, facial implants, and

long bones and joints. A nice summary on

what’s hot in 3D printing (no pun intended)

can be found in a recent article in The

Guardian.2

Given the nature of this column, we

should really think about where the

technology might fit into the analytical

laboratory, but I do feel that a discussion

on plastic egg flippers is needed first. I

recently argued with a colleague when

they suggested that soon every home will

have a 3D printer as it would be really cool

to simply print out a new egg flipper when

your previous one breaks. No, clearly you

would travel to your nearest kitchenware

shop or go on-line and buy one — it’s

not that you absolutely have to have an

egg flipper. You can use pretty much

anything else for flipping eggs in a pan.

My contention is that 3D printers will be

used to produce items which are highly

personal (prosthetics/orthotics/”selfies”), or

when you absolutely need that item right

now and can’t get it from anywhere else

without major inconvenience or a

long wait.

Where does this leave the use of 3D

printing in the laboratory then? I’ve

thought of a few applications, but I’d love

to hear more from our readers.

• Piston seals for high performance liquid

chromatography (HPLC) pump heads —

Your consumables cupboard is fresh out

of them and you absolutely need to put

that campaign of samples on overnight.

By the way, for pump seal read (perhaps)

pistons or check valves — although this

would require us printing some pretty

special plastics, which will resist wear and

pH extremes and have certain mechanical

properties.

• Printing metal alloys is possible using a

number of techniques, including selective

laser sintering (SLS), but I can’t see this

coming to a laboratory near me soon

unless costs can be significantly reduced.

What would you use to construct the

ruby ball in the check valve?

Incognito

11


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Professor Lee Cronin at Glasgow University

(Glasgow, UK) leads a group who are

producing bespoke micro-reactor units, and

using the technology to test out new flow

through synthetic routes: A little out of

our area perhaps but could the technology

be used to print out new “chip” designs

for HPLC or gas chromatography (GC)

systems to which other simple components

are added? Or perhaps new systems

could be designed into which the printed

chips are integrated?3 Perhaps capillary

electrophoresis (CE) system cartridges

could be designed and printed — again to

“plug and play” into integrated systems.

There is once again the issue of “printing”

chemistry (stationary phases) onto the

inside of these devices, which is not

possible at the moment. That doesn’t

mean we could not come up with a way of

functionalizing plastic or ceramic feedstock

materials in the future. Perhaps plastic

support materials for chromatography may

have their day after all?

My feeling is that early applications of

3D printing will result in similar benefits

to that offered by ultrahigh-pressure

liquid chromatography (UHPLC) or FAST

GC, in that one can significantly speed up

development tasks. I’m now developing

chromatography methods overnight

(previously impossible) thanks to the speed

of my chromatography equipment and

columns. Perhaps 3D printing will make it

possible for us to adapt or develop new

instrumentation in the same way and I can

develop and print a fully integrated HPLC

system which has dead volume low enough

to really take advantage of small internal

diameter HPLC columns packed with sub

1-μm particles — but I guess that’s another

installment altogether!

If you haven’t yet had the opportunity

to contemplate how 3D printing will affect

your lives, both personal and working,

I would urge you to do so now. The

revolution is coming, and there will be no

stopping it.

References

1. 3D Printing, Wikipedia: bit.ly/19C48fJ

2. 30 things being 3D printed right now (and

none of them are guns), The Guardian: bit.ly/

L8AR2p

3. P.J. Kitson, M.H. Rosnes, V. Sans, V. Dragone, and

L. Cronin, Lab Chip 12, 3267–3271 (2012).

Contact author : IncognitoE-mail : [email protected]

Incognito

12


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Evaluating the Temperature Shift in Analytical Temperature Rising Elution Fractionation

This article presents a new method to evaluate the temperature shift observed in analytical temperature rising elution fractionation

(ATREF). The evaluation is based on the dependence of the measured peak temperature as a function of heating rates. Application of the

proposed method does not require any knowledge of the f uid circuit characteristics geometry and avoids the use of narrow preparative

TREF standards. The results are found to be more accurate than the method that is usually applied.

Adrian Boborodea and André Luciani, Certech ASBL, Seneffe, Belgium.

Since the publication of the Wild article in

1982,1 analytical temperature rising elution

fractionation (ATREF) has been a method

of choice for polyolef n characterization. In

1991 Exxon introduced the f rst patents in

which polyethylenes were described by their

compositional distribution breadth index

(CDBI), an ATREF parameter equivalent to

the polydispersity index of the molecular

weight distribution. Exxon was followed

by other material producers like Dow

and Cryovac, and the number of patents

incorporating claims covered by ATREF

analysis steadily increased to about 30

patents per year. At the beginning, the

ATREF systems were built by modifying

gel permeation chromatography (GPC)

systems. The increasing demand for ATREF

measurements fostered the introduction of

the f rst automated commercial ATREF system

by Polymer Char in 1996. No standardized

method was published for this technique,

and most of the patents make reference to

Wild’s publication.1

To calculate the CDBI, several standards

of polyethylene with narrow compositions

and known short-chain branching

characteristics are injected into the ATREF

unit and chromatograms are recorded as

a function of time while monitoring the

column temperature. These data are used

to construct thermograms, in which the

detector signal is plotted as a function of the

column temperature. The recorded column

temperature is usually higher than the real

melting temperature of the eluted fraction

because of the delay between melting in the

column and detection. This delay depends on

several experimental parameters, including

column dimensions, tubing characteristics

between the column and detector,

and heating and elution rates. These

equipment-specif c parameters prevent the

use of any short chain branches (SCB)-elution

temperature calibration curves available in

the literature.1–3 Actually, as recommended

by the commercial ATREF manufacturer,4 the

use of an identical procedure and identical

equipment is mandatory to be able to

compare samples.

The procedure would be greatly simplif ed

with a robust method to calculate the

temperature shift, allowing for the correlation

between results obtained with different

instruments and methods.

To circumvent that problem, the

temperature shift can be calculated based

on the tubing volume between the column

and the detector. As the heating rate and

the f ow rate during the heating step are

usually constants, the tubing volume can be

converted into temperature shift using the

following relationship:

temperature shift (°C) = heating rate

(°C/min) × tubing volume (mL)/

f ow rate (mL/min) [1]

As this method assumes a detailed knowledge

of the tubing and does not take into

consideration the heat transfer delay in the

column heating system, it is expected that the

calculated temperature shift will be lower than

the actual one observed.

An alternative method to evaluate the

temperature shift is to analyze narrow

preparative TREF standards.5,6 In this

case, the temperature shift is given by the

difference between the measured ATREF

peak temperature and the preparative TREF

temperature of the standard; the diff culty

of the method is the fact that narrow TREF

standards, isothermally eluted in a temperature

interval of about 3 °C, are not easily available.

In addition, the high preparative column

13




volume introduces measurement errors close

to the temperature interval used to elute the

preparative fraction (3 °C). Another potential

issue with this method is the fact that usually

the analytical TREF solvent (trichlorobenzene) is

different from the solvent used in preparative

TREF (xylene).

In this article, we present a new method to

evaluate the temperature shift for ATREF that

does not require either the detailed knowledge

of the tubing between the column and the

detector or the availability of narrow standards

obtained by preparative TREF. To apply the

method, it is possible to use any unimodal

sample such as a commercial metallocene

polyethylene or high-density polyethylene.

The method is based on the idea that the

temperature shift is zero for isothermal elution

steps. Because f nite heating rates are used

with ATREF, we propose to f nd the real

melting temperature by analyzing the same

sample with different heating rates and to

extrapolate the measured elution temperatures

to isothermal conditions. After f nding the

real melting temperature by extrapolation, we

can then use a general rule to calculate the

temperature shift for a given heating rate.

Experimental

ATREF Apparatus: The analyses were done

using a TREF-Crystaf model 300 instrument

(Polymer Char). The TREF unit is integrated

into an Agilent Technologies model 7890

GC system oven, which is kept as originally

supplied by the manufacturer with the

exception that the main switch also activates

the TREF dedicated power supply and all other

electronics. TREF control software is used to

set the oven temperature programme through

one of the computer serial ports. The elution

step was performed with an Intelligent Pump

Series 300 (IP300, Flom Corporation), located

close to the oven and controlled by the TREF

software. The concentrations of the eluted

polymer fractions were measured with the

built-in infrared detector, using wavelengths

between 2800 and 3000 cm-1.

Samples: Four commercial metallocene

polyethylenes were analyzed using the ATREF

system. The selected samples have narrow

short-chain branching distributions and the

following densities: 0.923, 0.934, 0.947, and

0.955 g/cm3.

ATREF Methods: Each sample was dissolved

in 1,2,4-trichlorobenzene (TCB Spectropure

dry, Biosolve Chimie, CAS 120-82-1) at

0.12

IR a

bso

rba

nce

0.10

0.08

0.06

0.04

0.02

Time (min)

0.00

0 100 200 300

0.2 °C/min

0.5 °C/min

1.0 °C/min

2.0 °C/min

400 500 600-0.02

Figure 1: Overlaid chromatograms (infrared detector signal as a function of time) recorded using different heating rates for the 0.947-g/cm3 polyethylene.

Boborodea and Luciani

14


Abstract submission deadlines:

Oral Presentations July 15, 2014

Poster Presentations August 15, 2014

CALL FOR PAPERS

SFC 2014 CONFERENCE

October 8 - 10

Basel, Switzerland



150 °C to obtain a 2 mg/mL solution.

The solvent was stabilized with 0.3% of

antioxidant (butylated hydroxytoluene, BHT,

CAS 128-37-0). About 300 µL of the hot

solution was injected at 150 °C in the ATREF

column (100 mm × 0.25 in.) f lled with

metallic shots provided by Polymer Char. The

column f lled with solution was fast cooled

(1 °C/min) to 100 °C, and then was further

cooled to 30 °C with a cooling rate of 0.1 °C/

min. After the cooling step, the polymer was

eluted from the column, using a f ow rate of

0.5 mL/min. Before starting the heating to

130 °C, the column was maintained at 30 °C

for 30 min. For each sample, four different

heating rates were used: 0.2, 0.5, 1.0, and

2.0 °C/min.

Results and Discussion

Figure 1 is an overlay of the four elution

prof les (trefograms) recorded for the 0.947-g/

cm3 density polyethylene. The total elution

step increases from 80 min for a heating rate

of 2 °C/min to 530 min for a heating rate of

0.2 °C/min. Trefograms of the other three

standards showed similar behaviour.

Figure 2 shows the same elution prof les

as a function of the recorded temperature

during the elution step. The overlay of

the four thermograms is representative of

the results. One can note that the peak

temperature and the peak width increase

with the heating rate.

The measured temperatures for each

experiment were automatically corrected by

the commercial TREF software based on the

tubing lines between the TREF column and

detector cell, and the elution rate. The results

are given in Table 1. The differences between

the peak temperatures obtained with 0.2 and

2.0 °C/min are about 3 °C.

In Figure 3 the measured peak temperatures

are plotted as a function of the heating rates

for the different polyethylenes used in this

work. The extrapolated elution temperatures

at zero heating rates represent the real melting

temperatures. The obtained parallel lines of the

linear dependence of the elution temperatures

vs. heating rates for the different densities

implies the existence of a unique parameter to

110

105

100

95

90

85

80

0.0 0.5 1.0

Heating rate (¡C/min)

Measu

red

elu

tio

n t

em

pera

ture

(¡C

)

1.5 2.0 2.5

0.955

0.947

0.934

0.923

Figure 3: Plot of measured peak temperatures as a function of heating rates for four metallocene polyethylenes.

0.12

IR a

bso

rba

nce

0.10

0.08

0.06

0.04

0.02

0.00

-0.0280 90 100

Temperature (°C)

110 120 130

0.2 °C/min

0.5 °C/min

1.0 °C/min

2.0 °C/min

Figure 2: Overlaid thermograms (infrared detector signal as a function of recorded temperature) for the 0.947-g/cm3 polyethylene.


15




calculate the temperature shifts. For a f ow rate

of 0.5 mL/min, the observed average slope of

5.1 min corresponds to the time needed for

a polymer fraction to be melted and travel to

the detector. This time delay takes into account

the tubing volume as well as other parameters

such as the heat transfer between the oven

and the column, and the necessary time to

solubilize the melted polymer.

With this parameter, the temperature shift

can be calculated for any heating rate with

the following relationship:

temperature shift (°C) = heating rate

(°C/min) × time delay (min) [2]

With this equation, the measured

temperatures for the 16 ATREF experiments

were corrected; the results are presented in

Table 2. A maximum difference of 0.4 °C

was noted, which is about 10% of the error

recorded with the previous method. This

difference is close to the variability of the

ATREF method when determining the peak

temperature by repeated measurements (for

six measurements using a heating rate of 1

°C/min done on HDPE NIST 1475, the average

uncorrected elution temperature is 101.3 °C,

and the standard deviation is 0.2 °C).

Conclusion

A new and simple method is proposed

that allows for evaluating the temperature

shift in analytical temperature rising elution

fractionation (ATREF). This technique is

based on the variation of the measured

peak temperature as a function of the

heating rate during the elution step and

does not require the detailed knowledge of

the instrument tubing or the availability of

standards obtained by preparative TREF. To

apply the method, it is possible to use any

E-mail: [email protected]: www.certech.be

polyolef n sample, as long as it has a narrow

composition distribution.

Apart from measuring the correct melting

temperature with higher precision, the

method opens the possibility of creating

“universal” calibration curves (SCB vs.

temperature) for each type of comonomer.

Such a procedure would improve the

interlaboratory reproducibility for the

calculation of CDBI and other parameters

characterizing the chemical composition

distribution of different types of polyethylenes.

The method was initially developed on a

commercial ATREF system, but should be

valid with in-house ATREF instruments with

different conf gurations.

References

1. L. Wild, T.R. Ryle, D.C. Knobeloch, and I.R. Peat,

J. Polym. Sci.: Polym. Phys. Ed. 20,

441–455 (1982).

2. F.M. Mirabella, J. Polym. Sci: Part B: Polymer

Physics 39, 2819–2832 (2001).

3. F. Chen, R.A. Shanks, and G. Amarasinghe,

Polymer 42, 4579–4587 (2001).

4. Polymer Char TREF User Manual, pages 2–17

(2011).

5. A.G. Boborodea, D. Daoust, A. Jonas, and C.

Bailly, LCGC North Amer. 22(1), 52–57 (2004).

6. US 7389678, A.G. Boborodea, C. Bailly, D.

Daoust, A. Jonas, and B. Nysten, Column

for analytical temperature rising elution

fractionation (ATREF).

Adrian Boborodea and André Luciani

are with Certech ASBL in Seneffe,

Belgium.

Table 2: Corrected temperatures for all samples, calculated using the average slope of the linear dependence between the measured elution temperatures vs. heating rates

Heating rate

(°C/min)

Peak temperature for

0.923 g/cm3 (°C)


0.934 g/cm3 (°C)


0.947 g/cm3 (°C)


0.955 g/cm3 (°C)

0.2 84.2 90.1 94.6 96.4

0.5 84.0 90.2 94.7 96.5

1.0 84.0 90.2 94.6 96.4

2.0 84.0 89.8 94.8 96.5

Table 1: Automatically corrected temperatures for all experiments, calculated by TREF software based on the tubing lines between the TREF column and detector cell

Heating rate

(°C/min)


0.923 g/cm3 (°C)


0.934 g/cm3 (°C)


0.947 g/cm3 (°C)


0.955 g/cm3 (°C)

0.2 84.6 90.4 95.0 96.8

0.5 84.9 91.0 95.6 97.4

1.0 85.8 91.9 96.4 98.2

2.0 87.6 93.4 98.4 100.1


16



The 2014 LCGC Awards (Part 2)

The seventh annual LCGC Awards continue the time-honoured tradition of celebrating the careers of outstanding chromatographers.

We are proud to announce that the 2014 Emerging Leader in Chromatography Award is presented to André de Villiers. This is the

second of a two-part feature in The Column charting the career and accomplishments of the awardees.

Megan L’Heureux, Managing Editor, LCGC North America

LCGC’s 2014 Emerging Leader in

Chromatography award winner, André de

Villiers, received his Bachelor of Science

degree in chemistry and biochemistry (1997),

his Honours Bachelor of Science degree in

chemistry (cum laude, 1998), his Masters

of Science degree in analytical chemistry

(cum laude, 2000), and his doctoral

degree in analytical chemistry (2004) from

Stellenbosch University in South Africa. De

Villiers says his interest in analytical science

began with the start of his postgraduate

studies. In 1999 he was very unsure of his

future plans, but decided to meet with two

professors, Henk Lauer and Pat Sandra. At

that meeting, de Villiers decided to pursue

analytical chemistry. “Looking back now, it

seems a highly fortuitous conglomeration of

circumstances that made this possible,” said

de Villiers. “Essentially, my career path was

determined by a 30-minute discussion with

Pat Sandra and Henk Lauer.”

From the point of view of Lauer, who is

currently the managing director of HLCE

and was one of de Villiers’s supervisors of

his masters and PhD theses, the timing of

de Villiers’s decision was perfect, given the

changes that were going on at Stellenbosch

University at the time. Ben Burger, a professor

and the director of the Laboratory for

Ecological Chemistry (LECUS), had decided

that Stellenbosch University needed a

chemistry department with a focus on

separation science, so he enlisted Pat Sandra,

who was then at the University of Ghent

and also a director of his own Research

Institute of Chromatography (RIC), to set

up such a department. According to Lauer,

Sandra made sure that a strong programme

was established, and he imported a lot of

instrumentation from Europe and helped

secure funding for the new department. “De

Villiers brought his talent at the right time

and the right place,” said Lauer. “He showed

André de Villiers

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his talent with excellent study results, a great

understanding of the analytical problems, and

use of the instrumentation available then.”

De Villiers did his postdoctoral studies at

the Pfi zer Analytical Research Centre (PARC)

at Ghent University in Belgium, from 2004

to 2006. From there he decided to return

to Stellenbosch University as a lecturer in

chemistry, a position he held from August

2006 to July 2008. In August 2008, de

Villiers was promoted to senior lecturer of

chemistry, and he remained in that position

until December 2012. In January 2013,

he was promoted to associate professor

of chemistry and continues to hold that

position today.

Contributions to the Field: De Villiers’s

research interests include fundamental

studies that push the boundaries

of the chemical characterization of

complex mixtures using state-of-the-art

techniques such as multidimensional

liquid chromatography (LC) and gas

chromatography (GC) combined with mass

spectrometry (MS), and the application of

these methods, primarily to natural product

analysis. He has published 50 papers in

peer-reviewed journals, and his papers

have been cited 925 times — quite an

accomplishment for such a young scientist!

Sandra, who is now an emeritus

professor with the Research Institute for

Chromatography and was de Villiers’s

professor and thesis supervisor at

Stellenbosch, feels that de Villiers’s greatest

contribution to the fi eld of separation

science so far is his work developing new

LC methods and techniques, including

comprehensive LC×LC for the analysis of

Figure 1: Pat Sandra with his former coworkers in April 2008 in Stellenbosch, South Africa. From left to right: Pat Sandra, Frédéric Lynen, Andreas Tredoux, André de Villiers, Deirdre Cabooter, and Martina Sandra.

Figure 2: De Villiers at Cape Point, South Africa.

LCGC Awards 2014

18


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Bioanalysis: LC–MS-MS, Sample Prep, and Dried Blood Spot Analysis Bioanalysis uses a variety of separation techniques to analyze samples ranging from plasma and urine to dried blood spots. Participants in this Technology Forum are Ling Bei, Patrik Appelblad, and Dave Lentz of EMD Millipore; Nadine Boudreau of PharmaNet Canada; Diab Elmashni, Jeff Zonderman, and Simon Szwandt of Thermo Fisher Scientific; and Debadeep Bhattacharya of Waters Corporation. More...

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Health Sciences Unit LaunchedLGC has launched a new business unit, Health Sciences, which combines the group's sport, food, consumer safety and pharmaceutical testing activities within a single entity. More...

KNAUER is now offering its PLATINblue UHPLC systems with the MSQ Plus mass detector in a special package deal. PLATINblue systems and the MSQ Plus are an ideal combination for high-throughput applications. For a limited-time only, the UHPLC-MS package is being offered at a very special price. Don’t miss out!

Performance Materials Supplier AcquiredAmerican-based Avantor Performance Materials will acquire Polish Performance Materials Supplier (POCH S.A.). More...

Your brilliance. Our know-how. Collaborative Life Science. It all joins forces at EMD Millipore. Now your organization can leverage the combined synergies of two leading Life Science companies – for deep insight and know-how along every step of the biotherapeutic value chain. Find out how at www.emdmillipore.com

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Using LC–MS with Online Sample Preparation to Survey Metabolites Formed In VitrolAn Interview with Samuel Yang, University of Texas at Arlington. More...

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Featured Application Note:Screening and Quantification of Multiple Drugs in Urine Using Automated Online Sample Preparation

and Tandem Mass Spectrometry Barbora Brazdova and Marta Kozak, Thermo Fisher Scientific

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Evaluation of the Ultra Inert Liner Deactivation for Active Compounds Analysis by GCLimian Zhao, David Mao, and Allen Vickers, Agilent Technologies

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Screening and Identification with High Confidence Based on High Resolution and Accurate Mass LC–MS-MSAndre Schreiber and David Cox, AB SciexThis note describes a workflow and tools to identify targeted and nontargeted pesticides in fruits and vegetables. High resolution, accurate mass LC–MS-MS data is mined using advanced

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Highly Sensitive UV Analysis with the Agilent 1290 Infinity LC System for Fast and Reliable Cleaning Validation – Part 1Edgar Naegele and Katja Kornetzky, Agilent TechnologiesThis application note demonstrates high sensitivity measurement of pharmaceutical compounds

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Which GPC Column First? Bruce Kempf, Tosoh BioscienceMost manufacturers recommend the installation of SEC columns in order of decreasing pore size when running columns in series. Scientists at Tosoh Bioscience tested the validity of this recommendation.

Ultra-Fast Analytical Method for the Sample Cleanup and LC–MS-MS Analysis of

Chloramphenicol in Shrimp and Other Marine Food ProductsPhilip J. Koerner, Matthew Trass, Liming Peng, and Jeff Layne, PhenomenexA method for the analysis of chloramphenicol in shrimp has been developed with a limit of quantitation (LOQ) of 0.001 ng/g in shrimp (0.001 ppb) based on the calibration standards. This is 300 times lower than the current U. S. Food and Drug Administration (USFDA) method. The method described uses Strata-X solid phase extraction (SPE) cartridges for sample cleanup and

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Topics and categories include: HPLC • GC • Sample Prep • LC-MS and GC-MS • Emerging techniques

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ES438482_LCTC052214_V18.pgs 05.14.2014 23:57 ADV blackyellowmagentacyan


natural products, such as South African

wines.

Lauer agrees, citing “his endeavour to

understand and nail down the complexity of

molecules that def ne the colour, taste, and

bitterness of South African wines with all the

available separation techniques he could lay

his hands on”.

Tadeusz Górecki, a professor at the

University of Waterloo in Ontario, Canada,

said he values de Villiers’s contributions

to the area of multidimensional LC “from

his early work with Isabelle François to his

recent foray into hydrophilic interaction

reversed-phase LC×LC”. Górecki also

mentioned that de Villiers’s more theoretical

work, such as his papers on kinetic

optimization of LC separations, are also of

high quality.

Emily Hilder, a professor of chemistry and

director of the ARC Training Centre

for Portable Analytical Separation

Technologies at the University of Tasmania

(as well as the 2012 LCGC Emerging Leader

in Chromatography award winner), agreed

with Górecki’s thoughts on de Villiers’s

contributions in multidimensional LC. “His

work has demonstrated how 2D LC can

be applied to the analysis of very complex

samples from natural products (wine,

food, and so on),” said Hilder. “Such

practical applications of this technology

are what is needed to guide future

developments.”

Scientif c Accolades: De Villiers has

received a number of awards from the

separation science community, including

the 2009 Csaba Horváth Memorial Award

from the International Symposium on

High-Performance Liquid Phase Separations

and Related Techniques (HPLC) and the

2012 Chromatographer of the Year award

from the Chromatographic Society of

South Africa. He has also been invited to

deliver lectures at prestigious international

conferences, such as HPLC and the

International Symposium on Hyphenated

Techniques in Chromatography.

De Villiers is currently chairing the Western

Cape board of the Chromatographic Society

of South Africa. He was also responsible

for the organization of two successful

conferences that took place in Stellenbosch:

the 39th National South African Chemical

Institute convention in 2008 and Analitika

2010.

Future Contributions: Given what de

Villiers has already accomplished in his career,

we asked several of his mentors and peers

where they thought his work might take him.

Sandra expects de Villiers to make

contributions to the fundamental

understanding of chromatographic processes,

because he has a very strong theoretical

LCGC Awards 2014

19


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roof of his car,” said Górecki. “This earned

him the nickname ‘Professor Dude’ among

his students.”

Górecki shared a story about de Villiers’s

participating in a grueling bike race around

Cape Peninsula that is more than 100 km

long and has numerous climbs and strong

winds. Several years ago de Villiers crashed

and injured himself rather badly, so he could

not complete the race. It took several hours

for an ambulance to reach him. “De Villiers

declared that he would never do that race

again and stopped biking entirely. After a

few months, though, he registered for the

next year’s race and started to train again,”

said Górecki. “This dedication is what drives

his career as well.”

Lynen paraphrased one of de Villiers’s

favorite quotes: Something worth doing

is worth doing well. “I always found this

phrase very characteristic of his personality,”

said Lynen. He also mentioned a research

problem that de Villiers worked on in 2003

to address peak distortion problems when

drawing the calibration lines of organic

acids in capillary electrophoresis. Lynen

explained that the problem looked very

strange and diff cult to solve, but de Villiers

was able to correctly deduce that the

increasing concentrations of each organic

acid calibrant were effectively lowering the

pH of the migrating zone and, as a result,

creating an electrodispersion phenomenon

that could be f xed by adjusting the sample

pH.1 “The meticulous approach with

which he addressed that problem (also by

studying a lot of literature on the topic)

was very impressive and demonstrated the

eye for detail that is characteristic of a true

scientist,” said Lynen.

More About the Winners

In-depth interviews with Fred E. Regnier and

André de Villiers, focused on their research,

challenges, and accomplishments will be

published in upcoming editions of the LCGC

North America newsletter, E-Separations

Solutions.

Reference

1. A. de Villiers, F. Lynen, A. Crouch, and P.

Sandra, Eur. Food Res. Technol. 217(6),

535–540 (2003).

Megan L’Heureux is the managing editor

of LCGC North America.

E-mail: [email protected]: www.chromatographyonline.com

background. He also thinks de Villiers

will play an important role in education.

“He will def nitely have a great impact

in the education of students in Africa

on state-of-the-art analytical techniques

and, more specif cally, chromatography

and electrophoresis combined with

high-resolution mass spectrometry,” said

Sandra.

Górecki feels de Villiers will continue

the legacy of other great South African

separation scientists, like Victor Pretorius or

Ben Burger. “He has already made his mark

on separation science, and the trajectory

from here can only be up, especially knowing

de Villiers’s talent and work ethic,” Górecki

said.

Barend (Ben) V. Burger, an emeritus

professor from Stellenbosch University,

thinks that de Villiers might branch into GC,

even though that is not his primary area of

research. “I think there is still much scope

for the development of more affordable

two-dimensional instrumentation,” said

Burger.

Frédéric Lynen, an associate professor

at Ghent University, said de Villiers’s

research would continue in natural product

analysis with the “discovery of new, thus

far, biologically active compounds via the

combination of high-end chromatography

and the elucidation of structures of

unknown natural solutes”.

A Testament to De Villiers’s Character:

Other scientists describe de Villiers as

friendly and down to earth. Hilder recalls

f rst meeting him at an HPLC conference

in 2006, and said their friendship has

grown since then. “The separation science

community is very supportive and there is

now a good group of young people all at a

similar stage in their careers,” she said. “We

catch up at meetings, and this makes for

a great, fun support network of scientists

for bouncing off ideas and sharing advice.”

Hilder also said that she shares a love of

cricket with de Villiers.

De Villiers is indeed a big sports enthusiast

— not just as a fan, according to Górecki. De

Villiers regularly plays pickup football games,

and is an avid biker and an aspiring surfer.

“He often comes to the university in the

summer with a surfboard attached to the

Table 1: Winners of the LCGC Awards.

Year Emerging Leader

2008 Gert Desmet

2009 Kevin Schug

2010 Jared Anderson

2011 Dwight Stoll

2012 Emily Hilder

2013 Davy Guillarme

2014 André de Villiers

LCGC Awards 2014

20



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Hands-on GC Theory and Methods9 June 2014

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14–18 September 201430th International Symposium on Chromatography (ISC 2014)

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Chairpersons: Wolfgang Buchberger, Michael Laemmerhofer, and Wolfgang Lindner

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21–24 September 2014 5th International Conference on Polyolefin Characterization (ICPC)

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EDITORS’ SERIES

Contaminant Characterization in Drinking Water:

Green Methodologies

EVENT OVERVIEW:

In this web seminar, several aspects of sample prepara-

tion as well as analytical techniques including GC–MS,

GC with comprehensive MS, and LC–MS will be presented

for several categories of organic contaminants in water,

including:

n Disinfection byproducts

n Emerging disinfection byproducts

n Odorous compounds and precursors

n Hormones and pharmaceuticals

n Pesticides and their byproducts

This discussion will place special emphasis on green meth-

ods for analysis of these compounds.

Key Learning Objectives:

n Green methods for sample preparation and LC-MS and GC-MS analysis of organic compounds in drinking water.

n How to use these methods for the analysis of many categories of contaminants, such as disinfection byproducts, hormones and pharmaceuticals, and pesticides and their byproducts.

Who Should Attend:

n Environmental scientists, regulators, and scientists in the water industry interested in learning about advanced methods for the analysis of current and emerging contaminants in drinking water

n Analysts interested in adopting greener analytical methods for analyzing water quality

Presented by Sponsored by

O N - D E M A N D W E B C A S T :

Register free at

www.chromatographyonline.com/Water_Analysis_2

For questions, contact Kristen Moore at [email protected]

Presenters:

David BenanouExpert in Research Analytical Chemistry, Environment & Health Department, Veolia Environment Research & Innovation Maisons Laftte, France

Moderator:

Laura BushEditorial Director, LCGC


22 may 2014 volume 10 issue 9 cover story...

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