19 January 2023 Volume 19 Issue 1

Cover Story
2 Comparing the Flavour Profiles of Soft Drink
Brands Using Immersive Sorptive Extraction,
GC×GC–TOF-MS, and Chemometrics
L aura McGregor, James Ogden, and Elinor Hughes, SepSolve Analytical
Immersive
 sorptive extraction coupled with GC×GC–TOF-MS
was used to compare flavour profiles between popular
brand soft drinks and imitation products.

Features
13 Rising Stars of Separation Science: Martina Catani
This
 month we interview Martina Catani from the University of Ferrara, Italy,
about her work on the separation of enantiomers by HPLC/UHPLC and the
study of the kinetic and thermodynamic properties of chiral stationary phases,
and the separation and purification using LC analysis (single-column and
continuous multi-column) of biomolecules of high therapeutic interest.
17 Playing with Selectivity for Optimal Chiral Separation
Ramkumar Dhandapani and Bryan Tackett, Phenomenex
To optimize challenging separations, it is important to explore all parameters
through a chiral screening process across various chiral column chemistries.
21 A Focus on Forensic Taphonomy and Chromatography
The Column interviewed Maiken Ueland, an ARC discovery early career

research fellow at the Centre for Forensic Science and deputy director of
The Australian Facility for Taphonomic Experimental Research (AFTER) at
the University of Technology Sydney in Australia, on her work in forensic
taphonomy, where she uses analytical, biochemical, and spectroscopic
techniques to conduct human post-mortem investigations.
25 SCM-10 Event Preview
Peter Schoenmakers, University of Amsterdam
The
 Tenth International Symposium on the Separation and Characterization of
Natural and Synthetic Macromolecules, SCM-10, will take place 1–3 February
2023 in Amsterdam, the Netherlands.

Regulars
The Real Thing?
7 News
The latest research news and news in brief
9 Tips & Tricks GPC/SEC: A Look at Molar Mass Averages and How
Using GC×GC–TOF-MS to identify soft drinks They Are Affected By SEC Baseline and Integration Settings
Wolfgang Radke, PSS Polymer Standards Servive GmbH
What is the meaning of these averages?
 Comparing the Flavour
Profiles of Soft Drink
Brands Using Immersive
Sorptive Extraction,
GC×GC–TOF-MS,
and Chemometrics
Laura McGregor, James Ogden, and Elinor Hughes, SepSolve Analytical, Peterborough, UK

Immersive sorptive extraction coupled with comprehensive two-dimensional

gas chromatography and time-of-flight mass spectrometry (GC×GC–TOF-MS)
was used to compare flavour profiles from popular brand soft drinks with
those of imitation products.

Unlike counterfeit goods, replica and imitation
products are legal because they do not use
the branded product’s trademark. In the food
and beverage industry, imitation products
attempt to mimic the taste experience of
popular branded products.
However, flavour profiles are extremely
Jag_cz/stock.adobe.com
preference for a particular brand. It
is important to be able to confidently
identify these volatiles during product
development, as well as in quality and
authenticity studies.
Traditional sample preparation methods,
such as headspace and solid-phase

complex and consist of a broad range of microextraction (SPME), are widely used
chemical classes, the combination of which but are often limited in terms of sensitivity.
ultimately determines the consumer’s Additionally, sampling is generally restricted

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 The Column www.chromatographyonline.com McGregor et al.

Figure 1: Example GC×GC–TOF-MS chromatogram for immersive sampling of a cola-flavoured soft

drink (brand D). Spectral examples are provided in Figure 2.

3
4

1. Benzenepropanal 3
2. Perilla aldehyde

tR (s)
3. α-Curcumene
4. trans-α-Bisabolene 2

2
2
1

3 25 26
1
tR (min)
tR (s)
2

1
10 10 30 40 50 60
1
tR (min)

to the headspace volatiles, meaning that two-dimensional gas chromatography

there is a bias towards nonpolar analytes.
This article demonstrates the use of
high‑capacity sorptive extraction with novel
trap-based focusing to provide enhanced
sensitivity and improved chromatographic
performance, as well as the ability to
perform both headspace and immersive
sampling for compatibility with a wider
range of polar and semi-volatile analytes.
This improved performance, coupled with
improved separation by comprehensive
(GC×GC) and detection by time-of-flight
mass spectrometry (TOF-MS), gains greater
insight into sample composition.
However, sampling, separation, and
detection is just the beginning—the
resulting datasets must then be reduced
to discover significant differences and
ultimately allow meaningful conclusions
to be reached. This article will also
demonstrate the use of chemometrics
to help transform complex datasets

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 The Column www.chromatographyonline.com McGregor et al.

Shimadzu_TheColumn_TQ-8050:Layout 1 10.04.19 11:25 Seite 1

Figure 2: Identification of the four components in Figure 1 using comparison of TOF-MS spectra
against the NIST 20 commercial library with additional confirmation through mass accuracy of the
system, with all examples <10 ppm mass accuracy in this case.

Benzenepropanal Perilla aldehyde

68.0596
91.057

79.0533
BenchTOF2 BenchTOF2

107.0545
67.0525
92.063
100 100

134.074

150.1030
135.0871
121.0989
53.0357
41.0338
78.050

105.070
65.037
51.021

50 50

0 0
39.000

65.000

121.0000

135.0000
51.000

105.000

150.0000
50 50

41.0000

107.0000
53.0000
134.000
78.000

67.0000

79.0000
92.000

100 100

68.0000
91.000

NIST20 NIST20

α-Curcumene trans-α-Bisabolene
93.0666
119.089
132.095

BenchTOF2 BenchTOF2
100 100 119.0829
105.071

202.173

109.0975

204.1890
145.108

41.0359

136.1125
91.050

133.099
41.038

50 50

0 0
136.0000

204.0000
41.0000
145.000

202.000

109.0000
119.0000

50 50
105.000
41.000

Coupling powers
132.000

100 100
119.000

93.0000

NIST20 NIST20

Pioneering new fields in ultra-trace analysis, the new Comfortable, easy change of accessories
GCMS-TQ 8050 NX triple quadrupole couples the powers through the advanced, illuminated GC oven
of a world-leading GC and a newly designed detector.
into usable results. The approach used were purchased from different suppliers. Both provide outstanding sensitivity at femtogram and The GCMS-TQ 8050 NX complements the Shimadzu NX
even sub-femtogram levels. family, coupling the Nexis GC-2030 with the quadrupole
ensures that trace peaks are not ignored Sampling and analysis were performed in series TQ-8050, TQ-8040 or QP-2020. Shimadzu’s NX series
Superior performance of NX technologies provides high-end GCMS solutions for every analytical
and enables automated workflows to be triplicate for each soft drink. e.g. new flow controller and time management challenge.
adopted,. This workflow will be presented The following experimental conditions for maintenance

for the comparison of brand and imitation and instrumentation were used. For A wide variety of optional products supports
trace analysis
cola‑flavoured soft drinks. immersive sampling, HiSorb high-capacity such as autosamplers and inlets
sorptive extraction probes were used
Experimental (PDMS/CWR/DVB, Markes International). www.shimadzu.eu /coupling-powers

Five store-bought cola-flavoured soft drinks Full automation was performed on the

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 The Column www.chromatographyonline.com McGregor et al.

Figure 3: Principal component analysis (PCA) score plot of the top 50 significant differences

QUATTRO CPC, CCC + 2D HPLC

between soft drink brands.

0.30

0.20
Brand B Brand D PREPARATIVE CHROMATOGRAPHY
IS OUR SPECIALITY. PROTEINS, RNA’S,
0.10 Brand E API’S, MEDICINAL CANNABINOIDS,
0.00
NATURAL & SYNTHETIC PRODUCTS, ETC.
Brand C PROCESS, MILLIGRAMS UP TO
Presenter Name
MULTIPLE TONNES PER ANNUM
PC2 (19.78%)

-0.10

-0.20

Brand A PC1 (72.19%) Save ~50-90% Process

-0.30
0.4 Chromatography / Extraction,
-0.40 Solvent / Column Costs

PC3
0.2

(4.8
-0.50 0

3%)
Ionic Liquid Based CPC + CCC
-0.60
0.25 0.20
0.15 0.10
0.05 0.00
-0.4
Allow Green Processing And
-0.05 -0.10
PC1 (72.
-0.15 -0.20
-0.25 -0.30
Unique Opportunities For
19%) -0.4
-0.35 -0.40
-0.45 Forcing Equilibria For Ultra
Efficient API etc. Synthesis.

Figure 4: Box and whisker plots showing two key differentiators of the soft drink brands (1).
Website : https://www.quattroprep.com
Benzyl benzoate trans-Cinnamaldehyde
Balsamic, fruity1 Spicy, sweet, cinnamon1

ionization energy: tandem ionization evident in Figure 1. Four compounds are

at 70 eV and 14 eV. Software:
ChromSpace software (SepSolve
Analytical) for full instrument control and
processing was used, with chemometric
comparisons by ChromCompare+
(SepSolve Analytical)
Results and Discussion
Coupling high-capacity sorptive extraction
with GC×GC–TOF-MS ensured that a wide
range of chemical classes were efficiently
Centri extraction and enrichment platform modulation period (PM) = 2.5 s. TOF-MS: separated and confidently identified.
(Markes International). GC×GC: Insight BenchTOF2 mass spectrometer (SepSolve The additional chemical detail provided
flow modulator (SepSolve Analytical); Analytical); mass range: m/z = 40–350; on the composition of soft drinks is clearly
highlighted that would have coeluted in a
conventional one-dimensional (1D) analysis
but have been separated in the second
dimension and identified confidently using
the spectral quality and mass accuracy of
TOF-MS (Figure 2).
In this study, five “cola” soft drinks from
different manufacturers were analyzed
using this workflow. To determine the
differences in composition between
each brand of soft drink, a nontargeted,
tile‑based workflow was then applied in
the software to compare all of the raw
data, and the most significant differences

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17 Dhandapani and Tackett 21 Q&A Ueland 25 SCM-10 Event Preview 27 Training and Events
 The Column www.chromatographyonline.com McGregor et al.

Figure 5: Volcano plot in the software brands. Interestingly, brands B and E are using high-capacity sorptive extraction. GC×GC−TOF-MS. In her current role at
showing comparison of an imitation cola both “diet” soft drinks from different Enhanced separation by GC×GC using SepSolve Analytical, she specializes in
(brand C) against a popular brand (brand A),
with two significant differences highlighted manufacturers and cluster separately from consumable-free flow modulation obtained the application of GC×GC and TOF-MS
and identified (1). the “zero sugar” brands. comprehensive aroma profiles. Sensitive to challenging applications.
2-Ethylhexanol
Benzyl benzoate was found to be a key detection and confident identification of Elinor Hughes obtained her B.Sc.
Sweet, fatty1
differentiator of brand D (Figure 4, left) analytes were achieved using TOF‑MS. in chemistry and Ph.D. in organic
57.069

100
and likely contributed a balsamic, fruity Chemometrics offered automated chemistry at Bangor University,
70.074
43.054

83.083

50
112.119

flavour. Additionally, trans-cinnamaldehyde workflows for alignment and comparison UK. After working for a chemical
98.016

Brand C
0

(Figure 4, right) was found to differentiate of complex chromatograms. manufacturing company for three
112.000
98.000
83.000
70.000

Brand A
43.000

50

100
the “diet” and “zero sugar” classes. years, she moved to the Royal Society
57.000

Volcano plots in the software were also
used to directly compare the imitation
Increased in imitation brand C Increased in popular brand A colas against popular brand products—an
example of which is shown in Figure 5.
Figure 5 also shows the reference-quality
spectra of the MS analysis. This helped to
ensure confident identification of potential
markers of brand and/or imitation cola
products. It should also be noted that
tandem ionization data were acquired in
Caryophyllene alcohol
Spicy1 this study at 70 eV and 14 eV. The tandem
111.067
Reference
1. The Good Scents Company Information
System (search facility). http://www.
thegoodscentscompany.com/search2.html
(accessed 2022-06-20).
2. SepSolve, SepSolve Analytical White Paper
041: Improving discovery workflows using
Tandem Ionisation data. https://www.sepsolve.
com/white-papers/overview/technical-note-
improving-discovery-workflows-using-tandem-
ionisation-data.aspx (accessed 2022-12-19).
of Chemistry where she worked in
journals publishing for six years and
on Chemistry World magazine for four
years. This was followed by five years
as a freelance copyeditor and science
writer. Her current role is technical
copywriter at Markes International.
James Ogden spent nine years working
within an environmental analytical
laboratory, where he was responsible
for method testing, development,

100 data were used to confirm positive hits, and implementation. In his current
161.137
123.083

Laura McGregor received an M.Chem. role at SepSolve Analytical, James

55.037

81.065
41.030

improving the discovery of subtle, trace

189.172
95.074

Brand C
204.191
121.097
105.067
69.059

50
222.203

Brand A 0
differences by reducing the frequency of in chemistry from the University supports customers through the
189.000
204.000
67.000

95.000

161.000

of St Andrews, UK, followed by development, demonstration, and

123.000

false positives (2).

81.000

50
55.000
41.000

100
an M.Sc. in forensic science at the handover of analytical methods across
111.000

Conclusions University of Strathclyde, UK. Her the company’s portfolio of instruments

between the sample classes were This study has demonstrated an end‑to‑end Ph.D. in environmental forensics, and software.
identified automatically. nontarget workflow for aroma profiling also at the University of Strathclyde,
The resulting principal component of food and beverages comprising focused on the chemical fingerprinting E-mail: hello@sepsolve.com
analysis (PCA) score plot (Figure 3) fully automated immersive sampling of environmental contamination Website: www.sepsolve.com
shows distinct clustering of the different of wide‑ranging aroma-active volatiles using advanced techniques such as

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From the CEO
Welcome to the first issue of 2023! Our cover story for the January issue focuses
on assessing the flavour profiles of branded and imitation cola drinks using
two‑dimensional gas chromatography and time-of-flight mass spectrometry
(GC×GC–TOF-MS). Unlike counterfeit goods, replica and imitation products are legal
because they do not use the branded product’s trademark. The complexity of flavour
profiles mean that a robust method must be used to identify popular brands from
their imitators.
This month we spoke to “rising star” Martina Catani from the University of Ferrara
in Italy, about her work on the separation of enantiomers by high performance
liquid chromatography/ultrahigh-pressure liquid chromatography (HPLC/UHPLC) and
the study of the kinetic and thermodynamic properties of chiral stationary phases,
and the separation and purification using LC analysis (single-column and continuous
multi‑column) of biomolecules of high therapeutic interest.
Keeping with the chiral focus, we look at how to optimize chiral separations and
why it is important to explore all parameters through a chiral screening process across
a variety of chiral column chemistries.
Do you know what forensic taphonomy involves? We spoke to Maiken Ueland from
The Australian Facility for Taphonomic Experimental Research (AFTER) at the University
of Technology Sydney in Australia, about her work in this area, and how and where
she uses analytical techniques to conduct human post-mortem investigations.
As we hopefully embrace a full year of face-to-face conferences and events, Peter
Schoenmakers takes us behind the scenes at the upcoming SCM-10 conference, due to
be held 1–3 February in Amsterdam.
Our regular feature “Tips & Tricks GPC/SEC” kicks off with a focus on molar mass
distributions and the molar mass averages. What is the meaning of these averages and
how are they influenced by the setting of baselines and integration limits?
All the best to you and yours for 2023 and happy reading!
Mike Hennessy Jr.,
President and CEO, MJH Life Sciences®
2 McGregor et al. 7 News
7
17 Dhandapani and Tackett 21 Q&A Ueland
News
Agilent Achieves “Angel”
Level Sponsorship of My
Green Lab
Agilent Technologies Inc. (Santa Clara, California, USA) has announced that it has
become the first “Angel” level sponsor of My Green Lab, a nonprofit organization
devoted to building a global culture of sustainability in science.
Agilent is also sponsor of the My Green Lab Certification programme, and is
working hard to ensure its global on-site customer demonstration laboratories are
Green Lab Certified. The company has already achieved the highest level certification for
the Waldbronn, Germany, Cheadle, UK, and Santa Clara, USA, sites.
“Sustainability and environmental, social, and governance (ESG) standards are
central to our mission to advance the quality of life, and our sponsorship of My
Green Lab helps us to drive this forward,” said Darlene Solomon, senior vice
president, and chief technology officer at Agilent. “Being an ‘Angel’ level sponsor
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Recognized by the United Nations Race to Zero campaign as a measure of
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strong sustainability practices in energy use, water consumption, recycling,
and waste production.
For more information, please visit: www.agilent.com
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9 Tips & Tricks 13 Rising Stars
25 SCM-10 Event Preview 27 Training and Events
 The Column www.chromatographyonline.com News

Peaks of the Month News In Brief

• The LCGC Blog: How Well Do You Pipette?—At the heart of most chromatographic analyses are Thermo Opens New Site in China
the gravimetric and volumetric operations used to prepare samples, standards, and HPLC mobile
phases. Read Here>>
Thermo Fisher Scientific (Waltham, Massachusetts,
USA) is opening a site in Hangzhou, China,
as part of its global effort to help companies
• Separation Science: The State of the Art—In this extended special feature to celebrate the 35th provide therapies to patients faster. The
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SPECIAL ANNIVERSARY FEATURE

The state of the art in separation science
PEER REVIEW

anniversary edition of LCGC Europe, key opinion leaders from the separation science community
Reversed-phase separations of peptides
LIQUID CHROMATOGRAPHY

80,000-square‑metre current Good Manufacturing

Avoiding problems in proteomics
GAS CHROMATOGRAPHY
A look at electron capture detection
APPLICATION NOTES
November/December 2022 | Volume 35 Number 10
An update on practical applications
www.chromatographyonline.com
EVENTS

explore contemporary trends in separation science and identify possible future developments.
The must-attend events for chromatographers

Practices (cGMP) facility will offer integrated clinical

Read Here>>
and commercial drug substance and drug product
ANNIVERSARY ISSUE

capabilities, including process development,

35 Years Of
Innovation cell line development, biologics drug substance
manufacturing, and sterile fill-finish.
Christian Müller/stock.adobe.com

• W
 hat Will Be Happening at HPLC 2023?—As HPLC 2023 approaches, The Column spoke with the “Hangzhou is a strategic addition to our global
organizers, Oliver Schmitz and Michael Lämmerhofer, to find out what attendees can expect from pharma services network and the latest example
the conference, which is a highlight in the chromatography calendar. Read Here>> of our commitment to meeting customer demands
globally,” said Mike Shafer, senior vice president and
• Analytically Speaking Podcast: The Future of Chemometrics—Data-Driven Measurements and president, pharma services. “We continue to invest
Instruments for Chemistry—We spoke to Professor Karl Booksh of the Department of Chemistry in capabilities to support the manufacturing needs
and Biochemistry at the University of Delaware, Newark, USA, about his organization of a National of our customers, enabling them to serve more
Science Foundation workshop to explore research on the development of portable chemical sensors patients throughout the world.”
for environmental, biomedical, and industrial process monitoring. Karl’s own research is predicated “Thermo Fisher has been in China for 40 years,
on the belief that it is better to build small chemical sensors capable of reliable measurements in the
field or in the process than to collect samples for future laboratory analysis. Read Here>> serving this market through its bioproduction,
clinical research and pharma services businesses,”
said Hann Pang, president, Thermo Fisher China.
• T
 he LCGC 2023 Winners of the Lifetime Achievement and Emerging Leader in Chromatography “Committed to our ‘in China, for China’ localization
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For more information, please visit:
Like us Join us Follow Us www.thermofisher.com

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 Tips & Tricks GPC/SEC:
A Look at Molar Mass
Averages and How They Are
Affected By SEC Baseline
and Integration Settings
Wolfgang Radke, PSS Polymer Standards Service GmbH, Mainz, Germany

Determination of molar mass distributions and the molar mass

averages derived therefrom are the main objectives of gel permeation
chromatography/size-exclusion chromatography (GPC/SEC) analysis. But
what is the meaning of these averages and how are they influenced by
the setting of baselines and integration limits? This instalment of Tips and
Tricks in GPC/SEC will try to provide a better understanding.

Gel permeation chromatography/ molecules elute later from the column.

size‑exclusion chromatography (GPC/SEC)
has become the main technique used
to determine molar mass distributions
and molar mass averages. In GPC/
SEC, macromolecules are separated
according to their size, which for most
miiko/stock.adobe.com
The use of a calibration curve allows
the chromatogram to be translated into
a molar mass distribution from which
average molar masses, such as the number
(Mn) and weight average molar masses
(Mw), can be obtained. These average molar

polymers correlates to molar mass. At the masses can then be used for quality control
appropriate conditions the high molar or to release products for further use. For
mass species elute first, while smaller example, pharmacopoeias often request

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 The Column www.chromatographyonline.com Tips & Tricks

Figure 1: Left: Logarithmic molar mass distribution with indicated Mw. Right: Linear molar mass
distribution. Support corresponds to centre of gravity, equal to Mw.
1,0 1,0
1.0x105 2.0x105 3.0x105 4.0x105
Mw Molar Mass (g/mol)

0,8 0,8

0,6 0,6
w(logM)

Mw

w(M)
0,4 0,4

0,2 0,2

0,0 0,0
103 104 105 106
Molar Mass (g/mol)

Figure 2: Chromatogram of a sample containing a low amount of low molar mass material.

95

90

85

80

75

70

65

60

A pack size for

Normalized Detector Response

55

your application.
50

45

40

35

30
Whatever your application, we can supply
you with a pack size for successful testing.
25

20
lgcstandards.com/mikromol
15 And with no further preparation needed, mikromol@lgcgroup.com
10
you save on time and waste.
5

0
21.0 21.5 22.0 22.5 23.0 23.5 24.0 24.5 25.0 25.5 26.0 26.5
Elution Volume (mL)
27.0 27.5 28.0 28.5 29.0 29.5 30.0 30.5 31.0 31.5 32.0
Our high-quality ISO 17025 and 17034 products
come with extensive Certificates of Analysis
containing measurement uncertainty values
molar mass averages to lie within defined mass averages? Most of us know how to and can be used with pharmacopoeial
limits before the materials can be used for calculate these averages, but what do these methods and beyond.
pharmaceutical applications. values tell us?
But what is the meaning of these molar According to reference books (1), the Mn

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Figure 3: Molar mass distributions (left) and number distribution of a sample containing a low amount of low molar mass species (right).

95 95
95 95

90 90
90 90

85 85
85 85

80 80
80 80

75 75
75 75

70 70
70 70

65 65
65 65

60 60
60 60

55 55
55 55

50 50
50 50

45 45
45 45

40 40
40 40

35 35
35 35

30 30
30 30

25 25
25 25

20 20
20 20

15 15
15 15

10 10
10 10

5 5
5 5

0 0
0 0
1*103 1*104 1*105 1*106 1*103 1*104 1*105 1*106
Molar Mass (Da) Molar Mass (Da)

and Mw can be calculated according to:
[1]
[2]
Where ci, Ni, and wi are the weight
concentration, number, and weight fraction
of molecules with molar mass Mi. The sums
must be evaluated over all species present
in the sample. Other versions of the above
formulas that use integrals do exist. The
difference between the integrating versions
and the summations above result from the
assumption of the existence of discrete molar
masses (summation) or a continuum of molar
masses (integrations) present in the sample.
The latter assumption is often applied in
theoretical calculations.
Descriptive Meanings of Mn and Mw
While the formulas given above are mostly
known, their physical meaning and the
consequences are often not clear.
Let us first have a look at the number
average molar mass, Mn.
The right side of equation 1 provides
a simple rule for the determination of
the number average molar mass. As the
numerator is equal to the total mass of the
sample, while the denominator equals the
total number of molecules in the sample,
the rule reads “weigh in your sample and
determine the number of molecules within
the sample”. The number average molar
mass is therefore what we would typically
provide if someone asked us to calculate
“an average”.
While the meaning of the number average
molar mass is largely clear, a descriptive
meaning of the weight average molar mass is
often lacking. The weight average molar mass
neither provides the median of the weight
distribution where above and below exists the
same amount of sample molecules, nor does
the weight average molar mass correspond to
the maximum of the molar mass distribution
or to the most abundant molar mass.
To get a better understanding of Mw, we
can rearrange equation 2 to yield:
[3]
Equation 3 resembles the equation defining
the centre of gravity, with (Mi–Mw) interpreted
as the distance at which a force proportional
to wi is acting. Thus, Mw can be regarded
as the centre of gravity of the linear molar
mass distribution, w(M). This is schematically

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Figure 4: Left: Chromatogram of sample with a low amount of a high molar mass impurity. of polymer chains when including the small • The number average molar mass is obtained
Right: Linear molar mass distribution of a sample containing a low amount of high molar mass
impurity with indicated position of Mw. peak. Figure 3 shows a comparison of the by determining the number of molecules
corresponding weight distribution (the weight within a given amount of sample.
1,0
70 1x105 2x105 3x105 4x105 5x105 fraction of molecules of a given molar mass) • The weight average molar mass corresponds
Molar Mass (g/mol)
60 and the number or frequency distribution (the to the centre of gravity of the linear mass
0,8

50
molar fraction of molecules of a given molar distribution.
Signa Intensity (a.u)

0,6 mass). While the small peak contributes to less • Small variations in the setting of the baseline
40
than 1% of the total sample weight, it contains or integration may significantly affect the

30
20
10
0 0,0
15 20
Elution Volume (mL)
shown in Figure 1, where the position of Mw
is indicated in the logarithmic and the linear
molar mass distribution. Note: To convert a
logarithmic molar mass distribution into a linear
one it is not sufficient to simply replot the
x-axis, the y-values also need to be recalculated.
If a support (black triangle) is placed below
the linear weight distribution at Mw, the
distribution is in balance. Any displacement of
the support from Mw results in an imbalance.
Origin of the Effect of Setting of
Baseline and Integration Limits on
Mn and Mw
The descriptive meaning of Mn given above
helps us to understand the effect of setting
baseline and integration limits in GPC/SEC
on Mn.
w(M)
0,4
Mw
0,2
25 30 35 40
The chromatogram in Figure 2 shows a small
peak at high elution volume. Although this
peak contributes by less than 1% of the total
peak area, its inclusion or exclusion significantly
alters the number average molar mass—from
22200 g/mol when including the peak to
24100 g/mol when the peak is excluded. The
weight average molar mass remains nearly
unaffected (36100 g/mol vs. 36000 g/mol).
Knowing that determination of number average
molar mass is based on the total weight of the
sample in relation to the number of molecules
within the sample, and keeping in mind that
the total area under the curve is proportional
to the total mass of the sample, which varies
by less than 1%, the origin of the variation in
number average molar mass must be due to
the presence of a significantly higher number
nearly 8% of all polymer molecules. Excluding
this peak will therefore substantially alter the
number average molar mass. The settings of
baseline and integration limits, particularly at
high GPC/SEC elution volumes, is therefore
of great importance to help achieve reliable
number average molar masses.
Similarly, a small peak of very high molar
mass species, for example, from aggregates,
will move the weight average up substantially to
higher values. In the example shown in Figure 4,
the weight average molar mass corresponding
to the complete chromatogram is 102000 g/
mol. Ignoring the small peak at low elution
volume—which accounts for only 3% of the
total mass—results in a weight average molar
mass of 68200 g/mol. This large difference is
because the weight fraction of the high molar
mass species though low is multiplied with its
distance from the centre of gravity (Mw).
Summary
• The number average molar mass
corresponds to the usual arithmetic average.
number of molecules in a sample, and
therefore may have a strong impact on the
determination of the number average molar
mass by GPC/SEC.
• Ignoring small fractions of high molar mass
will alter Mw as determined by GPC/SEC
because the weight faction is multiplied
with its deviation from the centre of gravity.
Reference
1. Odian, G. Principles of Polymerization, Fourth
Edition; John Wiley & Sons, 2004.
Wolfgang Radke studied polymer chemistry
in Mainz, Germany, and Amherst,
Massachusetts, USA, and is head of the
application development department at
PSS—now a part of Agilent. He is also
responsible for instrument evaluation and
for customized trainings.
E-mail: WRadke@pss-polymer.com
Website: www.pss-polymer.com

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17 Dhandapani and Tackett 21 Q&A Ueland 25 SCM-10 Event Preview 27 Training and Events
 Rising Stars of
Separation Science:
Martina Catani
 his month we interview Martina Catani from the University of Ferrara, Italy,
T
about her work on the separation of enantiomers by high performance liquid
chromatography/ultrahigh-pressure liquid chromatography (HPLC/UHPLC) and
the study of the kinetic and thermodynamic properties of chiral stationary
phases, and the separation and purification using LC analysis (single-column
and continuous multi-column) of biomolecules of high therapeutic interest.

—Interview by Kate Jones

Q. When did you first encounter
chromatography and what attracted
you to the subject?
A: The first time I encountered
chromatography was during my bachelor’s
degree in chemistry. Even though at
that time I was only studying it from
a theoretical point of view, I became
fascinated by the great versatility of
this technique and its capability for
understanding the composition of
Designec/stock.adobe.com
my life” was during my master’s thesis.
My work was focused on the design of
enzymatic packed‑bed microreactors for
the enantioselective formation of C–C
bonds in aqueous environment. I had the
opportunity to functionalize silica particles
with the enzyme, to pack the columns
with this adsorbent, and to fully
characterize the performance of the
microreactor not only in terms of
catalytic conversion but also from a

unknown complex mixtures. However, chromatographic point of view. In addition,

the moment at which I said, “OK, this since the scope of the work included the
is what I would like to do for the rest of evaluation of the enantiomeric excess of

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 The Column www.chromatographyonline.com Rising Stars

media commonly used for ultrafast, high separations where the enantiomers of
Martina Catani is an assistant professor of analytical chemistry efficiency separations. I considered not some compounds of pharmaceutical
in the Department of Chemical, Pharmaceutical, and Agricultural only achiral adsorbents (such as C18 or interest were separated in less than one
Sciences at the University of Ferrara (Italy), and she received her perfluorinated stationary phases) but also second with extremely high efficiency
Ph.D. in chemical sciences in 2018 from the same university. (and especially, I would say) chiral ones. should be noted (1).
During her career, she has spent research periods at VUB Brussels In this context, I performed innovative
(Belgium) in the group of Gert Desmet, and at the University of studies to unravel the complex mass Q. What chromatographic techniques
Pécs (Hungary) under the supervision of Attila Felinger. In 2019 transfer phenomena dominating have you worked with?
she was a postdoctoral research fellow at ETH Zurich (Switzerland) in the group kinetics of chiral chromatography and I A: I have mainly worked with high
of Massimo Morbidelli. Martina works in the field of LC for both analytical and also investigated the thermodynamics performance liquid chromatography/
preparative purposes. Her main research activities are focused on the purification of adsorption through nonlinear ultrahigh-pressure liquid chromatography
of polypeptides, oligonucleotides, and proteins by means of single-column and chromatographic means (namely, (HPLC/UHPLC) coupled with different
continuous countercurrent multi-column preparative LC; separation of natural adsorption isotherm determination). detection techniques such as UV–vis diode
cannabinoids; and investigation of kinetic and thermodynamic phenomena in In particular, these investigations were array, mass spectrometry, fluorescence,
chiral and achiral HPLC and SFC. She is co-author of almost 60 papers in devoted to the understanding of the or refractive index detection. I have
peer‑reviewed journals and has presented her research at more than 20 national impact of kinetic aspects (adsorption– applied different chromatographic modes
and international scientific meetings. She has been the recipient of many awards, desorption kinetics and eddy dispersion), such as normal phase, reversed phase
including the “Csaba Horváth Young Scientist Award” conferred at HPLC2018 particle geometry (fully porous vs. core– (including separations with perfluorinated
Washington (USA) and the “2021 Young Researcher Award” conferred by the shell), and physicochemical characteristics and mixed-mode adsorbents), hydrophilic
Analytical Chemistry Division of the Italian Chemical Society. (specific loading of chiral selector, pore interaction liquid chromatography
size) on the performance of these columns. (HILIC), ion-exchange, affinity, and chiral
These studies not only opened up new chromatography.
the products, I also performed several Q. Can you tell us more about your perspectives in this field by reviving the I have also worked with preparative liquid
enantioseparations by using a chiral Ph.D. thesis? debate about the best particle format chromatography—both single‑column
column in gas chromatography (GC) and I A: I started my Ph.D. in 2014 at for ultrafast separation, but they have and multi-column countercurrent.
started developing a passion for separating the University of Ferrara under the also been the basis for the design, More recently, I have also worked with
enantiomers (a topic that is still dominating supervision of Alberto Cavazzini. My characterization, and operation of supercritical fluid chromatography (SFC)
a great part of my work). Ph.D. project was mainly focused on the high‑efficient columns for ultrafast chiral coupled with UV–vis diode array detection,
During that period, I started seriously investigation of theoretical aspects of chromatography. The paper (written in and, as I mentioned earlier, I had the
thinking about the idea of pursuing a Ph.D. liquid chromatography (LC), including mass collaboration with the group of Francesco chance to work with GC coupled with
in analytical chemistry—especially based transfer characteristics and thermodynamic Gasparrini of the University of Rome “La flame ionization detection (FID) during my
on chromatography. properties of different kinds of porous Sapienza”) for the realization of ultrafast master’s thesis.

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 The Column www.chromatographyonline.com
Q. Your research focus currently lies in
the separation of enantiomers by HPLC/
UHPLC and the study of the kinetic and
thermodynamic properties of chiral
stationary phases—what specifically
attracted you to this area of research?
A: As I mentioned earlier, I have always been
fascinated by the power of chromatography
to separate enantiomers. In particular, I am
stimulated by the fact that, experimentally
speaking, the separation of enantiomers could
be a very easy operation (once the optimal
experimental conditions are found), but if
we think about the mechanism that leads to
their separation, this is anything but easy, and
many aspects related to enantiorecognition
are not yet fully understood. This is why I am
very interested in widening the knowledge
of mass transfer phenomena and the
thermodynamic mechanisms affecting
chiral chromatography.
Q. You are also active in the field
of separation and purification
using LC analysis (single-column
and continuous multi-column) of
biomolecules of high therapeutic
interest, such as polypeptides,
proteins, and oligonucleotides.
You recently published a paper
on the enrichment and recovery
of oligonucleotide impurities by
2 McGregor et al.
15
17 Dhandapani and Tackett
N-Rich twin-column continuous
chromatography (2). What is continuous
chromatography? What challenges did
you face during this analysis?
A: Yes, this is another research field that
we began a few years ago, after a research
period at ETH Zurich in the group of
Massimo Morbidelli.
Continuous chromatography is considered
to be the most suitable solution for process
intensification in the (bio)pharmaceutical
industry, and to partially overcome intrinsic
limitations of single-column (batch)
preparative chromatography—the most
widely used approach for the industrial
purification of (bio)pharmaceuticals. When
only one column is used, the loading of
the feed is a discontinuous operation. In a
continuous (or semi-continuous) process,
the feed is continuously (or following a time
cycle) loaded into the purification unit, which
is usually made by (at least) two identical
columns. Multi-column processes typically
exploit the concept of countercurrent
chromatography, where the columns are
connected by a series of valves that allow
simulation of the movement of the stationary
phase in the opposite direction to the mobile
phase. Such countercurrent movement
increases the interphase mass-transfer rates,
therefore making the process more efficient.
In addition, the overlapping regions, eluting
7 News
21 Q&A Ueland
Rising Stars
from one column and containing both the
target product and impurities, are recycled
into the other column with increased
powered by
yield of the purification process and less
solvent consumption.
This kind of approach can also be applied We have 1000’s of
to the enrichment and isolation of species
present in very low amounts by applying
eLearning topics
the so-called N-Rich process, which is
CHROMacademy is the
particularly beneficial in the isolation of
critical impurities or minor compounds,
world’s largest eLearning
as in the paper mentioned. website for analytical
The most difficult part in setting up scientists, containing
mult‑column continuous purification is the 1000’s of interactive
choice of characteristic times defining the eLearning topics.
recycling and collection windows, which
requires much experimental work to find the Lite members have
optimal conditions. access to less than 5%
of our content.
Q. Another of your recent papers Premier members get
presents the use of multi-column so much more!
countercurrent chromatography for the
purification of either target peptides/
oligonucleotides or their impurities (3).
HPLC GC MS SPE IR
What benefits does this method offer
over existing ones? To find out more about CHROMacademy
A: As mentioned before, single-column Premier membership, contact:
chromatography has several intrinsic
Vito Laudati: (609) 819-5794
limitations. In particular, when several
e-mail: vlaudati@mjhlifesciences.com
product-related impurities are present in
the mixture, they can coelute with the www.chromacademy.com
target product both in the front and in
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25 SCM-10 Event Preview 27 Training and Events
 The Column www.chromatographyonline.com
the end of the peak, generating critical
peak overlapping. This situation, called
centre‑cut (or ternary) separation, leads to
a yield-purity trade-off, which means that
to achieve high purity, overlapping portions
of the chromatogram need to be discarded,
thus the collection window is narrowed at
the cost of yield. Vice versa, higher yield
can be obtained by enlarging the collection
window (therefore also collecting impurities),
but purity unavoidably decreases. The
employment of multi-column countercurrent
techniques allows this limitation to be
partially overcome by automating the
recycling of the overlapping portion of
the chromatogram into the system. As a
consequence, higher yield can be obtained
at the same purity constraint, solvent
consumption decreases, and the purification
process can be completely automated.
Q. What are your next steps with
this research?
A: We are working on an ambitious project
aimed at finding green solutions to increase
the sustainability of liquid chromatography,
particularly in preparative conditions (4).
Q. What advice would you give to
anyone wanting to enter this field?
A: Well, the very first step to becoming a
researcher is to get a Ph.D. degree. One
2 McGregor et al.
16
17 Dhandapani and Tackett
should be aware that this track can be
characterized by many ups and downs, but
my advice is to never give up and keep on
rocking, especially during the “downs”. I
have personally grown a lot during my Ph.D.
degree, not only from the scientific aspect
but also from a more personal perspective.
Connected to this, I also encourage people
to spend some time abroad since it helps
a lot in the development of skills that are
fundamental for researchers and helps
to create a network with other scientists.
Indeed, my final advice is that, in my opinion,
it is of fundamental importance to remember
that science is not an individual activity and
teamwork can make science work better.
Q. Is there any other recent research
or work you have been doing lately
that you think is important and want
to highlight?
A: Yes, I would like to mention that our
group is also active in the separation of
natural cannabinoids through HPLC and
SFC. Specifically, we have recently published
some interesting data regarding chirality of
cannabinoids, about which almost nothing
is known (5). Indeed, many cannabinoids are
chiral but they are preferentially produced as
(-)-L-trans form by the plant. For this reason,
common separation methods are based on
the use of achiral columns, irrespective of
7 News
21 Q&A Ueland
cannabinoids stereochemistry, and there are
only a few papers in the literature where
chiral cannabinoids were separated—
mostly dating back to the beginning of the
1990s. However, in recent years, due to the
increasing popularity of cannabis products,
there has been a growing interest towards
the characterization of cannabinoids in
terms of enantiomeric purity. For instance,
cannabichromene (CBC) is one of the rare
examples of natural racemic (or scalemic)
mixtures, as also demonstrated by our
work, where both CBC enantiomers from
a real sample have been separated in LC
with two polysaccharide-based chiral
stationary phases.
References
1. Ismail, O. H.; Pasti, L.; Ciogli, A.; et al. Pirkle-Type
Chiral Stationary Phase on Core–Shell and Fully
Porous Particles: Are Superficially Porous Particles
Always the Better Choice Toward Ultrafast
High‑Performance Enantioseparations? Journal of
Chrom. A 2016, 1466, 96–104. DOI: 10.1016/j.
chroma.2016.09.001
2. Lievore, G.; Weldon, R.; Catani, M.; Cavazzini,
A.; Müller-Späth, T. Enrichment and Recovery
of Oligonucleotide Impurities by N-Rich Twin-
Column Continuous Chromatography. Journal of
Chrom. B 2022, 1209, 123439. DOI: 10.1016/j.
jchromb.2022.123439
3. De Luca, C.; Felletti, S.; Bozza, D.; et al.
9 Tips & Tricks
25 SCM-10 Event Preview
Rising Stars
Process Intensification for the Purification of
Peptidomimetics: The Case of Icatibant through
Multicolumn Countercurrent Solvent Gradient
Purification (MCSGP). Ind. Eng. Chem. Res. 2021,
60, 6826−6834. DOI: 10.1021/acs.iecr.1c00520
4. Ferrazzano, L.; Catani, M.; Cavazzini, A.; et al.
Sustainability in Peptide Chemistry: Current
Synthesis and Purification Technologies and
Future Challenges. Green Chem. 2022, 24,
975–1020. DOI: 10.1039/D1GC04387K
5. De Luca, C.; Buratti, A.; Umstead, W.; et al.
Investigation of Retention Behavior of Natural
Cannabinoids on Differently Substituted
Polysaccharide-Based Chiral Stationary Phases
Under Reversed-Phase Liquid Chromatographic
Conditions. Journal of Chrom. A 2022, 1762,
463076. DOI: 10.1016/j.chroma.2022.463076
Rising Stars of Separation Science
The Column will be running a
series of interviews in 2023,
featuring the next generation of
separation scientists. If you would
like to nominate a “rising star” for
consideration, please send the name
of the candidate and why they
deserve recognition to Kate Jones,
Managing Editor, LCGC Europe
and The Column at
kjones@mjhlifesciences.com
6.
13 Rising Stars
27 Training and Events
 Playing with Selectivity
for Optimal Chiral
Separation
Ramkumar Dhandapani and Bryan Tackett, Phenomenex, Torrance, California, USA

Chiral separation is challenging and requires thorough method

optimization. Selectivity plays a critical role in chiral separation. In this
study, various ways of achieving resolution by playing with selectivity are
presented. This resolution improvement can be realized from the stationary
phase—the type of polysaccharide, nature of the chiral selector, position of
the functional group in the chiral selector—the mobile phase composition,
and from temperature. To optimize such challenging separations, it is
important to explore all of these parameters through a chiral screening
process across various chiral column chemistries.

Chirality is the potential of a molecule to effects of a chiral isomer is described in the

occur in two asymmetric forms (enantiomers)
that are non-superimposable mirror images
of each other without changing the atomic
composition, atom­-atom connections, or bond
orders. Chirality is a property of the whole
molecule, but the cause of chirality is the
chirality centre (asymmetric centre) within the
molecule. It is important for an active chiral
martinjanecek/stock.adobe.com
case of thalidomide (1–7), where pregnant
women treated with this drug gave birth
to children with birth defects due to the
(S)‑enantiomer being teratogenic. To separate
chiral isomers and quantify the amount of
each isomer in a given mixture, a column
that can recognize small spatial differences
between one chiral isomer and the other

ingredient to be in one chiral form for a few is required. Many column phases have
reasons—the other chiral isomer can be been developed to attempt to separate
inactive in the biological system or can have enantiomers and stereoisomers. This includes
adverse effects. A good example of the adverse ligand exchange, protein-based, chiral crown

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 The Column www.chromatographyonline.com Dhandapani and Tackett

Figure 1: Structure of cellulose vs. amylose polysaccharide. Figure 2: Selectivity from the type of polysaccharide.

OH OH
CH3
1000000
OH OH
O HO O HO H3C
HO O OH
O O

Intensity
HO O HO O NH 500000
OH OH O N
HO
O
OH n OH 0 N
N
Cellulose

Lux Cellulose-1 0.0 0.2 0.4 0.6 0.8 1.0 1.2 1.4 1.6 1.8 2.0 2.2 2.4 2.6 2.8 3.0 Cl
Retention time (min)
Cellulose : Layered Structure Cellulose tris(3,5-dimethylphenylcarbamate)
Analyte:
OH OH Tebucconazol
500000
CH3
400000
O O
HO O OH H3C
HO OH HO

Intensity
OH O HO OH O HO NH 200000
O
O O
O

Amylose : Helical Structure Amylose 0

OH n OH Lux Amylose-1
Amylose tris(3,5-dimethylphenylcarbamate) 0.0 0.2 0.4 0.6 0.8 1.0 1.2 1.4 1.6 1.8 2.0 2.2 2.4 2.6 2.8 3.0
Retention time (min)

ether‑based, cyclodextrin‑based, pirkle‑type,
polysaccharide‑based, macrocyclic
glycopeptide‑based, and cinchona
alkaloid‑based phases. The most popular
column currently used for this purpose is a
polysaccharide-based chiral stationary phase
(CSP). There are several advantages to using
a polysaccharide-based CSP. Polysaccharide
CSPs have a wide chiral recognition and
selectivity, multiple chiral selectors, can be
easily manipulated to reverse elution order,
high pressure stability, high loading capacity,
high efficiency, and—most importantly—
scalability from analytical to preparative
separation. Polysaccharide-based CSPs are
also amenable to various separation modes
such as reversed phase, normal phase, polar
organic, and supercritical fluid. For the
scope of this study, only chiral separation
from polysaccharide stationary phases will
be discussed.
Resolution Improvement
Resolution improvement in chromatography
can be brought about by three different
factors: efficiency term, selectivity term,
and retention term. Out of these three,
selectivity is the most influential term to
bring drastic improvement in resolution.
Selectivity for chiral separation can be
realized from three main sources: the
stationary phase, the mobile phase, and
temperature. Within the stationary phase,
selectivity differences arise from the type
of polysaccharide and type of selector.
Amylose or cellulose are the common
polysaccharides used in chiral stationary
phases and bring a difference in selectivity.
The nature and the position of the functional
group can have a huge impact on the
selectivity and eventual resolution of chiral
isomers. This will be discussed in detail in
the following sections.
Selectivity from the Stationary Phase depending on the analyte. As an example,
Selectivity can come from the polysaccharide fenarimol was analyzed using four cellulose
chosen for the stationary phase. Cellulose stationary phases, each with a different chiral
produces a tight, layered structure, whereas selector. Figure 3 shows how, depending on
amylose has a more helical structure to help which chiral selector is used, the separation and
separate enantiomeric analytes (Figure 1). retention times can be very different.
For example, tebuconazol was run on both a The enantiomers of diniconazole have very
cellulose- and amylose-based chiral column with different separation or no separation at all
the same chiral selector. Figure 2 shows that depending on which stationary phase and
enantiomers of tebuconazol can be separated chiral selector are used. Finally, the position of
depending on the polysaccharide. On the the side groups can also affect the selectivity
cellulose-based column, there was only a single of analytes. When analyzing myclobutanil, a
peak that eluted. However, the helical structure simple position change of the methyl group
of amylose allowed for the separation of the allowed for the enantiomers to be separated
enantiomeric pair of tebuconazol and resulted in completely (Figure 4).
two peaks.
The chiral selector that is attached to Selectivity from the Mobile Phase
the polysaccharide base can also affect the The content and composition of the mobile
selectivity of a column. The chiral selector phase can also affect the selectivity of a chiral
directs analytes towards the sugar moiety, separation. The addition of an additive or
thereby allowing for more specific selectivity adjusting the concentration of an additive can

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 The Column www.chromatographyonline.com Dhandapani and Tackett

Figure 3: Selectivity from the chiral selector. Figure 4: Selectivity from position of chiral selector.

CH3 CH3
900000 Cl
H3C 750000
NH 600000
Intensity
500000 Analyte:

Intensity
O
O 400000 O NH
0 Myclobutanil
Analyte: Fenarimol
Cellulose

Lux Cellulose-1 200000 O

0.0 0.2 0.4 0.6 0.8 1.0 1.2 1.4 1.6 1.8 2.0 2.2 2.4 2.6 2.8 3.0 3.2 Cellulose tris(3,5-dimethylphenylcarbamate)
Retention time (min) Cl 0 Cellulose

CH3 0.0 0.2 0.4 0.6 0.8 1.0 1.2 1.4 1.6 1.8 2.0 2.2 2.4 2.6 2.8 3.0 3.2 Lux Cellulose-2
Cl Retention time (min) Cellulose tris(3-chloro-4-methylphenylcarbamate)
900000
Intensity

Cl 500000
O NH H3C
OH C Cl
0 O N 600000
H3C
Cl 0.0 0.2 0.4 0.6 0.8 1.0 1.2 1.4 1.6 1.8 2.0 2.2 2.4 2.6 2.8 3.0 3.2 Cellulose
N N
Retention time (min) Lux Cellulose-2 400000
Cellulose tris(3-chloro-4-methylphenylcarbamate) N
NH

Intensity
800000 CH3
O
200000
Intensity

600000
400000
O
N N 200000 0
O Cellulose
0 O
0.0 0.2 0.4 0.6 0.8 1.0 1.2 1.4 1.6 1.8 2.0 2.2 2.4 2.6 2.8 3.0 3.2 Cellulose
0.0 0.2 0.4 0.6 0.8 1.0 1.2 1.4 1.6 1.8 2.0 2.2 2.4 2.6 2.8 3.0 3.2 Lux Cellulose-4
Retention time (min) Lux Cellulose-3 Retention time (min)
Cellulose tris(4-chloro-3-methylphenylcarbamate)
Cellulose tris(4-methylphenylcarbamate)

800000 Cl
H3C
Intensity

600000
400000
200000
0
O
O
NH
D-form eluting first. As the temperature was specific polysaccharide column does not resolve
0.0 0.2 0.4 0.6 0.8 1.0 1.2 1.4 1.6 1.8 2.0 2.2 2.4 2.6 2.8 3.0 3.2 Cellulose

Retention time (min) Lux Cellulose-4

Cellulose tris(4-chloro-3-methylphenylcarbamate)
increased, the enantiomers were coeluted, and the enantiomers; however, with reversed‑phase
then finally a reversal of elution order was seen mode, the compounds were separated on
not only affect the separation of enantiomers was analyzed. Under the current conditions with at 50 °C. The peak shape and peak efficiency the same column. Chiral separation is a
but can also change the elution order. This can a two-component mobile phase, one of the were higher at higher temperatures (8). three‑dimensional separation, unlike traditional
be seen with Fmoc-N-Isoleucine separation. As critical pairs was not separated. However, with reversed-phase separation. In reversed-phase
the concentration of formic acid (FA) increased the addition of a third component to the mobile Complementary Selectivities chromatography using a C18 column, highly
in the mobile phase, the two enantiomers phase—the addition of 6% dichloromethane to Finally, the combination of the mobile phase probable predictions can be made by looking at
eluted together to form one peak, and then the water and acetonitrile mixture—brought the and the chiral stationary phase can affect the structure of molecules, their hydrophobicity
changed elution order, with the L- form eluting critical pair resolution. The use of a chlorinated selectivity of enantiomers. The same mobile difference, and log P (octanol-water partition
first with 0.5% FA in the mobile phase. This solvent such as chloroform or dichloromethane phase was used with different stationary phases coefficient). However, with chiral separation,
same shift was seen when FA was maintained are allowed only on specialized phases (Figure 6). to analyze Fmoc-N-Isoleucine, giving different one should screen analytes through multiple
and the ratio of the other two components peak shapes, separation, and elution order. It phases and multiple modes to get the optimal
of the mobile phase were changed. As the Temperature-Based Selectivity is advantageous to screen multiple phases for resolution (9). Once the screening results are
percentage composition of hexane increased, Temperature change can bring a drastic a higher chance of success and to obtain the explored and the best separation is identified,
the selectivity change was clearly visible from change in selectivity in chiral separation. desired order of elution. further fine-tuning can then be done to
the flip in elution order of enantiomers of Fmoc-N‑Isoleucine was further explored for optimize the method for the ideal separation (8).
Fmoc‑n-Isoleucine (Figure 5). the effect of temperature. The stationary Chiral Screening
Selectivity can be enhanced by the addition phase and mobile phase were maintained, When screening a specific chiral isomer by Conclusion
of a mobile phase additive. For example, an but the temperature was increased. At 5 °C normal, reverse, and polar organic mode, the Chiral separation is three-dimensional, and
analyte with three chiral centres (eight isomers) the enantiomers were fully resolved, with the initial screening in normal phase with one the resolution of a given separation can

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17 Dhandapani and Tackett 21 Q&A Ueland 25 SCM-10 Event Preview 27 Training and Events
 The Column www.chromatographyonline.com Dhandapani and Tackett

Figure 5: Selectivity from mobile phase composition. Figure 6: Selectivity from adding a third component to the mobile phase.
Fmoc-L-isoleucine and Fmoc-D-isoleucine with Lux® Cellulose-1 A = 0.1% TFA-water
D L n-Hex/IPA/FA = 60/40/0.1
B = Acetonitrile Critical Pair
C = Dichloromethane Resolved
CH3

H3C
Gradient
O
NH Min %A %B %C
O 0 43 51 6
n-Hex/IPA/FA = 85/15/0.1 Cellulose 15 40 54 6
Lux Cellulose-1
D+L Cellulose tris(3,5-dimethylphenylcarbamate)

3rd component DCM in

mobile phase brings the
selectivity
n-Hex/IPA/FA = 95/5/0.1 Need special
“immobilized columns
D
L in order to use harsh
chlorinated solvent like
DCM

be improved by optimizing the selectivity.
Selectivity in chiral separation can come from
the mobile phase, the stationary phase, and
the temperature of the analysis. To obtain
high resolution and the right peak order
that meets the goal of the analysis, it is
important to consider optimizing all these
aspects of selectivity before settling on the
separation. Chiral screening is a prudent
approach to explore the selectivity aspects
of chiral separation.
References
1. Burley, D. M.; Lenz, W. Thalidomide and Congenital
Abnormalities. Lancet 1962, 279, 271–272. DOI:
10.1016/S0140-6736(62)91217-5
2. Lenz, W. A Short History of Thalidomide
Embryopathy. Teratology 1988, 38, 203–215. DOI:
10.1002/tera.1420380303
3. Diggle, G. E. Thalidomide: 40 Years On. Int. J. Clin.
Pract. 2001, 55, 627–631.
4. Stephens, T. D. Proposed Mechanisms of Action in
Thalidomide Embryopathy. Teratology 1988, 38,
229–239. DOI: 10.1002/tera.1420380307
5. Bartlett, J. B.; Dredge, K.; Dalgleish, A. G. The
Evolution of Thalidomide and its IMiD Derivatives as
Anticancer Agents. Nat. Rev. Cancer 2004, 4, 314.
DOI: 10.1038/nrc1323
6. Wu, J. J.; Huang, D. B.; Pang, K. R.; Hsu, S.; Tyring,
S. K. Thalidomide: Dermatological Indications,
Mechanisms of Action and Side-Effects. Br. J.
Dermatol. 2005, 153, 254–273. DOI: 10.1111/j.1365-
2133.2005.06747.x
7. Melchert, M.; List, A. The Thalidomide Saga. Int.
J. Biochem. Cell Biol. 2007, 39, 1489–1499. DOI:
10.1016/j.biocel.2007.01.022
8. Chankvetadze, B.; Farkas, T.; Peng, L.; et
al. Enantiomer Elution Order Reversal of
Fluorenylmethoxycarbonyl-Isoleucine in High-
Performance Liquid Chromatography by Changing
the Mobile Phase Temperature and Composition.
J. Chromatogr. A 2011, 1218, 6554–6560. DOI:
10.1016/j.chroma.2011.06.068
9. Phenomenex, Chiral HPLC/SFC Method
Development Poster. https://www.phenomenex.
com/ViewDocument?id=chiral+hplc_
sfc+method+development+poster&fsr=1 (accessed
2022-12-19).
Ramkumar Dhandapani has been
in the chromatography industry
for over 20 years and has hands-on
and troubleshooting experience in
chromatography. He has earned a
master’s degree and doctoral degree
in analytical chemistry from Seton
Hall University, New Jersey, USA, with
specialization in microextractions,
multidimensional gas chromatography,
and tandem mass spec techniques. He
has developed and validated several
regulatory compliant methods in the
pharmaceutical, food, and environmental
industries. He joined Phenomenex
in August 2014 and serves as Global
Product Manager-Gas Chromatography at
Phenomenex, USA.
Bryan Tackett earned a bachelor’s degree
in biochemistry and genetics from Texas
A&M University (Texas, USA) and a
doctoral degree in translational biology
and molecular medicine from Baylor
College of Medicine (Texas, USA). He
joined Phenomenex in September of 2020
and he serves as Technical Marketing
Writer at Phenomenex, USA.
E-mail: ramkumard@phenomenex.com
Website: www.phenomenex.com/

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 A Focus on Forensic
Taphonomy and
Chromatography
The Column interviewed Maiken Ueland, an ARC discovery early career
research fellow at the Centre for Forensic Science and deputy director of
The Australian Facility for Taphonomic Experimental Research (AFTER) at
the University of Technology Sydney in Australia, on her work in forensic
taphonomy, where she uses analytical, biochemical, and spectroscopic
techniques to conduct human post-mortem investigations.

—Interview by Kate Jones

Q. What are your main research my area is called—allows me to try and

interests, and what led you to the field
of forensic taphonomy?
A: My main research interest lies within the
intersection between analytical chemistry,
technology, and forensic science. One of
the main areas I am involved in is creating
new technology and methods to find
victims in mass disasters.
I loved solving puzzles as a kid and I
was always very curious; then when I got
freshidea/stock.adobe.com
understand how the human body works
and I can use that knowledge to help law
enforcement agencies. I want to be a voice
for victims, and also prevent future crimes.
Q. What exactly is forensic taphonomy?
A: Forensic taphonomy is the study of
everything that happens to an organism
from the moment it dies until it is found.
We use the information gained as the

older I found science and I realized how body breaks down to find missing persons
much science could help people. I realized and victims of crime. We also use the
how forensic taphonomy—which is what information to determine how long ago

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 The Column www.chromatographyonline.com
and how a person died, and also how to
identify individuals so that we can help
assist law enforcement.
Q. How do you develop
chromatographic strategies for this
research? What chromatographic
techniques do you use in
your research?
A: In my area we deal with very complex
samples. For example, the volatilomes
(volatile organic compound [VOC] profiles)
of humans—which is something I analyze
for both living and deceased individuals—
are incredibly complex and can be made
up of 1000s of compounds. The biggest
challenge is to have sufficient tools to
both capture and analyze these complex
profiles (odours).
For the volatile work this is mainly
untargeted, which means we do not know
the identity of everything we are looking
for. We use in-built libraries and external
standards to try and identify these, but
since there are so many this is a major
challenge. We also do not know until we
have done a lot of statistical analysis which
compound(s) are going to be important for
our research.
After years of research across the globe,
we do know some of the biomarkers in
the scent of decomposing human remains,
2 McGregor et al.
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17 Dhandapani and Tackett
Q&A Ueland
which allows us to use a more targeted
approach for those. However, because
a lot of research has been done using Maximize LC-MS/MS
pigs or with different analytical
instrumentation, we are still not fully
Performance with Restek
aware of all of the compounds that
Why choose Restek LC columns? Because exceptional LC-MS/MS performance is
will be important. The goal is to use built into them every step of the way. From new product R&D to manufacturing to
these biomarkers to develop field-based applications, the pillars for LC-MS/MS success are ingrained in every Restek LC column.
technologies.
• Stable retention times for tight MRM windows so you
I use more sophisticated analytical can analyze large analyte lists with confidence.
instruments such as comprehensive • Rugged manufacturing and rigorous LC-MS/MS testing
two‑dimensional gas chromatography ensure consistent column-to-column performance.
time‑of-flight mass spectrometry (GC×GC– • Wide range of formats and phases provides the
speed and selectivity you need.
TOF-MS) for VOC/volatilome analysis,
• Raptor SPP columns deliver the speed
which is beneficial over GC–MS for of core-shell technology.
complex profiles. • Force FPP columns are fully scalable
from HPLC to UHPLC.
For a more targeted approach we
have used GC–inductively coupled plasma
(ICP)‑MS, as mentioned below.
I also work on targeted approaches
when analyzing lipids from textiles or
tissue samples. This work is focused on Built for LC-MS/MS
High-Performance Raptor and Force Columns from Restek
attempting to create better ways
to determine time since death (how
long someone has been deceased for),
and is performed mostly using
GC–MS/MS.
Q. You use a range of analytical,
biochemical, and spectroscopic Recharge your LC-MS/MS Methods
techniques to conduct human www.restek.com/LC
post‑mortem investigations, and
7 News 9 Tips & Tricks 13 Rising Stars
21 Q&A Ueland 25 SCM-10 Event Preview 27 Training and Events
 The Column www.chromatographyonline.com
in a recent paper you analyzed the
decomposition of human remains in
a simulated building collapse using
GC×GC–TOF-MS (1). Please can you
talk about your findings.
A: One of my main areas involves
the use of VOCs to find missing
persons in mass disasters. In this
paper we had two large simulated
mass disaster events using six donated
humans. One of the major findings
was the degree of differential
decomposition we saw upon recovery
of the victims. The donors were
purposely placed in single or commingled
configurations, as this is something
that can occur when disasters happen
in densely populated areas. This was
suspected to affect the decomposition,
but it had not been studied until
this time.
The VOC profiles in the area were also
examined and we could use them to
determine at which point of decomposition
the majority of the remains were in. This
is particularly important in the training of
scent detection dogs.
The VOC profiles also add to the
database and search for biomarkers for
victim localization. The ultimate aim is to
develop portable technology for field use
that uses these biomarkers.
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17 Dhandapani and Tackett
Q. What are the advantages of GC×GC–
TOF-MS over existing techniques? Are
there any disadvantages?
A: The primary difference between GC–
MS (a common method for VOC analysis)
and GC×GC–TOF-MS is the addition
of a secondary column in GC×GC,
which enables further separation of
compounds with similar sizes and chemical
properties. This additional column is
generally of a different polarity than
the first, which increases the separation
ability and reduces coelution of chemical
compounds. GC×GC–TOF-MS is preferred
when analyzing biological volatilomes
due to its ability to accurately detect
more compounds through its enhanced
separation ability and its increased peak
capacity when analyzing complex samples.
A disadvantage of using GC×GC–TOF-MS
is that it does not have high-resolution
(HR) MS. It is possible to get a GC×GC–
HR–TOF-MS system to help improve the
identification of unknowns, but this is a
very costly instrument.
Q. Where do you source the
human cadavers?
A: We have a body donation programme
at the university. People donate their body
to science and the donors we get are
individuals who have specifically requested
7 News
21 Q&A Ueland
for their body to be used for taphonomic
experiments. Without these absolutely
amazing donations we would not be able
to do any of the work we do. We are
incredibly grateful for their gift.
Q. What are the challenges that you
face when planning and performing
these studies?
A: One of the biggest challenges is
the number of variables that we are
dealing with when analyzing living and
deceased individuals. These variables
include differences amongst individuals
and sampling during different seasons
and temperatures. We are also reliant on
receiving donors. We need a good breadth
of donors over all seasons in our specific
scenarios, which means that the work
might span over years.
Q. In another recent study, you
used GC–ICP-MS for the first time to
investigate VOCs from human remains
(2). What led you to consider this
technique in this research?
A: This work came about due to a
collaboration with David Clases. We were
looking for a way to accurately quantify
the level of mainly sulfur components
from decomposing human remains over
time. Sulfur components are known to
9 Tips & Tricks
25 SCM-10 Event Preview
Q&A Ueland
be important biomarkers present in the
odour emitted from human remains. A
lot of work is involved in attempting to
determine the constituents in the human
volatilome, but this does not really look
at the concentrations. We therefore used
ICP-MS after separation by GC to get the
concentrations as an improved alternative
to GC–MS. The concentrations were
found to be time-dependent and showed
potential as forensic markers to determine
post-mortem intervals.
Q. The development of an electronic
nose to detect human cadavers came
about after its initial development
in detecting illegally traded wildlife.
Please can you discuss how
this evolved.
A: The development of the electronic
nose was a collaboration between Steven
Su, Shari Forbes, and Wentian Zhang.
This initially started as colleagues noted
a smell when preparing illegally traded
horn samples for DNA sampling. We were
therefore curious to see whether scent
could be used to detect and potentially
give a preliminary identification of wildlife
species. This work was initially performed
using GC×GC–TOF-MS and showed
promising results (3). We therefore got
together and decided to develop electronic
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 The Column www.chromatographyonline.com
nose technology to allow for fast and
on‑site detection of illegally traded wildlife.
This work has evolved over the last six
years and we are now also developing
technology to detect human remains using
the same principles.
Q. You have also been involved
in another interesting forensic
analysis project involving “reptile
volatilome profiling optimization” (4).
Can you elaborate on what this
project involves and what your
main findings were?
A: This project is part of the work on
the illegal wildlife trade. We have
previously analyzed various commonly
encountered traded items such as ivory
and rhino horns using GC×GC–TOF-MS.
But being from Australia, our collaborator,
the Australian Museum, was also very
interested in testing whether VOC analysis
could be used to locate a commonly
traded lizard species and stop items
before they are illegally traded out of
Australia. Amber Brown, Greta Frankham,
Barbara Stuart, and I therefore initiated
a project where instead of looking
at wildlife parts we analyzed living
lizards. In the published work, the VOC
collection and analysis was optimized.
This was the first time that VOCs of
2 McGregor et al.
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17 Dhandapani and Tackett
lizards were collected, and we therefore
needed to create a new method. This
work demonstrated the complex profiles
of reptiles and also showed by using
the VOC profiles how to distinguish
shingleback lizard, eastern blue tongue
lizard, and Children’s python from
each other.
The aim of this work was to also
standardize a volatilome collection method
with limited influences from secondary
or tertiary factors that may reflect the
biological status of the animal (metabolism,
health status). This was to ensure that in
the future we select biomarkers that will
encompass all of the lizards within the test
species rather than the biological factors
that may separate young from old or the
different sexes.
Q. What analytical challenges did
you encounter and how did you solve
them?
A: There was no other work profiling
the volatilome of lizards on this scale.
We therefore had to take a broad
screening approach, as we did not know
which compounds to expect. To create
the best possible method, we also had to
optimize a number of parameters.
The biggest challenge in any optimization
is to optimize parameters that affect
7 News
21 Q&A Ueland
each other. This usually results in many
differing combinations having to be
tested. The work also demonstrated
the need for improved statistical analysis
when dealing with biological specimens.
References
1. Ueland, M.; Harris, S.; Forbes, S. L.
Detecting Volatile Organic Compounds
to Locate Human Remains in a Simulated
Collapsed Building. Forensic Science
International 2021, 323, 110781. DOI:
10.1016/j.forsciint.2021.110781
2. Clases, D.; Ueland, M.; de Vega, R. G.;
Doble, P.; Pröfrock, D. Quantitative
Speciation of Volatile Sulphur Compounds
from Human Cadavers by GC-ICP-MS.
Talanta 2021, 221, 121424. DOI: 10.1016/j.
talanta.2020.121424
3. Ueland, M.; Ewart, K.; Troobnikoff, A. N.; et al.
A Rapid Chemical Odour Profiling Method for
the Identification of Rhinoceros Horns.
Forensic Science International 2016, 266,
e99–e102. DOI: 10.1016/j.forsciint.2016.05.
011
4. Brown, A. O.; Frankham, G. J.; Stuart,
B. H.; Ueland, M. Reptile Volatilome
Profiling Optimisation: A Pathway
Towards Forensic Applications. Forensic
Science International: Animals and
Environments 2021, 1, 100024. DOI: 10.1016/j.
fsiae.2021.100024
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Q&A Ueland
Maiken Ueland is currently
an ARC discovery early
career research fellow at
the Centre for Forensic
Science and deputy
director of The Australian Facility for
Taphonomic Experimental Research
(AFTER) at the University of Technology
Sydney in Australia. She is an emerging
leader in the field of forensic
taphonomy, where she uses analytical,
biochemical, and spectroscopic
techniques to conduct human
post‑mortem investigations. Her main
research areas are human
decomposition chemistry, with a special
focus on markers in tissue and odour
and their use in criminal investigations,
including locating missing persons and
estimating time since death. Her
interest lies in the interface between
forensic science and analytical
chemistry. Her current work focuses on
developing methods for the successful
location and recovery of victims in
mass disaster scenarios using electronic
nose technology and sophisticated
analytical instrumentation.
E-mail: maiken.ueland@uts.edu.au
Website: www.uts.edu.au/about/
faculty-science/after-facility/about-us
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 SCM-10 Event Preview
Peter Schoenmakers, University of Amsterdam, Amsterdam, the Netherlands

The Tenth International Symposium on the Separation and Characterization

of Natural and Synthetic Macromolecules, SCM-10, will take place 1–3
February 2023 in Amsterdam, the Netherlands. The organizing committee
reveal what attendees can look forward to.

There is a great deal of interest in the
subject of large-molecule characterization,
be it synthetic polymers—where we have
learned that conventional characterization
methods fall short in guiding innovation;
recycled polymers—the safety and efficacy
of which cannot yet be properly assessed;
biopolymers, such as cellulose and lignin,
that are still notoriously difficult to
characterize; or proteins—the behaviour of
which is affected by the smallest details in
their structure. Scientists working on these
various types of macromolecules rarely meet,
because they are anchored in different fields.
However, they share a number of common
techniques, or at least technology with a
common basis. On this communal ground
scientists will meet for the tenth time at
the SCM conference on the Separation
and Characterization of Macromolecules in
gnoparus/stock.adobe.com
many forms, but especially size‑exclusion
chromatography (SEC), and mass
spectrometry (MS). One-day short courses will
be presented on these key subjects on the
two days preceding the conference. Laurence
Charles, from the Institute for Radical
Chemistry at the Aix-Marseille University
(Marseille, France), will discuss various aspects
of polymer mass spectrometry on Monday 30
January. She will present the basics but also
some of her own eye-catching results. On
Tuesday 31 January, André Striegel, from the
National Institute of Standards & Technology
(NIST) in Gaithersburg (Maryland, USA), will
discuss liquid chromatographic techniques
for polymer separation and characterization.
André is arguably the leading expert in
this field. Both courses offer fantastic
opportunities for students and professionals
to refresh the basics and to catch up with

Amsterdam on 1–3 February 2023. developments in the field.

Common technology for all the above Other liquid-phase separation
fields include liquid chromatography (LC) in techniques will be discussed at the

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17 Dhandapani and Tackett 21 Q&A Ueland 25 SCM-10 Event Preview 27 Training and Events
 The Column www.chromatographyonline.com
conference, including gradient LC,
hydrophobic‑interaction chromatography
(HIC), hydrodynamic chromatography
(HDC), capillary electrophoresis (CE), and
various two‑dimensional techniques,
including comprehensive two-dimensional
LC (LC×LC). There is a great deal of interest
in field-flow fractionation (FFF) techniques,
a field that has been steadily growing
during the 20 years of existence of the SCM
conference. FFF offers exciting possibilities
for characterizing very large molecules and
nanoparticles, as encountered, for example,
in drug formulations.
Separation methods go hand in hand
with characterization methods, although
it can be argued that MS is a bit of
both. SCM will feature presentations on
combinations of MS with LC and SEC,
pyrolysis-gas chromatography (Py-GC),
2 McGregor et al.
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17 Dhandapani and Tackett
ion-mobility spectrometry (IMS), and
even FFF. However, MS is not always the
answer. MS provides accurate masses,
but molecules need to be brought into
the gas (or near-vacuum) phase and the
quest for measurements under “native”
conditions has not come to a final
conclusion—measurements in the liquid
phase, using spectroscopic techniques,
such as nuclear magnetic resonance (NMR)
(is it really quantitative?), viscometry, and
light‑scattering detectors. The latter
are still as tightly interwoven with the
SCM field as MS, and the application
domains of MS and light scattering
overlap only partially.
All these tools bring new insights in the
many applications that come together at
SCM, including materials and coatings,
recycling, bio(-based)-materials, (bio-)
7 News
21 Q&A Ueland
pharmaceuticals, and drug formulations.
An eye-catching application of the many
technologies being discussed is the
characterization of objects of cultural
heritage, which will be discussed in at least
a handful of presentations, ranging from
protein binders to Asian lacquers.
People involved in the separation and
characterization of natural and synthetic
macromolecules share a great deal of
common interest and they have a lot
to discuss. The SCM community has
not been together for four years—for
obvious reasons—and the opportunity
to have a live meeting again is exciting.
The community has responded with a
record number of submitted presentations,
resulting in what is possibly the strongest
ever SCM scientific programme. The SCM
conference also offers a unique atmosphere
9 Tips & Tricks
25 SCM-10 Event Preview
SCM Event Preview
that encourages interactions between all
scientists, students, exhibitors, and other
delegates. It is the perfect place to meet
and to get to know colleagues, to renew
friendships, and to make new friends. For
science and for life.
Peter Schoenmakers has been affiliated
with the University of Amsterdam as
a professor of analytical chemistry for
nearly 25 years. His research has had
a strong focus on the separation (and
characterization) of macromolecules and
on multidimensional and hyphenated
techniques. He is the scientific chairman
of the SCM-10 conference.
E-mail: info@scm-10.nl
Website: https://scm-10.nl
13 Rising Stars
27 Training and Events
 The Column www.chromatographyonline.com Training & Events

Training Courses
GC
GC Introduction
Website: www.chromacademy.
com/channels/gc-training-courses/
principles/gc-introduction
GC Troubleshooter
Website: www.chromacademy.
com/channels/gc-training-courses/
troubleshooting/gc-troubleshooter
Operating and Understanding GC
Website: www.crawfordscientific.
com/training-consultancy/gc-training/
gc-fundamentals
GC Headspace
Website: www.crawfordscientific.
com/training-consultancy/gc-training/
gc-headspace
Hands-On GC×GC
18–19 May 2023
The Open University, Milton Keynes, UK
Website: www.anthias.co.uk/training-
courses/hands-on-GCxGC
HPLC/LC–MS
Understanding HPLC
Website: www.crawfordscientific.com/
training-consultancy/hplc-training/hplc-
fundamentals
HPLC Troubleshooter
Website: www.chromacademy.
com/channels/hplc-training-courses/
troubleshooting/hplc-troubleshooter
Fundamentals of LC–MS
Website: www.chromacademy.com/
channels/lc-ms/principles/fundamentals-
of-lc-ms-video-training-course
LC–MS Introduction
Onsite training
Website: www.chromacademy.
com/channels/lc-ms/principles/lc-ms-
introduction
Complete HPLC and LC–MS
27–31 March 2023
Virtual
Website: www.anthias.co.uk/training-
courses/completeLC
SAMPLE PREPARATION
Fundamentals of Solid-Phase
Extraction (SPE) Mechanisms
Online training
Website: www.chromacademy.com/
channels/sample-preparation/technique/
fundamentals-of-spe-mechanisms
Hands-On Solid-Phase
Microextraction
17 March 2023
The Open University, Milton Keynes,
UK
Website: www.anthias.co.uk/training-
courses/hands-on-SPME
MISCELLANEOUS
Introduction to Infrared (IR)
Spectroscopy
Online webcast from
CHROMacademy
Website: www.chromacademy.
com/channels/infrared/principles/
introduction-to-infrared-spectroscopy
Validation, Verification, and
Transfer of Methods for
Biopharmaceutical Analysis
6–9 March 2023
Online—virtual
Website: https://
mournetrainingservices.com/method-
validation-biopharma-course/
Please send your event and training
course information to Kate Jones
kjones@mjhlifesciences.com

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 The Column www.chromatographyonline.com Training & Events

Event News
11–14 April 2023
ANAKON 2023

Vienna, Austria

Email: office@anakon2023.at

Website: www.anakon2023.at

21–24 May 2023

8th International Conference on Polyolefin Characterization (ICPC)

Valencia, Spain

Email: Raquel.ubeda@icpc-conference.org

Website: www.icpc-conference.org

18–22 June 2023

HPLC 2023

Düsseldorf, Germany

Email: michael.laemmerhofer@uni-tuebingen.de / oliver.schmitz@uni-due.de

Website: www.hplc2023.com

17–22 September 2023

6th International Mass Spectrometry School

Cagliaria, Sardinia

Email: gianluca.giorgi@unisi.it

Website: www.spettrometriadimassa.it/imss2023/#hero

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17 Dhandapani and Tackett 21 Q&A Ueland 25 SCM-10 Event Preview 27 Training and Events
 Contact Information
Group Publisher Editor-in-Chief Headquarters: Custom Projects
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2 McGregor et al. 7 News 9 Tips & Tricks 13 Rising Stars

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17 Dhandapani and Tackett 21 Q&A Ueland 25 SCM-10 Event Preview 27 Training and Events