download pdf version - Separation Science

MS Solutions 

Separation Science ‘MS Solutions’ is the premier online resource for analytical scientists working with mass spectrometry across 

Europe, the USA and the Middle East. Covering MS method fundamentals, practicalities and troubleshooting it offers 

chromatographers and analytical chemists a genuine e-learning platform and searchable archive resource. 

Tech Tip 

Peptide Sequencing with Electrospray LC/MS 

Part 2: Interpretation of a Simple Spectrum 

Featured Applications 

The Scheduled MRM Algorithm Enables Intelligent 

Use of Retention Time During Multiple Reaction 

Monitoring 

Using Core-Shell Kinetex XB-C18 HPLC Columns as a 

Solution for Analyzing Fluoroquinolone Antibiotics by 

LC/MS 

Rapid Tea Analysis on Poroshell 120 SB-C18 with LC/MS 

Analysis of Water-Soluble Polymers Using Linear Size 

Exclusion HPLC Columns and a Semi-Micro SEC System 

In last month’s MS Solutions we discussed MS/MS fragmentation of 

polypeptides, the types of ions formed, and the mechanism of their 

formation. This month we will examine a tandem mass spectrum of 

a simple polypeptide and step through an interpretation strategy 

leading to the complete sequence determination. 

GC/MS Analysis of Phthalates in Children’s Products 

Accurate and reliable analysis of beer using time-offlight 

technology for gas chromatography 

Click titles to learn more 

Click here to read more... 

e-learn.sepscience.com/mssolutions Issue 10

Peptide Sequencing with Electrospray LC/MS 

Part 2: Interpretation of a Simple Spectrum 

Frederick Klink 

In last month’s MS Solutions we discussed MS/MS fragmentation of polypeptides, the types 

of ions formed, and the mechanism of their formation. This month we will examine a tandem 

mass spectrum of a simple polypeptide and step through an interpretation strategy leading 

to the complete sequence determination. 

To help you in understanding the interpretation 

process we have provided a table of the 20 

naturally occuring amino acids. Click on this link 

(Table 1) and the table will open as a PDF file in a 

separate window so you can refer to it as you read 

the rest of this article. 

The headers on the table, reading from left to 

right, are: 

1. Amino acid name and residue composition (a 

“residue” is the amino acid minus water) 

2. The two commonly used abbreviations, i.e. the 

three-letter and the single-letter forms 

3. The amino acid residue nominal (integer) mass, 

monoisotopic mass, and average mass 

4. The immonium ion nominal mass; immonium 

ions have this form 

where R SC 

is the side chain. 

5. Side chain nominal mass 

6. Structure of the amino acid 

The table is divided into three sections according 

to the polarity/ionic nature of the amino acid sidechain. 

MS/MS experiments with tandem transmission 

quadrupole (“triple quad”), instruments, 

quadrupole ion traps, and Q-ToF or IT-ToF hybrids 

employ low-energy CAD (collisionally-activated 

dissociation). Low-energy CAD of polypeptides 

results largely in formation of b and y ions, i.e., ions 

formed by cleavage of the peptide bonds. b ions 

are often accompanied by their corresponding a 

ion as well. (High-energy CAD will be discussed in 

a later instalment in this series.) 

Our example peptide is the same one we looked 

at last month with the sequence methioninetyrosine-glycine-alanine-valine, 

or MYGAV. The 

protonated molecule of this polypeptide has a 

nominal mass and mass-to-charge ratio (m/z) of 

540 and will be used as the precursor ion in our 

MS/MS experiment. Figure 1 is the mass spectrum 

resulting from this experiment. 

In a low-energy CAD experiment we start by 

looking for the y and b series of ions. The first,

Figure 1 

Figure 1: Mass spectrum resulting from the MS/MS fragmentation of a single-charge peptide precursor ion of m/z 

540. Red arrows indicate identification of y ion series; blue arrows are the b ion series. 

or highest mass, y ion is identified by finding the neutral loss is of a single 

amino-acid and residue mass from the precursor ion. In other words this 

neutral loss must be between 57 (glycine) and 186 (tryptophan). Because 

the y ions are C-terminal product ions, this neutral loss represents the first 

N-terminal amino acid residue. 

The first b ion results from the loss of a single amino-acid residue mass 

plus 18 from the precursor. Because b ions are N-terminal product ions, the 

neutral loss of the first C-terminal amino acid will contain the C-terminal – 

OH plus an additional hydrogen which accounts for the 18 additional mass 

units over the residue mass. 

Once the first ion in each series is identified, the subsequent ions in 

the series are identified by finding a peak in the mass spectrum which 

represents the neutral loss of an amino acid residue mass from the 

immediately preceeding peak in the series. 

This differs from classic mass spectral interpretation in which we usually 

look for neutral losses from the precursor ion. 

The interpretation of the mass spectrum is shown graphically in 

Figure 1 and in tabular form in Table 2. Looking first at the y ion series: 

the first y ion is identified by noting a peak at m/z 409 which represents 

a neutral loss of 131 from the precursor ion. Refering to Table 1, we find 

that 131 is the residue mass of methionine. So we can tentatively assign 

methionine as the N-terminal amino acid in this polypeptide. Realizing 

that these assignments are tentative at this early stage in the process is 

important. We may find that things do not add up and that we are forced to 

Discover TsKgel HiLic soLuTions for HPLc 

TSKgel aMIde-80 - no.1 for gLyCo-MappIng 

SMaLL parTICLe SIzeS for hIgh effICIenCy 

TSKgel nh2-100 - robuST aMIno bonded phaSe 

vIrTuaL abSenCe of bLeedIng, IdeaL for MS 

The TSKgel hILIC porTfoLIo IS a SeLeCTIon of STabLe, SILICa baSed hILIC phaSeS SuITed for a varIeTy 

of appLICaTIonS. vISIT our webSITe or TaLK To one of our TeChnICaL SpeCIaLISTS aT hpLC June 19-23,2011, 

budapeST, booTh M18, To Learn how our produCTS CouLd SoLve your SeparaTIon needS. 

STuTTgarT, gerMany | phone + 49 [0]711 · 13257 - 0 

fax + 49 [0] 711 · 13257 - 89 | info.tbg@tosoh.com 

KIng of pruSSIa, uSa | phone + 1 [800] 366 - 4875 

fax + 1 [610] 272 - 3028 | info.tbl@tosoh.com 

www.tosohbioscience.com

Table 1 

Ion m/z Neutral loss 

(from previous ion in 

the series) 

Precursor [M+H] + 540 

Amino Acid 

Residue 

y 4 409 131 M 

y 3 246 163 Y 

y 2 189 57 G 

y 1 118 71 A 

b 4 423 117 (99+18) V 

b 3 352 71 A 

b 2 295 57 G 

b 1 132 163 Y 

ADVANCE YOUR KNOWLEDGE IN 

SEPARATION SCIENCE WITH 

CHROMATOGRAPHY FORUM 

What is Chromatography Forum? 

Chromatography Forum is a public discussion group where you can post questions, news, or 

messages of interest to chromatographers everywhere. Please join in! Discussion forums exist for LC, GC, 

hyphenated systems, sample prep and data systems. Anyone can read messages and discussions, but 

you must be a registered member to post messages (registration is FREE). 

Find out more>> 

a 4 395? 

a 3 324 

a 2 267 

a 1 104 

Table 2: Tabular summary for the interpretation of the mass spectrum shown in Figure 1. 

start over. 

Subsequent y ions are identified in the same way, i.e., observing a neutral 

loss which is equivalent to an amino acid residue mass. Referring to 

Figure 1 and Table 2, we find the y series of ions gives us the sequence 

M-Y-G-A-. The y 1 ion represents the C-terminal amino acid residue which is 

identified by subtracting 19 from the y 1 ion mass (17 for the C-terminal –OH, 

1 for the N-terminal –H and 1 for the proton which imparts the +1 charge). 

This results in a residue mass of 99 which we can determine from Table 1 is 

valine. We are fortunate to observe a complete series of y ions which give us 

the complete polypeptide sequence of MYGAV. 

We identify the first b ion at m/z 423 by looking for the neutral loss of a 

residue mass +18. This residue mass is 99 or valine which corresponds to the 

identification of the C-terminal amino acid which we made based on the 

y 1 ion. That’s good news; we are on the right track! The subsequent b ions 

are found by looking for neutral losses which correspond to residue masses 

and thus we find (reading from the C-terminus), V-A-G-Y-. The b 1 ion at 132 

represents the N-terminal amino acid residue mass plus one (to account for 

ADVICE 

ANSWERS 

SAMPLE PREP 

EMPLOYMENT 

MASS SPEC DATA SYSTEMS 

GAS CHROMATOGRAPHY 

REVIEWS 

GC-MS 

LIQUID CHROMATOGRAPHY 

ANOUNCMENTS 

PROBLEM SOLVING 

LC-MS 

QUESTIONS 

www.chromforum.org

J ap de Z euw 

Uwe D. Neue 

Non 

Con 

Ben 

Cl 

the proton), which is 131 or methionine, giving us 

a complete polypeptide sequence, MYGAV. This 

corresponds exactly with the results from the y 

series. 

Note in Table 2 that a nearly complete series of 

a ions are also present. These are easily identified 

as being 28 m/z units (and 28 u), less than the 

corresponding b ions. The only questionable one 

is the a 4 ion which should be at 395. The only 

peak observed in this mass spectrum in that 

region is labelled as 394. This will require us to 

look at the tabular data and/or expand this region 

of the spectrum to see if a 395 peak is present 

or if the peak is labelled incorrectly because of 

rounding. 

The last and extremely important step makes 

certain that the mass of the precursor ion and 

the mass of our tentatively determined sequence 

match up. To complete this step, add up all of the 

residue masses in the sequence, add 17 for the 

C-terminal –OH, add one for the N-terminal –H 

and another one for the proton of the [M+H] + ion. 

We find: 131 +163 +57 +71 +99 +17 +1 +1 = 540 

which matches the selected precursor ion m/z. 

We’re done! 

This example is designed to be simple and to 

work out perfectly as is appropriate for a learning 

environment. In the “real world”, of course most 

of your sequencing work will not be so straightforward. 

We have found that the probability of 

producing any given structurally significant ion 

in peptide analysis is highly dependent on the 

sequence itself. The presence of incomplete ion 

series makes the interpretation more complex. 

Better results can often be obtained with 

multiple-charge precursor ions but this also 

adds complexity to the interpretation because 

product ions will be produced with varying 

charge numbers and thus m/z values which 

may be both higher and lower than that of the 

precursor ion. These and other advanced topics 

in polypeptide sequencing will be discussed in 

future instalments of MS Solutions. 

Fred Klink is a trainer and consultant to the 

pharmaceutical, biotech, and chemical industries 

as well as law enforcement and other government 

laboratories. Fred’s specialty is HPLC, LC/MS, and solidphase 

extraction technologies. 

Fred received a degree in biochemistry from 

Northwestern University and completed graduate 

studies and an internship in forensic chemistry at the 

University of Illinois. After graduation, Fred entered 

the analytical instruments industry where he spent 

seventeen years in varying positions from applications 

chemist, development project manager, and manager 

for strategic planning. Fred has been teaching highly 

regarded MS and LC/MS courses and providing 

consulting services since 1996. 

Fred is the author of several journal articles and 

book chapters including the LC/MS entry in the Wiley 

Encyclopedia of Analytical Chemistry. He is a member of 

the American Chemical Society and American Society 

for Mass Spectrometry 

Volume 3, I sue 3 

February 2011 

Europe 

Accurate and 

Rapid Metabolomics 

Strategies Using 

GC-MS/MS-MRM 

Technology 

www.sepscience.com 

Clinical Edition 

Accurate and Rapid Metabolomics 

Strategies Using GC-MS/MS-MRM Technology 

Lena Fragner, Wolfram Weckwerth, Hans-Joachim Huebschmann 

Simultaneous Measurement of Reduced and 

Oxidized Coenzymes Q9 and Q10 in Rat Tissues 

by LC-MS 

A.V. Pirogov, E.B. Pashkova, A.A. Bendryshev and O.A. Shpigun 

An 

Glycopeptide Analysis of Antibodies by 

Capi lary Electrophoresis and Q-TOF Ma s 

Spectrometry 

GC Solutions 

GC-TOFMS Analysis of Urine Extract Samples 

Used for a Liver Drug-Induced Injury Study 

In-depth Characterization of Neutral and 

Acidic Glycopeptides by ZIC-HILIC Enrichment 

and Ma s Spectrometry 

Separation of Substituted Methoxybenzene 

Isomers 

Tu 

Optimizing the Analysis of Sugar Alcohol 

Excipients in Pharmaceutical Tablet 

Formulations Using Ion Exclusion HPLC 

Columns 

Isolera Flash Purification System 

Agilent Instruments & A ce sories Catalogue 

Separation Science ‘GC Solutions’ is the premier online resource for GC and GC/MS users working acro s the Asia Pacific region. 

Covering GC method fundamentals, practicalities and troubleshooting it offers chromatographers and analytical chemists a genuine 

e-learning platform and searchable archive resource. 

Tech Tip 

Capi lary Column Backflush 

This month we introduce the concepts of backflushing capi lary columns. 

The technique of reversing the flow in GC columns to remove highly retained 

components from the front of the column has b en used for decades with packed 

columns in valved configurations. Packed columns are usua ly used in isothermal 

analyses that require the use of several different columns in order to cover the 

boiling point range of a sample. 

Click here to read more . 


Fast Screening of Reca led Tylenol for 

Tribromoanisole and Related Adulterants Using 

QuEChERS and GC-TOFMS 

Qualitative Comparison of Whisky Samples Using 

Fast GC/TOFMS 

Determination of Dioxin-Like and Non-Dioxin-Like 

Polychlorinated Biphenyl Congeners in Foodstuffs 

and Animal Feed Using Triple Quadrupole GC/MS 


www.sepscienceasia.com Issue 13: March 2011 

MS Solutions 

Separation Science ‘MS Solutions’ is the premier online resource for analytical scientists working with mass spectrometry acro s 



Tech Tip 

Confusion Resulting from Molecular Weight 

and the Nominal Mass, Monoisotopic Mass, 

and Average Molar Mass 


e-learn.sepscience.com/m solutions I sue 8: February 2011 

63 

16 March 2011 

John Dolan’s 

HPLC Solutions 

Click here for 

FREE ACCESS 

to Separation 

Science learning 

platforms>> 

John Dolan is best known as one of the world’s foremost troubleshooting authorities. Separation Science and John Dolan have co laborated to 

offer this weekly digita learning platform providing valuable advice on everyday i sues, problems and cha lenges faced by LC practitioners. 

Importantly, you wi l also have the opportunity to interact with John through our online questions submi sion system. 

Tech Tip 

PEEK Fo low-up 

PEEK (poly-ether-ether-ketone) tubing and fi tings have become 

a standard part of most HPLC systems today. The flexibility and 

ease of cu ting the tubing, coupled with the convenience of fingertightened 

fi tings make a combination that is hard to beat for 

most applications. In at least two of the previous HPLC Solutions 

discu sions (#9 and #55), I’ve discu sed various aspects of PEEK 

products used in the HPLC environment. 


MS Solutions 

Separation Science ‘MS Solutions’ is the premier online resource for analytical scientists working with ma s spectrometry acro s 



Tech Tip 

Adjusting Electrospray Voltage for 

Optimum Results 


Determining Pesticides in Dietary Supplements with 

QuEChERS Extraction, Cartridge SPE, and GCxGC-TOFMS 

Metabolic Profiling of A curate Mass LC-MS/MS Data to 

Identify Unexpected Environmetal Pollutants 


Products 

Click here to read more .. 

Xevo G2 QTof 

AB SCIEX QTRAP® 5500 LC/MS/MS System 


e-learn.sepscience.com/m solutions I sue 7: December 2010 


December 2010 

Workshops 

Separation Science, in conjunction with John Dolan 

and Tom Jupi le (LC Resources), offers a “Advanced 

HPLC Method Development” Master Cla s in 

Switzerland. 

Click here to learn more 

Ask the Doctor 


Through ‘John Dolan’s HPLC Solutions’ you wi l be 

able to ask questions directly. So if you have 

problems with ca ryover, ghost peaks or any other 

HPLC i sues then click here to contact John. 

www.sepscience.com www.lcresources.com 

MS Solutions 

64 

23 March 2011 



High-Speed, High-Resolution Analysis of Catechins in 

Green Tea 

Metabolic Profiling of A curate Mass LC-MS/MS Data to 

Identify Unexpected Environmental Po lutants 


Products 

Xevo G2 QTof 

UltrafleXtreme MALDI 


Enviro Edition 

Cd 

Developing fast GC methods – avoiding 

changesin elution order or separation efficiency 

GC Solutions 

62 

2 March 2011 



John Dolan is best known as one of the world’s foremost troubleshooting authorities. Separation Science and John Dolan have collaborated to 



Tech Tip 

Method Adjustment vs Change 

Part 7: Fo low-up 

In the past instalments of HPLC Solutions (#56-61), we have explored the concept 

of the adjustment of HPLC methods. In particular, we have tried to determine the 

difference betw en method adjustment and method change. The reason for this 

spli ting of hairs is the interpretation of regulatory guidelines that adjustments can 

be made to m et system suitability requirements, but changes wi l require some 

level of revalidation. As a reference point, we used the European Pharmacopoeia (EP) 

and United States Pharmacopoeia (USP) recommendations. The problem with any 

discu sion of method alteration is tha there is a lot of interpretation that goes on. 





Tech Tip 

What is a good selection for an 

unretained peak in 

reversed-phase chromatography? 

An 


Alternative Procedure for Extraction and 

Analysis of PAHs in Seafood by QuEChERS- 

SBSE-GC-MS 

Tu 

Sensitive Determination of Microcystins in 

Drinking and Environmental Waters 

7000B Triple Quadrupole GC/MS/MS 

The Flame Ionization Detector 

Method to Enable Compliance with Updated 

Federal Workplace Drug Testing Guidelines 

This month we initiate a discu sion on the workhorse detector of gas 

chromatography: the flame ionization detector. The flame ionization detector 

(FID) is the premier detector in gas chromatography. It has unique properties and 

performance that puts it above and beyond a l other general-use detectors in gas 

chromatography (or any other form of chromatography, for that ma ter). 


GC Solutions 




Tech Tip 

Dionex Introduces New Capabilities for the Fast 

Separation of Common Anions and Cations 

Developing fast 

GC methods 

The Flame Ionization Detector - Part 2 

Continuing last month’s discussion of the most popular GC 

detector, the FID, this month we cover deviations from typical unit carbon 

response, optimization and troubleshooting. 





Tech Tip 

Why Acid 

Recently, I received an “Ask the Doctor” email from a reader asking 

why formic acid was specified as an additive for the mobile phase in 

an HPLC method he was using. Formic or trifluoroacetic acid at 0.1% 

concentrations are common, especia ly for LC-MS work. There are a 

number of reasons for adding an acid at low concentration to the mobile 

phase. Let’s look at two of these: the influence on the column and the 

sample. 





Tech Tip 

Confusion Resulting from Molecular Weight 

and the Nominal Mass, Monoisotopic Mass, 

and Average Molar Mass 



February 2011 

North America 

GC 

Accurate and 

Rapid Metabolomics 

Strategies Using 

GC-MS/MS-MRM 

Technology 


Workshops 

www.sepscience.com I sue 12: February 2011 


and Tom Jupille (LC Resources), offers a “Advanced 


Chicago, USA. 








Determining Pesticides in Dietary Supplements with 

QuEChERS Extraction, Cartridge SPE, and GCxGC-TOFMS 

Metabolic Profiling of A curate Ma s LC-MS/MS Data to 

Identify Unexpected Environmetal Po lutants 


Products 

Xevo G2 QTof 

AB SCIEX QTRAP® 5500 LC/MS/MS System 



e-learn.sepscience.com/m solutions I sue 8: February 2011 



Lena Fragner, Wolfram Weckwerth, Hans-Joachim Huebschma n 



by LC-MS 


An 

Tu 


www.sepscienceasia.com I sue 11: January 2011 

64 

23 March 2011 



Spectrometry 




Acidic Glycopeptides by ZIC-HILIC Enrichment 


Separation of Substituted Methoxybenzene 

Isomers 

Optimizing the Analysis of Sugar Alcohol 



Columns 



Agilent Instruments & A ce sories Catalogue 


MS Solutions 




Tech Tip 

Workshops 


and Tom Jupi le (LC Resources), offers a “Advanced 


Chicago, USA. 


Peptide Sequencing with Electrospray LC/ 

MS Part 1: Ion Types and Nomenclature 

One of the most significant and important applications for ma s 

spectrometry is the sequencing of polypeptides by electrospray LC/ 

MS. An e ror in the sequence or the substitution of one amino acid 

with another can completely alter the biological function of a peptide 

molecule. Determination of sequences is therefore a vital part of 

biomedical research, proteomics, and the manufacture of peptide-based 

drug substances. 








Solid Phase Extraction for Low Cutoff Drugs of 

Abuse Panel 

Rapid Analysis of Pesticides in Difficult Matrices 

Using GC/MS/MS 


Products 

Micro-Chamber/Thermal Extractor: µ-CTE 

GC/MS Melamine Analyser 



May 2010 

Solutions 


Screening PAHs in Soil Using RTL Database with 

QuEChERS Extraction Kits and GC/MS 

The Analysis of 16 EPA PAHs by GC/MS using 

Hydrogen Ca rier Gas 

Determining Pesticides in Dietary Supplements 

with QuEChERS Extraction, Cartridge SPE, and 

GCxGC-TOFMS 

Automated Static and Dynamic Headspace Analysis 

www.sepscienceasia.com 




Tech Tip 

Why Acid 

Recently, I received an “Ask the Doctor” email from a reader asking 

why formic acid was specified as an additive for the mobile phase in 

an HPLC method he was using. Formic or trifluoroacetic acid at 0.1% 

concentrations are common, especia ly for LC-MS work. There are a 

number of reasons for adding an acid at low concentration to the mobile 

phase. Let’s look at two of these: the influence on the column and the 

sample. 


with GC-MS for Determination of Abundant and 

Trace Flavour Compounds in Alcoholic Beverages 

Containing Dry Extract 

Using Classifications and Ma s Spectral Filtering 

to Process GCxGC-TOFMS Data for Polychlorinated 

Biphenyls 


Determination of 

losartan, atenolol and 

hydrochlorothiazide 

in tablet dosage form 




Tech Tip 

The Flame Ionization Detector 

Workshops 

This month we initiate a discu sion on the workhorse detector of gas 

chromatography: the flame ionization detector. The flame ionization detector 

(FID) is the premier detector in gas chromatography. It has unique properties and 

performance that puts it above and beyond a l other general-use detectors in gas 

chromatography (or any other form of chromatography, for that ma ter). 



and Tom Jupille (LC Resources), offers a “Advanced 


Chicago, USA. 









Pharmaceutical Edition 

The Development Phase of an LC 

Method Using QbD Principles 

Phil Borman, John Roberts, Chris Jones, Meli sa Ha na-Brown, Roman Szucs and 

Simon Bale 

Method Validation Following Multiple 

Adjustments to the Compendial 

Monograph for Atenolol and Related Impurities 

Sky Countryman and Phil Koerner 

www.sepscienceasia.com I sue 11: January 2011 

Development and validation of stability 

indicating HPLC method for the simultaneous 

determination of losartan, atenolol and 

hydrochlorothiazide in tablet dosage form 

Sweta B. Shah, Vijay. G. Bhave, Lakshmi Narasimham Y.S and Ramesh Yamgar 

Quantitative Analysis of Busulfan in Human Plasma by LC-MS–MS 

Systematic evaluation of new chiral stationary phases for 

supercritical fluid chromatography 

Identification of adulterants in a Chinese herbal medicine by 

LC–HRMS and LC–MS–SPE/NMR 

Analysis of drug interactions with high-density lipoprotein by 

high-performance affinity chromatography 

Liquid chromatographic determination of lumiracoxib in 

pharmaceutical formulations 

Tu An 


Comprehensive analysis of crude oil by twodimensional 

GC (GCxGC) and time-of-flight (TOF) MS 

Food Safety Analysis: LC/MS/MS Applications Using 

Core-She l Technology HPLC Columns 

Non-targeted Screening and A curate Mass 

Confirmation of 510 Pesticides Uaing High Resolution 

Benchtop LC/MS 


e-learn.sepscience.com/m solutions I sue 9: March 2011 

Solid Phase Extraction for Low Cutoff Drugs of 

Abuse Panel 

Rapid Analysis of Pesticides in Difficult Matrices 

Using GC/MS/MS 


Products 

Micro-Chamber/Thermal Extractor: µ-CTE 

GC/MS Melamine Analyser 



February 2011 

Asia Pacific 


Strategies Using GC-MS/MS-MRM 

Technology 

www.sepscienceasia.com 

MS Solutions 

Separation Science ‘MS Solutions’ is the premier online resource for analytical scientists working with m 

Europe, the USA and the Middle East. Covering MS method fundamentals, practicalities and troublesho 

chromatographers and analytical chemists a genuine e-learning platform and searchable archive reso 

Tech Tip 

Peptide Sequencing with Electrospray LC/ 

MS Part 1: Ion Types and Nomenclature 

One of the most significant and important applications for mass 

spectrometry is the sequencing of polypeptides by electrospray LC/ 

MS. An e ror in the sequence or the substitution of one amino acid 

with another can completely alter the biological function of a peptide 

molecule. Determination of sequences is therefore a vital part of 

biomedical research, proteomics, and the manufacture of peptide-based 

drug substances. 


e-learn.sepscience.com/m solutions 




Lena Fragner, Wolfram Weckwerth, Hans-Joachim Huebschma n 



by LC-MS 


An 



Spectrometry 




Acidic Glycopeptides by ZIC-HILIC Enrichme 


Separation of Substituted Methoxybenzen 

Isomers 

Tu 

Optimizing the Analysis of Sugar Alcoho 



Columns 


Agilent Instruments & A ce sorie 

Feature 

Compre 

dimens 

Food S 

Core- 

www.sepscience.com

Application Note: ANBT11 

Comprehensive analysis of crude oil by two-dimensional GC 

(GCxGC) and time-of-ight (TOF) MS 

Introduction 

non-polar column. The resulting 

reservoirs containing “heavier” crudes, 

chromatograms are highly convoluted bituminous shales and tar sands. These 

Crude oil is the generic term for the and usually characterised by a matrix of contain higher proportions of involatile, 

unrened ammable liquid that is mined unresolved material that appears as a polar compounds, exacerbating the 

from the ground. It contains vast 

signicant background “hump” beneath problem of incomplete analysis 1 and 

amounts of organic compounds ranging the partially resolved non-polar 

such constituents are thought to 

from light hydrocarbons to complex compound peaks (Figure 1). 

contribute to the process of 

biomolecules, derived from the remains 

“solidication” of oil in transmission 

of ancient marine organisms and 

A background subtraction/library search 

pipes. Others might have relevance to 

bacteria. 

method is able to identify the non-polar 

the environmental impact of oil, a factor 

compound peaks, but for those lost in 

that will gain further attention with the 

the matrix, there is little hope for a 

One-dimensional gas 

introduction of programs such as 

positive identication. Incomplete 

chromatography 

European REACH (Registration Evaluation 

characterisation limits the understanding 

Authorisation and Restriction of 

Within the complexity of crude oil, the 

of the oil sample’s geochemistry and 

Chemicals), which focuses on the 

compounds of most interest to the 

provides no insight into issues that might 

identication of chemicals in order to 

petroleum industry are relatively volatile 

arise during extraction, transport, or 

assess their toxicological impact. 

(boiling points generally below 400°C) 

rening. In addition, as the more easily 

and non-polar, therefore separations are 

extracted “light” crude oils become 

Comprehensive (2D) gas 

predominantly performed by GC with a 

depleted, oil companies are moving to 


“Comprehensive” two-dimensional 

chromatography (GCxGC) is a technique 

which has benetted the petrochemical 

industry signicantly due to its ability to 

separate very complex mixtures 2,3 . This 

technique requires a conventional nonpolar 

column to be connected to a short 

length of a polar column with a GCxGC 

modulator. The modulator collects time 

slices of efuent from the rst column 

(typically 5 seconds wide) and re-injects 

them onto the short polar column. The 

result is a separation that details both 

polar and non-polar elements along two 

planes. With two levels of separation, the 

complexity of the matrix is pulled apart, 

increasing the resolution and allowing 

the identication of far more 

Figure 1. GC/MS total ion chromatogram of a sample of lubricating oil (a distillate fraction of a crude 

components. 

oil) 

T: +44 (0)1443 233920 | F: +44 (0)1443 231531 | E: enquiries@almsco.com 

Food Safety Analysis: LC/MS/MS Applications Using New Kinetex ® Core-Shell 

Technology HPLC Columns 

Philip J. Koerner, Terrell Mathews, and Jeff Layne 

Phenomenex, Inc., 411 Madrid Ave., Torrance, CA 90501 USA 

The availability of alternative bonded phases based on the ultra-high 

efficiency Kinetex core-shell technology provides orthogonal selectivity 

that is shown to be useful for the separation challenges presented 

in food safety analysis. 

Introduction 

The safety of our food supply has come under increasingly intense 

scrutiny with recent episodes of food products found tainted 

with melamine, antifreeze, salmonella, and potentially harmful 

antibiotics, to name a few of the higher profile examples. 

The chemistry and structure of these potential food contaminants 

and the potential for the presence of multiple contaminants 

presents a significant separation challenge. A single bonded 

phase, such as C18, is unlikely to offer the selectivity required to 

chromatographically resolve these potentially complex mixtures. 

Therefore, the availability of orthogonal bonded phases that 

provide alternative selectivity through additional modes of 

interaction is important for the separation of this broad spectrum 

of analytes. Increased testing mandated by government 

regulations for an ever expanding list of contaminants in food and 

beverages has driven the need for increased sample throughput. 

Additionally, the very complex sample matrices present unique 

sample preparation challenges. 

Over the last several years, smaller fully-porous LC particles 

(sub-2 μm diameter) have been introduced and sparked much 

interest because they provide higher efficiency and resolution, 

which results in significantly shorter analysis times and 

increased sensitivity. However, the widespread adoption of sub- 

2 μm HPLC column technology has been slow because these 

smaller particle size columns generate system backpressures 

that require the use of specialized ultra-high pressure capable 

LC instrumentation. 

A newly developed, commercialized Kinetex 2.6 μm core-shell 

chromatographic particle offers the performance benefits of 

fully-porous sub-2 μm particles (increased chromatographic 

efficiency and resolution, shorter analysis times, and increased 

sensitivity) but at substantially lower operating pressures. The 

benefits provided by the core-shell technology are illustrated 

in three food safety LC/MS applications (antibiotics in meat, 

aflatoxins in peanut butter, and melamine and cyanuric acid in 

baby formula) on the three Kinetex phases currently available. 

Kinetex Core-Shell Technology 

The Kinetex technology comprises a nearly monodisperse 1.9 μm 

solid silica core and a 0.35 μm porous silica shell (Figure 1). This 

particle design results in a very stable and nearly homogeneous 

packed column bed that significantly reduces peak dispersion 

due to eddy diffusion (the “A” term of the van Deemter equation). 

Additionally, the short diffusion path of the 0.35 μm porous silica 

shell allows for faster kinetics of diffusion, thereby minimizing 

peak dispersion due to resistance to mass transfer (the “C” term 

in the van Deemter equation). Figure 2A shows a FE-SEM of 

2.6 μm Kinetex particles under 2,500x magnification highlighting 

the monodisperse nature of the porous shell particle, and Figure 

2B shows a FE-SEM of a single 2.6 μm particle under 100,000x 

magnification highlighting the 100 Å porous surface of the shell. 

Figure 1. 

Kinetex 2.6 µm Core-Shell Technology 

0.35 µm Porous Shell 

1.9 µm Solid Core 

2.6 µm Core-Shell 

Particle 

Figure 2. 


A 

min 

A. FE-SEM of 2.6 µm Kinetex particle 2500x B. FE-SEM of a single 2.6 µm particle 

magnification showing the monodisperse 100,000x magnification showing the 100Å 

nature of the porous shell particle. 

porous surface of the shell. 

The core-shell technology columns provide an increase in 

chromatographic efficiency which allows faster analysis through 

the use of shorter columns without compromising resolution. 

This will significantly improve sample throughput for food safety 

laboratories where government regulations mandate increased 

sample testing. In addition, the sharper chromatographic peaks 

obtained with core-shell columns result in increased sensitivity, 

making it easier to achieve the required lower limits of detection. 

Kinetex core-shell columns are currently available in three 

different (orthogonal) bonded phases and each is highlighted 

here to illustrate the benefits of the core-shell technology for 

specific food safety applications. Antibiotics in meat were 

analyzed using Kinetex C18, aflatoxins in peanut butter were 

analyzed using Kinetex PFP, and melamine and cynauric acid in 

baby formula were analyzed using Kinetex HILIC. 

For additional technical notes, visit www.phenomenex.com Page 1 of 4 

Application 

Note: 51878 

Allen Zhang, James S. Chang, Christine Gu, Mark Sanders, Thermo Fisher Scientific, San Jose, CA, USA 

Overview 

Key Words 

As agricultural trade grows and food safety concerns 

• Exactive 

mount, stricter pesticide regulations are being enforced 

around the world. Increased pesticide testing and 

• High Mass 

reductions in maximum permissible residue levels have 

Accuracy 

driven demand for fast, sensitive and cost-effective 

• High Resolution analytical methods for high-throughput screening of 

multi-class pesticides in food. Detection of 510 pesticides 

• Orbitrap 

at low ppb levels was achieved within 12 minutes using 

Technology 

the Thermo Scientific Exactive benchtop LC/MS system 

• Pesticide Analysis powered by Orbitrap technology. The high resolving 

power of the Thermo Scientific Orbitrap platform enables 

accurate mass confirmation of all compounds, including 

isobaric pesticides. Accurate, robust, easy to use and costefficient, 

the Exactive LC/MS is ideally suited for routine, 

Pesticides in food were traditionally monitored and 

quantified using gas chromatography (GC) coupled with 

comprehensive screening of targeted and non-targeted 

either selective detectors (e.g. electron capture) or mass 

pesticides at or below the 0.01 mg/kg (10 ppb) default 

spectrometry (MS). GC/MS continues to be widely used in 

limit set by EU and Japanese legislation. 

pesticide analysis because it is highly selective, provides 

confirmation of multiple classes of pesticides in a single 

Introduction 

analytical run, and is relatively inexpensive and easy to 

operate. However, GC/MS cannot detect polar, thermally 

In 2007, the United States Environmental Protection 

unstable or low volatility compounds without derivatization. 

Agency (EPA) completed a ten-year reassessment of 9,721 

Recent improvements in liquid chromatography (LC) 

pesticide tolerances to meet more stringent safety standards 

throughput and MS detection capabilities have led to a 

and recommended the revocation or modification of 

surge in the use of LC/MS-based techniques for screening, 

thousands of uses of pesticides in food. 1 China published 

confirmation and quantitation of ultra-trace levels of 

national standard GB 2763-2005 in 2005, which 

multi-class pesticide residues, including those that are not 

established 478 maximum residue levels (MRLs) for 136 

GC-amenable. LC-triple quadrupole tandem MS 

pesticides. 2 Japan’s Positive List System, introduced in 2006, 

(LC/MS/MS) enables highly selective and sensitive 

established MRLs for hundreds of agricultural chemicals, 

quantification and confirmation of hundreds of target 

including approximately 400 pesticides, in food and set a 

pesticides in a single run, but this approach requires extensive 

uniform limit of 10 ppb to chemicals for which MRLs 

compound-dependent parameter optimization and cannot 

have not been determined. 3 Regulation (EC) No. 396/2005 

be used to screen for untargeted pesticides. Full scan 

of the European Parliament, implemented in 2008, 

approaches using high performance time-of-flight (TOF) 

harmonized all pesticide MRLs for European Union (EU) 

or Orbitrap mass spectrometers coupled to ultra-high 

member states and set default limits of 0.01 mg/kg for all 

pressure LC (U-HPLC) facilitate rapid and sensitive 

pesticide/commodity combinations for which no MRLs 

screening and detection of LC-amenable pesticide residues 

have been set. 4 A pesticide safety review of about 1,000 

present in a sample. The superior resolving power of the 

active substances on the market was mandated by EU 

Orbitrap mass spectrometer (up to 100,000 FWHM) 

Directive 91/414/EEC and, upon completion in 2009, led 

compared to TOF instruments (10,000–20,000) ensures 

to the approval of only about 250 substances, effectively 

the high mass accuracy required for complex sample 

setting the permissible levels of over 700 de-listed pesticides 

analysis. 6 High resolution LC/MS instrumentation, 

to the default limit. 5 The EU and Japanese regulations are 

however, can be cost-prohibitive for many routine 

among the most stringent in the world and have fueled the 

monitoring laboratories. 

need for faster and more sensitive analytical methods for 

cost-efficient, high-throughput screening of multi-class 

pesticide residues. 

B 

 


WWW.ALMSCO.COM 

Gwaun Elai Medi Science Campus | Llantrisant | RCT | CF72 8XL | United Kingdom 

Download 

The Scheduled MRM Algorithm Enables Intelligent Use of Retention Time During 

Multiple Reaction Monitoring 

Company: AB SCIEX 

The use of MRM for targeted protein quantitfication and biomarker verification/ 

validation studies on triple quadrupole based MS systems is an active research area 

driven by the well known sensitivity and selectivity attributes of MRM. As more 

extensive protein panels need to be monitored in a targetted way across multiple 

samples, higher MRM multiplxing is becoming essential for throughput. This 

application describes how the Scheduled MRM Algorithm improves this process. 

TN-1080 

APPLICATIONS 

Download 

Using Core-Shell Kinetex XB-C18 HPLC Columns as a Solution for Analyzing 

Fluoroquinolone Antibiotics by LC/MS 

Company: Phenomenex 

Additional Kinetex core-shell media research and product development has 

culminated in the introduction of unique bonded phases, such as Kinetex XB- 

C18. Kinetex XB-C18 is used for developing a rapid LC/MS method for separating 

fluoroquinolones. Data from this technical note demonstrate the utility of the 

Kinetex XB-C18, 2.6 µm column for separating fluoroquinolones, a difficult class of 

basic pharmaceutical compounds. The excellent peak shape for basic compounds in 

volatile buffers makes the Kinetex XB-C18 phase an ideal choice for separating basic 

compounds for LC/MS applications where non-ion pairing buffers are used. 

Non-targeted Screening and Accurate Mass 

Confirmation of 510 Pesticides on the High 

Resolution Exactive Benchtop LC/MS Orbitrap 

Mass Spectrometer 

Download 

Rapid Tea Analysis on Poroshell 120 SB-C18 with LC/MS 

Company: Agilent 

An analysis of ten compounds (9 catechins + caffeine) commonly found in green tea 

demonstrates similar selectivity on Agilent ZORBAX SB-C18 and Agilent Poroshell 120 

SB-C18. The 1.4 min gradient analysis on Poroshell 120 generates linear calibration 

curves for all ten compounds through LC/MS. Several bottled and brewed tea samples 

are quantified and compared. An unfiltered, undiluted brewed green tea sample 

demonstrates a lifetime of more than 1500 injections on the Poroshell 120 column 

with a dirty sample at high pressure.

Application Note: ANBT11 

Comprehensive analysis of crude oil by two-dimensional GC 

(GCxGC) and time-of-ight (TOF) MS 

Introduction 

non-polar column. The resulting 

reservoirs containing “heavier” crudes, 

chromatograms are highly convoluted bituminous shales and tar sands. These 

Crude oil is the generic term for the and usually characterised by a matrix of contain higher proportions of involatile, 

unrened ammable liquid that is mined unresolved material that appears as a polar compounds, exacerbating the 

from the ground. It contains vast 

signicant background “hump” beneath problem of incomplete analysis 1 and 

amounts of organic compounds ranging the partially resolved non-polar 

such constituents are thought to 

from light hydrocarbons to complex compound peaks (Figure 1). 

contribute to the process of 

biomolecules, derived from the remains 

“solidication” of oil in transmission 

of ancient marine organisms and 

A background subtraction/library search 

pipes. Others might have relevance to 

bacteria. 

method is able to identify the non-polar 

the environmental impact of oil, a factor 

compound peaks, but for those lost in 

that will gain further attention with the 

the matrix, there is little hope for a 

One-dimensional gas 

introduction of programs such as 

positive identication. Incomplete 


European REACH (Registration Evaluation 

characterisation limits the understanding 

Authorisation and Restriction of 

Within the complexity of crude oil, the 

of the oil sample’s geochemistry and 

Chemicals), which focuses on the 

compounds of most interest to the 

provides no insight into issues that might 

identication of chemicals in order to 

petroleum industry are relatively volatile 

arise during extraction, transport, or 

assess their toxicological impact. 

(boiling points generally below 400°C) 

rening. In addition, as the more easily 

and non-polar, therefore separations are 

extracted “light” crude oils become 

Comprehensive (2D) gas 

predominantly performed by GC with a 

depleted, oil companies are moving to 


“Comprehensive” two-dimensional 

chromatography (GCxGC) is a technique 

which has benetted the petrochemical 

industry signicantly due to its ability to 

separate very complex mixtures 2,3 . This 

technique requires a conventional nonpolar 

column to be connected to a short 

length of a polar column with a GCxGC 

modulator. The modulator collects time 

slices of efuent from the rst column 

(typically 5 seconds wide) and re-injects 

them onto the short polar column. The 

result is a separation that details both 

polar and non-polar elements along two 

planes. With two levels of separation, the 

complexity of the matrix is pulled apart, 

increasing the resolution and allowing 

the identication of far more 

Figure 1. GC/MS total ion chromatogram of a sample of lubricating oil (a distillate fraction of a crude 

components. 

oil) 

T: +44 (0)1443 233920 | F: +44 (0)1443 231531 | E: enquiries@almsco.com 

Food Safety Analysis: LC/MS/MS Applications Using New Kinetex ® Core-Shell 

Technology HPLC Columns 

Philip J. Koerner, Terrell Mathews, and Jeff Layne 

Phenomenex, Inc., 411 Madrid Ave., Torrance, CA 90501 USA 

The availability of alternative bonded phases based on the ultra-high 

efficiency Kinetex core-shell technology provides orthogonal selectivity 

that is shown to be useful for the separation challenges presented 

in food safety analysis. 

Introduction 

The safety of our food supply has come under increasingly intense 

scrutiny with recent episodes of food products found tainted 

with melamine, antifreeze, salmonella, and potentially harmful 

antibiotics, to name a few of the higher profile examples. 

The chemistry and structure of these potential food contaminants 

and the potential for the presence of multiple contaminants 

presents a significant separation challenge. A single bonded 

phase, such as C18, is unlikely to offer the selectivity required to 

chromatographically resolve these potentially complex mixtures. 

Therefore, the availability of orthogonal bonded phases that 

provide alternative selectivity through additional modes of 

interaction is important for the separation of this broad spectrum 

of analytes. Increased testing mandated by government 

regulations for an ever expanding list of contaminants in food and 

beverages has driven the need for increased sample throughput. 

Additionally, the very complex sample matrices present unique 

sample preparation challenges. 

Over the last several years, smaller fully-porous LC particles 

(sub-2 μm diameter) have been introduced and sparked much 

interest because they provide higher efficiency and resolution, 

which results in significantly shorter analysis times and 

increased sensitivity. However, the widespread adoption of sub- 

2 μm HPLC column technology has been slow because these 

smaller particle size columns generate system backpressures 

that require the use of specialized ultra-high pressure capable 

LC instrumentation. 

A newly developed, commercialized Kinetex 2.6 μm core-shell 

chromatographic particle offers the performance benefits of 

fully-porous sub-2 μm particles (increased chromatographic 

efficiency and resolution, shorter analysis times, and increased 

sensitivity) but at substantially lower operating pressures. The 

benefits provided by the core-shell technology are illustrated 

in three food safety LC/MS applications (antibiotics in meat, 

aflatoxins in peanut butter, and melamine and cyanuric acid in 

baby formula) on the three Kinetex phases currently available. 

Kinetex Core-Shell Technology 

The Kinetex technology comprises a nearly monodisperse 1.9 μm 

solid silica core and a 0.35 μm porous silica shell (Figure 1). This 

particle design results in a very stable and nearly homogeneous 

packed column bed that significantly reduces peak dispersion 

due to eddy diffusion (the “A” term of the van Deemter equation). 

Additionally, the short diffusion path of the 0.35 μm porous silica 

shell allows for faster kinetics of diffusion, thereby minimizing 

peak dispersion due to resistance to mass transfer (the “C” term 

in the van Deemter equation). Figure 2A shows a FE-SEM of 

2.6 μm Kinetex particles under 2,500x magnification highlighting 

the monodisperse nature of the porous shell particle, and Figure 

2B shows a FE-SEM of a single 2.6 μm particle under 100,000x 

magnification highlighting the 100 Å porous surface of the shell. 

Figure 1. 


0.35 µm Porous Shell 

1.9 µm Solid Core 

2.6 µm Core-Shell 

Particle 

Figure 2. 


A 

min 

A. FE-SEM of 2.6 µm Kinetex particle 2500x B. FE-SEM of a single 2.6 µm particle 

magnification showing the monodisperse 100,000x magnification showing the 100Å 

nature of the porous shell particle. 

porous surface of the shell. 

The core-shell technology columns provide an increase in 

chromatographic efficiency which allows faster analysis through 

the use of shorter columns without compromising resolution. 

This will significantly improve sample throughput for food safety 

laboratories where government regulations mandate increased 

sample testing. In addition, the sharper chromatographic peaks 

obtained with core-shell columns result in increased sensitivity, 

making it easier to achieve the required lower limits of detection. 

Kinetex core-shell columns are currently available in three 

different (orthogonal) bonded phases and each is highlighted 

here to illustrate the benefits of the core-shell technology for 

specific food safety applications. Antibiotics in meat were 

analyzed using Kinetex C18, aflatoxins in peanut butter were 

analyzed using Kinetex PFP, and melamine and cynauric acid in 

baby formula were analyzed using Kinetex HILIC. 

For additional technical notes, visit www.phenomenex.com Page 1 of 4 

Application 

Note: 51878 

Allen Zhang, James S. Chang, Christine Gu, Mark Sanders, Thermo Fisher Scientific, San Jose, CA, USA 

Overview 

Key Words 

As agricultural trade grows and food safety concerns 

• Exactive 

mount, stricter pesticide regulations are being enforced 

around the world. Increased pesticide testing and 

• High Mass 

reductions in maximum permissible residue levels have 

Accuracy 

driven demand for fast, sensitive and cost-effective 

• High Resolution analytical methods for high-throughput screening of 

multi-class pesticides in food. Detection of 510 pesticides 

• Orbitrap 

at low ppb levels was achieved within 12 minutes using 

Technology 

the Thermo Scientific Exactive benchtop LC/MS system 

• Pesticide Analysis powered by Orbitrap technology. The high resolving 

power of the Thermo Scientific Orbitrap platform enables 

accurate mass confirmation of all compounds, including 

isobaric pesticides. Accurate, robust, easy to use and costefficient, 

the Exactive LC/MS is ideally suited for routine, 

Pesticides in food were traditionally monitored and 

quantified using gas chromatography (GC) coupled with 

comprehensive screening of targeted and non-targeted 

either selective detectors (e.g. electron capture) or mass 

pesticides at or below the 0.01 mg/kg (10 ppb) default 

spectrometry (MS). GC/MS continues to be widely used in 

limit set by EU and Japanese legislation. 

pesticide analysis because it is highly selective, provides 

confirmation of multiple classes of pesticides in a single 

Introduction 

analytical run, and is relatively inexpensive and easy to 

operate. However, GC/MS cannot detect polar, thermally 

In 2007, the United States Environmental Protection 

unstable or low volatility compounds without derivatization. 

Agency (EPA) completed a ten-year reassessment of 9,721 

Recent improvements in liquid chromatography (LC) 

pesticide tolerances to meet more stringent safety standards 

throughput and MS detection capabilities have led to a 

and recommended the revocation or modification of 

surge in the use of LC/MS-based techniques for screening, 

thousands of uses of pesticides in food. 1 China published 

confirmation and quantitation of ultra-trace levels of 

national standard GB 2763-2005 in 2005, which 

multi-class pesticide residues, including those that are not 

established 478 maximum residue levels (MRLs) for 136 

GC-amenable. LC-triple quadrupole tandem MS 

pesticides. 2 Japan’s Positive List System, introduced in 2006, 

(LC/MS/MS) enables highly selective and sensitive 

established MRLs for hundreds of agricultural chemicals, 

quantification and confirmation of hundreds of target 

including approximately 400 pesticides, in food and set a 

pesticides in a single run, but this approach requires extensive 

uniform limit of 10 ppb to chemicals for which MRLs 

compound-dependent parameter optimization and cannot 

have not been determined. 3 Regulation (EC) No. 396/2005 

be used to screen for untargeted pesticides. Full scan 

of the European Parliament, implemented in 2008, 

approaches using high performance time-of-flight (TOF) 

harmonized all pesticide MRLs for European Union (EU) 

or Orbitrap mass spectrometers coupled to ultra-high 

member states and set default limits of 0.01 mg/kg for all 

pressure LC (U-HPLC) facilitate rapid and sensitive 

pesticide/commodity combinations for which no MRLs 

screening and detection of LC-amenable pesticide residues 

have been set. 4 A pesticide safety review of about 1,000 

present in a sample. The superior resolving power of the 

active substances on the market was mandated by EU 

Orbitrap mass spectrometer (up to 100,000 FWHM) 

Directive 91/414/EEC and, upon completion in 2009, led 

compared to TOF instruments (10,000–20,000) ensures 

to the approval of only about 250 substances, effectively 

the high mass accuracy required for complex sample 

setting the permissible levels of over 700 de-listed pesticides 

analysis. 6 High resolution LC/MS instrumentation, 

to the default limit. 5 The EU and Japanese regulations are 

however, can be cost-prohibitive for many routine 

among the most stringent in the world and have fueled the 

monitoring laboratories. 

need for faster and more sensitive analytical methods for 

cost-efficient, high-throughput screening of multi-class 

pesticide residues. 

B 

 

Europe 

Volume 3, Issue 5 


Highly Sensitive Mass Spectrometry 

without Compromising Speed or 

Selectivity 

Determining saccharidic tracers in 

atmospheric aerosols 

S. Rick, A. Wille, and A. Steinbach 

Stereolithography Rapid Prototyping to Drive 

the Development of a new Chromatographic 

Technique, Dynamic Field Gradient Focusing 

Thomas Wray and Peter Myers. 

Analysis of Water-Soluble Polymers Using Linear 

Size Exclusion HPLC Columns and a Semi-Micro 

SEC System 

Faster Real-time Response to Bacterial Infection 

of Bioethanol Fermentation using a Short Rezex 

ROA Column 

Rapid Tea Analysis on Poroshell 120 SB-C18 with 

LC/MS 

240 Ion Trap GC/MS 

VOC analysis of air sampled using canisters 

Clarus 600 GC/Mass Spectrometers 

WWW.ALMSCO.COM 

Gwaun Elai Medi Science Campus | Llantrisant | RCT | CF72 8XL | United Kingdom 

Download 

Analysis of Water-Soluble Polymers Using Linear Size Exclusion HPLC Columns and a Semi-Micro 

SEC System 

Company: Tosoh 

In recent years water-soluble polymers are gaining more and more interest in different applications. 

The molecular weight distribution of polymers is usually characterized by size exclusion 

chromatography (SEC) coupled with refractive index, viscometric or laser light scattering detection. 

Recent advances in SEC comprise semi-micro SEC and the design of linear columns providing wide 

molecular weight separation ranges and near-linear calibrations. We describe the separation of 

water-soluble polymers with a new generation of linear, polymer-based SEC columns using the 

compact EcoSEC SEC system. 

TN-1080 

APPLICATIONS 

Download 

GC/MS Analysis of Phthalates in Children’s Products 

Company: PerkinElmer 

This application note presents a robust and accurate GC/MS analysis which determines the 

phthalate content of plastic-toy components. Important aspects of sample preparation – cleanup 

and laboratory practices – are presented. Sample preparation for surface and total phthalate 

measurements are discussed. The GC/MS analysis demonstrates separation and detection of 

common phthalates. This measurement achieved detection at levels considerably lower than the 

regulations. Finally, the power of GC/MS analysis was used to screen the samples for other common 

additives including anti-oxidants and PAHs. 

Non-targeted Screening and Accurate Mass 

Confirmation of 510 Pesticides on the High 

Resolution Exactive Benchtop LC/MS Orbitrap 

Mass Spectrometer 

Download 

Accurate and reliable analysis of beer using time-of-flight technology for gas chromatography 

Company: ALMSCO 

SPE-tD offers a simple but very sensitive means of extracting volatile components from highly 

complex aqueous products such as beer. The introduction of a novel time-of-flight MS provides a 

detector which can fully characterise a sample of wide dynamic range, providing sensitivity even 

for very low level compounds. The generation of classical spectra for compound peaks means that 

compound identification is possible with proprietary databases. 

Recommend a Colleague 

Read the latest 

Enviro Edition 

Cd 

If you have a work colleague, collaborator or staff member who would benefit from 

this monthly publication then send us their details below. 

Recommend 

Determining saccharidic tracers in atmospheric aerosols 

Highly Sensitive Mass Spectrometry without 

Compromising Speed or Selectivity 

Determining saccharidic 

tracers in atmospheric 

aerosols 

An 

Tu

download pdf version - Separation Science

You also want an ePaper? Increase the reach of your titles

Delete template?

Save as template?